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ADVANCES IN CHEMISTRY RESEARCH

ADVANCES IN
CHEMISTRY RESEARCH
VOLUME 37

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ADVANCES IN CHEMISTRY RESEARCH

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ADVANCES IN CHEMISTRY RESEARCH

ADVANCES IN
CHEMISTRY RESEARCH
VOLUME 37

JAMES C. TAYLOR
EDITOR

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CONTENTS

Preface vii
Chapter 1 The Electrochemical Determination of the
Synthetic Antioxidants Used as Stabilizers in
Mineral Oils and Biofuel 1
Jaromíra Chýlková, Markéta Tomášková,
Vladimír Jehlička, Ivan Švancara and
Renáta Šelešovská
Chapter 2 Cholinesterase-Inhibitory Activity of Essential Oils 15
Franko Burčul, Mila Radan, Olivera Politeo and
Ivica Blažević
Chapter 3 Phytochemical Functionalized Metal Nanocatalyst
(Ag, Au, Fe, Zn and Pd) for Remediation of
Organic Dyes 87
Brajesh Kumar, Kumari Smita and Brajendra Kumar
Chapter 4 Ionic Liquid-Promoted Synthesis of Phosphinates
and Bisphosphonic Acid Derivatives 121
Nóra Zsuzsa Kiss, Dávid Illés Nagy
and György Keglevich
Chapter 5 Low Molecular Mass Reactive Acrylamide
Copolymers as Carriers of Biologically
Active Compounds 141
M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina,
A. I. Amirova and N. V. Zakharova

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vi Contents

Chapter 6 The Phenomenon of Mass Concentration


Periodicity in Amorphous Matter:
A New Kind of Periodicity in Nature 163
K. Zubow, A. Zubow and V. A. Zubow
Chapter 7 Use of Furan Derivatives Acting as Electrophilic
Dienophiles: An Experimental and Theoretical
Analysis of Polar Cycloaddition Reactions 177
Pedro M. E. Mancini, Mauro Cainelli,
Carla M. Ormachea and María N. Kneeteman
Chapter 8 The Use of Surfactants in the Chemical Treatment
of a Crude Case Oriental Zone in Venezuela for
Asphaltene Mitigation 207
Juan Pereira, Henry Labrador, Víctor Perez,
Ivan Villanueva, José Bustamante, Sofia Guevara,
Maria Mannello and Miguel Parra
Index 235

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PREFACE

The authors of this latest volume discuss recent advances in chemistry


research. Chapter One summarizes the latest achievements on the
electrochemical determination of synthetic antioxidants. Chapter Two provides
an overview on the activity of essential oils with defined chemical
composition, tested against cholinesterases and identifies the ones that could
be potentially used in Alzheimer’s treatment. Chapter Three describes a
simple, cost-effective and ecofriendly approach for the fabrication of different
metal nanoparticles (MNPs) including silver, gold, iron, zinc and palladium by
using different plant phytochemicals as potential reducers and stabilizers.
Chapter Four studies ionic liquid-promoted synthesis of phosphinates and
biphosphonic acid derivatives. Chapter Five focuses on low molecular mass
reactive acrylamide copolymers as carriers of biologically active compounds.
Chapter Six discusses the phenomenon of mass concentration periodicity in
amorphous matter. Chapter Seven studies the use of furan derivatives acting as
electrophilic dienophiles. Chapter Eight examines the use of surfactants in the
chemical treatment of crude case oriental zone in Venezuela for asphaltene
mitigation.
Chapter 1 - In this chapter, the latest achievements on the electrochemical
determination of synthetic antioxidants are summarized concerning mainly the
substances based on phenolic compounds and used to stabilize the mineral oils
or to prevent undesirable oxidation processes in biofuel/biodiesel. This
overview is mostly based on the authors’ own results, and of interest is a group
of the most frequently used antioxidants; namely, 2,6-di-tert-butyl-4-
methylphenole (BHT), 2-tert-butyl phenol, 2- and 3-terc-butylhydroxy-
anisoles (mixture of isomers, BHA), terc-butylhydroquinone (TBHQ),
4,4´-methylene-bis-(2,6-di-tert-butylphenol) (MBP), propyl gallate (PG), and

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viii James C. Taylor

pyrogallol (PY). In order to determine these compounds electrochemically, the


principal step is the anodic oxidation at solid electrodes of the Au- or BDDE
type, followed by detection of the corresponding signal(s) in the linear scan
voltammetric mode. The respective method utilizes 0.18 M H2SO4 as the
supporting electrolyte – either as such (for substances soluble in aqueous
solutions) or with suitable organic solvent ensuring sufficient solubility of the
target analyte(s). From mineral oils, the antioxidants are being isolated by
extraction with ethanol, whereas biodiesel can be analyzed directly. In the
case, when the voltammetric signals are well developed and separated enough
one from another, the antioxidants of interest can also be determined in
mixtures and even simultaneously. Due to overall simplicity of the
corresponding procedure(s) and thanks to favorable prices of electrochemical
instrumentation, such determinations may compete with those based on
otherwise prevailing spectral measurements in tribodiagnostic laboratories.
The use of electrochemical methods described herein can be recommended to
control the degree of fuel degradation, thus offering a way to watch the actual
condition of some industrial machines and similar devices with respect to their
possible wearing out.
Chapter 2 - Essential oils (EOs) are a complex mixtures containing
volatile secondary metabolites with various structures (terpenes, terpenoids,
phenylpropanoids, and others). Their roles in plants vary from attracting
pollinators to a defense against insects, food storage, and adaptations to harsh
climate. Since ancient times, EOs are recognized for their medicinal value,
representing a valuable source of biologically active compounds. EOs possess
various activities including: antimicrobial, antiviral, antioxidant, antitumor,
etc. They have also been used as perfumes, flavors for foods and beverages,
remedies for the body and mind, and continue to be of paramount importance
to date.
Having a small molecular weight and lipid solubility, EO constituents
possess the ability to pass the blood-brain barrier. Given the complexity of
their chemical composition and ability to enter the brain, EOs and their
constituents offer a promising strategy for the treatment of various
neurological disorders. Alzheimer’s disease has been responsible for more
than 50% of neurological diseases among persons over the age of 65 years.
EOs are found to be potential acetylcholinesterase and butyrylcholinesterase
inhibitors, two enzymes which still represent the only pharmacoterapeutic
targets against Alzheimer’s disease. The purpose of this chapter is to provide
an overview of the current knowledge about the activity of the EOs with

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Preface ix

defined chemical composition, tested against cholinesterases and to identify


the ones that could be potentially used in Alzheimer’s treatment.
Chapter 3 - In this chapter, the authors describe a simple, cost-effective
and ecofriendly approach for the fabrication of different metal nanoparticles
(MNPs) including silver, gold, iron, zinc and palladium by using different
plant phytochemicals as a potential reducer and stabilizers. It can be used as a
catalyst, photocatalyst, adsorbent or an alternative agent for removal of
different organic dyes. The kinetic enhancement of MNPs on
degradation/removal of dyes can possibly provide significant and valuable
insight for the engineering application of phytochemical functionalized MNPs.
This review article supports the environmental protection and adequately
attractive compared to other techniques.
Chapter 4 - These days, the ionic liquids (ILs) are in the focus as green
and tunable solvents in organic syntheses. However, another application of ILs
has also emerged based on their potential as catalysts or additives in different
reactions. IL additives may enhance the rate of the reactions, and promote
complete conversions. Literature examples are shortly overviewed in this
paper. The authors found that the microwave (MW)-assisted esterification of
phosphinic acids may also be facilitated by the presence of 10% of a suitable
IL. The presence of e.g., [bmim][PF6] allowed a lower temperature of ca. 160–
180°C (instead of ca. 220°C), shorter reaction times, and higher
conversions/yields. Hence, a novel method was developed for the syntheses of
cyclic and acyclic phosphinates. In another field, the synthesis of dronic acid
derivatives, such as pamidronic acid and alendronate useful in the treatment of
osteoporosis was influenced positively by the presence of 30% of
[bmim][BF4]. Both in the case of using sulfolane as a solvent, and in the
solvent-free variation, significant increase in the yields up to 80% could be
observed that means a record in the dronic acid discipline.
Chapter 5 - Low molecular mass linear statistic copolymers of acrylamide
(AA) are promising as carriers for drugs and biologically active compounds
(BAC). Minor toxicity of polyacrylamide (PAA) for warm-blooded animals
makes it possible to use AA copolymers with reactive comonomers for the
purpose of drug delivery. Since they are not biodegradable, their molecular
mass should be restricted to (2–5) × 104 for their complete removal from the
organism.
The aim of this chapter is to synthesize low molar mass ionic AA
copolymers for drug delivery and to study their behavior in water solutions.
As anionic systems, non-toxic statistical AA copolymers with acrylic
(AAc), methacrylic (MAAc) and 2-acrylamido-2-methylpropansulfonic

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x James C. Taylor

(AAMPSAc) acids having different comonomer composition were synthesized


by means of radical copolymerization using AIBN as initiator. AA with AAc
copolymers were also obtained by means of alkaline hydrolysis. The structure
of the obtained copolymers was confirmed using IR and NMR spectroscopy.
It is shown that AA is more active when copolymerizing with MAAc than
with AAc. Different pattern of functional group distribution along the chains
of AA with MAAc and AA with AAc copolymers results in their different
complexing ability with respect to cationic BAC. The synthesized anionic
copolymers were used as drug carriers in complex with aminoglycoside
antibiotics to reduce the toxicity of the latter. The complexation was confirmed
using the methods of equilibrium dialysis, potentiometric titration and
molecular hydrodynamics. It was found that the toxicity and antimicrobial
activity of the resulting aminoglycoside complexes depend on their stability.
The toxicity of aminoglycoside antibiotics in polymeric form proved to be
reduced 3-4 times on average as compared with corresponding intact
aminoglycosides, while completely retaining their antimicrobial activity. The
antiviral activity of AA with AAMPSAc copolymers was also discovered.
These copolymers were successively used to obtain water-soluble nontoxic
polymer complexes of antiviral arbidol drug.
As cationic carriers, AA copolymers with 2-aminoethylmethacrylate
hydrochloride (2-AEMH) were synthesized. A method of ion exchange
conversion of AA with 2-AEMH copolymers into AA with 2-
aminoethylmethacrylate (2-AEM) copolymers containing highly reactive
primary amino groups was developed. Polymeric ketimin derivatives of
doxycycline antibiotic were obtained. They proved to be nontoxic and showed
a marked antimicrobial activity. They influenced the antibody response at
intraperitoneal injection with antigen, demonstrating immunosuppressant
properties depending on the dose. The obtained polymeric conjugates of
doxycycline can be used as antibacterial formulations after tissue and organ
transplantations.
Chapter 6 - Periodic structures in some amorphous substances (water,
starch, heart biomatrix, styrene-maleic anhydride copolymer (1:1) esterified by
bis-tri-n-butyltinoxide and gelatin) were investigated by the gravitational mass
spectroscopy method (GMS). There are periodic sub ensembles (PSE) of
masses in the interval from 30 mio. to 4 billion Daltons. PSE were
characterized to have 5 mass peaks, at log m = 7.62; 8.57; 9.02; 9.31 and 9.54
(m in Daltons), they were represented as sub micelle and micelle structures in
the expanded form. In white gravitational noises, PSE were observed to be
stable, in colored gravitational noises, however, these periodic structures were

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Preface xi

destroyed with the emergence of new sub ensembles both in the expanded and
the collapsed forms. It was tried to find is there any correlation between the
principles of periodic structures in the amorphous molecular matter and those
in the universe.
Chapter 7 - Furans have, fundamentally, biological and chemical uses. For
several years, the authors have been working with substituted furans as
eletrophilic dienophiles joint to different nucleophilic dienes in Polar Diels-
Alder Reactions (P-DA). The principal objective of this analysis was the
preparation of benzofurans and dibenzofurans. To develop these reactions, the
authors used conventional thermal conditions and microwave irradiation. The
solvents employed were organic ones and protic ionic liquids (PILs). Also, the
authors worked in free solvent conditions. The authors demonstrated that
substituted furans result good electrophiles in cycloaddition processes. The
best experimental condition was the combination of microwave irradiation in
presence of PILs like ethylamonium nitrate (NEA) and N-methylimidazolium
tetrafluoroborate ([HMIM][BF4]). When the nitro group is used as substituent
in the electrophile, the process is irreversible due to the loss of nitrous acid.
Then this substituent is convenient for this transformation. Moreover, a
computational theoretical study of furan reactivity as dienophile in Polar
Diels-Alder reactions (P-DA) was performed. For this purpose, the Density
Functional Theory (DFT) method was employed. The principal aim of this
theoretical study was to analyze the reactivity, regioselectivity, solvent effect
(organics and ionic liquids) and reaction mechanisms of these reactions.
Gaussian 09 is the software used to develop the theoretical calculations. The
functional applied was B3LYP and the basis set 6-31G(d). The structures of
reactants, products and transition states were optimized and validated through
the calculation of the vibrational frequencies. For the analysis of reactivity and
regioselectivity, the nucleophilic and electrophilic global and local indexes
were used, respectively. The solvent effect was considered employing two
different solvatation models: the Polarizable Continuum Model (PCM) and the
Supermolecular Approach. For the mechanistic analysis, the stationary points
were located in the Potential Energy Surface (PES) and then optimized and
validated. The activation energy values and reactions paths were analyzed for
each reaction.
Chapter 8 - Petroleum asphaltenes are interesting molecules that have
been studied extensively due to various problems caused by the petroleum
industry. The high tendency of self association of asphaltene behavior is the
main strength of this complex behavior of crude oil. The phenomenon of
aggregation is the association of asphaltene molecules in particles of different

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xii James C. Taylor

shape and size. This chapter of book discusses the use of surfactants in oil
chemical treatment to remedy the problems caused by asphaltenes during
production of crude oil. In this particular and specific case it will be presented
of Furrial area in eastern Venezuela, which is one of the most important
operational areas of the oil industry at Venezuela. Different changes in
pressure, temperature, and secondary recovery methods may induce
aggregation asphaltenes. These changes affect their interfacial properties
because they can alter the physicochemical properties in the medium (crude oil
or solvent) where they are present. The adsorption process of asphaltenes in
liquid-liquid aggregates interfaces, it is slower than the adsorption of
asphaltene molecules. The aggregation state of asphaltene also depend on the
nature of crude oil. Furrial crude oil has always been characterized by the high
tendency to precipitation of asphaltenes.
Asphaltenes aggregation significantly affects the stability of emulsions, oil
viscosity and asphaltene precipitation. Recently, Acevedo showed the colloidal
nature of asphaltenes focused on the point of view of its fractions A1 and A2,
the first insoluble in toluene. The A1 fraction is responsible for the
aggregation behavior due to its solubility behavior. This model can explain in
detail their aggregation in crude oil and other organic solvents. Surfactants are
commonly used in the solution to the problem of asphaltenes in crude oil
production, and stability of water in oil and asphaltene precipitation.
Surfactants are the main asset of the formulation components (a chemical
cocktail) used in the chemical treatment. Also the other formulation
components and additives such as polar solvents, play a key role. A good
formulation should increase surfactant´s properties such as, surface tension,
adsorption, interfacial activity, solubility, etc. The appropriate combination
between surfactants in solvent is important to achieve high dispersion thereof.
Besides the proper structure and conformation they are key in the work of
chemical treatment. For this reason the addition of molecules that help
disperse the surfactant is very important. There are still many things that
explain the action of surfactants in the chemical treatment. However, this
chapter aims to provide an efficient contribution to this important issue related
to the oil industry.

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 1

THE ELECTROCHEMICAL DETERMINATION


OF THE SYNTHETIC ANTIOXIDANTS
USED AS STABILIZERS IN
MINERAL OILS AND BIOFUEL

Jaromíra Chýlková1,, Markéta Tomášková1,


Vladimír Jehlička2, Ivan Švancara3
and Renáta Šelešovská1
1
Institute of Environmental and Chemical Engineering,
University of Pardubice, Pardubice, Czech Republic
2
Department of Informatics in Transport,
University of Pardubice, Pardubice, Czech Republic
3
Department of Analytical Chemistry,
University of Pardubice, Pardubice, Czech Republic

ABSTRACT
In this chapter, the latest achievements on the electrochemical
determination of synthetic antioxidants are summarized concerning
mainly the substances based on phenolic compounds and used to stabilize
the mineral oils or to prevent undesirable oxidation processes in


Corresponding Author Address: Jaromíra Chýlková, University of Pardubice, Studentská 573,
532 10 Pardubice, Czech Republic. Email: Jaromira.Chylkova@upce.cz.

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2 Jaromíra Chýlková, Markéta Tomášková, Vladimír Jehlička et al.

biofuel/biodiesel. In this overview, mostly based on the authors’ own


results, of interest is a group of the most frequently used antioxidants;
namely, 2,6-di-tert-butyl-4-methylphenole (BHT), 2-tert-butyl phenol, 2-
and 3-terc-butylhydroxy-anisoles (mixture of isomers, BHA), terc-
butylhydroquinone (TBHQ), 4,4´-methylene-bis-(2,6-di-tert-butylphenol)
(MBP), propyl gallate (PG), and pyrogallol (PY). In order to determine
these compounds electrochemically, the principal step is the anodic
oxidation at solid electrodes of the Au- or BDDE type, followed by
detection of the corresponding signal(s) in the linear scan voltammetric
mode. The respective method utilizes 0.18 M H2SO4 as the supporting
electrolyte – either as such (for substances soluble in aqueous solutions)
or with suitable organic solvent ensuring sufficient solubility of the target
analyte(s). From mineral oils, the antioxidants are being isolated by
extraction with ethanol, whereas biodiesel can be analyzed directly. In the
case, when the voltammetric signals are well developed and separated
enough one from another, the antioxidants of interest can also be
determined in mixtures and even simultaneously. Due to overall
simplicity of the corresponding procedure(s) and thanks to favorable
prices of electrochemical instrumentation, such determinations may
compete with those based on otherwise prevailing spectral measurements
in tribodiagnostic laboratories. The use of electrochemical methods
described herein can be recommended to control the degree of fuel
degradation, thus offering a way to watch the actual condition of some
industrial machines and similar devices with respect to their possible
wearing out.

Keywords: electrochemistry, voltammetry, antioxidants, oils and fuels


(biofuels)

INTRODUCTION
The main reason for degradation of the mineral oils applied for lubricating
of metallic parts is their oxidation caused by air oxygen. This spontaneous
process is more pronounced at higher temperatures and in the presence of the
so-called wear metals that act as specific catalysts. The oxidation products are
various oxygen-containing compounds, such as aldehydes, ketones, ethers, or
carboxylic acids and their esters that subsequently undergo condensation and
polymerisation reactions, giving rise to a higher acidity and enhanced viscosity
of lubricating oils [1].
It is obvious that the oils after such a degradation are inevitably losing
their useful properties, which - if not being revealed in the right time - may

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The Electrochemical Determination of the Synthetic Antioxidants … 3

result in damage of lubricated parts of each machines or any other device


preserved in this way. In order to define the actual stability of industrial oils
(with respect to degradative oxidation), the procedures utilise the Rotation
Pressure Vessel Oxidation Test (RPVOT) specified by the ASTM 2272
standard [2] and being based on the monitoring of oxygen consumption under
exactly defined conditions. To perform the test in its entirety, one has to
consider from 3 up to 20 h according to the type and stability of the oil
examined.
Among the newest approaches on how to define the oxidation resistivity
of an oil, a testing method known as PetroOxy [3, 4] is often used. It can be
applied to a wide variety of samples covering numerous representatives of
fuels, traditional lubricating oils, or even some plastic lubricants. Regarding
the proper procedure, the sample is being oxidised in a special closed unit at a
temperature of 160 °C and a pressure (of oxygen) of about 600 kPa as long as
the actual pressure decreases to 90% of the initial value. The measure of the
oxidation stability is then a time period needed for such a test with oxygen
consumption.
Yet another possibility of how to control the oxidation stability of
industrial oils is based on examining the samples with respect to the
concentration of typical antioxidants contained in. Usually, it is being
performed with Fourier Transform Infra Red (FTIR) spectrometry by means
of which the intensity of absorption band belonging to the phenolic OH
group can be measured at a frequency of about 3600 cm-1. Alternatively, the
intensity of the carbonyl group of the respective oxidation product can also be
detected, offering to evaluate the signal in an interval of 1700-1750 cm-1.
However, despite this choice, the monitoring of aminic antioxidants with the
aid of FTIR is quite problematic.
In general, the occurrence of the oxidation processes should also be
controlled in the case of fuels represented by mixtures with methylesters of
fatty acids (FAME [5]). Here, the oxidation stability is given mainly by the
presence of FAME and the oxidation processes can be identified again via the
enhanced acidity, moreover, with appearance of typical precipitates formed in
these fuel systems. In common practice, the oxidation stability can be
evaluated by means of time-consuming methods, such as the above-discussed
PetroOxy method, differential scanning calorimetry (DSC [6, 7]) or the so-
called Rancimat test.
The approach with DSC utilises the detection and quantification of
temperature effects, the Rancimat test [8] is based on the absorption of
oxidation products in distilled water whose conductivity is continuously

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4 Jaromíra Chýlková, Markéta Tomášková, Vladimír Jehlička et al.

measured; the experiment being stopped when having achieved a value of 200
µS.cm-1.
Besides these rather common procedures, there are some less-known
possibilities and one is represented by a voltammetric method that enables to
define the content of antioxidants with the aid of their electrochemical
oxidation and whose details can be found in the ASTM D6971-09 standard,
when also a special RULER® apparatus is operated via such principles. When
going into some details, this voltammetric analysis requires a set of coloured
solutions with unspecified composition that have to be purchased as
commercially available formulations. Afterwards, the oily phase is isolated
with the aid of a sand-like material and the content of antioxidants indicated
with a carbonaceous electrode. The proper evaluation is then based on direct
comparison of two different media and of the respective responses – the first
one belonging to the used oil and the second to the new one.
In the framework of our long-time research, we have been developing and
testing various voltammetric procedures with a goal to obtain a simple but
reliable tool for determining some selected - but frequently used - antioxidants
in lubricating oils and biofuel. Usually, our effort has also been focused on
feasibility to discriminate among the individual substances. The working
electrodes have been chosen in accordance of a demand for the widest possible
applicability and without a necessity to mechanically (pre)treat the electrode
surface as a time-consuming and often not easy-to-perform operation that may
principally affect the electrochemical reactions taking place at the electrode or
in its vicinity and, subsequently, the corresponding current-flow response(s).

VOLTAMMETRIC DETERMINATION OF BHT IN


MINERAL OILS AND ECODIESEL OR IN
SOME MIXTURES WITH OTHER ANTIOXIDANTS
Voltammetric determination of synthetic antioxidants from the phenol
family [e.g., 2,6-di-tert-butyl-4-methylphenole (generally known as “BHT”),
2-tert-butylphenol, 2- and 3-tert-butylhydroxyanisoles (“BHA”), tert-
butylhydroquinone (“TBHQ”), 4,4´-methylene-bis(2,6-di-tert-butylphenol),
propyl gallate (“PG”) and pyrogallol (“PY”)], or aromatic amines [N-phenyl-
1-naphthylamine (“PNA”)], respectively, utilises the anodic oxidation in the
positive potential range, for which e.g., the (compact) gold disc electrode
(“AuE”) or boron-doped diamond electrode (“BDDE”) are the proper choice;

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The Electrochemical Determination of the Synthetic Antioxidants … 5

both being chemically inert, requiring only a short pretreatment/regeneration


prior to use. The respective experiments can then be performed by using the
ordinary and widely available electrochemical analysers, like that illustrated in
Figure 1. From voltammetric techniques, already traditional linear scan
voltammetry (LSV) offers a sufficiently sensitive measurement of the current
response in dependence of continuously changing potential (or voltage,
respectively) applied to the working electrode with constant surface.
The result is typically broad peak with a maximum whose position versus
E-axis reflects the quality; i.e., belongs to the respective species or a
compound, whereas its height corresponds to the quantity of such species or
compound to be determined; see Figure 2.
A typical example of wisely chosen electrolyte is a mixture of 0.1 M
H2SO4 with isopropanol which quantitatively dissolves biofuels so that the
direct analysis can then be performed - without any additional and time-
consuming sample pretreatment. A detailed procedure for the determination of
2,6-di-tert-butyl-4-methylfenolu (BHT) in real biofuels, including a
specification of all other conditions obeying the above-stated criteria is then
available in an ultimate report published recently [9]. Among others, this
article also offers a comparison of voltammetric results with those obtained by
the reference FT-IR method.
Herein, it should be added that the supporting electrolyte based on 0.1 M
H2SO4 and isopropanol is also applicable in other related analyses. Namely, it
can be used to determine BHA [10], mixtures of BHT and BHA [11], and,
eventually, BHT and TBHQ [12] in samples, such as mineral and synthetic
oils, ecodiesel and similar “green” fuels like diesel mixed with methylesters of
fatty acids.

Figure 1. Multipurpose electrochemical analyser (model “AUTOLAB”, with courtesy


of Metrohm).

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6 Jaromíra Chýlková, Markéta Tomášková, Vladimír Jehlička et al.

Figure 2. The current-potential dependence at the working solid electrode used in the
LSV mode.

In oil analyses, the antioxidants have to be first isolated from the matrix
by extracting with ethanol; best, when assisted by a short (ultra)sonication.
Then, the upper ethanolic layer - that will serve for later voltammetric analysis
- is purified from the oily residua by adding solid Na2SO4 with subsequent
filtrating through a dry filter paper.
For the BHA antioxidant, the current response is not linear with
concentration, which has been confirmed repeatedly. Due to this, the
quantification of this substance can be made with the aid of a special
programme called “Nonlinear” that has been proposed for this purpose by our
group [10]. The phenomenon of the peak overlay with respect to the function
of this programme can be described as follows. If one analyses the mixture of
BHA and BHT antioxidants, their signals appears quite close to each other
being partially overlapped (the peak maximum for the first one is +895 mV
and +1075 mV for the second; both versus Ag/AgCl. As found out, this
proximity affects mainly the peak shape of the follow-up substance, i.e., BHT.
The proper evaluation to obtain the correct peak height (or area, respectively)
then relies on a procedure depicted in Figure 3. Here, the part of the
voltammogram - namely, the curve between the maximum for BHA and the
following minimum - is linearized by approximating it with tangent in the
respective inflection point. Afterwards, a perpendicular from the maximum for
BHT (point “T”) is constructed, ending in point “P”; in other words, as an
intersection with the previously set tangent. The actual peak height for BHT is
then represented by the distance between points “P” and “T”.
For comfort use, the Nonlinear programme offers an algorithm comprising
all the steps for performing the above-described evaluation [11]. The user just
inputs the respective experimental data that characterise the I-E curve in the

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The Electrochemical Determination of the Synthetic Antioxidants … 7

range of interest and which have been read-out prior and automatically by the
analyser (software). Finally, the quantification of BHT is accomplished by the
standard addition method with linear I-vs-c relation.
In practical samples of crude-oil products, mixtures of TBHQ and BHT
antioxidants are often being the case; usually, at comparable concentrations.
The determination of both substances in the supporting electrolyte of choice is
normally feasible, due to the fact that the corresponding signals are sufficiently
separated and therefore fine evaluable. In this constellation, the maximum for
TBHQ can be found at a potential of +755 mV and at +1070 mV for BHT
(again, both versus Ag/AgCl). However, if an ethanolic extract with real oil
matrix is to be analysed, there is a particular effect evident in noticeable shift
of both peaks towards more positive potentials. If this occurs, the response for
BHT oxidation is already superposed upon the high background indicating the
decomposition of the supporting electrolyte and so deformed peaks are
difficult to compute.
Then it is possible to use again a mathematic evaluation via special
algorithm that is also integrated in the software. Its function is based on
subtraction of the base-line followed by derivation of the increasing part of the
respective voltammogram(s); the respective procedure being sketched in
Figure 4. Within, the part “A” illustrates the original voltammograms obtained
in the model sample spiked with a mixture of BHT and TBHQ antioxidants.
The part “B” represents the already processed curves serving for quantitative
evaluation of the content of TBHQ from the peak height, whereas BHT is
quantified by evaluating shown by the set “C”.

Figure 3. Graphical evaluation of the peak-height of the BHT signal.

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8 Jaromíra Chýlková, Markéta Tomášková, Vladimír Jehlička et al.

Figure 4. Typical linear scan voltammograms obtained by oxidising the TBHQ a BHT
mixture. A) Original recordings, B) and C) curves after processing by mathematic
transformation. Legend: 1 - base-line (supporting electrolyte) for comparison, 2 -
sample, 3 and 4 – standard additions of BHT, 5,6 - standard additions of TBHQ.

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The Electrochemical Determination of the Synthetic Antioxidants … 9

VOLTAMMETRIC DETERMINATION OF THE


PHENOLIC ANTIOXIDANTS SOLUBLE IN WATER –
PYROGALLOL AND PROPYL GALLATE
Oxidation stability of biofuels is often being ensured by means of
antioxidants from the family of multi-substituted phenols, such as pyrogallol
(PY) and propyl gallate (PG). These substances are readily (PY) or partially
(PG) soluble in aqueous solutions, which is also the case of the sulphuric-acic
based media; the optimal supporting electrolyte being 0.18 M H2SO4. The
oxidation of PY gives rise to a single voltammetric signal with a maximum of
+1270 mV (vs. Ag/AgCl), which is rather high positive value, requiring the
properly chosen working electrode; e.g., the boron doped diamond electrode
(BDDE). The oxidation of PG is more complicated, corresponding to a two-
step reaction pathway and reflected in two subsequent maxima with potentials
+880 and +1050 mV, respectively. In this case, the electrode from compact
gold (AuE) and designed in common disc configuration can be selected;
however, its surface is more sensitive to oxidation, thus requiring the adequate
care with regular control and occasional electrolytic or even mechanical
regeneration.
In contrast to the previously discussed determinations, the I-vs-c
dependence is fairly linear for both PY and PG, which allows one to use the
standard addition method in traditional way, without any experimental or
mathematic treatments. Samples of the ecodiesel type can be again analysed
directly – by adding them into the selected supporting electrolyte. If needed,
the heterogeneous mixtures can be intensively stirred to accomplish
quantitative transfer of the analytes into the aqueous phase. Alternatively, the
samples can also be extracted with water phase yet before the voltammetric
analysis [13, 14].

VOLTAMMETRIC DETERMINATION OF
PHENOLIC AND AMINIC ANTIOXIDANTS
In order to enhance the antioxidation effect, some oils are enriched by
selectively chosen mixtures made of phenols and aromatic amines and the
respective analyses should be able of distinguishing the individual items. And
voltammetry is one of the approaches that may do so this under certain
circumstances.

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10 Jaromíra Chýlková, Markéta Tomášková, Vladimír Jehlička et al.

Figure 5. Anodic oxidation of PNA + BHT mixture (1:1). Set of typical


voltammograms 1 – supporting electrolyte, 2 … 9: increasying concentration of PNA +
BHT in the interval of 5-40 mg L-1 of each.

For instance, if one analyses the mixture of N-phenyl-1-naphthylamine


(PNA) and 2,6-di-tert-butyl-4-methylphenole (BHT) in a supporting
electrolyte containing 0.1 M H2SO4 and a mixture of ethanol and acetonitrile
(2.5:1, v/v), the respective anodic signals are well developed and separated.
For PNA, it represents an interval from +855 to +880 mV vs. Ag/AgCl with
the maximum shifted to lower potentials with the increasing concentration of
phenyl-naphthyl amine. For BHT, both analogical limits are +1110 and +1140
mV according to the concentration level chosen; however, oppositely to
observations with PNA, the increasing concentration of BHT leads to more
positive potentials. A set of voltammograms obtained by analysing such a
mixture - here, PNA a BHT, 1:1 (v/v) is shown in Figure 5.
This phenomenon, when the distance between the two signals changes
unequally with the increased concentration, then enables the direct
determination of both substances by applying standard additions without any
sample (pre)treatment.
This is possible up to the ratio of PNA:BHT = 10:1, but yet higher content
of PNA will already affect the second response making the results of analysis
negatively erratic. In these case, the content of PNA is determined first and
then, its overall amount has to be lowered chemically - with the aid of reaction
with temporarily existing nitrous acid. The whole removal is aimed by adding
solid NaNO2 into the mixture to be analysed and based upon the ethanolic
solution of 0.02 M H2SO4. As nitrite is being added in considerable excess, the
reaction is quite rapid and only about 5 minutes is needed to eliminate the

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The Electrochemical Determination of the Synthetic Antioxidants … 11

interfering aminic antioxidant. Afterwards, the determination of BHT may


follow. The corresponding procedures, experimental and instrumental settings
plus further details can already be found in the original report [15].

CONCLUSION
In this text, it has been shown that electrochemical measurements, namely,
linear scan voltammetry in combination with some solid electrodes are finely
applicable to determine a number of antioxidants used for stabilising mineral
oils and fuels against the undesirable oxidation. Moreover, in some cases, such
antioxidants can also be identified and quantified simultaneously. Typical
conditions for these analyses involve the use of acidic solutions based 0.10-
0.18 M H2SO4 and usually the presence of an organic solvent enabling to
dissolve properly the analytes and/or some matrix constituents. As shown and
discussed for the individual examples (mostly based on the authors’ own
results), one can analyse the new as well as the already used oils. Whenever
needed, the respective procedures may also incorporate a pre-extraction step in
combination with sonication.
It can be stated that this concise overview that summarises mainly some
original methods from our research [9-12, 15] demonstrates well the
possibilities of electrochemical measurements for rather atypical employment
like the analysis of oily and fuel materials. The use of voltammetric
procedures discussed herein can be recommended to control the degree of
oxidative degradation of selected oils and fuels.
Compared to the standard methods and instrumentation used for these
purposes, electrochemical experiments presented above offer an interesting
alternative with inexpensive equipment and relatively rapid and simple
procedures. Regarding the electroanalysis itself, the methodology surveyed
herein can be considered as another contribution into the database of already
existing practical applications with a similar focus, such as (i) electroanalysis
in non-aqueous media (see e.g., [16, 17]) and references therein, (ii)
electroanalytical tensammetry of oils and fuels [16, 17], or (iii) determination
of some trace metals in crude oil homogenised with a mixture of 65% HNO3 +
30% H2O2 by sonication and subsequently decomposed in a microwave
mineralisation unit [20].

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12 Jaromíra Chýlková, Markéta Tomášková, Vladimír Jehlička et al.

REFERENCES
[1] Černý, J., (2001). Principles of low-temperature and high-temperature
oxidation of hydrocarbon mixtures. In: Černý, J. (ed) Reotrieb, 7th
conference. Velké Losiny, CZ, p. 45.
[2] “ASTM D2272-14a”, (2014). Standard Test Method for Oxidation
Stability of Steam Turbine Oils by Rotating Pressure Vessel. Prague, CZ:
Czech Office for Standards, Technology and Testing.
[3] Mužíková, Z., Černý, J., (2011). Possibilities and limitations of the
determination of oxidation stability of gasoline fuels. In: Černý, J. (ed)
Reotrieb, 17th conference. Velké Losiny, CZ, p. 43.
[4] Ladyka, N., Černý, J., (2012). Laboratorní testování oxidační stability
turbínových olejů/Laboratory testing of the oxidation stability of turbine
oils. Paliva 4, 61-65.
[5] Waynick, J. A., (2005). Characterization of biodiesel oxidation and
oxidation products. New York, USA: CRC Project NO.AVFL-2b.
[6] “ASTM D6186”, (2013). Standard Test Method for Oxidation Induction
Time of Lubricating Oils by Pressure Differential Scanning Calorimetry
(PDSC). Prague, CZ: Czech Office for Standards, Technology and
Testing.
[7] Tomašíková, S., Pospíšil, M., Mužíková, Z., Černý, J., (2013) Stanovení
oxidační stability metylesterů mastných kyselin pomocí tlakové
diferenciální skenovací kalorimetrie/Determination of oxidation stability
of the fattry acid methylesters with the aid of differential scanning
calorimetry. Paliva 5, 87-90.
[8] “EN 14112”, (2012). Fat and oil derivatives. Fatty acid methyl esters
(FAME). Determination of oxidation stability (accelerated oxidation
test). Prague, CZ: Czech Office for Standards, Technology and Testing.
[9] Chýlková, J., Tomášková, M., Mikysek, T., Šelešovská, R., Jehlička, J.,
(2012). Voltammetric Determination of BHT Antioxidant at Gold
Electrode in Biodiesel. Electroanalysis 24, 1374-1379.
[10] Tomášková, M., Chýlková, J., Jehlička, V., Navrátil, T., Šelešovská, R.,
(2013). Voltammetric determination of butylated hydroxyanisol in
biodiesel, mineral and synthetic oils using gold electrode. Sci. Pap.
Univ. Pardubice, Ser. A 19, 155-172.
[11] Tomášková, M., Chýlková, J., Jehlička, V., Navrátil, T., Švancara, I.,
Šelešovská, R., (2014). Simultaneous determination of BHT and BHA in
mineral and synthetic oils using linear scan voltammetry with a gold disc
electrode. Fuel 123, 107-112.

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The Electrochemical Determination of the Synthetic Antioxidants … 13

[12] Tomášková, M., Chýlková, J., Navrátil, T., Šelešovská, R., (2014).
Voltammetric determination of antioxidant TBHQ individually and
mixed with BHT in petroleum products using gold disc electrode.
Energy fuels 28, 4731-4736.
[13] Chýlková, J., Tomášková, M., Janíková, L., Šelešovská, R., Navrátil, T.,
Chudobová, P., (2016). Sensitive voltammetric method for the fast
analysis of the antioxidant pyrogallol using a boron-doped diamond
electrode in biofuels. Chem. Pap. (in press) DOI: 10.1007/s11696-016-
0025-3.
[14] Tomášková, M., Chýlková, J., Šelešovská, R., Janíková, L., (2016).
Voltammetric method for rapid determination of propyl gallate and its
application for monitoring of biofuels quality. Monatfh. Chem. (in
press). DOI: 10.1007/s00706-016-1860-1.
[15] Tomášková, M., Chýlková, J., Machalický, O., Šelešovská, R., Navrátil,
T., (2013). Voltammetric determination of different antioxidants in
petroleum products by working gold electrode. Int. J. Electrochem. Sci.
8, 1664-1677.
[16] Dahmen, E.A.M.F., (1986). Electroanalysis: Theory and applications in
aqueous and non-aqueous media and in automated chemical control (1st
ed.). Amsterdam, NL: Elsevier.
[17] Izutsu, K., (2009). Electrochemistry in non-aqueous solutions (2nd ed.).
New York, USA: Wiley.
[18] Kalvoda, R., Novotný, L., (1986). Polarographic behavior of petroleum
components in aqueous solutions: A study usig DPP and convective
adsorption accumulation. Collect. Czechoslov. Chem. Commun. 51,
1587-1594.
[19] Thomas, F. G., Henze, G., (2001). Introduction to voltammetric
analysis: Theory and practice. Collingwood VIC, Australia: CSIRO
Publishing; 51-52.
[20] Švancara, I., Fairouz, M., Ismail, Kh., Šrámková, J., Metelka, R., Vytřas,
K., (2004). Applicability of electrochemical stripping analysis at
mercury- and bismuth film plated carbon paste electrodes to crude oil
digests. Sci. Pap. Univ. Pardubice, Ser. A 10, 5-20.

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 2

CHOLINESTERASE-INHIBITORY ACTIVITY
OF ESSENTIAL OILS

Franko Burčul, Mila Radan,


Olivera Politeo and Ivica Blažević*
University of Split, Faculty of Chemistry and Technology,
Division of Chemistry, Split, Croatia

ABSTRACT
Essential oils (EOs) are a complex mixtures containing volatile
secondary metabolites with various structures (terpenes, terpenoids,
phenylpropanoids, and others). Their roles in plants vary from attracting
pollinators to a defense against insects, food storage, and adaptations to
harsh climate. Since ancient times, EOs are recognized for their medicinal
value, representing a valuable source of biologically active compounds.
EOs possess various activities including: antimicrobial, antiviral,
antioxidant, antitumor, etc. They have also been used as perfumes, flavors
for foods and beverages, remedies for the body and mind, and continue to
be of paramount importance to date.
Having a small molecular weight and lipid solubility, EO
constituents possess the ability to pass the blood-brain barrier. Given the
complexity of their chemical composition and ability to enter the brain,

*
Corresponding author address: University of Split, Faculty of Chemistry and Technology,
Division of Chemistry, Ruđera Boškovića 35, 21000, Split, Croatia, Europe. E-mail:
blazevic@ktf-split.hr.

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16 Franko Burčul, Mila Radan, Olivera Politeo et al.

EOs and their constituents offer a promising strategy for the treatment of
various neurological disorders. Alzheimer’s disease has been responsible
for more than 50% of neurological diseases among persons over the age
of 65 years. EOs are found to be potential acetylcholinesterase and
butyrylcholinesterase inhibitors, two enzymes which still represent the
only pharmacoterapeutic targets against Alzheimer’s disease. The
purpose of this chapter is to provide an overview of the current
knowledge about the activity of the EOs with defined chemical
composition, tested against cholinesterases and to identify the ones that
could be potentially used in Alzheimer’s treatment.

Keywords: essential oils, terpenes, terpenoids, phenylpropanoids,


acetylcholinesterase, butyrylcholinesterase

1. INTRODUCTION
Essential oils (EOs) are a mixture of volatile lipophilic constituents
produced in 17500 aromatic species of higher plants belonging mostly to
Asteraceae, Lamiaceae, Lauraceae, and Myrtaceae families [1]. EOs are
isolated from different plant organs, e.g., flowers (bergamot orange, Citrus
bergamia), leaves (lemon grass, Citronella spp.; eucalyptus, Eucalyptus spp.),
fruits (anise, Pimpinella anisum), seeds (nutmeg, Myristica fragrans), wood
(sandalwood, Santalum spp.), roots (vetiver grass, Chrysopogon zizanioides),
rhizomes (ginger, Zingiber officinale); turmeric (curcuma, Curcuma longa).
They are obtained by distillation process or cold pressing (from the citrus
peels). EOs possess various activities including: antimicrobial, antiviral,
antioxidant, antitumor, etc. During the last fifteen years, EOs have also been
recognized as cholinesterases inhibitors that represent the main target in
treatment of Alzheimer’s disease. Table 1 includes all the EOs reported with
their constituents (over 4%) that were tested against acetylcholinesterase
and/or butyrylcholinesterase.
Constituents of EOs belong mainly to terpenes (monoterpenes and
sesquiterpenes), terpenoids (monoterpenoids and sesquiterpenoids) of low
molecular weight and, to a lesser extent phenylpropanoids and others.
According to their functional groups they include saturated and unsaturated
hydrocarbons, alcohols, aldehydes, ketones, esters, ethers, oxides, phenols etc.
Structures of some common constituents found in EOs are given in Figure 1.

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Cholinesterase-Inhibitory Activity of Essential Oils 17

Terpenes

1 2 3 4 5 6

E
E

E E
H
H H

7 8 9 10

Terpeneoids

OH OH

OH OH O
OH

11 12 13 14 15 16

O Ac
O
O HO Ac O

17 18 19 20 21 22

Figure 1. (Continued)

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18 Franko Burčul, Mila Radan, Olivera Politeo et al.

O HO H
H

H OH
H H

23 24 25

Phenylpropanoids

OCH 3 OH O O
O O
O O
O
O

E E
O

26 27 28 29 30 31

Others

32 33

Figure 1. Structures of the main constitutents from essential oils: Monoterpenes: 1 - α-


pinene, 2 - β-pinene, 3 - limonene, 4 - γ-terpinene, 5 - δ-3-carene; Sesquiterpenes: 6 -
β-caryophyllene, 7 - α-humulene (α- caryophyllene), 8 - germacrene D, 9 - δ-cadinene,
10 - β-cubebene; Monoterpenoids: 11 - thymol, 12 - carvacrol, 13 - menthol, 14 -
terpinen-4-ol, 15 - linalool, 16 - pulegone, 17 - carvone, 18 - menthone, 19 - 1,8-
cineole, 20 - linalyl acetate, 21 - borneol, 22 - bornyl acetate; Sesquiterpenoids: 23 -
caryophyllene oxide, 24 - spathulenol, 25 - β-eudesmol; Phenylpropanoids: 26 - (E)-
anethole, 27 - eugenol, 28 - (E)-methyl isoeugenol, 29 - safrole, 30 - estragole, 31 - β-
asarone; Others: 32 - capillene, 33 - capillin.

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Table 1. Overview of the essential oils and their ability to inhibit cholinesterases (AChE and BuChE)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
ACORACEAE
Acorus calamus L. (AC) India/Rhizome β-Asarone (79.5%), α-asarone (8.5%) 10.7 - [8]
ALTINGIACEAE
Liquidambar orientalis purchased from AYUS trans-Cinnamyl alcohol (45.1%), hydrocinnamyl <10% at 1000 µg/mL - [9]
GmbH (Weinstrasse, alcohol (41.1%)
Bühl/Baden, Germany)
ANACARDIACEAE
Pistacia lentiscus L. Tunisia/Leaf α-Pinene (14.4%), germacrene D (12.1%), terpinen-4- 55.3 - [11]
(14 population) ol (9.6%), δ–cadinene (8.1%), limonene (6.3%), β-
caryophyllene (4.9%), β-pinene (4.5%), γ-terpinene
(4.5%), sabinene (4.4%)
Tunisia/Leaf β-Caryophyllene (12.1%), α-pinene (10.4%), δ– 129.2 - [11]
cadinene (9.9%), germacrene D (8.2%), terpinen-4-ol
(7.2%), sabinene, (6.3%), limonene (6.0%)
Tunisia/Leaf Germacrene D (15.9%), δ–cadinene (10.2%), 148.3 - [11]
limonene (7.6%), α-pinene (7.2%), β-caryophyllene
(5.7%), sabinene, (4.8%), α-phellandrene (4.4%), β-
pinene (4.1%), δ–cadinol (4.0%)
Tunisia/Leaf α-Pinene (14.0%), germacrene D (8.2%), limonene 181.7 - [11]
(8.0%), δ–cadinene (7.4%), β-caryophyllene (7.0%),
terpinen-4-ol (4.6%), β-myrcene (4.5%), α-cadinol
(4.0%)
Tunisia/Leaf Germacrene D (13.1%), β-caryophyllene (11.7%), δ– 199.0 - [11]
cadinene (9.1%), limonene (6.8%), α-pinene (5.1%),
terpinen-4-ol (4.5%)

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Tunisia/Leaf Germacrene D (18.6%), δ–cadinene (15.2%), β- 226.7 - [11]
caryophyllene (12.3%), δ–cadinol (5.6%), α-cadinol
(5.0%), γ-muurolene (4.2%), α-humulene (4.0%)
Tunisia/Leaf Germacrene D (18.6%), δ–cadinene (11.3%), 233.7 - [11]
limonene (10.6%), α-pinene (9.4%), β-caryophyllene
(6.5%), sabinene (4.1%)
Tunisia/Leaf α-Pinene (11.0%), germacrene D (10.4%), limonene 238.7 - [11]
(9.5%), β-myrcene (7.8%), β-caryophyllene (7.4%), δ–
cadinene (7.0%), terpinen-4-ol (5.0%)
Tunisia/Leaf Germacrene D (11.7%), limonene (10.2%), δ– 310.3 - [11]
cadinene (9.2%), β-caryophyllene (8.7%), α-pinene
(6.5%), terpinen-4-ol (5.2%), β-pinene (4.6%), β-
myrcene (4.6%)
Tunisia/Leaf Germacrene D (13.2%), β-caryophyllene (11.4%), δ– 318.3 - [11]
cadinene (8.6%), limonene (7.8%), α-pinene (6.6%),
β-myrcene (6.3%), terpinen-4-ol (5.6%)
Tunisia/Leaf Limonene (20.7%), α-pinene (13.0%), germacrene D 383.3 - [11]
(8.4%), β-caryophyllene (7.1%), α-phellandrene
(6.7%), sabinene (6.1%), δ–cadinene (4.0%)
Pistacia lentiscus L. Tunisia/Leaf α-Pinene (11.6%), limonene (10.5%), germacrene D 400.3 - [11]
(14 population) (10.1%), β-caryophyllene (8.2%), δ–cadinene (7.2%),
β–myrcene (6.0%), terpinen-4-ol (4.3%)
Tunisia/Leaf α-Pinene (16.2%), germacrene D (11.2%), δ–cadinene 470.0 - [11]
(7.1%), β-pinene (5.9%), limonene (5.5%), sabinene
(5.1%), β-caryophyllene (4.8%), β–myrcene (4.7%),
terpinen-4-ol (4.5%)

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FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Tunisia/Leaf α-Pinene (10.5%), sabinene (9.1%), β–myrcene 483.7 - [11]
(8.2%), terpinen-4-ol (7.9%), germacrene D (7.0%), β-
caryophyllene (6.8%), limonene (6.6%), β-pinene
(5.4%), γ-terpinene (4.7%), δ-cadinene (4.5%)
Schinus areira L. Argentina/Leaf β-Phellandrene (17.6%), α-phellandrene (16.2%), 233.8 - [10]
camphene (6.3%), guaiol (6.2%), α-pinene (5.4%), p-
cymene (5.1%), γ-muurolene (5.1%), sabinene (4.3%)
Argentina/Fruit α-Phellandrene (31.8%), β-phellandrene (19.9%), β- 487.9 - [10]
myrcene (19.3%), δ-cadinene (4.6%)
Schinus longifolia Argentina/Aerial part Globulol (14.2%), viridiflorol (10.5%), β- 20.0 - [10]
(Lindl.) Speg. caryophyllene (9.0%), β-selinene (7.1%), δ-cadinene
(6.3%), spathulenol (5.1%), γ-muurolene (5.0%), δ-
cadinol (4.1%), cedrol (4.0%)
ANNONACEAE
Monodora myristica Nigeria/Seed Germocrene-D-4-ol (25.5%), tricycle dec-2-ene 14.9 - [13]
(Gaertn) (6.7%)
Nigeria/Stem γ-Cadinene (31.3%), α–elemene (8.0%) 15.6 - [13]
Xylopia aethiopica Nigeria/Fruit Eugenol (35.0%), terpinen-4-ol (7.2%), β- 0.02 µL/mL 0.03 µL/mL [12]
(Dun.) A. Rich caryophyllene (6.0%), germacrene D (5.5%), (-)
spathulenol (4.6%)
APIACEAE
Crithmum maritimum L. Cyprus/Leaf γ-Terpinene (39.3%), β-phellandrene (22.6%), 50.3% at 121 μg/mL 59.8% at 121 [82]
carvacrol methylether (10.5%), (Z)-β-ocimene (8.2%), μg/mL
p-cymene (6.4%), sabinene (4.4%)
Croatia/Flower Limonene (62.2%), γ-terpinene (13.8%), sabinene 44.4% at 45.5 μg/mL 16.8% at 45.5 [83]
(12.0%), α-pinene (4.9%) μg/mL
Croatia/Stem Limonene (74.2%), sabinene (8.1%), terpinen-4-ol 65.2% at 45.5 μg/mL 24.8% at 45.5 [83]
(5.9%), γ-terpinene (4.6%) μg/mL
Croatia/Leaf Limonene (57.5%), sabinene (13.4%), γ-terpinene 38.3% at 45.5 μg/mL 24.5% at 45.5 [83]
(12.0%), terpinen-4-ol (6.9%) μg/mL

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Ferula communis L. Tunisia/Flower Camphor (18.3%), α-pinene (15.3%), β-eudesmol - 64.6% at 10000 [84]
(9.3%), caryophyllene oxide (8.0%), myrcene (5.0%), μg/mL
γ-eudesmol (4.6%), β-pinene (4.4%)
Tunisia/Stem β-Eudesmol (28.1%), γ-eudesmol (11.1%), α- - 54.7% at 20000 [84]
eudesmol (9.6%) μg/mL
Tunisia/Leaf α-Eudesmol (25.2%), β-eudesmol (20.7%), γ- - 55.7% at 20000 [84]
eudesmol (10.1%), caryophyllene oxide (7.2%) μg/mL
Ferula lutea L. Tunisia/Root δ-3-Carene (72.6%), α-pinene (5.8%), myrcene 28.6 - [85]
(5.1%), α-phellandrene (4.0%)
Tunisia/Flower δ-3-Carene (31.2%), α-pinene (25.8%), 2,3,6- 70.3 - [86]
trimethylbenzaldehyde (10.9%), limonene (6.3%),
myrcene (5.1%), α-phellandrene (4.0%)
Ferulago carduchorum Iran/Aerial part (Z)-β-Ocimene (43.3%), α-pinene (18.2%), bornyl 23.6 µL/mL - [87]
Boiss and Hausskn acetate (4.0%)
Foeniculum vulgare Portugal/Aerial part α-Pinene (25.8%), limonene (16.6%), trans-Anethole 215.0 - [16]
Mill. (11.8%), p-cymene (11.5%), α-phellandrene (6.9%), β-
pinene (6.8%), fenchone (6.3%), β-myrcene (5.4%)
Portugal/Aerial part (E)-Anethole (70.2%), (Z)-anethole (18.6%), α- 252.0 - [88]
thujone (6.0%)
provided by Zaraphyt trans-Anethole (75.0%) 1187.7 - [32]
(Rabat, Morocco)/Aerial
part
ASTERACEAE
Artemisia absinthium Algeria/Aerial part Chamazulene (31.3%), camphor (16.5%), 4-terpineol 18.0% at 100 µg/mL 12.8% at 100 [20]
(4.3%) µg/mL
Artemisia annua L. China/Flower - β-Myrcene (37.7%), 1,8-cineole (16.1%), camphor 1250.0 - [19]
preflowering stage (15.0%)

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FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
China/Flower – full β-Caryophyllene (19.4%), germacrene D (18.1%), 2920.0 - [19]
flowering stage camphor (15.8%), 1,8-cineole (10.6%), β-farnesene
(9.4%)
Chine/Flower - post Camphor (16.6%), β-caryophyllene (16.3%), β- 130.0 - [19]
flowering stage caryophyllene oxide (15.8%), β-farnesene (9.1%), (-)-
spathulenol (7.2%), hexadecanoic acid (4.4%)
Artemisia dracunculus purchased (origin: Italy, Estragole (77.0 - 89.0%), limonene (0.3 -4.9%) 58.0-1900.0 - [21]
L. USA, Hungary, France)
Artemisia fragrans Algeria/Aerial part α-Thujone (39.8%), camphor (27.4%), β-thujone 18.6% at 100 µg/mL Nd at 100 µg/mL [20]
(13.2%), 1,8-cineole (10.3%)
Artemisia herba-alba Tunis/Aerial part β-Thujone (41.9%), α-thujone (18.4%), camphor 165.0 - [17]
(Asso.) (13.2%), germacrene D (4.8%)
Artemisia judaica Egypt/Aerial part β-Thujone (49.8%), chrysanthenone (10.9%), α- 16.1 - [15]
thujone (8.2%), 1,8-cineole (4.9%)
Artemisia monosperma Egypt/Leaf Capillene (36.9%), capillin (14.7%), γ-terpinene 194.1 - [23]
Del (12.5%), β-pinene (7.9%)
Egypt/Leaf Capillene (36.9%), capillin (14.7%), γ-terpinene 120.0 - [15]
(12.5%), β-pinene (7.9%)
Artemisia macrocephala Pakistan/Aerial part α-Thujone (56.2%), 3-thujanone (11.7%), 1,8-cineol 40.0 30.0 [24]
(10.8%), 1-terpinen-4-ol (5.5%)
Artemisia India/Aerial part α-Humulene (46.3%), β-caryophyllene (9.3%), α- 31.3 - [89]
maderaspatana L. copaene (8.2%), β-myrcene (4.3%)
Asteriscus maritimus Tunisia/Root Tetradecanol (12.2%), α-ylangene (8.3%), 11.0 - [90]
(L.) Less caryophyllene alcohol (8.3%), hexadecylacetate
(7.0%), hexyl benzoate (4.4%), 1-heneicosene (4.2%),
nonadecane (4.0%)
Chrysanthemum Korea/Flower Camphor (42.4%), cis-verbenol (26.9%), 1,8-cineole 39.4% at 166 µg/mL 42.2% at 166 [91]
indicum L. (7.5%), borneol (6.1%) µg/mL

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Centaurea lycopifolia Turkey/Whole plant Caryophyllene oxide (9.7%), spathulenol (7.3%), 63.5% at 200 µg/mL 65.8% at 200 [25]
Boiss. et Kotschy arachidic acid (5.0%), widdrol (4.3%), α-pinene µg/mL
(4.0%)
Centaurea balsamita Turkey/Whole plant α-Selinene (8.5%), hexatriacontane (8.3%), 2,5-di-tert 44.9% at 200 µg/mL 51.6% at 200 [25]
Lam. octyl-p-benzoquinone (7.4%), tetracosane (6.0%), 1,3- µg/mL
di-tertbutyl benzene (4.3%), (Z)-8-octadecen-1-ol
acetate (4.1%), arachidic acid (4.0%)
Centaurea iberica Trev. Turkey/Whole plant Arachidic acid (25.3%), hexadecanoic acid (5.9%), 49.6% at 200 µg/mL 59.0% at 200 [25]
ex Sprengel choleic acid (5.5%), isononane (5.0%), nonacosane µg/mL
(4.6%)
Crassocephalum Nigeria/Leaf β-Cubebene (13.8%), α-humulene (10.3%) 11.0 - [13]
crepidioides (Benth S. Nigeria/Stem Thymol (43.9%), 4-cyclohexabutyramide (20.9%) 12.2 - [13]
More)
Dittrichia viscosa Portugal/Aerial part trans-Nerolidol (8.4%), β-oplopenone (7.2%), δ- 916.9 - [16]
cadinene (5.7%), 1,8-cineole (5.6%), τ-cadinol (5.5%),
α-cadinol (5.3%), 10-epi-γ-eudesmol (4.2%)
Gynura bicolor DC Japan/Leaf (E)-β-Caryophyllene (31.4%), α-pinene (17.1%), α- 85.0 - [18]
humulene (9.7%), bicyclo-germacrene (8.1%),
phenylacetaldehyde (5.5%)
Japan/Stem α-Pinene (61.4%), β-pinene (14.4%), myrcene (5.1%) 92.0 - [18]
Inula graveolens L. purchased (origin: Bornyl acetate (54.0%), borneol (20.0%), camphene 270.0 - [21]
France) (4.9%)
Pluchea lanceolata India/Aerial part Linalool (32.2%), β-caryophyllene (8.5%), α-terpineol 2.5 - [22]
(DC.) Oliv. and Hiern (8.0%) spathulenol (7.4%), linalyl acetate (5.6%), 1,6-
dimethyl-4-(1-methylethyl)naphthalene (4.3%)

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FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Rhanterium suaveolens Alegria/Aerial part Perillaldehyde (45.8%), caryophyllene oxide (24.8%), Nd at 200 µg/mL Nd at 200 µg/mL [92]
β-cadinol (5.6%), β-caryophyllene (5.2%), 8-cedrene-
13-ol (5.0%)
Senecio ventanensis Argentina/Aerial part α-Terpinene (12.2%), limonene (11.9%), α-humulene 8.9% at 166 µg/mL - [93]
Cabrera (10.5%), sabinene (9.1%), terpinolene (8.8%), p-
cymene (8.1%), α-ocimene (7.3%), β-pinene (6.0%),
spathulenol (5.1%)
Tanacetum Turkey/Flower Camphor (35.2%), (E)-sesquilavandulol (19.0%), 1,8- 85.3% at 121 µg/mL - [94]
abrotanifolium cineole (13.5%)
Druce.
Tanacetum densum Turkey/Stem 1,8-Cineole (28.3%), camphor (16.4%), borneol 16.1% at 121 µg/mL - [95]
(Labill.) Heywood ssp. (6.4%)
sivasicum Hub. - Mor.
and Grierson
Tanacetum densum Turkey/Leaf Camphor (27.7%), camphene (7.0%), bornyl acetate Nd at 121 µg/mL - [95]
(Labill.) Heywood ssp. (11.8%), α-pinene (5.3%), borneol (5.2%)
eginense Heywood
Tanacetum Turkey/Flower 1,8-Cineole (21.9%), camphor (6.4%) 13.8% at 121 µg/mL - [95]
mucroniferum Hub. -
Mor. and Grierson
Tragopogon latifolius Turkey/Whole plant α-Selinene (10.5%), 2,5-di-tert-octyl-p-benzoquinone 55.4% at 200 μg/mL 46.5% at 200 [26]
var. angustifolius (9.5%), valencene (7.0%), 1,3-di-tert-butylbenzene μg/mL
(4.9%)
BORAGINACEAE
Lycopsis orientalis Turkey/Whole plant Heptacosane (10.5%), τ-muurolene (9.6%), 48.5% at 200 μg/mL 43.7% at 200 [26]
tetratetracontane (9.4%), α–selinene (9.2%), μg/mL
hexatriacontane (8.5%), 2,5-di-tert octyl-p-
benzoquinone (5.6%), mint furanone (5.4%)

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Cordia gilletii Congo/Leaf Phytol (20.4%), hexadecanoic acid (15.2%), (E)-2- 105.6 369.3 [27]
hexenal (9.6%), nonacosane (9.2%), nonanal (5.8%),
eicosane (4.7%), heptacosane (4.5%)
BUDDLEJACEAE
Buddleja asiatica Lour. Pakistan/Leaf 1,8-Cineole (38.1%), β-sinensal (11.8%), 1,10-seco-1- 5.2 μM 27.9 μM [29]
hydroxy-calamenen-10-one (10.2%), α-phellandrene
(5.8%), α-pinene (4.9%)
BURSERACEAE
Boswellia ameero Balf. Yemen, Soqotra Island/ (3E,5E)-2,6-dimethyl-1,3,5,7-octatetraene (34.9%), 1- 41.6% at 200 µg/mL - [30]
f, Oleogum resins (2,4-dimethylphenyl)ethanol (20.3%), 3,4-
dimethylstyrene (17.3%), α-campholenal (13.4%), α-
terpineol (12.4%)
Boswellia elongata Balf. Yemen, Soqotra Island/ Verticiol (52.4%), caryophellene (39.1%), 29.6% at 200 µg/mL - [30]
f Oleogum resins methylcycloundecane-carboxylate (7.9%)
Boswellia socotrana Yemen, Soqotra Island/ (E)-2,3-Epoxycarene (51.8%), 5-isopropyl-2- 59.3% at 200 µg/mL - [30]
Balf. f, Oleogum resins methylbicyclo[3.1.0]hex-3-en-2-ol (31.3%), α-cymene
(7.1%)
CISTACEAE
Cistus creticus Italy/Leaf Vitispirane I (17.4%), manoyloxide (14.0%), 12.9% at 1000 μg/mL 29.1 [31]
hexahydrofarnesylacetone (6.8%), viridiflorol (5.2%),
(E)-phytol (5.1%)
Cistus libanotis Tunisia/Leaf Camphene (25.2%), α-pinene (21.6%), β-pinene 71.2 23.7 [31]
(10.8%), bornyl acetate (9.8%)
Cistus monspeliensis Tunisia/Leaf 13-epi-Manoyl oxide (17.7%), manoyloxide (11.1%), Nd at 1000 μg/mL 180.4 [31]
carvacrol (8.5%), borneol (6.3%)
Cistus salvifolius Italy/Leaf Germacrene D (9.1%), elemicin (8.7%), caryophyllene 58.1 34.2 [31]
oxide (6.7%), viridiflorol (4.6%), manoyloxide (4.5%)

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FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Cistus villosus Tunisia/Leaf Hexadecanoic acid (11.7%), undecan-2-one (7.5%), Nd at 1000 μg/mL 123.2 [31]
hexahydrofarnesylacetone (6.9%), heptacosane
(6.5%), nonacosane (5.6%), pentacosane (4.2%), (E)-
β-ionone (4.1%)
CUPRESSACEAE
Cupressus macrocarpa Egypt/Leaf Terpinen-4-ol (20.3%), sabinene (18.7%), β-citronellol 12.3 - [23]
Hartw. Ex Gordon (13.0%)
Cupressus provided by Zaraphyt α-Pinene (49.0%), limonene (32.0%), -3-carene 283.7 - [32]
sempervirens L. (Rabat, Morocco)/Aerial (18.0%)
part
EUPHORBIACEAE
Croton zehntneri Pax. Brazil/Leaf1 (E)-anethole (89.1%) 0.6 cm at 2000 - [33]
& K. Hoffm. µg/mL
Brazil/Stalk1 (E)-anethole (70.5%), (Z)-methyl isoeugenol (6.3%), 0.6 cm at 2000 - [33]
1,8-cineole (5.2%) µg/mL
Brazil/Root1 (E)-anethole (33.7%), (E)-metyl isoeugenol (29.6%), 0.7 cm at 2000 - [33]
methyl eugenol (28.4%) µg/mL
Brazil/Leaf2 Eugenol (84.2%), eugenol acetate (5.7%), Nd at 2000 µg/mL - [33]
bicyclogermacrene (4.2%)
Brazil/Stalk2 Eugenol (49.1%), (E)-methyl isoeugenol (12.2%), 1,8- 0.5 cm at 2000 - [33]
cineole (11.0%), (E)-caryophyllene (6.8%) µg/mL
Brazil/Root2 (Z)-Methyl isoeugenol (81.5%), stragol (12.6%), (E)- 0.6 cm at 2000 - [33]
caryophyllene (4.4%) µg/mL
Brazil/Leaf3 Stragol (90.2%) Nd at 2000 µg/mL - [33]
Brazil/Stalk3 (Z)-Methyl isoeugenol (53.4%), (E)-anethole (14.7%), 0.5 cm at 2000 - [33]
β-elemene (11.4%), α-copaene (6.0%), (E)- µg/mL
caryophyllene (4.5%)
Brazil/Root3 (E)-Methyl isoeugenol (91.5%) 0.6 cm at 2000 - [33]
µg/mL

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
FABACEAE
Bauhinia ungulata L. Brazil/Leaf β-Caryophyllene (15.9%), caryophyllene oxide 96.0% at 1000 - [35]
(9.2%), α-humulene (8.1%), epi-γ-eudesmol (7.5%), α- µg/mL
bisabolol (4.7%)
Peltophorum dasyrachis Thailand/Bark (+)-(S)-ar-turmerone (22.3%), octanoic acid (16.5%), 83.2 - [36]
Kurz ex Bakar hexadecanoic acid (6.6%), (+)-camphor (6.0%),
hexanoic acid (4.7%)
ILLICIACEAE
Illicium verum Hook. f. India/Fruit Anethole (major compound) 39.9 75.4 [37]
LAMIACEAE
Clinopodium México/Leaf Menthone (35.3%), piperitone oxide (31.2%), linalool 320.0 - [96]
macrostemum var. (5.2%)
laevigatum (Standl.) B.
L. Turner
Cyclotrichium niveum Turkey/Aerial plant Isomenthone (56.2%), pulegone (19.8%) Nd for 2000 µg/mL - [97]
Lavandula dentata L. Saudi Arabia/Leaf and Camphor (61.4%), fenchone (24.3%) 9.7 µL/mL - [50]
flower
Lavandula officinalis purchased (origin: Linalyl acetate (36.0%), linalool (34.0%) β- 820.0 - [21]
Chaix France) caryophyllene (4.5%)
Lavandula pedunculata Portugal/Aerial part Camphor (40.6%), fenchone (38.0%) 57.2% at 2500 48.2% at 2500 [98]
ssp. lusitanica (Chaytor) µg/mL µg/mL
Franco
Lavandula viridis Portugal/Aerial part Camphor (31.6%), 1,8-cineole (21.3%) 411.3 748.7 [99]
L’Hér

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FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Mentha aquatica Brasil/Leaf Carvone D (58.8%), limonene (28.3%) 0.7 cm at 20 µg EO - [38]
Data not available Menthol (24.0%), (-)-methyl acetate (17.0%), (-)- 64.0 - [2]
menthone (17.0%), (+)-menthofuran (17.0%), (+)-
pulegone (11.0%)
Data not available (-)-Menthone (33.0%), (+)-neomenthol (21.0%), (+)- 28.0 - [2]
neomenthyl acetate (16.0%), (+)-menthofuran
(11.0%), (-)-limonene (7.0%)
Data not available Elemol (22.0%), (+)-menthofurane (21.0%), 26.0 - [2]
viridiflorol (13.0%), 1,8-cineole (10.0%), β-
caryophyllene (7.0%), germacrene D (7.0%)
Mentha arvensis Brasil/Leaf Linalyl acetate (39.7%), linalool (34.6%), 1,8-cineole 0.6 cm at 20 µg EO - [38]
(10.0%)
Data not available (-)-Menthol (66.0%), (-)-menthyl acetate (15.0%), (-)- 32.0 - [2]
menthone (8.0%)
Data not available (-)-Piperitone oxide (33.0%), (+)-piperitenone oxide 49.0 - [2]
(26.0%)
Mentha canadensis Brasil/Leaf 1-Menthol (47.0%), isomenthone (29.1%), limonene Nd at 20 µg EO - [38]
(4.3%), menthyl acetate (4.3%)
Mentha citrata Data not available (-)-Linalyl acetate (51.0%), (-)-linalool (39.0%) 38.0 - [2]
Mentha gentilis Data not available (+)-Pulegone (48.0%), (+)-piperitenone oxide 30.0 - [2]
(29.0%), menthone (5.0%)
Data not available (-)-Menthol (59.0%), (-)-menthone (22.0%) 58.0 - [2]
Data not available (-)-Menthone (44.0%), (-)-linalool (39.0%) 40.0 - [2]
Data not available (-)-Linalool (45.0%), -terpinene (9.0%), β-pinene 164.0 - [2]
(8.0%), β-caryopyllene (7.0%), germacrene D (4.0%)
Data not available (+)-1,2-Epoxyneomenthyl acetate (50.0%), menthol 56.0 - [2]
(19.0%), menthyl acetate (12.0%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Data not available (-)-Methyl acetate (19.0%), (+)-pulegone (18.0%), (-)- 56.0 - [2]
menthol (15.0%), (+)-piperitone (14.0%), (-)-
menthone (12.0%), piperitenone (7.0%)
Data not available (-)-Menthol (69.0%), (-)-menthone (18.0%) 88.0 - [2]
Data not available (-)-Menthone (36.0%), (+)-pulegone (35.0%), (+)- 30.0 - [2]
neomenthol (11.0%)
Data not available (-)-Menthol (96.0%) 54.0 - [2]
Data not available (-)-Menthone (32.0%), (-)-menthol (16.0%), (+)- 36.0 - [2]
piperitone (14.0%), (+)-pulegone (11.0%), menthyl
acetate (10.0%), piperitenone (4.0%)
Data not available (+)-Pulegone (84.0%), Piperitenone (4.0%) 52.0 - [2]
Data not available (-)-Carvone (73.0%), (-)-limonene (16.0%) 58.0 - [2]
Data not available (+)-Pulegone (81.0%), (-)-borneol (6.0%) 42.0 - [2]
Data not available (-)-Menthol (65.0%), (-)-menthone (14.0%), 154.0 - [2]
isomenthone (6.0%)
Data not available (-)-Menthone (30.0%), (+)-neomenthol (22.0%), (+)- 40.0 - [2]
pulegone (13.0%), (+)-neomenthyl acetate (13.0%),
borneol (5.0%)
Mentha japonica Data not available (-)-Menthone (57.0%), (+)-pulegone (29.0%), 120.0 - [2]
limonene (4.0%)
Mentha longifolia L. Brasil/Leaf Piperitenone oxide (60.8%), limonene (13.8%), 0.7 cm at 20 µg EO - [38]
carvone D (5.2%), germacrene D (5.2%), trans-β-
caryophyllene (4.7%)
Saudi Arabia/Leaf Pulegone (75.0%), 1,8-cineole (7.4%), l-menthone 1.0 µL/mL - [50]
(6.6%)

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Mentha piperita Brasil/Leaf Menthofuran (23.7%), menthone (17.3%), D- 1.0 cm at 20 µg EO - [38]
neoisomenthol (14.4%), pulegone (10.7%), iso-
menthyl acetate (6.4%), germacrene D (5.5%), 1,8-
cineole (4.8%), trans-β-caryophyllene (4.6%)
Brasil/Leaf Linalyl acetate (51.4%), linalool (25.4%), Nd at 20 µg EO - [38]
menthofuran (4.0%)
Data not available (-)-Menthol (32.0%), (-)-menthone (31.0%), 1,8- 74.0 - [2]
cineole (7.0%), germacrene D (4.0%)
Mentha x piperita Brasil/Leaf Carvone D (49.3%), limonene (37.2%) Nd at 20 µg EO - [38]
Mentha pulegium Portugal/Aerial part Pulegone (35.1%), piperitenone (27.4%) 324.0 - [88]
Data not available (+)-Pulegone (48.0%), menthone (41.0%) 38.0% at 164 µg/mL - [2]
Mentha requienii Data not available (+)-Pulegone (83.0%), (-)-menthone (6.0%), 130.0 - [2]
isomenthol (4.0%)
Mentha rotundifolia Data not available (+)-Piperitenone oxide (46.0%), germacrene D 39.0% at 164 µg/mL - [2]
(21.0%), β-caryophyllene (6.0%)
Data not available Germacrene D (43.0%), (+)-piperitenone oxide 43.0% at 164 µg/mL - [2]
(17.0%), β-farnesene (8.0%)
Mentha spicata Brasil/Leaf Carvone D (60.1%), limonene (19.9%), 1,8-cineole 0.9 cm at 20 µg EO - [38]
(7.4%)
Brasil/Leaf Carvone D (31.4%), limonene (22.1%), pulegone 0.7 cm at 20 µg EO - [38]
(6.7%), β-bourbonene (5.4%), β-pinene (4.3%),
Brasil/Leaf Carvone D (54.9%), limonene (28.8%) 0.6 cm at 20 µg EO - [38]
Brasil/Leaf Carvone D (54.0%), menthone (11.6%), 1,8-cineole Nd at 20 µg EO - [38]
(8.1%), isomenthone (4.5%)
Portugal/Aerial part Carvone (75.9%) 357.0 - [88]
Data not available (-)-Carvone (62.0%), (-)-limonene (7.0%), 88.0 - [2]
dihydrocarvone (6.0%), 1,8-cineole (5.0%)
Data not available (-)-Carvone (69.0%), 1,8-cineole (11.0%), (-)- 57.0 - [2]
dihydrocarvone (9.0%), limonene (8.0%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Data not available (+)-Piperitenone oxide (57.0%), 1,8-cineole (13.0%), 37.0 - [2]
β-myrcene (7.0%)
Data not available (+)-Linalool (98.0%) 130.0 - [2]
Nepeta menthoides Iran/Aerial part 4a-α,7β,7a-α-Nepetalactone (18.4%), 4a-α,7α,7a-α- 64.9 - [100]
Boiss and Buhse Nepetalactone (17.6%), 1,8-cineol (16.7%), geranyl
acetate (7.0%), α–terpineol (5.3%), β-pinene (4.1%)
Ocimum basilicum L. Burkina Faso/Leaf Linalool (48.7%), eugenol (27.5%), trans-α- 32.4% at 100 µL/mL - [45]
bergamotene (5.4%)
Ocimum canum Sims Burkina Faso/Leaf 1,8-Cineole (59.9%), camphor (8.1%), β-pinene 36.2 - [45]
(5.8%), α-terpineol (4.6%), α-pinene (4.5%)
Ocimum gratissimum Nigeria/Leaf -Terpinene (52.9%), (Z)-tert-butyl-4-hydroxy anisol 6.5 - [13]
(Linn) (13.9%), caryophyllene (10.4%)
Nigeria/Seed α–Pinene (48.2%), caryophyllene (10.7%) 6.7 - [13]
Ocimum sanctum L. purchased (origin: India) Eugenol (59.0%), β–caryophyllene (33.0%) 1600.0 - [21]
Origanum ehrenbergii Lebanon/Aerial part Thymol (19.6%), p-cymene (16.1%), 2-isopropyl-1- 0.3 0.3 [46]
Boiss methoxy-4-methylbenzene (14.9%), -terpinene
(11.8%), carvacrol (6.7%)
Origanum majorana L. Tunisia/Aerial part Terpinen-4-ol (23.2%), cis-sabinene hydrate (17.5%), 150.3 - [101]
γ-terpinene (10.5%), p-cymene (9.0%), sabinene
(7.5%), α–terpinene (5.6%), α–terpineol (5.6%), α-
terpineol (4.7%), trans-sabinene hydrate (4.0%)
Egypt/Leaf 4-Terpineol (30.0%), γ-terpinene (15.4%), trans- 36.4 - [102]
sabinene hydrate (10.9%), α-terpinene (6.9%), (S)-
methanal,a,a4-trimethyl-3-cycolohexene-1-1 (6.5%)

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
purchased from Thai- 1,8-Cineole (63.5%), α-terpineol (13.9%), limonene 0.24 - [47]
China Flavour and (5.6%), α-pinene (4.7%), β-pinene (4.2%)
Fragrances Industry Co.,
Ltd. and Perfumesworld,
(Bangkok, Thailand)/
Aerial part
Origanum syriacum L. Lebanon/Aerial part Thymol (24.7%) carvacrol (17.6%), -terpinene 1.7 1.6 [46]
(12.6%), p-cymene (8.7%), 2-isopropyl-1-methoxy-4-
methylbenzene (7.9%)
Origanum vulgare L. Egypt/Aerial part Pulegone (77.5%), menthone (4.9%) 24.4 - [23]
Egypt/Aerial part Pulegone (77.5%), menthone (4.9%) 61.3 - [15]
Spain/Aerial part Carvacrol (58.7%), γ-terpinene (10.7%), p-cymene 73.7 µL EO/L - [103]
(8.2%), thymol (5.8%), (E)-β-caryophyllene (4.5%)
Spain/Aerial part Carvacrol (77.4%) 56.7 µL EO/L - [103]
Spain/Aerial part Carvacrol (66.3%), γ-terpinene (9.1%), p-cymene 61.5 µL EO/L - [103]
(6.8%), thymol (4.6%)
Portugal/Aerial part γ-Terpinene (49.1%), thymol (14.7%), p-cymene 699.3 - [16]
(14.1%), α-terpinene (4.7%)
Origanum vulgare ssp. Turkey/Aerial part Thymol (58.3%), carvacrol (16.1%), p-cymene 1.57 mg 1.74 mg [104]
vulgare (13.5%), –terpinene (4.6%) galanthamine galanthamine
equivalent/g oil equivalent/g oil
Origanum vulgare ssp. Turkey/Aerial part Linalool (96.3%) 1.64 mg 1.75 mg [104]
hirtum galanthamine galanthamine
equivalent/g oil equivalent/g oil
Phlomis armeniaca Turkey/Aerial part Germacrene D (24.1%), n-hexadecanoic acid (21.8%), 0.67 mg 1.88 mg [105]
Willd. hexahydrofarnesyl acetone (13.7%), spathulenol galanthamine galanthamine
(6.0%), (Z)-β-farnesene (5.9%), β-caryophyllene equivalent/g oil equivalent/g oil
(4.8%)

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Phlomis nissolii L. Turkey/Aerial part Germacrene D (15.1%), β-caryophyllene (12.7%), 1.09 mg 3.03 mg galanthamine [105]
hexahydrofarnesyl acetone (11.9%), linalool (11.3%), galanthamine equivalent/g oil
caryophyllene oxide (7.1%), allo-aromadendrene equivalent/g oil
(6.9%), spathulenol (4.0%)
Phlomis pungens Turkey/Aerial part n-Hexadecanoic acid (68.1%), germacrene D (7.2%), 1.23 mg 2.52 mg galanthamine [105]
Willd. var. pungens hexahydrofarnesyl acetone (4.0%) galanthamine equivalent/g oil
Willd. equivalent/g oil
Rosmarinus Egypt/Aerial part 1,8-Cineole (19.6%), camphor (17.0%), α-pinene 20.8 - [23]
officinalis L. (15.1%), verbenon (9.6%), endoborneol (8.2%),
linalool (5.2%)
Portugal/Aerial part Verbenone (35.4%), α-terpineol (7.2%), camphor 69.8 - [88]
(5.5%)
Tunisia/Aerial part 1,8-Cineole (52.6%), camphor (7.8%), α-pinene 498.9 924.2 [48]
(7.1%), borneol (4.1%), trans-caryophyllene (4.1%)
1,8-Cineole (44.9%), α-pinene (9.5%), camphor 98.2 346.7 [48]
(8.0%), trans-caryophyllene (7.7%), borneol (6.2%)
1,8-Cineole (51.4%), α-pinene (8.3%), camphor 200.5 697.8 [48]
(7.2%), borneol (5.9%)
1,8-Cineole (39.1%), camphor (12.0%), borneol 122.8 29.5 [48]
(10.0%), α-pinene (8.0%), trans-caryophyllene (4.1%)
1,8-Cineole (50.3%), camphor (10.5%), α-pinene 478.0 918.2 [48]
(9.1%), borneol (5.9%), trans-caryophyllene (4.3%)
1,8-Cineole (23.2%), camphor (27.5%), camphene 108.8 122.7 [48]
(11.1%), α-pinene (10.4%)

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Tunisia/Aerial part 1,8-Cineole (25.9%), camphor (21.1%), camphene 64.7 353.1 [48]
(10.4%), α-pinene (10.1%), borneol (6.0%), bornyl
acetate (4.2%)

1,8-Cineole (26.0%), camphor (24.3%), α-pinene 227.2 647.4 [48]


(9.4%), camphene (9.1%), borneol (4.4%)
purchased from Thai- 1,8-Cineole (23.2%), camphor (19.2%), o-cymene 51.6% at 120.0 - [47]
China Flavour and (4.1%) µg/mL
Fragrances Industry Co.,
Ltd. and Perfumesworld,
(Bangkok, Thailand)/
Aerial part
Salvia ballsiana Turkey/Aerial part Caryophyllene oxide (34.1%), β-caryophyllene 43.2% at 80 Nd at 80 µg/mL [106]
(8.2%), α-pinene (7.5%) µg/mL
Salvia chionantha Turkey/Aerial part Germacrene D (25.0%), β-caryophyllene (8.7%), 264.1 41.7% at 500 µg/mL [107]
Boiss spathulenol (5.9%), α-humulene (4.8%), α-elemene (calculated)
(4.2%), bornyl acetate (4.2%)
Salvia chrysophylla Turkey/Aerial part α-Terpinenyl acetate (36.3%), β-caryophyllene 838.8 96.6 [108]
Staph (15.3%), linalool (8.1%), β-elemene (4.3%)
Salvia cyanescens Turkey/Aerial part Spathulenol (23.2%), p-cymene (10.3%), 1,8-cineole 15.5% at 80 41.9% at 80 µg/mL [106]
(9.1%), α-pinene (6.4%), β-pinene (6.2%), µg/mL
caryophyllene oxide (4.0%)
Salvia dichroantha Turkey/Aerial part Caryophyllene oxide (22.4%), phytol (5.6%), 43.3% at 400 µM 47.1% at 400 µM [109]
Stapf. caryophyllenol II (5.5%), eudesma-4(15),7-dien-1β-ol
(4.7%)
Salvia divaricata Turkey/Aerial part 1,8-Cineole (30.9%), α-pinene (17.1%), camphor 64.7 - [106]
(10.1%), camphene (7.7%)
Salvia fruticosa Mill. Turkey/Aerial part 1,8-Cineol (36.3%), camphor (19.1%), thujon (7.8%), 49.1% at 100 22.7% at 100 µg/mL [110]
β-pinene (6.4%), camphene (5.7%), α-pinene (5.3%), µg/mL
caryophyllene (4.8%)

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil AChE assay BuChE assay Ref
(percentages) IC50 (µg/mL)* IC50 (µg/mL)*
Turkey/Aerial part 1,8-Cineole (58.9%), α-pinene (5.6%), β- 53.7% at 25 μg/mL 51.2% at 100 μg/mL [111]
pinene (5.2%), β-myrcene (5.2%), camphor
(4.5%), β-caryophyllene (4.2%)
Cyprus/Leaf Camphor (49.2%), 1,8-cineole (17.6%), 50.0 if incubated 5 min; 150.0 if incubated 5 [6]
caryophyllene (11.9%), borneol (4.6%) 60.0 if incubated 30 min; min;
50.0 if incubated 60 min; 60.0 if incubated 30
60.0 if incubated 90 min. min;
40.0 if incubated 60
min;
35.0 if incubated 90
min
Cyprus/Leaf Camphor (49.3%), 1,8-cineole (21.5%), 40.0 if incubated 5 min; 10.0% at 300 µg/mL if [6]
caryophyllene (6.6%), camphene (5.0%) 55.0 if incubated 30 min; incubated 5 min;
60.0 if incubated 60 min; 14.0% at 300 µg/mL if
60.0 if incubated 90 min incubated 30 min;
17.0% at 300 µg/mL if
incubated 60 min;
21.0% at 300 µg/mL if
incubated 90 min
Salvia fruticosa Mill. Italy/Aerial part β-Pinene (13.7%), 1,8-cineole (10.6%), 37.0% at 1000 μg/mL Nd at 1000 µg/mL [112]
ssp. thomasii viridiflorol (9.7%), camphor (8.5%), α-pinene
(7.6%), β-caryophyllene (7.3%), α-humulene
(6.5%), myrcene (5.3%)
1,8-Cineole (25.7%), β-pinene (12.5%), 39.2% at 1000 μg/mL Nd at 1000 µg/mL [112]
viridiflorol (9.7%), β-caryophyllene (8.8%),
camphor (7.7%), α-pinene (6.8%), α-
humulene (6.1%), myrcene (5.3%)

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil AChE assay BuChE assay Ref
(percentages) IC50 (µg/mL)* IC50 (µg/mL)*
Salvia hydrangea Turkey/Aerial part Camphor (46.9%), camphene (9.4%), 1,8- 40.0 8.0% at 80 µg/mL [106]
cineole (7.4%)
Salvia kronenburgii Turkey/Aerial part Geranyl acetate (16.0%), 1,8-cineole 41.5% at 80 µg/mL 12.8% at 80 µg/mL [106]
(12.5%), carvone (11.9%), linalyl acetate
(6.7%), limonene (6.2%), hexadecanoic acid
(4.5%)
Salvia purchased from Camphor (27.0%), 1,8-cineole (17.0%), β- 0.03 - [4]
lavandulaefolia Vahl. Baldwins, London pinene (12.0%), α-pinene (5.0%),
Data not available Camphor (37.0%), 1,8-cineole (36.4%), 3.0 22.0% for 500 µg/mL [39]
camphene (5.7%), α-pinene (5.6%), β-
pinene (5.1%), limonene (4.2%)
Spain/Leaf Camphor (42.5%), 1,8-cineole (17.4%), (+)- 53.0 if incubated 5 min; 26.0% at 200 µg/mL if [6]
2-carene (8.8%), borneol (6.1%) 60.0 if incubated 30 min; incubated 5 min;
80.0 if incubated 60 min; 23.0% at 200 µg/mL if
120.0 if incubated 90 min incubated 30 min;
26.0% at 200 µg/mL if
incubated 60 min;
18.0% at 200 µg/mL if
incubated 90 min
purchased from 1,8-Cineole (26.8%), camphor (24.7%), α- 50.0 - [5]
Baldwins, London pinene (6.6%), β-pinene (5.4%)
Salvia leriifolia Iran/Aerial part Camphor (10.5%), 1,8-cineole (8.6%), 0.3 µL/mL 0.3 µL/mL [40]
Benth camphene (6.2%), β-gurjunene (4.9%), α-
pinene (4.7%), viridiflorol (4.1%), globulol
(4.0%)
Salvia macrochlamys Turkey/Aerial part β-Caryophyllene (26.4%), caryophyllene 42.7% at 80 µg/mL Nd at 80 µg/mL [106]
oxide (22.2%), β-pinene (5.1%), δ-elemene
(4.3%), germacrene D (4.3%)

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Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Salvia nydeggeri Turkey/Aerial part α-Pinene (15.8%), β-pinene (9.2%), cubebol (6.2%), 49.4% at 80 6.6% at 80 µg/mL [106]
caryophyllene oxide (5.6%), 1,8-cineole (4.8%) µg/mL
Salvia officinalis L. Romania/Leaf 1,8-Cineole (13.9%), thujone and camphor (12.2%), α- 0.48 μL/mL - [41]
and β-pinene (4.6%)
Portugal/Aerial part 1,8-Cineole (59.1%), α-pinene (8.4%), camphor 50.8 - [16]
(5.7%)
UK/Leaf α-Humulene (23.2%), camphor (11.0%), borneol 70.0 if incubated 330.0 if incubated 5 [6]
(8.7%), thujone isomers (6.2%), 1,8-cineole (5.4%), 5 min; min;
1H-cycloprop(e)-azulene,deca-hydro 50.0 if incubated 240.0 if incubated 30
1H-cycloprop(e)-azulene,deca-hydro 1,1,7-trimethyl- 30 min; min;
4-methylene-, (1a,a) (5.3%) 65.0 if incubated 210.0 if incubated 60
60 min; min;
80.0 if incubated 230.0 if incubated 90
90 min min
Salvia officinalis L. UK/Leaf α-Humulene (32.5%), ledol isomers (9.4%), thujone 100.0 if 35.0% at 300 µg/mL if [6]
purpurea isomers (8.4%), camphor (4.8%) incubated 5 min; incubated 5 min;
140.0 if 220.0 if incubated 30
incubated 30 min;
min; 240.0 if incubated 60
180.0 if min;
incubated 60 200.0 if incubated 90
min; min
240.0 if
incubated 90 min

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
UK/Leaf α-Humulene (24.2%), thujone isomers (12.7%), 40.0 if incubated 140.0 if incubated 5 [6]
camphor (12.3%), ledol isomers (6.8%) 5 min; min;
30.0 if incubated 90.0 if incubated 30
30 min; min;
20.0 if incubated 74.0 if incubated 60
60 min; min;
30.0 if incubated 60.0 if incubated 90
90 min min
Salvia pachystachys Turkey/Aerial part β-Pinene (24.0%), α-pinene (12.2%), spathulenol 35.4% at 80 Nd at 80 µg/mL [106]
(10.4%), viridiflorol (7.7%), 1,8-cineole (6.5%), µg/mL
caryophyllene oxide (4.5%), limonene (4.3%)
Salvia potentillifolia Turkey/Aerial part α-Pinene (29.2% and 31.3%), β-pinene (14.9% and 21.9% at 200 105.0 [113]
Boiss. and Heldr. ex 14.6%), 1,8-cineole (7.4% and 7.3%) µg/mL
Bentham
Salvia Turkey/Aerial part Camphor (53.6%), 1,8-cineole (18.2 17.4%), cryptone 26.0 49.2% at 80 µg/mL [106]
pseudeuphratica (5.2%)
Salvia russellii Turkey/Aerial part β-Pinene (20.4%), 1,8-cineole (9.3%), α-copaene 43.9% at 80 24.7% at 80 µg/mL [106]
(8.7%), valeranone (8.7%), α-gurjunene (4.8%) µg/mL
Salvia tomentosa Mill. Bulgaria/Leaf Borneol (10.3%), β-pinene (9.0%), camphor (7.9%), 0.3 - [42]
α-pinene (6.0%), camphene (4.0%)
Salvia verticillata L. Turkey/Aerial part Germacrene D (36.6%), β-caryophyllene (7.6%), 20.4% for 400 1.8% for 400 µM [109]
ssp. amasiaca (Freyn hexadecanoic acid (6.7%), β-copaene (5.7%), µM
and Bornm.) Bornm. spathulenol (4.5%)
Salvia wiedemannii Turkey/Aerial part α-Pinene (36.2%), β-pinene (13.3%), 1,8-cineole 350.2 µM 380.1 µM [109]
Boiss. (14.2%), camphor (7.4%), p-cymene (5.4%),
camphene (4.3%)
Satureja thymbra L. Turkey/Aerial part Carvacrol (34.6%), -terpinene (22.9%), p-cymene 150.0 166.0 [114]
(13.0%), thymol (12.8%), thymyl methyl ether (4.4%)
Sideritis galatica Turkey/Aerial part β-Pinene (32.2%), α-pinene (23.0%), β-caryophyllene 618.0 632.0 [115]
Bornm. (16.9%), (Z)-β-ocimene (9.5%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Thymbra capitata (L.) Spain/Aerial part Carvacrol (69.4%), γ-terpinene (7.7%), p-cymene 75.1 µL EO/L - [103]
Cav. (7.1%), (E)-β-caryophyllene (4.0%)
Spain/Aerial part Carvacrol (75.5%), γ-terpinene (5.5%), p-cymene 62.2 µL EO/L - [103]
(6.6%)
Spain/Aerial part Carvacrol (70.4%), p-cymene (7.3%), γ-terpinene 67.6 µL EO/L - [103]
(7.0%), (E)-β-caryophyllene (4.1%)
Portugal/Aerial part Carvacrol (68.1%), p-cymene (12.7%), γ-terpinene 51.9 - [16]
(6.1%)
Thymus caespititius Portugal/Aerial part α-Terpineol (40.3%), p-cymene (13.8%), γ-terpinene 5898.0 - [44]
Brot. (5.4%), τ-cadinol (5.2%)
Portugal/Aerial part α-Terpineol (35.2%), p-cymene (17.3%), γ-terpinene 4647.2 - [44]
(9.1%), τ-cadinol (6.2%)
Portugal/Aerial part α-Terpineol (23.5%), p-cymene (15.9%), γ-terpinene 1766.5 - [44]
(11.7%), τ-cadinol (6.9%)
Portugal/Aerial part α-Terpineol (51.5%), p-cymene (14.5%), γ-terpinene 12448.0 - [44]
(6.5%), τ-cadinol (6.2%)
Portugal/Aerial part α-Terpineol (40.5%), p-cymene (13.7%), γ-terpinene 3000.6 - [44]
(8.7%)
Portugal, Azores/Aerial Carvacrol (50.7%), carvacryl acetate (18.7%), p- 567.4 - [44]
part cymene (5.7%)
Portugal, Azores/Aerial Carvacrol (32.2%), thymol (23.0%), carvacryl acetate 181.4 - [44]
part (7.0%), p-cymene (5.9%)
Portugal, Azores/Aerial Carvacrol (61.9%), carvacryl acetate (11.5%) 158.9 - [44]
part
Portugal, Azores/Aerial Thymol (24.9%), α-terpineol (19.1%), p-cymene 3601.6 - [44]
part (11.5%), γ-terpinene (9.6%)

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Portugal, Azores/Aerial Thymol (34.8%), carvacrol (12.7%), p-cymene 139.1 - [44]
part (8.2%), thymyl acetate (7.9%)
Portugal/Aerial part α-Terpineol (24.1%), γ-terpinene (13.8%), p-cymene 2579.1 - [44]
(12.3%), γ-eudesmol (6.2%)
Portugal/Aerial part α-Terpineol (42.4%), p-cymene (13.5%), γ-terpinene 4219.5 - [44]
(6.2%)
Thymus camphoratus Portugal/Aerial part 1,8-Cineole (46.7%), linalool (12.2%), 182.0 - [44]
Hoffmanns. Thymastra linalyl acetate (8.8%), α -pinene (4.3%)
and Link Portugal/Aerial part 1,8-Cineole (26.5%), borneol (15.0%), α-pinene 123.5 - [44]
(12.3%), camphene (11.6%)
Portugal/Aerial part 1,8-Cineole (37.0%), α -pinene (10.1%), terpinen-4-ol 115.0 - [44]
(9.8%), borneol (4.3%)
Portugal/Aerial part Borneol (23.2%), camphor (19.1%), 195.3 - [44]
camphene (17.2%), linalool (9.5%)
Portugal/Aerial part 1,8-Cineole (21.3%), borneol (13.3%), α-pinene 137.1 - [16]
(11.9%), camphene (10.6%), camphor (8.1%)
Thymus capitellatus Portugal/Aerial part 1,8-Cineole (35.0%), borneol (16.2%), α-pinene 450.3 - [44]
Hoffmanns. and Link (12.4%), camphene (11.5%)
Portugal/Aerial part Borneol (20.3%), camphene (18.2%), 561.4 - [44]
camphor (17.8%), α-pinene (12.4%)
Portugal/Aerial part 1,8-Cineole (33.7%), borneol (16.9%), α-pinene 125.1 - [44]
(13.6%), camphene (11.2%)
Portugal/Aerial part Borneol (22.4%), 1,8-cineole (21.1%), 178.5 - [44]
camphene (16.9%), camphor (10.5%)
Portugal/Aerial part 1,8-Cineole (25.8%), borneol (21.0%), camphene 148.9 - [44]
(12.9%), α-pinene (11.1%)
Thymus carnosus Portugal/Aerial part Borneol (22.9%), camphene (21.1%), 130.1 - [44]
Boiss. terpinen-4-ol (11.1%), α-pinene (9.7%)

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Portugal/Aerial part Borneol (22.9%), terpinen-4-ol (18.3%), 117.9 - [44]
camphene (16.9%), bornyl acetate (6.0%)
Portugal/Aerial part Borneol (26.0%), camphene (18.7%), terpinen-4-ol 86.9 - [44]
(11.1%), bornyl acetate (10.2%)
Portugal/Aerial part Camphene (22.5%), borneol (20.1%), α-pinene 136.0 - [44]
(10.0%), bornyl acetate (9.6%)
Portugal/Aerial part Borneol (21.1%), camphene (19.8%), terpinen-4-ol 1022.9 - [44]
(13.6%), bornyl acetate (8.0%)
Portugal/Aerial part Borneol (24.8%), camphene (23.7%), bornyl acetate 159.0 - [44]
(9.5%), terpinen-4-ol (8.1%)
Portugal/Aerial part Borneol (20.2%), terpinen-4-ol (13.1%), camphene 721.7 - [16]
(11.4%), cis-sabinene hydrate (7.3%), γ-terpinene
(6.3%), α-thujene (5.1%), α-pinene (4.9%), trans-
sabinene hydrate (4.2%), bornyl acetate (4.1%)
Thymus mastichina Portugal/Aerial part 1,8-Cineole (49.4%), α-pinene (7.0%), camphene 45.8 - [16]
(6.9%), camphor (5.8%), β-pinene (5.3%)
Thymus haussknechtii Turkey/Aerial part Thymol (52.2%), p-cymene (16.6%), -terpinene 57.3% at 25 40.1% at 25 µg/mL [116]
Velen. (6.6%), 1,8-cineole (4.6%) µg/mL
Thymus lotocephalus Portugal/Aerial part Linalool (10.4%), caryophyllene oxide (8.7%), 900.0 500.0 [117]
G. López and R. camphor (8.0%), borneol (5.6%), α-terpineol (4.5%)
Morales
Thymus serpyllum Portugal/Aerial part Carvacrol (56.0%), α-terpineol (4.9%) 190.0 - [88]
Thymus spathulifolius Turkey/Aerial part Thymol (50.5%), borneol (16.7%), carvacrol (7.7%), 950.0 1050.0 [118]
Hausskn. et Velen β-caryophyllene (6.1%), 1,8-cineole (4.2%)
Thymus vulgaris L. Croatia/Aerial part Thymol (35.3%), p-cymene (34.7%), terpinene 540.0 - [119]
(8.4%), carvacrol (5.8%)
provided by Zaraphyt p-Cymene (24.0%), carvacrol (16.0%), borneol 216.9 - [32]
(Rabat, Morocco)/Aerial (16.0%), thymol (12.0%)
part

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Thymus zygis Loefl. ex Portugal/Aerial part Carvacrol (31.8%), p-cymene (23.5%), 980.3 - [44]
L. ssp. Sylvestris γ -terpinene (7.7%), borneol (6.4%)
(Hoffm. and Link) Portugal/Aerial part p-Cymene (35.9%), thymol (24.2%), γ-terpinene 1352.5 - [44]
Brot. Ex Couthino (7.1%), borneol (6.4%)
Portugal/Aerial part Carvacrol (34.6%), p-cymene (24.6%), 1479.3 - [44]
borneol (9.7%), camphene (5.3%)
Portugal/Aerial part p-Cymene (39.4%), thymol (21.6%), γ-terpinene 1766.6 - [44]
(10.7%), linalool (4.3%)
Thymus zygis Loefl. ex Portugal/Aerial part Carvacrol (43.6%), p-cymene (24.1%), 1.1 - [44]
L. ssp. zygis γ-terpinene (15.8%)
Zataria multiflora Iran/Aerial part Thymol (37.6%), carvacrol (33.7%), p-cymene (7.7%) 0.97 - [51]
Boiss.
LAURACEAE
Aniba canelilla Brasil/Trunk wood 1-Nitro-2-phenylethane (70.2%), methyl-eugenol Detection limit - [120]
(H.B.K.) Mez (25.8%) 0.01 ng/spot
Beilschmiedia glabra Leaf β-Eudesmol (15.4%), β-selinene (12.2%), 48.1% at 1000 - [121]
caryophyllene oxide (8.1%), γ-gurjunene (5.2%) µg/mL
Bark β-Eudesmol (19.3%), β-selinene (16.9%), δ-cadinene 45.2% at 1000 - [121]
(15.8%), germacrene D (9.8%), β-caryophyllene µg/mL
(5.5%)
Beilschmiedia Malaysia/Leaf β-Caryophyllene (12.1%), germacrene B (11.2%), α- 42.8% at 95 - [54]
kunstleri cadinol (10.4%), τ-muurolol (7.2%), caryophyllene µg/mL
oxide (7.0%), δ-cadinene (5.9%), ledol (4.9%)
Malaysia/Bark δ-Cadinene (13.4%), β-caryophyllene (10.6%), α- 50.3% at 95 - [54]
cadinol (9.0%), germacrene B (8.5%), caryophyllene µg/mL
oxide (5.4%), dehyroaromadendrene (4.4%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Beilschmiedia madang Malaysia/Leaf δ-Cadinene (17.0%), β-caryophyllene (10.3%), α- 55.2% at 95 60.4% at 95 µg/mL [55]
Blume cubebene (11.3%), bicyclogermacrene (6.7%), α- µg/mL
cadinol (5.8%), (E)-nerolidol (5.0%), germacrene D
(4.7%), α-humulene (4.3%)
Malaysia/Bark δ-Cadinene (20.5%), β-caryophyllene (6.7%), α- 48.2% at 95 50.2% at 95 µg/mL [55]
cubebene (15.6%), α-cadinol (10.6%) µg/mL
Beilschmiedia Malaysia/Leaf β-Eudesmol (24.1%), caryophyllene oxide (11.0%), β- 66.6% at 95 - [54]
maingayi panasinsene (10.2%), α-ylangene (4.3%) µg/mL
Malaysia/Bark β-Eudesmol (17.5%), caryophyllene oxide (12.8%), α- 53.3% at 95 - [54]
eudesmol (12.2%), β-panasinsene (11.6%) µg/mL
Beilschmiedia Malaysia/Leaf δ-Cadinene (28.7%), germacrene D (20.7%), β- 51.8% at 95 - [54]
penangiana caryophyllene (10.4%), α-copaene (7.7%), germacrene µg/mL
B (5.9%)
Malaysia/Bark δ-Cadinene (17.5%), germacrene D (14.6%), β- 47.1% at 95 - [54]
caryophyllene (12.6%), germacrene B (10.7%), µg/mL
viridiflorol (8.0%), τ-muurolol (7.5%), α-guaiene
(4.4%)
Beilschmiedia Malaysia/Leaf and stem Eugenol (45.3%), eugenol acetate (5.6%) 56.5% at 95 48.2% at 95 µg/mL [53]
pulverulenta Kosterm bark µg/mL
Cinnamomum purchased from Thai- 1,8-Cineole (39.9%), limonene (23.7%), o-cymene 60.6% at 120 - [47]
camphora (L.) J.Presl China Flavour and (9.1%), α-terpineol (5.0%), α-pinene (4.9%) µg/mL
Fragrances Industry Co.,
Ltd. and Perfumes world,
(Bangkok, Thailand)/
Leaves

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Cinnamomum griffithii Malaysia/Leaf Methyl eugenol (38.5%), safrole (6.4%), p-cymene 42.5% at 95 50.4% at 95 μg/mL [52]
Meisn. (4.9%), caryophyllene oxide (4.8%) μg/mL
Malaysia/Bark Methyl eugenol (43.8%), safrole (7.0%), 52.2% at 95 63.2% at 95 μg/mL [52]
aromadendrene (4.5%) μg/mL
Cinnamomum Malaysia/Leaf Safrole (59.5%), methyl eugenol (11.1%) 25.8% at 95 36.3% at 95 μg/mL [52]
macrocarpum Hook.f. μg/mL
Malaysia/Bark Safrole (54.5%), methyl eugenol (12.0%), 55.8% at 95 66.1% at 95 μg/mL [52]
aromadendrene (5.2%) μg/mL
Cinnamomum India/Leaf 2-Methoxy-3-(2-propenyl)-phenol, (74.7%), 8.3 μL/mL 3.3 μL/mL [122]
zeylanicum Blume caryophyllene (8.5%)
MYRTACEAE
Callistemon viminals Egypt/Leaf 1,8-Cineole (71.8%), α-pinene (11.5%) 290.2 - [23]
(Sol.ex Gaertn.) G. Egypt/Leaf 1,8-Cineole (71.8%), α-pinene (11.5%) 28.5 - [15]
Don
Eucalyptus Burkina Faso/Leaf 1,8-Cineole (33.9%), α-pinene (12.5%), p-cymene 19.0 - [45]
camaldulensis (12.3%), limonene (11.5%)
Dehnhardt
Eucalyptus globulus provided by Zaraphyt Limonene (55.0%), 1,8-cineole (38.0%) 129.8 - [32]
Labill. (Rabat, Morocco)/Aerial 1,8-Cineole (83.3%), α-pinene (6.8%), limonene 0.21 - [47]
part (4.6%)
purchased from Thai-
China Flavour and
Fragrances Industry Co.,
Ltd. and Perfumesworld,
(Bangkok, Thailand)/
Branches and leaves
Eugenia hiemalis Brazil/Leaf Spathulenol (5.4–16.1%), δ-cadinene (7.5–15.9%), Nd - [58]
Cambess. bicyclogermacrene (5.7–14.2%), β-caryophyllene at 1000 μg/mL
(4.8–9.4%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Eugenia pruniformis Brazil/Leaf β-Caryophyllene (46.9%), bicyclogermacrene 1798.0 - [60]
Cambess. (14.9%), globulol (8.9%), α–copaene (5.5%), α–
humulene (4.4%)
Eugenia riedeliana O. Brazil/Leaf Valerianol (28.1%), 10-epi-γ-eudesmol 67.3 - [62]
Berg (12.6%), β-caryophyllene (10.9%), α-
selinene (6.1%), δ-cadinene (5.6%), β-selinene (5.2%)
Eugenia sulcata Spring Brazil/Leaf β–Caryophyllene (24.6%), α–pinene (17.2%), β- 4.7 - [123]
ex Mart. pinene (10.9%), 1,8-cineole (5.6%), α–humulene
(5.1%), trans-calamenene (4.4%), γ-cadinene (4.3%)
Marlierea racemosa Brazil/Leaf Spathulenol (25.0%), muurola-4,10(14)-dien-1β-ol 35.0% at 600 - [63]
(13.2%), 14-hydroxi-9-epi-(E)-caryophyllene (5.5%) μg/mL
Brazil/Leaf Spathulenol (31.9%), p-mentha-1,5-dien-8-ol (6.6%), 65.2 - [63]
α-pinene (4.6%)
Melaleuca alternifolia Yamamoto Perfumery Terpinene-4-ol (35.6%), γ-terpinene (19.5%), α- 51.2 - [57]
Cheel Co. Ltd (Osaka, Japan) terpinene (8.3%), p-cymene (7.2%), 1,8-cineole
(4.4%)
supplied by Pranarom Terpinene-4-ol (46.9%), γ-terpinene (22.5%), α- 3.5 𝜇L/mL - [56]
terpinene (10.3%), p-cymene (5.9%)
Melaleuca cajuputi purchased from Thai- 1,8-Cineole (70.2%), α-terpineol (10.1%), limonene 0.63 - [47]
Powell China Flavour and (5.5%)
Fragrances Industry Co.,
Ltd. and Perfumes world,
(Bangkok,
Thailand)/Leaves

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Myrceugenia Brazil/Leaf α-Pinene (23.2%), limonene (18.3%), α-phellandrene 28.9% at 600 - [62]
myrcioides (Cambess.) (6.9%), (Z)-2-decenal (6.6%), spathulenol (6.2%), 1- μg/mL
O. Berg hydroxyundecan-3-one (5.9%), β-pinene (4.2%), o-
cymene (4.2%)
Myrciaria floribunda Brazil/Leaf 1,8-Cineole (38.4%), γ-himachalene (7.0%), α– 681.0 - [61]
(H.West ex Willd.) O. terpineol (5.5%), zonarene (4.6%), δ-amorphene
Berg (4.0%)
Brazil/Stem (2E,6E)-Farnesyl acetate (19.9%) (2E,6Z)-farnesol Nd at 1000 - [61]
(13.1%), linalool (7.0%), γ–himachalene (5.9%), μg/mL
zonarene (5.0%), (E)-caryophyllene (4.7%), selin-11-
en-4-α-ol (4.6%), selina-3,7(11)-diene (4.5%), δ-
amorphene (4.2%)
Brazil/Flower 1,8-Cineole (22.8%), (2E,6Z)-farnesol (16.1%), 1583.0 - [61]
(2E,6E)-farnesyl acetate (13.4%), linalool (12.7%),
(Z)-β–ocimene (7.6%), α–terpineol (5.4%)
Neomitranthes obscura Brazil/Leaf cis-Nerolidol (19.3%), trans-nerolidol (17.1%), β- 75.9 - [59]
(DC.) N. Silveira bisabolene (11.7%), (Z)-caryophyllene (7.5%),
dehydro-aromadendrene (7.2%), cis-eudesma–6,11–
diene (5.7%), β–longipinene (5.5%), (E)-
caryophyllene (5.5%)
Syzygium aromaticum Nigeria/Bud Eugenol (85.6%), β-caryophyllene (5.8%) 0.01 µL/mL 0.02 µL/mL [12]
(L.) Merrill and Perry India/Flower bud Eugenol (0.5 𝜇g/mL) 49.7 88.1 [64]
NELUMBONACEAE
Nelumbo nucifera Pakistan/Seed 1,8-Cineole (25.6%), α-terpeneol (11.1%), α-asarone 64.0% at 100 58.0% at 100 µL/mL [65]
(10.3%), borneol (7.7%), γ-gurjunene (6.8%), µL/mL
camphene (5.1%), bicycle-germacrene (5.1%),
geraniol (4.3%), α-pinene (4.2%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
PINACEAE
Pinus brutia Ten. Turkey/Needle β-Pinene (47.5%), α-pinene (17.0%), germacrene D 28% at 200 25% at 200 𝜇g/mL [66]
(7.5%), β-caryophyllene (4.6%), α-terpineol (4.4%) 𝜇g/mL
Turkey/Twig β-Pinene (19.2%), α-pinene (14.5%), δ-3-carene 3% at 200 1% at 200 𝜇g/mL [66]
(14.2%), myrcene (11.0%), β-caryophyllene (9.7%), 𝜇g/mL
germacrene D (5.5%)
Pinus halepensis Mill. Turkey/Needle β-Pinene (46.8%), α-pinene (18.4%), β-caryophyllene 58% at 200 52% at 200 𝜇g/mL [66]
(9.2%), germacrene D (8.8%) 𝜇g/mL
Turkey/Twig β-Pinene (18.7%), limonene (18.7%), α-pinene 83.9% at 200 80.6% at 200 𝜇g/mL [66]
(16.4%), -3-carene (16.3%), β-caryophyllene (9.5%) 𝜇g/mL
Pinus heldreichii Italy/Needle α-Pinene (24.2%), β-pinene (8.4%), limonene (7.8%), 51.1 80.6 [68]
Christ ssp. α-cubebene (7.6%), terpinolene (5.9%), trans-
leucodermis. (Antoine) caryophyllene (4.5%)
E. Murray
Pinus nigraArnold Turkey/Needle α-Pinene (44.2%), β-pinene (21.3%), germacrene D 41% at 200 22% at 200 𝜇g/mL [66]
(16.7%), β-caryophyllene (4.9%) 𝜇g/mL
Turkey/Twig α-Pinene (69.5%), β-pinene (8.5%) 22% at 200 36% at 200 𝜇g/mL [66]
𝜇g/mL
Pinus nigra Arnold Italy/Needle α-Pinene (24.6%), β-pinene (10.9%), γ-cadinene 101.5 128.0 [68]
var. calabrica C. K. (9.9%), β-phellandrene (6.3%), manoyl oxide (6.2%),
Scheid trans-caryophyllene (4.2%), (Z)-β-ocimene (4.0%)
Pinus nigra Arnold Croatia/Needle α-Pinene (24.4%), β-pinene (16.0%), germacrene D 42.7 - [67]
ssp. dalmatica (Vis.) (14.6%), β-caryophyllene (9.6%)
Franco
Pinus nigra Arnold Italy/Needle α-Pinene (25.3%), limonene (22.6%), sabinene 94.4 162.5 [68]
ssp. nigra (12.8%), α-terpineol (8.3%), β-pinene (4.8%),
terpinolene (4.5%)

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Pinus pineaL. Turkey/Needle β-Pinene (42.4%), α-pinene (15.1%), germacrene D 31% at 200 42% at 200 𝜇g/mL [66]
(10.4%), α-terpineol (6.3%), β-caryophyllene (5.5%), 𝜇g/mL
Turkey/Twig β-Pinene (29.9%), -3-carene (16.7%), α-pinene 46% at 200 32% at 200 𝜇g/mL [66]
(14.0%), myrcene (11.1%), β-caryophyllene (8.8%), 𝜇g/mL
germacrene D (4.7%)
Pinus sylvestris L. Turkey/Needle α-Pinene (39.1%), germacrene D (16.1%), β-pinene 8% at 200 25% at 200 𝜇g/mL [66]
(15.6%), β-caryophyllene (8.6%) 𝜇g/mL
Turkey/Twig α-Pinene (50.4%), β-caryophyllene (10.8%), 31% at 200 18% at 200 𝜇g/mL [66]
sandaracopimarinal (6.7%) 𝜇g/mL
PIPERACEAE
Piper nigrum L. Thailand/Fruit δ-3-Carene (30.8%), limonene (25.4%), (-)-β-pinene 5.97 - [69]
(12.2%), α-pinene (6.3%), (-)- caryophyllene oxide
(4.2%)
Piper aleyreanum C. Brazil/Aerial part β-Elemene (16.3%), bicyclogermacrene (9.2%), δ- Detection limit - [70]
DC. elemene (8.2%), germacrene D 10.0 ng/spot
(6.9%), β-caryophyllene (6.2%), spathulenol (5.2%)
Piper anonifolium Brazil/Aerial part Selin-11-en-4- α-ol (20.0%), β-selinene (12.7%), α- Detection limit - [70]
Kunth selinene (11.9%), α-pinene (8.8%) 0.01 ng/spot
Piper hispidum Sw. Brazil/Aerial part β-Caryophyllene (10.5%), α-humulene (9.5%), δ-3- Detection limit - [70]
carene (9.1%), α-copaene (7.3%), limonene (6.9%), 0.01 ng/spot
caryophyllene oxide (5.9%), β-selinene (5.1%)
POACEAE
Cymbopogon Tunisia/fresh leaf Limonene (24.2%), β-phellandrene (13.4%), - 670.0 - [71]
schoenanthus L. terpinene (9.6%), α-terpineol (9.1%), α-cadinene
Spreng. ssp. laniger (6.0%), α-eudesmol (5.5%), -patchoulene (5.3%),
(Hook) Maire et Weill junipercamphor (4.1%)
Tunisia/fresh leaf Limonene (27.3%), β-phellandrene (13.5%), - 260.0 - [71]
terpinene (21.2%), α-terpineol (9.1%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Tunisia/fresh leaf Limonene (24.6%), β-phellandrene (16.0%), α-terpineol 610.0 - [71]
(11.7%), -terpinene (8.4%), -cadinene (4.5%)
Tunisia/dried leaf Limonene (24.6%), β-phellandrene (15.7%), α-terpineol 520.0 - [71]
(9.6%), -terpinene (8.9%), α-eudesmol (5.1%), α-
cadinene (4.2%), α-selinene (4.1%)
Tunisia/dried leaf Limonene (23.5%), β-phellandrene (14.1%), -terpinene 440.0 - [71]
(11.3%), α-terpineol (10.1%), α-cadinene (5.5%), β-
eusdemol (5.3%)
Tunisia/dried leaf Limonene (22.1%), β-phellandrene (16.3%), α-terpineol 280.0 - [71]
(11.0%), -terpinene (7.4%), -cadinene (5.4%), β-
eusdemol (5.1%)
Tunisia/dried root Limonene (26.0%), β-phellandrene (15.9%), -terpinene 320.0 - [71]
(9.7%), α-terpineol (9.4%), -cadinene (4.9%), elemol
(4.3%), α-eusdemol (4.3%), valencene (4.1%)
Tunisia/dried root β-Eusdemol (14.2%), limonene (10.5%), β-phellandrene 270.0 - [71]
(8.2%), junipercamphor (8.2%), valencene (7.2%), α-
terpineol (6.8%), -cadinene (6.1%), elemol (4.6%),
germacrene B (4.5%), -terpinene (4.3%)
POLYGONACEAE
Polygonum hydropiper Pakistan/Leaf Decahydronaphthalene (38.3%), 1,2,3,6- 120.0 130.0 [73]
L. tetramethylbicyclo[2.2.2]oct-2-ene (36.3%), β-elemene
(6.8%)
Pakistan/Flower Caryophylene oxide (41.4%), β- caryophyllene epoxide 220.0 225.0 [73]
(18.2%), humulene oxide (16.1%), β-elemene (4.8%)
Polygonum minus Malaysia/Leaf; Stem; Dodecanal (24.5-59.5%), β-caryophyllene (8.9-19.6%), For all Nd at - [74]
Root decanal (10.3-11.6%) 10000 μg/mL

Complimentary Contributor Copy


FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
Rumex hastatus D. Pakistan/Aerial part 2,4,6-Trimethyloctane (28.2%), cyclooctanone (7.8%), 2- 32.5 97.4 [72]
Don (p-methylphenyl)-2-nitropropane (7.4%), trans-3-nonen-
2-one (6.3%), ethyl amyl carbinol (6.3%), 5-ethyl-2(5H)-
furanone (5.9%), trimethylacetic anhydride (5.9%), 5-
methyl-3-heptanol (5.6%)
ROSACEAE
Rosa damascena Mill. provided by “SEBAT Citronellol (30.6%), geraniol (27.9%), nerol (12.3%), 60.9% at 1000 μg/mL 51.1% at 1000 [75]
Rose Oil and nonadecane (7.3%) μg/mL
Essential Oils Ltd.”
located in Isparta
province (Turkey)
RUTACEAE
Citrus aurantifolia Italy/Peel Limonene (49.2%), β-pinene (14.1%), γ-terpinene (6.6%) 139.3 235.5 [124]
(Christm.) Swingle Egypt/Peel Limonene (40.2%), β-pinene (19.7%), α-citral (8.1%), γ- 105.8 - [23]
terpinene (6.3%)
Egypt/Peel Limonene (40.2%), β-pinene (19.7%), α-citral (8.1%), γ- 29.4 - [15]
terpinene (6.3%)
Thailand/Leaf Limonene (26.4%), L-camphor (19.9%), citronellol 140.0 42.0 [125]
(8.5%), o-cymene (5.8%), 1,8-cineole (5.3%), geranial
(4.2%)
Citrus aurantium L. Italy/Peel Limonene (65.8%) 147.5 266.6 [124]
Tunisia/Peel Limonene (87.5%) 2.94 mM - [126]
Citrus bergamia Risso Italy/Peel Limonene (38.1%), linalyl acetate (28.9%), γ-terpinene 161.6 243.6 [124]
and Poit. (7.3%), linalool (6.4%), β-pinene (5.4%)
Citrus limon (L.) Egypt/Peel Limonene (56.3%), β-pinene (8.8%), γ-terpinene (6.4%), 35.3 - [23]
Burm.f. α-citral (5.0%)
Egypt/Peel Limonene (56.3%), β-pinene (8.8%), γ-terpinene (6.4%), 20.2 - [15]
α-citral (5.0%)

Complimentary Contributor Copy


Table 1. (Continued)

FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
provided by Zaraphyt Limonene (99.0%) 849.9 - [32]
(Rabat, Morocco)/
Peel
Nigeria/Peel Limonene (53.1%), β-pinene (9.5%), borneol (5.6%), 0.2 μL/mL 0.2 μL/mL [77]
neral (4.7%), sabinene (4.2%)
Citrus medica L. cv. Italy/Peel Limonene (15.2%), citropten (12.6%), γ-terpinene 621.0 - [127]
Diamante (10.3%), 9,17-octadecadienal (9.3%), hexadecanoic acid
(5.2%), nerol (4.7%), geraniol (4.6%)
Italy/Peel Limonene, (35.4%), γ-terpinene (24.5%), geranial (5.5%), 171.3 154.6 [128]
neral (4.4%)
Italy/Peel Limonene (44.5%), γ-terpinene (26.2%) 298.8 Nd at 102 μL/mL [128]
Citrus sinensis (L.) Nigeria/Peel Limonene (92.1%) 2.6 2.6 [76]
Osbeck Nigeria/Seed Limonene (13.7%), tetradecanoic acid (11.9%), 3.5 3.5 [76]
hexadecanoic acid (10.6%), borneol (5.4%), 6-
hydroxyeicosane (5.3%), α-terpineol (4.6%), β-
caryophyllene (4.3%), 1,8-cineole (4.2%), 3-dihydroxy-2-
octadecene (4.0%)
Haplophyllum Saudi Arabia/Aerial trans-p-Menth-2-en-1-ol (19.2%), cis-p-menth-2-en-1-ol 26.7% at 80 μg/mL Nd at 80 μg/mL [129]
tuberculatum Juss. part (13.2%), myrcene (10.1%), δ-3-carene (8.8%), β-
phellandrene (6.9%), limonene (6.6%), cis-piperitol
(6.4%), piperitone (4.1%), trans-piperitol (4.0%)
VALERIANACEAE
Valeriana wallichii AYUS GmbH β-Asarone (88.8%) <10% at 1000 µg/mL - [9]
(Weinstrasse,
Bühl/Baden,
Germany)

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FAMILY/SPECIES Origin/Part used Major constituents of essential oil (percentages) AChE assay BuChE assay Ref
IC50 (µg/mL)* IC50 (µg/mL)*
VERBENACEAE
Lantana camara L. Brazil/Flower Germacrene D (23.7%), germacrene B (13.2%), β- 77.2% at 1000 µg/mL - [79]
caryophyllene (9.5%), longicyclene (8.4%), β-
sesquiphellandrene (4.9%), (E,E)-farnesene (4.9%)
ZINGIBERACEAE
Aframomum melegueta Nigeria/Seed α-Humulene (48.8%), 11.8 - [13]
(K. Schum) β-caryophyllene (32.5%)
Nigeria/Stem Caryophyllene oxide (19.7%), myrtenyl acetate (14.7%), 15.3 - [13]
β-eudesmene (10.8%)
Nigeria/Leaf Myrtenyl acetate (29.1%), limonene (19.5%), γ-elemene 16.0 - [13]
(8.8%)
Nigeria/Rhizome Myrtenyl acetate (22.7%), pinocarvyl acetate (11.5%), 29.0 [13]
cyperene (9.0%), caryophyllene (6.0%)
Hedychium Portugal, δ-Cadinene (0-16.5%), (-)-cedreanol (15.2-16.3%), α- 1030.0-1370.0 - [81]
gardnerianum Azores/Leaf calacorene (1.1-16.2%) 3,4-dimethyl-3-cyclohexen-1-
Sheppard ex Ker-Gawl carboxaldehyde (9.0-10.5%), cadalin (5.0-6.4%), 9,10-
dehydroisolongifolene (4.8-5.6%), trans-α-bisabolene
(4.0-4.5%)
Zingiber officinale purchased from α-Zingiberene (34.0%), β-sesquiphellandrene (15.3%), β- 35.0% at 1000 µg/mL - [80]
Roscoe Sigma-Aldrich, bisabolene (9.9%), α-curcumene (8.7%), camphene
Germany/Rhizome (6.7%), α-farnesene (4.8%), β-phellandrene (4.1%)
*If IC50 (µg/mL) was not determined or other method or type of concentrations was used, the units describing cholinesterase inhibitory activity are given as stated
originally; Nd, not detected; EO, essential oil; AChE, acetylcholinesterase; BuChE, butyrylcholinesterase; Ref., reference number.

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54 Franko Burčul, Mila Radan, Olivera Politeo et al.

2. ESSENTIAL OILS WITH


CHOLINESTERASE INHIBITORY ACTIVITY
Table 1 summarizes the results of studies published from 1998. onward on
EOs that have been investigated on acetylcholinesterase (AChE) and
butyrylcholinesterase (BuChE) inhibition, beginning with the research from
Myazawa et al. including Mentha [2] and Citrus [3] species and Perry et al.
and Savelev et al. research on Salvia species [4-6]. The most authors used
Ellman method to evaluate cholinesterases (ChEs) inhibitory value and
expressed the results as IC50 (representing the concentration that achieved 50%
of the enzyme inhibition). The values for EO are ranked as following: <50
µg/mL excellent; <100 µg/mL very good; <500 µg/mL good. However, some
authors tested only one concentration with adequate inhibition percentage. On
the other hand, some authors used TLC autobiography method to evaluate
ChEs inhibition as a novel approach and expressed the results as the diameter
of the inhibition spot. The following part reviews the EOs obtained from the
species of different families.

Acoraceae

Acoraceae family comprises a single genus called Acorus. It includes 2-4


species native to Northern Hemisphere, but perhaps it was naturalized in
Europe and America [7].
Acorus calamus is the only species which rhizome essential oil was tested
for AChE inhibition. EO, with the dominant, phenylpropanoid compound type,
β-asarone (ca. 80%) showed excellent inhibition with IC50 at 10.7 µg/mL [8].

Altingiaceae

Altingiaceae family comprises a single genus called Liquidambar


including 13 species found in East Mediterranean, East Asia to Malaysia, and
Central America [7].
EO of Liquidambar orientalis, with phenylpropanoid compounds, trans-
cinnamyl alcohol and hydrocinnamyl alcohol, representing ca. 85% of the oil,
showed very weak AChE inhibitory effect [9].

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Anacardiaceae

The Anacardiaceae is a family of flowering plants including ca. 80 genera


with about 873 species found in tropics, but also in temperate area. They are
generally divided in two groups: Spondiadoideae and Anacardioideae.
Spondiadoideae group, found mostly in tropical parts, includes 20 genera (138
species) with the largest being Lannea (40) [7]. Economically important
species from this group include pantropical Spondias fruits, the marula of
Africa (Sclerocarya birrea), and the Neotropical fruits of Antrocaryon
which are restricted to localized cultivation and consumption. Anacardioideae
group, found largely in tropical, but also in temperate parts, includes 60 genera
(735 species) with the largest genera being Searsia (120), Semecarpus (72),
Mangifera (68), Ozoroa (40) [7]. Several genera of this family are of
economic importance including mango (Mangifera indica), pistachio
(Pistacia vera), cashew (Anacardium occidentale), and pink peppercorn
(Schinus terebinthifolia). EOs tested against AChE from this family include
Pistacia lentiscus, Schinus areira, and Schinus longifolia.
The sesquiterpene type EO from S. longifolia, having oxygenated
sesquiterpenes (globulol, viridiflorol, spathulenol, cadinol, cedrol) as dominant
compounds (over 40%) elicited excellent inhibition with IC50 at 20.0 μg/mL
[10]. The monoterpene type EO from S. areira leaf, having monoterpene
hydrocarbons (β-phellandrene, α-phellandrene, camphene, α-pinene, p-
cymene, sabinene representing ca. 55% of the EO), and sesquiterpenes (guaiol
and γ-muurolene representing over 10% of the EO) showed good AChE
inhibition with IC50 at 233.8 μg/mL. The monoterpene type EO from S. areira
fruit, having over 70% monoterpene hydrocarbons (α-phellandrene, β-
phellandrene, β-myrcene), showed weaker activity with IC50 at 487.9 μg/mL
[10].
Essential oil leaves of 14 Tunisian P. lentiscus populations, growing wild
in three bioclimatic zones, were found to be mostly mixed monoterpene and
sesquiterpene types with IC50 ranging from 55.3 to 483.7 μg/mL. The EO with
highest AChE inhibition comprises of 42.6% monoterpene hydrocarbons,
14.2% oxygenated monoterpenes, 35.0% sesquiterpene hydrocarbons, and
4.1% oxygenated sesquiterpenes. Two EOs having high content of
monoterpene hydrocarbons i.e., 64.5% and 57.0% showed IC50 at 383.3 and
483.7 μg/mL respectively, while EO having 70.4% sesquiterpene
hydrocarbons showed IC50 at 226.7 μg/mL [11].

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Annonaceae

The Annonaceae family is the largest family in the Magnoliales which


includes 129 genera with 2120 species found largely in tropical areas [7].
Several genera produce edible fruit, most notably Annona, Anonidium,
Asimina, Rollinia, and Uvaria. Only two EOs tested on AChE were found
including Monodora myristica and Xylopia aethiopica.
The EO from Xylopia aethiopica fruit having phenylpropanoid eugenol as
the main compound (35.0%) was tested against AChE and BuChE showing
excellent inhibition activities (0.02 and 0.03 μL/mL, respectively) [12]. The
EOs from M. myristica seed and stem having sesquiterpenes germocrene-D-4-
ol, and γ-cadinene as the main compounds (25.5, and 31.3%, respectively) also
showed excellent inhibition against AChE with IC50 14.9, and 15.6 μg/mL,
respectively [13].

Apiaceae

The Apiaceae family (also called Umbelliferae) commonly known as the


celery, carrot or parsley family, includes 434 genera with 3780 species [7].
They are found world-wide, though most species are concentrated in the North
temperate areas. Many of them are widely used as vegetables and spices,
including parsley (Petroselinum crispum), carrot (Daucus carota), celery
(Apium graveolens), parsnip (Pastinaca sativa), fennel (Foeniculum vulgare),
anise (Pimpinella anisum), dill (Anethum graveolens), coriander (Coriandrum
sativum), caraway (Carum carvi), and cumin (Cuminum cyminum). Among
five tested EOs from this family, F. lutea, F. carduchorum and F. vulgare
were tested only on AChE inhibitory activity, while F. communis was tested
only on BuChE, and C. maritimum was tested on both AChE and BuChE
inhibitory activity.
The strongest activity on AChE was obtained with EOs from F. lutea root
and F. carduchorum having IC50 28.6 µg/mL and 23.6 µL/mL, respectively.
Both EOs have monoterpene hydrocarbons as main constituents were δ-3-
carene was the major compound in F. lutea, with 72.6%, while (Z)-β-ocimene
and α-pinene represented more than 60% of the EO. Other EOs containing
monoterpene hydrocarbons as the main compounds i.e., γ-terpinene, limonene,
δ-3-carene and α-pinene showed good AChE inhibition. On the other hand F.
vulgare EOs containing phenylpropanoid trans-anethole as the main

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Cholinesterase-Inhibitory Activity of Essential Oils 57

compound (over 70%) reported controversial IC50 with values of 252.0 and
1187.7 µg/mL.
BuChE inhibitory activity of C. maritimum EO from Cyprus showed good
activity (59.8% at 121 µg/mL) with γ-terpinene and β-phellandrene as major
constituents (over 60%), while EO from Croatia showed weak activity (>25%
at 45.5 µg/mL) with monoterpene hydrocarbon limonene as the major
compound (57-74%). F. communis EO showed good inhibition but at very
high concentrations (>10000 µg/mL).

Asteraceae

The Asteraceae family (also called Compositae) comprises of 1620 genera


and 25040 species distributed worldwide and represents the largest
angiosperm family [7]. Many of the species are known for their widespread
usage such as oils (sunflower, Helianthus annuus L.; safflower, Carthamus
tinctorius L.), food (artichoke, Cynara cardunculus L; lettuce, Lactuca sativa
L.; tarragon, Artemisia dracunculus L.), medicine (sweet wormwood,
Artemisia annua L.; Indian fleabane, Pluchea indica (L.) Less.), industry
(absinthe, Artemisia absinthium L.; pyrethrum, Tanacetum spp.; stevia, Stevia
rebaudiana (Bertoni) Bertoni;), and ornaments (chrysanthemums, marigolds,
zinnias) [14].
The most information on EOs AChE and BuChE inhibition were reported
for species from this family and Artemisia species were the most studied.
Oxygenated monoterpene EOs, A. judaica (α- and β-thujone,
chrysanthenone >70%.), and A. macrocephala (α-thujone, 3-thujanone, 1,8-
cineol >80%) showed excellent activity with IC50 being 16.1 and 40.0 µg/mL,
respectively [15].
Sesquiterpene type EOs of Crassocephalum crepidioides (β-cubebene, α-
humulene) and A. maderaspatana (α-humulene, β-caryophyllene, α-copaene
>65%) showed excellent AChE activity with IC50 of 11.0 and 31.3 µg/mL,
respectively, while Dittrichia viscosa (trans-nerolidol, β-oplopenone, δ-
cadinene, τ-cadinol, α-cadinol, 10-epi-γ-eudesmol) showed weak activity
916.9 µg/mL [13, 16].
EOs of A. herba-alba (β-thujone, 39.8%), A. fragrans (α-thujone, 39.8%),
and A. annua (β-myrcene, 37.7%) having monoterpenes as the main
constituents showed AChE inhibitory activities with IC50 ranging from 165 –
1250 µg/mL. Monoterpene type EOs of Gynura bicolor (α-pinene, 61.4%) and
Inula graveolens (bornyl acetate, 54.0%) showed very good activity with IC50

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58 Franko Burčul, Mila Radan, Olivera Politeo et al.

92 and 270 µg/mL, while C. crepidioides (thymol, 43.9%, 4-


cyclohexabutyramide, 20.9%) showed excellent activity with IC50 of 12.2
µg/mL [13, 17-21].
Two EOs of A. annua having mixed monoterpene and sesquiterpene
composition (camphor 16.6%, β-caryophyllene, 16.3%; β-caryophyllene
oxide, 15.8% and β-caryophyllene, 19.4%; germacrene D, 18.1%; camphor
15.8%, respectively) showed contradictory AChE inhibitory activity with IC50
of 130 and 2920 µg/mL, respectively [19].
P. lanceolata (linalool, 32.2%; β-caryophyllene, 8.5; α-terpineol, 8.0%;
spathulenol 7.4%) and G. bicolor (β-caryophyllene, 31.4%; α-pinene, 17.1%)
having mixed type EOs showed excellent and very good AChE inhibitory
activity with IC50 of 2.5 and 85.0 µg/mL, respectively [18, 22].
A. dracunculus, EO having phenylpropanoid estragole as the major
compound (77-89%) showed controversial AChE inhibitory activity with IC50
ranging from 58-1900 µg/mL [21].
Unusual polyacetylene derivatives, capillene and its oxygenated form
capillin, were reported to be the major compounds in A. monosperma EO
which was tested as an insecticide against two different insect species. AChE
was obtained from these species (rice weevil - Sitophilus oryzae and red flour
beetle - Tribolium castaneum) and it was shown that A. monosperma EO
showed very good inhibitory activity with IC50 of 120.0 and 194.1 µg/mL,
respectively [15, 23].
Investigation of BuChE inhibitory activity were scarce in comparison to
AChE, the only one IC50 of 30.0 µg/mL was given for A. macrocephala (α-
thujone, 3-thujanone, 1,8-cineol >80%) showing excellent BuChE activity
[24]. Other EOs were tested at 100 and 200 µg/mL and the strongest EOs were
that from A. absinthum (12.8% at 100 µg/mL) and Centaurea lycopifolia
(65.8% 200 µg/mL) [20, 25].

Boraginaceae

The Boraginaceae family comprises of 94 genera with 1973 species,


which are largely found in north (warm) temperate areas, and some of them in
mountains in tropical areas [7]. There are some well-known members of this
family like alkanet (Alkanna tinctoria) which is used as a red dye and
starflower (Borago officinalis) known for its culinary and medicinal uses.
Only two EOs from this family were tested for ChE inhibition. EO from
Lycopsis orientalis with high percentage of long-chain hydrocarbons showed

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Cholinesterase-Inhibitory Activity of Essential Oils 59

relatively good inhibitory activity on both AChE (48.5% at 200 μg/mL) and
BuChE (43.7% at 200 μg/mL) [26].
EO from Cordia gilletii with high-molecular semivolatile constituents
such as phytol, hexadecanoic acid, (E)-2-hexenal and nonacosane (20.4%,
15.2%, 9.6% and 9.2%, respectively) as main compounds showed good
inhibitory activity on both AChE with IC50 of 105.6 μg/mL and BuChE with
IC50 of 369.3 μg/mL [27].

Buddlejaceae

According to Angiosperm Phylogeny Group - APG IV, Buddlejaceae


family is not recognized as distinct family [7, 28], on the other hand it is being
used as a distinct family by many scientists while awaiting clarifications and
rearrangements of the Lamiales order. It is classified as one of 7 major groups
within Scrophulariaceae family (order Lamiales) [7, 28] comprising 10 genera
with 145 species. Buddleja genus is the largest within the group with 125
species [7].
Only one EO, that of Buddleja asiatica Lour., was tested on both AChE
and BuChE inhibitory activity. B. asiatica EO with oxygenated monoterpene
1,8-cineole (38.1%) and oxygenated sesquiterpene β-sinensal (11.8%) as
major compounds showed excellent inhibitory activity on both AChE and
BuChE with IC50 of 5.2 and 27.9 μg/mL, respectively [29].

Burseraceae

The Burseraceae family comprises of 19 genera with 755 species which


grow mostly in tropical areas [7]. Only one genus (Boswellia) from this family
was found to be tested on AChE inhibitory activity. Boswellia spp. are very
well known for resins that the plant produces which is widely used as an
incense, mostly from B. sacra, B. carterii, B. frereana etc.
EOs from three endemic Soqotra Boswellia species were tested only on
AChE inhibitory activity at 200 μg/mL. B. socotrana having unusally high
percentage of (E)-2,3-epoxycarene (51.8%) showed the best activity with
inhibition of 59.3% [30].

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60 Franko Burčul, Mila Radan, Olivera Politeo et al.

Cistaceae

The Cistaceae family comprises of 8 genera with 175 species, growing in


Eurasia, North Africa, North America, southern South America, and especially
in the Mediterranean region. Some of the Cistaceae species are used in
perfume industry for their aromatic resin. Cistus essential oil is approved by
the Food and Drugs Administration (FDA) as a food additive and flavoring
agent and it may be used as a dietary supplement.
Five EOs from this family were tested on both AChE and BuChE
inhibitory activity. C. salvifolius having sesquiterpene type EO and C.
libanotis monoterpene type EO showed very good AChE inhibitory activity
with IC50 of 58.1 and 71.2 μg/mL, respectively [31]. In addition these EOs
showed excellent inhibition of BuChE with IC50 of 34.2 and 23.7 µg/mL,
respectively. Other three tested species also showed excellent (C. creticus) and
good (C. monspeliensis and C. villosus) BuChE inhibitory activity while they
did not show AChE inhibitory activity at 1000 µg/mL [31].

Cupressaceae

The Cupressaceae family comprises of 30 genera with 133 species,


growing especially in the Northern hemisphere and are more scattered in south
temperate regions [7]. Species from this family are famous for being the
largest (Sequoiadendron giganteum), tallest (Sequoia sempervirens) and
stoutest (Taxodium mucronatum) trees, as well as among the most massive and
long-lived ones (Fitzroya cupressoides).
Two species EOs from this family were tested on AChE inhibitory
activity. Cupressus macrocarpa (terpinen-4-ol, 20.3%; sabinene, 18.7%; β-
citronellol, 13.0%) showed excellent inhibitory activity with IC50 of 12.3
μg/mL, while C. sempervirens (α-pinene, 49.0%; limonene, 32.0%; δ-3-
carene, 18.0%) showed good activity with IC50 of 283.7 μg/mL [23, 32].

Euphorbiaceae

The Euphorbiaceae family comprises of 218 genera with 6745 species


growing pantropically as well in temperate (cool) areas [7]. Some of the plants
from this family have economic importance (tapioca or yuca, Manihot
esculenta; castorbean, Ricinus communis; Barbados nut, Jatropha curcas;

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Cholinesterase-Inhibitory Activity of Essential Oils 61

rubber tree, Hevea brasiliensis) while other have been used as violent
purgative (Croton tiglium) and liquid bandage (Croton lechleri) and other have
very well-known ornamental usage (Poinsettia or Christmas star, Euphorbia
pulcherrima).
EOs from three chemotypes of Croton zehntneri having (E)-anethole
(33.7, 70.5, 89.1%), eugenol (49.1, 84.2%), (Z)-methyl isoeugenol (53.4,
81.5%), stragole (90.2%) and (E)-methyl isoeugenol (91.5%) as major
compounds were tested on AChE inhibitory activity via TLC-autobiography
test at 2000 μg/mL. All EOs showed inhibition of AChE with exception of
EOs having stragole (90.2%) and eugenol (89.1%) as major constituents [33].

Fabaceae

The Fabaceae family (also called Leguminosae or Papilionaceae)


comprises of 766 genera with 19580 species growing World-wide [7], it is the
third largest angiosperm family after Asteraceae and Orhidaceae [7, 34]. Many
of the Fabaceae family members are widely used for food (bean - Vicia faba,
lentil - Lens culinaris, pea - Pisum sativum, peanut - Arachis hypogaea),
forage (alfalfa - Medicago sativa, clover - Trifolium spp., vetches - Vicia spp.,
Arachis spp., Albizia spp., Leaucaena spp.), industry production (natural
gums: tragacanth - Astragalus gummifer; gum arabic - Acacia senegal; guar
gum - Cyamopsis tetragonoloba; as well as dyes: indigo - Indigofera
suffruticosa, Natal indigo - Indigofera arrecta, yellow dye - Butea
monosperma), ornamental: Bauhinia forficata, Vachellia caven, Clianthus
puniceus. One of the reasons for such a wide usage is their ability to fixate
atmospheric nitrogen hence reducing the usage of fertilizers. Physostigmine a
well-known AChE and BuChE reversible inhibitors also called eserine was
found in West African calabar bean plant (Physostigma venenosum) which is a
member of Fabaceae.
Although widely used, only two species from this family were tested on
AChE activity: Bauhinia ungulata leaf and Peltophorum dasyrachis bark. B.
ungulata having more than 45% sesquiterpene type EO showed high inhibition
(96%) at 1000 μg/mL, while P. dasyrachis with more than 22% sesquiterpene
ar-turmerone and more than 27% of carboxylic acids showed very good AChE
inhibition with IC50 of 83.2 μg/mL [35, 36].

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Illiciaceae

According to Angiosperm Phylogeny Group - APG IV, Illiciaceae family


is not recognized as distinct family [7, 28], on the other hand it is being used
as a distinct family by some other systems. It is classified as a genus Illicium
within the Schisandraceae family (order Austrobaileyales) with 42 species,
growing from South East Asia to West Malaysia, as well as South East USA,
East Mexico, and Greater Antilles [7, 28].
Only one species, I. verum with anethole (correct isomer not reported)
being the major compound, was tested on both AChE and BuChE inhibitory
activity showing inhibitory activity on both enzymes with IC50 of 39.9 and
75.4 μg/mL, respectively [37].

Lamiaceae

The Lamiaceae family (also known as Labiatae) comprises of 236 genera


with 7173 species growing World-wide. The largest genera are Salvia (900),
Scutellaria (360), Stachys (300), Plectranthus (300), Hyptis (280), Teucrium
(250), Vitex (250), Thymus (220), and Nepeta (200) [7].
Generally, the first investigation of essential oils as AChE inhibitors was
conducted by Miyazawa et al. in 1998 on 31 sample of EO from 10 Mentha
species [2]. All samples showed inhibitory activity of AChE with IC50 ranging
from 26 to164 µg/mL with M. aquatica having the highest IC50 and M. gentilis
having the lowest, while EOs from M. rotundifolia and M. pulegium did not
reach 50% of inhibition at 164 µg/mL. EOs composition was found to be
dominantly of monoterpene type where some compounds i.e., carvone,
menthol, linalyl actetate, 1,2-epoxyneomenthyl acetate, pulegone, menthone,
piperitenone oxide, linalool constituted more than 50% of EO. Mentha spp.
EOs were also investigated using TLC-autobiography method by de Sousa
Barros et al. [38] by applying 20µg of EO onto a TLC plate. Some compounds
i.e., carvone, linalyl acetate, piperitenone oxide constituted more than 50% of
EOs tested. Several EOs having carvone and linalyl acetate as major
compounds showed contradictory results, suggesting possible synergistic
and/or antagonistic effects of other constituents of the EO.
Following Miyazawa et al. investigation, Perry et al. [4], investigated EO
from Salvia lavandulaefolia species from the largest genus of Lamiaceae
family, of which 23 EOs, with full EO composition reported, are represented
in Table 1, most of them (21 EO) were tested on both AChE and BuChE. S.

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Cholinesterase-Inhibitory Activity of Essential Oils 63

lavandulaefolia [4, 39], S. leriifolia [40], S. officinalis [41] and S. tomentosa


[42] EOs were reported to show extreme inhibitory activity on AChE having
IC50 from 0.03 to 3.0 µg/mL, with variable monoterpene composition. Other
EOs tested showed good inhibition of AChE in general and along with
monoterpenes some of them also contained notable percentage of
sesquiterpenes such as caryophyllene oxide, α- and β-caryophyllene,
germacrene D, spathulenol. Next to the AChE S. leriifolia showed extremely
strong inhibitory activity on BuChE as well with IC50 of 0.3 µL/mL. In
contrast to AChE inhibitory activity other EOs did not show good inhibitory
potential on BuChE, in general.
Orhan et al. reported a screening of 55 Turkish Salvia species (of which
28 were endemic ones) extracts (i.e., CH2Cl2, EtOAc, MetOH) all tested on
AChE at three different concentrations (i.e., 25, 50 and 100 µg/mL). Only S.
fruticosa CH2Cl2 extract showed good inhibitory activity having 51.1% of
inhibition at 100 µg/mL [43].
Thymus spp EOs were mainly reported on AChE inhibitory activity.
Although having monoterpene composition when compared to Mentha and
Salvia spp., Thymus spp. EOs did not show inhibitory potential on AChE.
Notable exceptions were T. zygis and T. mastichina having carvacrol and 1,8-
cineole as major compounds with IC50 of 1.1 and 45.8 µg/mL, respectively
[16, 44]. T. caespititius EOs having over 40% of combination of α-terpineol,
p-cymene and/or γ-terpinene showed weak inhibitory activity with IC50 values
significantly over 1000 µg/mL [44], while most of the EOs containing
carvacrol, thymol, 1,8-cineole, and/or borneol showed relatively good
inhibitory activity on AChE [16, 44].
Ocimum spp. were tested on AChE inhibition among which O. canum
(leaf) and O. gratissimum (leaf and seed) showed excellent activity with IC50
of 36.2, 6.5 and 6.7 µg/mL, respectively [13, 45]. The major component of the
EO of O. canum leaf was 1,8-cineole (59.9%), while EO of O. gratissimum
leaf and seed were γ-terpinene (52.9%) and α-pinene (48.2%), respectively. On
the contrary EO of the O. sanctum, with eugenol being the major compound
(59.0%) showed weak activity (with IC50 of 1600 µg/mL) [21]. All Origanum
spp. showed excellent potential on AChE inhibitory activity in general.
O. ehrenbergii and O. syriacum having similar mixed composition of
thymol (19.6 and 24.7%), p-cymene (16.1 and 8.7%), 2-isopropyl-1-methoxy-
4-methylbenzene (14.9 and 7.9%), γ-terpinene (11.8 and 12.6%) and carvacrol
(6.7 and 17.6%) were tested on both AChE and BuChE and showed excellent
activity on both enzymes having IC50 lower than 1.7 µg/mL [46]. O. majorana

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64 Franko Burčul, Mila Radan, Olivera Politeo et al.

having 1,8-cineole (63.5%) as the major compound, also showed excellent


inhibitory activity on AChE with IC50 of 0.24 µg/mL [47].
O. vulgare EO having pulegone and menthone as major compounds (77.5
and 4.9%) was tested on two different AChEs isolated from Tribolium
castaneum (Herbst.) and Sitophilus oryzae L. showing IC50 of 24.4 and 61.3
µg/mL [15, 23]. Although EO showed pronounced activity on both AChEs the
higher activity observed in case of T. castaneum indicates the difference
between two AChEs active sites.
Several authors reported inhibition of both AChE and BuChE by
Rosmarinus officinalis EO which were composed from 1,8-cineole, camphor,
α-pinene and borneol mixture representing over 50% of EO. Tested EOs IC50
values for AChE ranged from 20.8 to 498.9 µg/mL, while for BuChE the
range was from 29.5 to 924.2 µg/mL [23, 48, 49].
Notable activities were also reported for Zataria multiflora and Lavandula
dentata having thymol and carvacrol (over 70%) and camphor (61.4%) as
major compounds with IC50 values of 0.97 µg/mL and 9.7 µL/mL, respectively
[50, 51].

Lauraceae

The Lauraceae family comprises of 50 genera with 2500 species, growing


pantropically (temperate areas), lowlands to mountains [7].
EO from Cinnamomum zeylanicum, having 2-methoxy-3-(2-
propenyl)phenol (74.7%) as the major compound showed excellent inhibitory
against both AChE and BuChE with IC50 of 8.3 and 3.3 μL/mL, respectively.
C. griffithii and C. macrocarpum (leaf and bark) EOs having also
phenylpropanoid composition with safrole and methyl eugenol as the typical
major compounds, showed good inhibitory activity ranging from 25.8 to
55.8% for AChE and from 36.3 to 66.1% for BuChE at 95 µg/mL [52].
EOs from Beilschmiedia spp. were the most investigated from this family
having sesquiterpene type composition, although IC50 values were not reported
good AChE inhibitory activity can be observed. B. kunstleri, B. madang, B.
maingayi, B. penangiana and B. pulverulenta (leaf and bark) showed
inhibitory activity ranging from 42.8 to 66.6% at 95 µg/mL, while only two
species were tested for BuChE activity (B. madang and B. pulverulenta) and
showed similar activities [53-55].

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Myrtaceae

The Myrtaceae family comprises of 131 genera with 4620 growing


worldwide, mostly in tropical-warm temperate areas [7], a large number of
Myrtaceae species are found in the wet tropics, particularly South America,
Tropical Asia and Australia. Myrtaceae family include many economically
important species e.g., eucalyptus (Eucalyptus globulus), Bay rum (Pimenta
racemosa) and cajeput (Melaleuca cajuputi) which provide oils for the
perfume industry; clove (Syzygium aromaticum) and allspice (Pimenta dioica)
are important in the spice industry; eucalyptus, tea tree (Melaleuca
alternifolia), Callistemon and Leptospermum are known sources of antiseptic
oils. Flesh-fruited Myrtaceae are almost all edible and include economically
important fruits such as: guava (Psidium guajava) and rose apple (Syzygium
aqiueum), with many lesser known species which are locally important for
juices, sweets and jams i.e., jaboticaba (Myrciaria cauliflora) and pitanga
(Eugenia uniflora).
EOs from Eucalyptus globulus and Melaleuca cajuputi having high
content of 1,8-cineole (over 70%) showed pronounced activity against AChE
with IC50 of 0.21 and 0.63 and µg/mL, respectively [47].
EO from Callistemon viminals having 1,8-cineole (71.8%) as the major
constituent was tested on two different AChEs isolated from T. castaneum and
S. oryzae showed significantly different activity with IC50 of 290.2 and 28.5
µg/mL, respectively. These results emphasize the difference between two
AChE active sites even more than in the case of O. vulgare EO mentioned
previously [15, 23].
Species having monoterpene type EOs containing over 45% of mixture of
1,8-cineole, terpinen-4-ol, α-pinene and γ-terpinene (i.e., E. camaldulensis, E.
globulus, M. alternifolia, M. cajuputi, C. viminals) generally showed very
good AChE inhibitory activity with IC50 ranging from 0.21 to 290.2 µg/mL.
Only exception was Myrceugenia myrcioides EO which showed weak activity
[15, 23, 32, 45, 47, 56, 57].
Species with sesquiterpene type EOs (i.e., Eugenia hiemalis, E.
pruniformis, Marlierea racemosa and Myrciaria floribunda stem) in
comparison to monoterpene type EOs generally did not show good AChE
inhibitory activity except for the Eugenia riedeliana and Neomitranthes
obscura having IC50 of 67.3 and 75.9 µg/mL [58-63].
Syzygium aromaticum EO having phenylpropanoid eugenol as the major
compound (over 80%) was tested on both AChE and BuChE and showed
excellent activity on both enzymes [12, 64]. It is worth to emphasize that S.

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66 Franko Burčul, Mila Radan, Olivera Politeo et al.

aromaticum EO can be obtained in very high percentage up to 20% from the


buds, representing a good source of eugenol.

Nelumbonaceae

The Nelumbonaceae family (sometimes called the sacred lotus family)


includes single genus Nelumbo with two species N. lutea and N. nucifera [7],
growing in North America and tropical Asia, respectively.
EO from N. nucifera seed containing monoterpenes (1,8-cineole, α-
terpeneol, borneol, camphene, geraniol and α-pinene), sesquiterpenes (γ-
gurjunene, bicycle-germacrene), and phenylpropanoid (α-asarone) showed
very good activity on both AChE and BuChE having 64% and 58% inhibitory
activity at 100 µg/mL, respectively [65].

Pinaceae

The Pinaceae family comprises of 11 genera with 231 species growing in


North temperate regions [7]. Most of the species are trees or shrubs, including
very well-known conifers many of which have commercial importance such
as: cedars (Cedrus spp.), firs (Abies spp.), hemlocks (Tsuga spp.), larches
(Larix spp.), pines (Pinus spp.) and spruces (Picea spp.).
From this family, only EOs from nine Pinus spp. needle and/or twigs,
were tested for ChE activity. All EOs contained α- and β-pinene as dominant
components and showed some activity on both enzymes at maximum
concentration of 200 µg/mL [66-68].
P. nigra Arnold spp. dalmatica (Vis.) Franco and P. helderichii Christ ssp.
leucodermis (Antoine) E. Murray showed the best activity for AChE with IC50
of 42.7 and 51.1 µg/mL respectively, while P. helderichii Christ ssp.
leucodermis (Antoine) E. Murray showed the best activity for BuChE with
IC50 of 80.6 µg/mL [67, 68].

Piperaceae

The family Piperaceae (also known as the pepper family) comprises of 5


genera with 3615 species growing pantropically [7]. Piperaceae spp. may be

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small trees, shrubs or herbs and the best know species is Piper nigrum L.
which yields the most peppercorns and is used as a spice.
P. nigrum monoterpene (δ-3-carene, limonene, β-pinene, α-pinene, over
70%) EO type showed excellent activity on AChE with IC50 of 5.97 µg/mL
[69]. Three other Piper spp. (P. aleyreanum, P. anonifolium, P. hispidum) EOs
highly represented by sesquiterpenes i.e., selin-11-en-4-α-ol, β-elemene, β-
selinene, α-selinene, bicyclogermacrene, β-caryophyllene, α-humulene, and δ-
elemene) were tested for AChE activity via TLC autobiography method where
P. anonifolium and P. hispidum showed 100 times better activity than
phystostigmine which was used as control sample [70].

Poaceae

The Poaceae family (also known as grasses) comprises of 707 genera with
11337 species growing worldwide [7]. The Poaceae family includes the cereal
grasses, bamboos and the grasses of natural grassland and cultivated lawns as
well as pastures.
EO from only one species was reported i.e., Cymbopogon schoenanthus L.
Spreng. ssp. laniger (Hook) Maire et Weill to have inhibitory activity on
AChE with IC50 ranging from 260 to 670 µg/mL [71].

Polygonaceae

The Polygonaceae family (also known as knotweed, smartweed or


buckwheat family) comprises of 55 genera with 1110 species growing
worldwide [7]. Several species are cultivated as popular ornamentals and
vegetables such as: Sea grape (Coccoloba uvifera) used as fruit, seeds from
Fagopyrum provide grain for dark flour, rhubarb (Rheum rhabarbarum)
petioles and sorrel (Rumex acetosa) leaves are used as vegetables.
Three species from this family were investigated on ChEs activity
Polygonum minus (leaf, stem, root), Polygonum hydropiper (leaf, flower) and
Rumex hastatus. EOs of P. hydropiper (leaf, flower) and R. hastatus were
tested on both AChE and BuChE.
R. hastatus EO having not common EO constituents with 2,4,6-
trimethyloctane (28.2%) as major compound, showed excellent activity on
AChE with IC50 of 32.5 µg/mL and very good activity for BuChE with IC50 of
97.4 µg/mL [72].

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68 Franko Burčul, Mila Radan, Olivera Politeo et al.

P. hydropiper leaf (decahydronaphthalene, 38.3%; 1,2,3,6-


tetramethylbicyclo[2.2.2]oct-2-ene, 36.3%) and flower (caryophylene oxide,
41.4%; β-caryophyllene epoxide, 18.2%; humulene oxide, 16.1%) EOs
showed good activity with IC50 of 120 and 220 µg/mL for AChE and 130 and
225 µg/mL for BuChE, respectively [73].
P. minus was tested for AChE activity and showed no activity at
concentration of 10000 µg/mL [74].

Rosaceae

The family Rosaceae (also called rose family) comprises of 90 genera with
2520 species growing worldwide, but especially in Northern hemisphere, often
not deserts or tropical rainforest [7]. The family includes herbs, shrubs, and
trees as well as some economically important products such are edible fruits:
apples, pears, quinces, apricots, plums, cherries, peaches, raspberries, loquats,
and strawberries, almonds, and ornamental trees and shrubs such as: roses,
meadowsweets, photinias, firethorns, rowans, and hawthorns.
Rosa damascena EO is the only one that was tested for both ChEs and
showed weak activity on both enzymes, having monoterpene alcohols as major
compounds (representing over 70% of EO, i.e., citronellol, geraniol, nerol)
[75].

Rutaceae

The family Rutaceae (also known as rue and citrus family) comprises of
161 genera with 2070 species growing largely in tropical areas [7]. Species in
this family generally contain herbs, shrubs and trees with flowers having
strong scent. The most economically known genus is Citrus which includes:
orange, lemon, grapefruit, lime, mandarin, and kumquat.
Seven species were reported of which six belong to Citrus and one to
Haplophyllum genus and all of them were tested on both ChEs. EO from C.
paradisi is one of the earliest reports of ChE inhibition by EO constituents,
namely nootkatone and auraptene showing 17-24% AChE inhibition at
concentration of 1.62 μg/mL [3]. Citrus EOs generally have limonene as the
major constituent ranging from 13.7% to 99.0% along with other
monoterpenes such as: β-pinene, γ-terpinene and others having excellent
activity reported for both ChEs in general [15, 23, 76, 77].

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EOs from C. aurantifolia and C. limon having limonene over 40% as the
major constituent were tested on two different AChEs isolated from T.
castaneum and S. oryzae showed difference in activity with IC50 of 105.8 and
29.4 µg/mL, which was significant, and 35.3 and 20.2 µg/mL, respectively.
These results suggest the similar conclusion for selectivity for different AChE
active sites as in the cases of Artemisia monosperma (Asteraceae), Origanum
vulgare (Lamiaceae), Callistemon viminals (Myrtaceae) EOs mentioned
previously [15, 23].

Valerianaceae

According to the APG system III the family Valerianaceae is now


considered as a part of Caprifoliaceae family [78]. The Caprifoliaceae family
comprises of 42 genera with 890 species growing largely in temperate and
warm temperate areas in the northern hemisphere, some mountain tropics
(except for Valerianoideae), but neither the Antipodes nor the Pacific [7].
The only species tested was Valeriana wallichii with β-asarone as the
main constituent (88.8%) and it showed weak activity with less than 10% at
1000 µg/mL on AChE [9]. These results are in contradiction with ones
reported for A. calamus (Acoraceae) [8].

Verbenaceae

The Verbenaceae family comprises of 31 genera with 918 species growing


pantropically (up to warm temperate areas), but mostly in New World [7].
Only Lantana camara EO was tested for AChE activity and showed
77.2% of inhibition at 1000 µg/mL [79].

Zingiberaceae

The Zingiberaceae family comprises of 46-52 genera with 1075-1340


species growing in sub-tropical areas, especially South East Asia-Malaysia [7].
Afromomum melegueta EOs (seed, stem, leaf, rhizome) having different
constituents, showed excellent activity against AChE with IC50 ranging from
11.8 to 29.0 µg/mL [13]. Hedychium gardnerianum and Zingiber officinale did
not show promising inhibitory activity [80, 81].

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70 Franko Burčul, Mila Radan, Olivera Politeo et al.

CONCLUSION
The essential oils with their main constituents represent adjuvant in the
treatment of neurological diseases such as Alzheimer’s due to the fact that they
show inhibition potential on ChEs which are target in treatment of such
diseases.
Most investigations on ChE inhibition were conducted on EOs obtained
from Lamiaceae (72 species), Asteraceae (25 species), Myrtaceae (14 species),
and Lauraceae (11 species) families. EOs which exhibited excellent inhibitory
activity on ChEs derive from: Lamiaceae (Lavandula dentata, Mentha
aquatica, M. arvensis, M. citrata, M. gentilis, M. spicata, Ocimum canum, O.
gratissimum, Origanum ehrenbergii, O. majorana, O. syracum, O. vulgare,
Rosmarinus officinalis, Salvia hydrangea, S. lavandulaefolia, S. leriifolia, S.
officinalis, S. pseudeuphratica, S. tomentosa, Thymus mastichina, T. zygis,
Zataria multiflora), following Asteraceae (Artemisia judaica, A.
macrocephala, A. maderaspatana, Asteriscus maritimus, Crassocephalum
crepidioides, Pluchea lanceolata), then Myrtaceae (Calistemon viminals,
Eucalyptus camaldulensis, E. globulus, E. sulcata, Melaleuca alternifolia, M.
cajuputi, Syzygium aromaticum), and Rutaceae (Citrus aurantifolia, C.
aurantum, C. limon, C. sinensis). Some other representatives of different
families that showed notable activity were: Acorus calamus (Acoraceae),
Xylopia aethiopica (Annonaceae), Buddleja asiatica (Buddlejaceae),
Cinnamomum zeylanicum (Lauraceae), Piper nigrum (Piperaceae).
Considering all the reports given in Table 1, it is clear that some EOs with
similar composition have different inhibitory effects against tested ChEs.
These investigations suggest that some constituents may have synergistic
effect on other components either in positive or negative manner. However
only few reports considered these effects which are quite reasonable in
complex mixtures such are EOs. In order to draw proper conclusions future
investigations should involve testing of those EOs that showed excellent
inhibitory activity using different combinations and ratios of the main
constituents. Such investigations of constituents should include monoterpenes
(i.e., 1,8-cineole, limonene, α-pinene, β-pinene, terpinene-4-ol, linalool, etc.),
sesquiterpenes (i.e., β-caryophyllene, α-humulene, caryophyllene oxide etc.),
phenylpropanoids (eugenol, β-asarone, etc.) and others.
Differences observed in the inhibitory activity between two studies
investigating the same EOs were, in some cases, very significant, (A.
macrocarpa, C. viminals, O. vulgare, C. limon, C. aurantifolia) towards two
AChEs isolated from different insects (S. oryzae, T. castaneum) which

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Cholinesterase-Inhibitory Activity of Essential Oils 71

indicates the importance of the AChE active site structure. Reported


composition of active EOs did not consider the stereochemistry of the
components present in the EOs which could account for the different activities
of some EOs with similar composition towards the same enzyme. When
analyzing the data from Table 1 it is notable that inhibitory activity towards
BuChE was scarcely investigated when compared to AChE.
Although there are many reports that include various types of EOs
(terpene, terpenoids, phenylpropanoids, and others) there are no reports
whatsoever of the so called mustard oils with isothiocyanates as the main
constituents known for their diversified and generally marked bioactivity.
These oils are characteristic for the Brassicaceae plant family which include
many species used in daily diet (cabbage, horseradish, cauliflower, turnip,
radish, etc.).
Considering all the above, the selection of the EO or possibly one or more
EO constituents which could be effectively used as alternative or
complementary to conventional treatments used in neurological diseases
represents a challenge in further research.

ACKNOWLEDGMENT
This work has been supported by Croatian Science Foundation grant
number IP-2014-09-6897.

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 3

PHYTOCHEMICAL FUNCTIONALIZED METAL


NANOCATALYST (AG, AU, FE, ZN AND PD)
FOR REMEDIATION OF ORGANIC DYES

Brajesh Kumar1,2,*, Kumari Smita2


and Brajendra Kumar3
1
Department of Chemistry, TATA College,
Kolhan University, Chaibasa, Jharkhand, India
2
Centro de Nanociencia y Nanotecnologia,
Universidad de las Fuerzas Armadas ESPE,
Av. Gral. Rumiñahui s/n, Sangolqui, Ecuador
3
Government Women Polytechnic College,
Bokaro Steel City, Bokaro, Jharkhand, India

ABSTRACT
In this chapter, we describe a simple, cost-effective and ecofriendly
approach for the fabrication of different metal nanoparticles (MNPs)
including silver, gold, iron, zinc and palladium by using different plant
phytochemicals as a potential reducer and stabilizers. It can be used as a
catalyst, photocatalyst, adsorbent or an alternative agent for removal of
different organic dyes. The kinetic enhancement of MNPs on
degradation/removal of dyes can possibly provide significant and

*
Corresponding author email: krmbraj@gmail.com; Tel: +91-6542243083; +91-8757618562.

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88 Brajesh Kumar, Kumari Smita and Brajendra Kumar

valuable insight for the engineering application of phytochemical


functionalized MNPs. This review article supports the environmental
protection and adequately attractive compared to other techniques.

Keywords: metal nanoparticles, phytochemicals, organic dyes, ecofriendly,


environment.

1. INTRODUCTION
For the last few decades, wastewaters containing dyes and organic
pollutants from various industries, factories, and laboratories discharged into
water reservoirs without any treatment and it represents an increasing
worldwide environmental hazard (Safavi and Momeni, 2012). These dyes are
being used in large quantities in several industries for different applications
such as textiles, papers, leathers, foodstuffs, cosmetics, laser materials,
xerography, laser printing, gasoline, additives, etc. In most of the cases, the
resultant by-products contain dyes and heavy metal ions or both (Sharma et al.,
2012). It has been estimated that over 15% of the total world production of
dyes is lost in their synthesis and dyeing process (Safavi and Momeni, 2012).
Most of these anthropogenic dyes are carcinogenic, harmful and reduce the
light penetration in aqueous systems. As a consequence, it causes a negative
effect on photosynthesis, harmful to human health and contributes a big share
to the overall imbalance of the ecosystem (Singla et al., 2014).
In recent years, several processes have been studied to reach partial or
complete degradation of organic pollutant compounds such as adsorption,
coagulation, biodegradation and chemical or photochemical degradation
(Wang et al., 2008). Although physical and chemical methods usually show
high dye-removal efficiencies, high operating costs are the main drawback due
to the large-scale application of these methods (Prasannan and Imae, 2013).
Furthermore, due to the high chemical stability of synthetic dyes, conventional
biological treatment using bacteria cannot remove the dyes efficiently (Huo et
al., 2013). So, it is necessary to develop new technologies enabling to use non-
toxic and easily available materials for the complete removal of the pollutant
from wastewater and favors the concept of environmental remediation using
natural material and its modified form. Therefore, a critical need in the field of
nanotechnology is the development of reliable and eco-friendly processes for
the synthesis of metallic nanoparticles.

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Phytochemical Functionalized Metal Nanocatalyst … 89

Nanoparticles (NPs) consist of atomic and molecular assemblies of


metals and inorganic semiconductor oxides or chancogenides although
macromolecules and aggregates of organic molecules having nanometer scale
(10-9 m) dimensions with shapes, sizes and surface properties that can often be
meticulously engineered (Alam et al., 2013). Heterogeneous photocatalytic
degradation process employing metal nanoparticles (MNPs) and solar light or
reducing agent has emerged as an efficient and a promising new route for the
degradation of these dyes and other toxic substances. Specially, noble metals
(Gold, Silver, Palladium etc.) exhibit photocatalytic activity upon irradiation
of light under UV-visible region due to its interband transition of electrons.
Therefore, the effective utilization of energy is limited, i.e., 3–5% of the total
solar energy (Ahmed et al., 2010).
Over the last one decade, various plant materials and their phytochemicals
have been exploited for the capability to bioreduce the metal salts or change in
valency with various global importance materials due to their availability, low
cost, and ecofriendliness over the conventional chemical/physical routes. The
plant phytochemicals contain various complex chemical substances of
different composition, including flavonoids, polyphenolic, carotenoids,
glucosinolates, alkaloids, glycosides, lipids, essential oils, etc., which are
found as secondary plant metabolite in one or more parts of the plants (Iravani,
2011; Njagi et al., 2010; Kumar et al., 2016a). The main aim of this review
chapter is to give a positive message that MNPs including silver (Ag), gold
(Au), iron (Fe), zinc (Zn) and palladium (Pd) NPs synthesized through
phytochemical approach are much safer, ecofriendly and further used for the
remediation of organic dyes, efficiently (Figure 1). It is very promising
technique to clean our environment using nature gifted material either in the
pure or modified form.

1
Figure 1. Scheme for the preparation and application of MNPs.

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90 Brajesh Kumar, Kumari Smita and Brajendra Kumar

2. CLASSIFICATION OF ORGANIC DYES


Dyes constitute a major class of organic compounds, which find several
applications in our daily life. In fact the art of applying color to fabric has been
known by mankind since 3500 BC, but the first use of synthetic dyes has been
discovered by WH Perkins in 1856. To better understand the complex
structure of organic dyes in terms of their treatment, a brief review of dyes in
general and their structures in particular are presented in Figure 2. Generally,
organic dyes contain two key components: the chromophores, delocalized
electron systems with conjugated double bonds that is responsible for
producing the color and the auxochromes, electron-withdrawing or electron-
donating substituents that intensify the color of the chromophore by altering
the overall energy of the electron systems. Usual chromophores are –C=C–, –
C=N–, –C=O, –N=N–, –NO2 and quinoid rings, while the auxochromes are –
NH3, –COOH, –SO3H and –OH groups. Depending on the chemical structure
or chromophore, a plentiful of different groups of dyes can be distinguished.
Each different dye is given a color index (CI) generic name determined by its
application characteristics and its color (Lam et al., 2012; Hunger 2003). The
classes of organic dyes and its applications are explained in Table 1.

1
Figure 2. The structural formula of different organic dyes.

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Phytochemical Functionalized Metal Nanocatalyst … 91

Table 1. Classification of organic dyes and its applications

Class Compounds Applications Ref.


Acid dyes Anionic and water soluble Dyes for wool, (Lam et al.,
(1st largest dye) Azo, anthraquinone or polyamide, silk, 2012; Christie
triarylmethane, azine, xanthene, modified acryl and to 2007; Hunger
nitro and nitroso compounds some extent for ink- 2003; Abrahart
jet printing, leather, 1977)
paper and cosmetics
Basic dyes Cationic and water soluble Dyes for modified
Diarylmethane, triarylmethane, nylon, modified
anthraquinone or azo compounds polyesters and
polyacrylonitrile
Reactive dyes Forming a covalent bond Dyes for cotton,
between dye and dying material wool, silk and nylon
(–OH, – NH or –SH groups)
Azo or metal complex azo
compounds, anthraquinone and
phthalocyanine compounds
Mordant dyes Fixed to the fabric by the Dyes for wool,
addition of a mordant leather, silk, paper
Azo, oxazine or triarylmethane and modified
compounds cellulose fibers
Direct dyes Anionic and water soluble Dyes for cotton,
(2nd largest dye) More than one azo bond or rayon, nylon and to
phthalocyanine, stilbene or some extent to
oxazine compounds leather and paper
(High affinity for
cellulose fibers)
Solvent dyes Non-ionic and water insoluble Plastics, varnishes,
Diazo, triarylmethane, inks, waxes and oils
anthraquinone and
phthalocyanine compounds
Disperse dyes Water insoluble Dyes for Cellulose
(3rd largest dye) Small azo or nitro compounds, acetate, polyester,
anthraquinones or metal complex polyamide and acryl
azo compounds
Vat dyes Water-insoluble Dyes for cellulose
Anthraquinones or indigoids, fibers, detergents,
stilbene, pyrazoles, coumarin and soaps, oils, paints,
naphthalimides groups fibers and plastics

Overall at present, azo dyes represent the largest class of organic dyes
listed in the CI (65 – 70% of the total dyes) and their relative share among
reactive, acid and direct dyes is even higher, it can be expected that they make
up the vast majority of the dyes discharged by textile processing industries
(Gupta and Suhas, 2009; Christie 2007). Anthraquinone dyes are the second
largest class, followed by triarylmethanes and phthalocyanines of the entries in
the CI. Moreover, reactive dyes are known to form a covalent bond with the
fiber in the dyeing process. This leads to favorable properties such as wash-

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92 Brajesh Kumar, Kumari Smita and Brajendra Kumar

fastness. However, the unfixed dye reacts with water to form hydrolyzed or
Oxo-dye intermediate that has lost its bonding capacity and thus cannot be
reused. Consequently, dye recovery is not an option with reactive dyes and the
treatment process must lead to final destruction or disposal of these organic
pollutants (Lam et al., 2012).

3. REMEDIATION OF ORGANIC COLORED DYES


The dye contaminant water or colored effluents coming out of the
industries are intensely colored and contaminated with high concentrations of
chemical oxygen demand (COD), suspended and dissolved salts and traces of
recalcitrant material. If improperly processed, these effluents not only
deteriorate the aesthetics of receiving waters and may hinder the penetration of
oxygen, but also pose significant threat to life forms upon hydrolysis of some
dyes in the wastewater to form toxic products (Tezcanli-Guyer and Ince,
2003). Effluents of textile dyeing processes are toxic and not only pollute
surface water but also groundwater systems. So, the remediation and
detoxification of dye molecules from industrial wastewater need to be suitable
treatments that produce harmless effluents and recyclable water before
discharging wastewater into natural water bodies and also essential for a clean
environment.

Figure 3. Selective pictures of phytochemical reduced nanoparticles with (a, b) and


without (a) phytochemical coating for the degradation of toxic organic dyes.

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Table 2. An overview of different MNPs and their role in organic dye degradation

Degradation (%)
Plant Processing Catalytic
Plant/Chemicals MNPs Organic dyes or rate of Ref.
Materials and Size reagents/Sunlight
reaction (k)
Leaf Mimosa pudica AuNPs 16 nm rhodamine B (Rh B) NaBH4 k = 0.6319 Devi et al.,
min-1 2015
Pogestemon AuNPs 10–50 nm MB NaBH4 k = 0.1758 min-1 Paul et al.,
benghalensis 2015
Cinnamomum Au/TiO2 8-20 nm MO Sunlight k = 0.346 h-1 Nick et al.,
tamala 2013
Coccinia grandis AgNPs 20-30 nm Coomassie Brilliant Fluorescent UV - Arunachalam
Blue G-250 light et al., 2012
Morinda tinctoria AgNPs 79-96 nm MB Sunlight 95.3% at 72 h Vanaja et al.,
2014
Mussaenda AgNPs 82-88 nm MO NaBH4 >50%, 45 mins Thivaharan
erythrophylla et al., 2016
Green, Oolong and FeNPs 40-50 nm MG - 81.6%, 75.6% Huang et al.,
black teas and 67.1% 2014
Green tea FeNPs 130-270 nm MB NaBH4 k = 0.0404 min-1 Lin et al.,
2015
Andean Blackberry FeNPs 54.5 ± 24.6 MB, CR, MO Sunlight k = 0.0105475, Kumar et al.,
nm 0.0043240, and 2016a
0.0028930 min-1
Grape leaf FeNPs 18-30 nm Orange II High temperature >92% Luo et al.,
2015
Cassia fistula ZnO NPs 5-15 nm MB UV and Sunlight >96% Suresh et al.,
2015
Plectranthus ZnO NPs 88 nm MR UV light 92.45%, k = Fua and Fu,
amboinicus 0.01421 min-1 2015

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Table 2. (Continued)

Degradation (%)
Plant Processing Catalytic
Plant/Chemicals MNPs Organic dyes or rate of Ref.
Materials and Size reagents/Sunlight
reaction (k)
Coleus aromaticus PdNPs 2.8 nm MO, MR, NaBH4 k = 0.1137, Vilas et al.,
Eriochrome black T, 0.1552, 0.2772, 2016
MB, Rhodamine B 1.0060, 0.3056
min-1
Myristica fragrans PdNPs 2.9 nm MO, MR, NaBH4 k = 0.1576, Vilas et al.,
Eriochrome black T, 0.0994, 0.4046, 2016
MB, Rhodamine B 1.2442, 0.1109
min-1
Andean Blackberry PdNPs 55-60 NM MB Sunlight >72%, k = Kumar et al.,
0.0021614 min-1 2015b
Plumeria alba AuNPs 28 ± 5.6 MB, Eosin Y, MR, NaBH4 - Mata et al.,
CR, Ethidium 2016
bromide
Flower Lantana camara AgNPs 33 ± 5 nm MB Sunlight k = 3.407 x 10-3 Kumar et al.,
min-1 2016b
Lantana camara AuNPs 10.6 ± 2.9 nm MB Sunlight >62% Kumar et al.,
2016d
Punica granatum AgNPs, 36 and MB, MO, Eosin Y NaBH4 83, 99, 96% MeenaKumari
AuNPs 18 nm 95, 94, 91% and Philip,
2015
Seed & Sacha Inchi Oil AgNPs 60 nm MB Sunlight 2.776 x 10−3 Kumar et al.,
Fruit min−1 2014a
Sacha Inchi Oil AuNPs 5–15 nm MB Sunlight 3.263 × 10−3 Kumar et al.,
min−1 2016e
Capsicum baccatum AuNPs 23.9 ± 9.7 nm MB Sunlight >50%, k = 1.9585 Kumar et al.,
x 10−3 min−1 2015a

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Degradation (%)
Plant Processing Catalytic
Plant/Chemicals MNPs Organic dyes or rate of Ref.
Materials and Size reagents/Sunlight
reaction (k)
Shora fruit AgNPs 10-30 nm MB Sunlight k= 0.0025293163 Vizuete et al,
Capparis petiolaris min-1 (>58%) 2016a
Mortino/Andean AgNPs 20.5 ± 1.5 MB Sunlight k= 0.00707788 Vizuete et al.,
Blueberry min-1 2016b
Garcinia ZnO-NPs 20–30 nm MB UV and Sunlight >75% Nethravathi
xanthochymus et al., 2015
Stem/Bark/ Terminalia cuneata AgNPs 25-50 nm Direct yellow-12 NaBH4 >95%, 40 mins Edison et al.,
Root and 2016
biomass Beet AgCl/Ag 100 nm MO Xe arc lamp Kou and
NPs Varma et al.,
2014
Cinchona species PdNPs 4 to 7 nm MB NaBH4 98.5% Trung
(Cinchonidine) et al., 2015
Sacha Inchi Shell AgNPs 7.2 nm MO Sunlight 60%, k = Kumar et al.,
biomass 0.0008898 min-1 2014b
Rambutan Peel Ag@TiO2 MB Sunlight k = 0.002495 min-1 Kumar et al.,
2016c
Citrus paradesi ZnO-NPs 12 to 72 nm MB Sunlight >56%, k = Kumar et al.,
biomass 0.002392 min−1 2014c
Tannic acid AgNPs and 10 nm MO NaBH4 0.5853 min−1 and Gupta et al.,
AuNPs 0.0049 min−1 2011
kAgNPs>> kAuNPs

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96 Brajesh Kumar, Kumari Smita and Brajendra Kumar

Photocatalytic treatment of the dye using Sunlight/solar radiation in the


presence of a suitable photocatalyst is the popular and economical method as
wastewater treatment over activated carbon for the degradation of dye
(Vinogdopal and Kamat, 1995; Muneer et al., 1997). In addition, to ensure full
use of the solar energy source, it is of great interest to develop photocatalysis
of organic dyes for expansion of the adsorption to the visible light range. In
this respect, Figure 3 and Table 2 summarizes the combination of solar light/
reducing agent and different metal nanoparticles (MNPs) synthesized using
non-toxic phytochemicals being clean, inexpensive and stable under ambient
condition would be ideal for use in green technology for the degradation of
organic dyes. The MNPs including silver (Ag), gold (Au), iron (Fe), zinc (Zn)
and palladium (Pd) have been used in engineering applications since first
colloidal syntheses of more than five centuries ago.

3.1. Silver Nanoparticles (AgNPs)

Metallic AgNPs exhibit photocatalytic activity upon irradiation of light


under UV- visible region due to its interband transition of electrons. Lantana
camara L. is a woody straggling plant with various flower colors, red, pink,
white, yellow and violet. Recently, we have synthesized silver nanoparticles
using an ethanolic extract of L. camara flower as both the reducing and
stabilizing agent. The synthesized silver nanoparticles are spherical, 33 ± 5 nm
average sized and appeared at 470 nm. It was found that the rate constant (k)
for the degradation of methylene blue (MB) was higher (k = 3.40736 x 10-3
min-1), when 1 mL of silver nanoparticles was used and optimized degradation
percentage was 70.20% for the concentration of 10 mg/L of MB in 6 h (Kumar
et al., 2016b). Vizuete et al., 2016a reported a rapid, facile and an ecofriendly
synthesis of silver nanoparticles (10-30 nm) at λmax = 423 nm by a rapid
reduction of silver ions using fruit extracts of Shora (Capparis petiolaris) and
sunlight. It showed that the Sunlight irradiation step is of prime importance
and more ecofriendly in comparison to pH, concentration or temperature
maintenance procedure for the rapid fabrication of silver nanoparticles.
Furthermore, as-synthesized nanoparticles showed photocatalytic activity for
the degradation of MB (>58%, 240 mins) and well fitted by a first order rate
law with degradation rate = 0.0025293163 min-1.
Sacha inchi (Plukentia volubilis L.) is a promising crop, which produces a
unique seed rich in oil omega 3,6,9 fatty acid (35 – 60%), proteins (27%) and
contain heat-labile substances with a bitter taste. We synthesized silver

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Phytochemical Functionalized Metal Nanocatalyst … 97

nanoparticles using Sacha inchi oil in the presence of sunlight and TEM
characterization revealed the nanostructure were highly dispersed, distorted
cubic in shape and 60 nm size. The stable and crystalline nanoparticles showed
photocatalytic activity in the decomposition of the MB (>65%) without using
any reducing agent and the calculated first-order rate constants were found to
be 2.776 x 10−3 min−1 (Kumar et al., 2014a). In another study, a new approach
for the synthesis of monodispersed silver nanoparticles was reported by using
cheaply available Sacha inchi shell biomass (SISB) and can be used as a
photocatalyst for the remediation of methyl orange (MO). The UV–vis
spectroscopy and Transmission electron microscopy (TEM) techniques
indicated the SPR at 420 nm (Figure 4a) corresponds to spherical shape with
7.2 nm sized silver nanoparticles. Further, the experimental evidence showed a
strong effect of the pH 2 for maximum removal of MO in the presence of
silver nanoparticles and sunlight (~60%, 5 h, 64 mg/L) from aquatic systems
the rate constant was found to be 0.0008898 min-1 (Kumar et al., 2014b).

Absorbance (a.u.)
4,0 (a) AgNPs
(b) AuNPs
3,5 (c) FeNPs
(d) ZnO-NPs
3,0 (e) PdNPs

2,5

2,0

1,5

1,0

0,5

0,0

300 400 500 600 700 800

Wavelength (nm)
Figure 4. UV-vis spectrum of different nanoparticles prepared using phytochemicals of
different sources.

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98 Brajesh Kumar, Kumari Smita and Brajendra Kumar

Arunachalam et al., 2012 presented a bioreductive synthesis of nano-sized


Ag particles using Coccina grandis leaf extract as a reducing as well as
capping agent. This work demonstrated the use of an ecofriendly and low cost
biological reducing agent to produce metal nanostructure in the size range of
20–30 nm and crystallized in face centered cubic symmetry in aqueous
solution at room temperature, avoid the input of hazardous and toxic solvents.
From the present study it was also found that the Ag nanoparticles exhibited
photocatalytic activity by degradation of Coomassie Brilliant Blue G-250
under UV light and can be used in water purification systems. Kou and Varma,
2014 explored a simple, green, and fast approach (complete within 5 min) for
the fabrication of hybrid AgCl/Ag plasmonic nanoparticles under microwave
(MW) irradiation. In this method, beet juice served as a reducing reagent,
which is an abundant sugar-rich agricultural product without using an
additional surfactant or reducing agent. Interestingly, the obtained AgCl/Ag
samples have a smaller size than the AgCl reaction precursor. This makes it an
unusual top-down hydrothermal synthesis. The as-prepared material displayed
good photocatalytic activity for the degradation of MO dye.
Recently, Thivaharan et al., 2016 reported a simple, instant and an
environmentally safe method for silver nanocatalyst synthesis using the leaf
extract of Mussaenda erythrophylla. It displayed a distinct peak at 414 nm
corresponding to the UV-visible spectrum of silver nanoparticle. SEM analysis
confirmed the formation sub-100 nm sized particles and EDAX proved the
existence of elemental silver in the sample. XRD analysis established the face-
centred cubic crystalline nature whereas zeta potential value of -47.7 mV,
defines the good stability of the silver nanoparticle solution. The synthesized
silver nanoparticles have remarkable catalytic activity and exploited to
degrade the MO using NaBH4 as a reductant. Vanaja et al., 2014 synthesized
AgNPs using Morinda tinctoria leaf extract under different pH conditions and
AgNPs were further characterized by different optical, microscopy and
spectroscopic techniques. They concluded that the size as well as the quantity
of the AgNPs formed is strongly dependent on the pH and basic pH supports
the biosynthesis of AgNPs whereas no AgNPs were detected in the acidic
medium. The spherical shape of the AgNPs with the size ranges from 79 to 96
nm were prepared and showed photocatalytic activity by degrading the MB
dye nearly 95% at 72 h of exposure time, efficiently.
Mortiño (Vaccinium floribundum Kunth) berry is an endemic fruit from
the Andes region and rich in vitamins, polyphenolic and anthocyanin
compounds. Although, green synthesis of nanoparticle is well known, but it
was the first report that describes the ecofriendly synthesis of silver

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Phytochemical Functionalized Metal Nanocatalyst … 99

nanoparticles using a berry extract of Mortiño (Vizuete et al., 2016b). All


characterization technique confirmed that the silver nanoparticles were stable,
non-aggregated, monodispersed, spherical shape with an average size of 20.5
± 1.5 nm and face centered cubic in nature. Further, It exhibited photocatalytic
activity for the degradation of methylene blue dye (5 mg/L, k = 0.00707788
min-1) in the presence of sunlight. Recently, we have synthesized silver-doped
titanium dioxide nanoparticles (Ag/TiO2 NPs) in an ecologically and
economically favorable way using rambutan (Nephelium lappaceum L.) peel
extracts along with the study of its photocatalytic activity on MB dye. A wide
range of analytical techniques including UV-vis spectroscopy, transmission
electron microscopy (TEM), dynamic light scattering (DLS), X-ray diffraction
(XRD), and Fourier transform infrared (FTIR) spectroscopy analysis were
performed to ascertain the synthesis of Ag/TiO2 NPs. The synthesized
Ag/TiO2 NPs enhanced the photocatalytic degradation of MB (81.6%,
k = 0.002495 min-1) under direct solar light irradiation. The results showed that
discarded agricultural waste as rambutan peel can be utilized as natural
bioreductant in future materials science applications (Kumar et al., 2016c).
Punica granatum or pomegranate fruit were characterized by its high
phenolic contents and antioxidant properties. MeenaKumari and Philip, 2015
presented first time biogenic reduction and stabilization of gold and silver ions
at room temperature using fruit juice of Punica granatum. The formation,
morphology and crystalline structure of the synthesized AgNPs and AuNPs
were determined using different analytical techniques, whereas FTIR
confirmed the partial role of phenolic hydroxyls in the reduction of Au3+ and
Ag+ to Ag0 and. The synthesized nanoparticles are used as potential catalysts
in the degradation of dyes and the rate constants from pseudo first order
kinetic data fit gives a comparative study on degradation of organic dyes in
presence of prepared Ag and AuNPs. The kinetic data plots fitted also suggest
the fast removal of MB in the presence of AuNPs and that of MO in presence
of AgNPs. It can be concluded that the catalytic activity for AuNPs is in the
order MB > MO > EY while that for AgNPs is as MO > EY > MB. Edison et
al., 2016 showed the efficacy of Terminalia cuneata bark extract synthesized
silver nanoparticles (AgNPs) in catalyzing the reduction of direct yellow-12.
The phytoconstituents (mostly tannins and poly phenols) present in the
extracts of T. cuneata act as reducing agents as well as capping agents
providing stability to AgNPs as evident from FT-IR study. The
phytosynthesized AgNPs were found to stable, have a crystalline structure
with face centered cubic geometry oriented in (111) plane and the size
ranging approximately 25-50 nm with distorted spherical shape. Complete

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100 Brajesh Kumar, Kumari Smita and Brajendra Kumar

decolorization of direct yellow-12 in presence of AgNPs and NaBH4 was


observed within 40 min after the reaction commenced.
Gupta et al., 2011 evaluated the nature of nanoparticles for the catalytic
degradation of MO in the presence of sodium borohydride (NaBH4). They
synthesized spherical and 10 nm size of Gold, silver and platinum (Pt)
nanoparticles by reducing the corresponding salt using tannic acid at room
temperature. The AgNPs have a drastic catalytic effect as compared to Au or
PtNPs on the degradation of MO and rate constant follows the order: kAgNPs>>
kAuNPs > kPtNPs > k uncatalyzed reaction. It is due to the effect of AgNPs has been
attributed to its low value of work function as compared to Au and Pt. Sodium
borohydride being such a strong reducing agent is not able to reduce methyl
orange in absence of catalyst, indicating the catalytic efficacy of metal
nanoparticles.

3.2. Gold Nanoparticles (AuNPs)

Gold nanoparticles display high potential for application in photocatalyst,


cell labeling, photothermal and drug delivery systems due to its biocompatible
properties. Kumar et al., 2016d investigated that the shape and size of gold
nanoparticles have been successfully controlled by introducing small amounts
of L. camara flower extract. The UV–vis absorption spectra for green
synthesized gold nanoparticles showed an absorption maximum, λmax at 530
nm (Figure 4b) corresponding to the transverse surface plasmon vibration. It
produced monodispersed and spherical nanogold of average size 10.6 ± 2.9 nm
without any aggregation and showed significant photocatalytic degradation
activity of the MB (>62%, 10 mg/L) in the presence of solar light. In addition,
the experimental approach is inexpensive, rapid, eco-friendly and can be used
as an alternative agent for remediation of MB from waste water. We have
also developed a simple and cost-effective methodology to obtain gold
nanoparticles using Sacha inchi (Plukenetia volubilis) oil in the presence
of sunlight. The spectroscopic and morphological properties of gold
nanoparticles revealed the surfaced plasmon resonance at 515–520 nm and is
almost spherical in shape with an average size of 5–15 nm. The as-synthesized
gold nanoparticles showed remarkable photocatalytic decomposition of the
MB (>75%) without using any reducing agent and sunlight exposure. The
experimental approach is promising and suggested that the sunlight is a good
source of energy for enhancement of AuNP synthesis via Sacha inchi oil and
its photocatalytic activity (Kumar et al., 2016e).

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Capsicum baccatum L. (Aji Amarillo) is a traditional vegetable crop in


Ecuador, Argentina, Bolivia, Brazil, Chile, Peru, Costa Rica, Hawaii, and is
also cultivated in many countries. It is believed that, Chilli peppers have been
a part of the human diet worldwide since at least 7500 BC. Kumar et al., 2015a
covers the importance of Andean Ajı′/Chilli (C. baccatum) mediated synthesis
of non-aggregated, spherical, and 23.9 ± 9.7 nm size gold nanoparticles at
λmax = 540 nm, which favors for green chemistry and escape us from the use
of hazardous chemicals. Further, as-synthesized gold nanoparticles, showed
enhanced photocatalytic degradation of methylene blue (>50%, k = 1.9585 ×
10−3 min−1) under direct solar light irradiation. Devi et al., 2015 reported
Mimosa pudica leaves extract mediated environmental benign synthesis
AuNPs in aqueous medium at room temperature. The synthesized particles are
spherical shape, monodispersed and average diameter is 16 nm. The ensuing
AuNPs were used as an efficient nanocatalyst in the degradation of
Rhodamine B in the presence of NaBH4 in aqueous medium at room
temperature. It showed that in the absence of a catalyst, the reactions occur
extremely slow but enhance abruptly upon addition of gold nanoparticles
indicating their first order catalytic efficacy (k = 0.6319 min-1). Such
significant catalytic behavior of the so-synthesis Au NPs can be attributed to
their large number of active sites for the reactant molecules to interact by
serving an electron relay effect.
Recently, Mata et al., 2016 synthesized two different sizes of AuNPs
using 1% and 5% concentrations of aqueous Plumeria alba flower extract. The
size-controlled formation of AuNPs showed surface plasmon resonance (SPR)
peaks at 552 and 536 nm corresponding to the spherical shape of nanoparticles
of 28 ± 5.6 and 15.6 ± 3.4 nm, respectively. Furthermore, the size dependent,
catalytic activity of both gold nanoparticles were analyzed on six hazardous
dyes and smaller sized AuNPs exhibited more pronounced catalytic activity
than larger sizes. Not only catalytic activity, but also the antibacterial activity
of the small-sized AuNPs exhibited better antibacterial activity with a 16-mm
zone of inhibition at a concentration of 400 μg/mL against Escherichia coli.
The mechanism of catalysis in the presence of both AuNPs can be described as
an electron transfer process from donor NaBH4 to an acceptor. Paul et al.,
2015 accomplished the biosynthesis of AuNPs via reduction of the aqueous
chloroauric acid solution with the leaf extract of Pogestemon benghalensis (B)
O. Ktz., as both reductant and stabilizer. The UV–visible spectrum of the
synthesized AuNPs showed surface plasmon resonance (SPR) around 555 nm
after 12 h. The shapes of synthesized AuNPs are mostly spherical and
triangular with sizes 10–50 nm, whereas the XRD pattern furnished evidence

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102 Brajesh Kumar, Kumari Smita and Brajendra Kumar

for the formation of face-centered cubic structure of gold having average


crystallite size 13.07 nm. The catalytic degradation of MB dye using AuNPs
was found to be 0.1758 min-1 as under visible light illumination. Naik et al.,
2013 prepared Nanocomposites of Au/TiO2 by green synthesis method using
the aqueous extract of Cinnamomum tamala leaves as the reductant. Analytical
characterization exhibited the formation of well dispersed AuNPs on the
surface of TiO2. Further, the photocatalytic activity of the catalysts prepared
by green method was evaluated for the degradation of MO as a model reaction.
It was found that the catalysts modified with 2 wt% AuNPs exhibited 2.5 times
higher degradation activity towards MO dye compared to TiO2 under solar
light irradiation. The effect of parameters such as pH and plant extract
concentration has been evaluated on the morphology of AuNPs and
photocatalytic degradation of MO. The dye degradation rate followed first
order kinetics with rate constant 0.346 h-1.

3.3. Iron Nanoparticles (FeNPs)

Nanoscale iron particles, FexOy and FeOOH (in total 16 polymorphic


forms) are of significant interest because of their rapidly developing
applications for disinfection of water and remediation of heavy metals from
soils (Kharisov et al., 2012). The Andean blackberry (Rubus glaucus Benth)
leaf contains a notable amount of flavonoids, ellagic acid, tannins etc. Keeping
the importance of these phytochemicals, we developed a simple, low cost, and
ecofriendly method for the synthesis of magnetite nanoparticles (Fe3O4 NPs).
The disappearance peak at 240 to 360 nm clearly indicated the involvement of
the blackberry extract in the synthesis process of iron nanoparticles (Figure
4c). Transmission electron microscopy (TEM) and Dynamic light scattering
(DLS) characterization indicated the formation of spherical Fe3O4 NPs of
average size 54.5 ± 24.6 nm. X-ray diffraction (XRD) and Fourier transform
infrared spectroscopy (FTIR) studies confirmed the existence of the cubic
spinel phase of Fe3O4 NPs and Fe-O peak at 570 cm-1, whereas Thermal
gravimetric (TG) analysis indicated that the nanoparticles contain 94% metal
and 6% capping ligand. It has been also observed that, as-synthesized Fe3O4
NPs exhibited photocatalytic activity for degradation of organic dyes such
as MB (k = 0.0105475 min-1), CR (k = 0.0043240 min-1), and MO (k =
0.0028930 min-1), efficiently (Kumar et al., 2016a). Huang et al., 2014
synthesized iron nanoparticles (Fe NPs) using green methods based on
tea extracts, including green, Oolong and black teas instead of using

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environmentally toxic sodium borohydride. They concluded that the best


method for degrading malachite green (MG) was Fe NPs synthesized by green
tea extracts because it contains a high concentration of caffeine/polyphenols
which act as both reducing and capping agents in the synthesis of Fe NPs.
Furthermore, kinetics for the degradation of MG using these different sizes
and concentrations of Fe NPs being synthesized by tea extracts, fitted well to
the pseudo first-order reaction kinetics model with more than 20 kJ/mol
activation energy, suggesting a chemically diffusion-controlled reaction. The
degradation mechanism using these Fe NPs included adsorption of MG to Fe
NPs, oxidation of iron, and cleaving the C=C- and =C=N- bond that was
connected to the benzene ring.
Lin et al., 2015 investigated the synthesis of iron-based nanoparticles (Fe
NPs) under various atmospheres (N2, O2 and air) to understand how
atmospheres impacting on the reactivity of Fe NPs, and Fe NPs were further
used for the degradation of MB. It is confirmed by the results of SEM and
FTIR that the morphology and change in size of iron-based nanoparticles
before and after reaction with MB, indicating that different Fe composition,
morphology and size were obtained under various atmospheres then resulted in
different reactivity of Fe NPs. Furthermore, the data fitted well to the pseudo-
second-order adsorption and pseudo-first- order reduction models, confirming
that the removing MB based on both adsorption and reduction. Finally, the
degraded products such as benzothiazole were identified by Gas
chromatography-mass spectrometry (GC-MS) after the degradation of MB.
Luo et al., 2015 proposed the single-step synthesis of Fe0 NPs using grape leaf
extract and quick and cost-effective for the in-situ remediation of effluent
containing Orange II. Batch experiments showed that more than 92% of
Orange II were removed by Fe NPs at high temperature based on adsorption
and reduction and confirmed by kinetic studies. Different spectroscopic and
microscopic analysis showing that the Fe NPs were composed of
biomolecules, hydrous iron oxides and Fe0, thus providing evidence for the
adsorption of Orange II onto hydrous iron oxides and its reduction by Fe0.
Degraded mechanism based on asymmetrical azo bond cleavage and products
such as 2-naphthol were identified using LC–MS analysis.

3.4. Zin Oxide Nanoparticles (ZnO NPs)

Besides TiO2, ZnO NPs has photoadsorption to the visible light range and
exhibit photocatalytic activity and decolorization of basic dyes in the visible

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light range owing to their lower band gap. Suresh et al., 2015 presented the
green synthesis of ZnO nanoparticles (Nps) using aqueous Cassia fistula plant
extract as fuel by solution combustion synthesis. The extract was found to
contain reducing components such as polyphenols (11%) and flavonoids
(12.5%). The Nps were found to have a hexagonal wurtzite structure. UV–
visible absorption of ZnONPs showed absorption band at 370 nm which can
be assigned to the intrinsic band-gap absorption of ZnO due to the electron
transitions from the valence band to the conduction band. TEM image
confirms the formation of nanoparticles and the average crystallite sizes were
found to be ~5–15 nm. The ZnO-NPs were evaluated for photodegradative,
antimicrobial and antioxidant activities. It showed efficient MB degradation
(>96%) under UV and Sun light illumination. Not only photocatalytic activity
but also ZnO NPs showed significant antioxidant activity against 1, 1-
Diphenyl-2-picrylhydrazyl (DPPH) free radicals and antibacterial activity
against Klebsiella aerogenes, Escherichia coli, Plasmodium desmolyticum and
Staphylococcus aureus. Kumar et al., 2014c demonstrated the application of
discarded agricultural waste for the fabrication of ZnO-NPs. They proved that
the aqueous peel extract of Citrus paradisi promotes the fabrication of the
ZnO-NPs with particle size ranging from 12 to 72 nm. The synthesized ZnO-
NPs exhibited strong UV absorption spectra with the absorption peak ranging
from 360 to 375 nm due to their excitonic transition (Figure 4d). The formed
ZnO-NPs are highly stable and exhibited more than 56% degradation of MB in
sunlight for 6.0 h. In addition, the current study has clearly demonstrated that
the ZnO-NPs are responsible for significant antioxidant activity (≥80% for 1.2
mM). Stan et al., 2015 successfully prepared the hexagonal wurtzite structure
of ZnO nanoparticles using aqueous extracts of Allium sativum (garlic), Allium
cepa (onion) and Petroselinum crispum (parsley). The biomolecules present in
the plant extract also influenced the particle size and obtained size varies
between 14 and 70 nm. The photodegradation studies conducted in the
presence of UV light irradiation indicated that ZnO nanoparticles prepared
using garlic extract exhibit the highest efficiency in the photodegradation of
MB dye.
Highly stable and spherical zinc oxide nanoparticles (25– 40 nm) are
produced by using zinc nitrate and Aloe barbadensis Miller leaf extract. As a
result, greater than 95% conversion to nanoparticles was achieved using aloe
leaf broth concentration greater than 25%. It was shown that the zinc oxide
nanoparticles were poly dispersed. The particle size could be controlled by
varying the concentrations of leaf broth solution (Sangeetha et al., 2011;
Kharissova et al., 2012). Fua and Fu, 2015 demonstrated the synthesis of ZnO

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Phytochemical Functionalized Metal Nanocatalyst … 105

nanoparticles using leaf extract of Plectranthus amboinicus. The


biosynthesized ZnO NPs exhibited a rod shape structure with an average size
of 88 nm and the band gap is 3.07 eV. Photocatalytic performance shows that
the biosynthesized ZnO NPs owing a superior photocatalytic activity towards
degradation of MR in comparison to hydrothermal synthesized ZnO NPs.
Nethravathi et al., 2015 synthesized multifunctional ZnO NPs employing
water extract of Garcinia xanthochymus by solution combustion synthesis.
The spectroscopic and microscopic studies indicated the formation of 20-30
nm spherical ZnO NPs that agglomerated to form spongy cave like pure
wurtzite structure with absorption maximum of 370 nm corresponding to band
gap energy of 3.33 eV. As-synthesized NPs not only exhibited remarkable
photodegradation of MB (>75%) in presence of UV and sun light but also
showed antioxidant activity by inhibiting the 1,1-diphenyl-2-picrylhydrazyl
(DPPH) free radicals.

3.5. Palladium Nanoparticles (PdNPs)

Palladium nanoparticles (PdNPs), which are of interest because of their


catalytic properties and affinity for hydrogen, have been phyto-synthesized
under moderate pH and room temperature (Kharissova et al., 2012). Recently,
Vilas et al., 2016 reported first the utilization of essential oils of Coleus
aromaticus and Myristica fragrans as bioreductant for the synthesis of PdNPs.
Their formation has been identified by the color change from pale yellow to
intense brown and is confirmed by an absorption continuum between 300 and
800 nm as revealed from the UV-vis spectral analysis. TEM and XRD
analyses disclosed the formation of clusters of 2.8 and 2.9 nm sized
monodispersed (fcc) structured Pd nanocrystals with spherical morphology. As
inferred from the FTIR spectra, terpenoids and phenolic ether derivatives, the
primary composition of the essential oils, are responsible for reduction and
further stabilization of the nanoparticles. The synthesized PdNPs showed
exceptional catalytic potential of PdNPs in the degradation of a wide spectrum
of organic pollutants, including methyl red, methyl orange, eriochrome black
T, methylene blue, rhodamine B. It is due to extremely small Pd NPs which
enhances the availability of a large number of active surface atoms. Similarly,
in another study, Kumar et al., 2015b reported ultrasonication assisted
fabrication of PdNPs using leaf extract of Andean blackberry. It showed the
reduction of K2PdCl6 solution to PdNPs (Pd0) by the change in their color
(yellow to brownish-black) of the reaction mixture and by appearance of

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106 Brajesh Kumar, Kumari Smita and Brajendra Kumar

absorbance peak around 360 nm, and 500 – 700 nm (Figure 4e). The obtained
PdNPs were crystalline in nature with decahedron morphology and size around
55-60 nm. It further used as photocatalyst for the decomposition of MB using
sunlight and an enhanced photocatalytic activity (>72%, k = 0.0021614 min-1)
for PdNPs was observed due to electron relay effect.
Trung et al., 2015 reported the facile and green synthesis of activated
carbon-supported palladium (Pd/AC) containing homogeneously dispersed Pd
nanoparticles (Pd NPs) by using eco-friendly and naturally available
Cinchonidine as the capping agent. The Pd NPs in the synthesized Pd/AC
hybrid are uniform with sizes predominantly in the range 4 to 7 nm. The
synthesized Pd/AC was characterized with various methods, such as TEM,
XRD, and XPS, and the influence of the synthetic conditions on its properties
was investigated. The advantages of cinchonidine over conventional capping
agents include its easy depletion after the synthesis with a simple rinsing
process. Owing to the ultrafine, well-dispersed and purified Pd NPs, the
synthesized hybrid exhibits excellent catalytic activities in the reduction of 4-
nitrophenol and methylene blue. These findings further the development of
novel stabilizing agents from naturally available sources for the preparation of
heterogeneous catalysts with enhanced performance.

4. GENERAL MECHANISM OF DEGRADATION


OF ORGANIC DYES

The mechanism of catalysis in the presence of NaBH4 and MNPs/AuNPs


can be described as an electron transfer process from donor NaBH4 to an
acceptor. MNPs act as an electron relay and initiate shifting of electron from
BH4- ion (donor B2H4/BH4-) to organic dye (acceptor) and thus causing a
reduction of dye. BH4- ion simultaneously adsorbed on the surface of NPs and
thus electron transfer occurs from BH4 - ion to organic dye through NPs (Paul
et al., 2015).
Using solar energy is an interesting aspect in photocatalyst technologies.
Solar photocatalysis has become an important area of research in which
sunlight is the source of illumination to perform various photocatalytic
reactions with regard to different kinds of dyes. In literature, photocatalysis
research reveals that photocatalytic activity can be strongly dependent on the
crystallographic structure, morphology, and size of the particles (Kamat,
1993). The photocatalytic mechanism of MNPs can be schematically

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Phytochemical Functionalized Metal Nanocatalyst … 107

illustrated in Figure 5. During the photocatalytic process, the absorption of


photons by the photo catalysts leads to the excitations of electrons from the
valance band (VB) to the conduction band (CB) generating electrons (e-) /hole
(h+) pairs. The holes can react to the H2O molecule and transform to •OH,
eventually the transformed active species and also h+ could effectively react to
the organic dye. The electrons in the conduction band is captured by oxygen
molecules dissolved in the suspension and in the valance band captured by
OH- or H2O species absorbed on the surface of the catalyst to produce the
hydroxyl radicals (·OH). Those hydroxyl radicals then oxidize the pollutants/
dye to small inorganic molecules (Bhattacharjee and Ahmaruzzaman, 2015;
Zhu et al., 2016). The photocatalytic degradation process may be represented
by the following reaction:

MNPs + hν → e- + h+ (1)

H2O + h+ → OH- + H+ (2)

OH- + h+ → ·OH (3)


e- + O2→ ·O2- (4)

·O2- + H+ → ·OOH/H2O2 (5)

Dye + hν → Dye* (6)

Dye* + O2 or ·OH or ·O2- → Degradation products (CO2 +H2O) (7)

Figure 5. General photocatalytic mechanism of different metal nanoparticles for the


degradation of various dyes.

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108 Brajesh Kumar, Kumari Smita and Brajendra Kumar

5. TOXICITY OF DYES
Use of synthetic dyes has an adverse effect on all forms of life. Nearly
50% of organic dyes are non-biodegradable, insoluble, toxic and their
persistence in wastewater. More than 79105 metric tonnes of dye stuffs are
produced worldwide annually, with 10 to 50% of this amount being released
into wastewater (Russ et al., 2000; Julkapli et al., 2014). These high
concentrations of dyes in effluents interfere with the penetration of visible
light into the water, resulting in a hindrance to photosynthesis and a decrease
in gas solubility, since less than 1 mgL−1 of dye is highly visible. Furthermore,
colorants having aromatic and heterocyclic rings containing oxygen, nitrogen
or sulfur, are regarded as toxic, carcinogenic, and xenobiotic compounds (Jie
et al., 2013). They cause toxicity not only to aquatic life and may be
mutagenic and carcinogenic and can cause intense damage to human beings,
including the reproductive system and dysfunction of the kidneys, brain, liver,
and central nervous system (He et al., 2011; Julkapli et al., 2014). According
to the criteria of the European Union for the classification of dangerous
substances, the acute toxicity of azo dyes, is low and the values of LD50
(median letal dose) are 250-2000 mg Kg-1 body weight (Clarke and Anliker,
1980). Dyes have various effects on human health depending on the
application area. Skin irritation and contact dermatitis have been reported for
some synthetic dyes and the use of azo dyes made from carcinogenic amines
has been banned by legislation in many countries. Some azobasic, acid and
direct dyes are classified into very toxic or toxic to fishes, crustaceans, algae
and bacteria, while reactive azo dyes are toxic only at very high concentrations
(>100 mg L-1), therefore, excluded from considering toxic for aquatic
organisms (Novotný et al., 2006). Nohynek et al., 2004 investigated that the
hair dye ingredients (p-aminophenol, Lawsone) have moderate to low acute
toxicity and human poisoning accidents are rare only due to oral ingestion.
The use of food additives in food products including the food food or
beverages colorants can cause toxic and carcinogenic effects. Several azo dyes
can cause the genesis of malignant tumors due to DNA damage (Gürses et al.,
2016). As a consequence, some food scientists try to use natural colorants in
order to hinder the negative effects of synthetic colorants.
It is very important to know whether biodegradation of a dye leads to
detoxification of the dye or not. Kalyani et al., 2009 and Parshetti et al., 2006
reported that that the phytotoxicity of the metabolites produced after the
biodegradation of Reactive Red 2 and Malachite Green were less toxic
compared to the original dye. Similarly, Jadhav et al., 2008 studied

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phytotoxicity of Methyl red and its metabolites formed after biodegradation by


Galactomyces geotrichum MTCC 1360 whereas, Shedbalkar et al., 2008
compared phytotoxicity of Cotton Blue and its extracted metabolites formed
after biodegradation by Penicillium ochrochloron MTCC 517. Both results
concluded that the phytotoxicity of degraded products are less than organic
dyes.

6. TOXICITY OF MNPS
With increasing the public knowledge about health care in the world,
people are increasingly concerned about the rise of possible subsequent
diseases caused by new technologies including nanotechnology and
application of nano-materials especially inhalation during manufacturing or
usage. Some evidence proved the safety of application of nano-structured
materials (Dastjerdi and Montazer, 2010). Living cells become more
permissive to chemical and physical compounds found in blood circulation,
including circulating nanoparticles depending of their material, size, charge,
surface engineering, and others characteristics (Leite et al., 2015). Toxicity are
caused by physical restraints or the release of toxic ions from metallic
nanoparticles or from the production of reactive oxygen species (ROS).
Further, the toxicity of some metallic nanoparticles has been found to be light-
dependent (e.g., TiO2), becoming more toxic under irradiation, 51, 110
whereas others (e.g., CeO2) have antioxidative effects (Quigg et al., 2013).
Silver is mentioned as an almost non-toxic to mammalian systems
(Dastjerdi and Montazer, 2010). Skin-innoxiousness of nano-silver colloidal
solution especially in the case of smaller nano-particles has been demonstrated
via the skin irritation test performed on the rabbits (Lee and Jeong, 2005). In
addition, rats orally exposed to AgNPs showed several synaptic structures
modifications and degeneration in hippocampus (Skalska et al., 2015). Toxic
effects of Ag-NP were also observed in cortical cell cultures inducing
oxidative stress and higher Ca2+ intracellular levels, triggering increased levels
of cell death. AgNPs are mostly internalized by astrocytes inducing
morphologic modifications, but in neuron cells these alterations require higher
concentrations of nanoparticles (Haase et al., 2012). Several studies have
emerged showing that AuNP can induce cellular damage by necrosis,
apoptosis, oxidative stress, inflammation, DNA damage, alterations in gene
expression, or through indirect mechanisms (Leite et al., 2015). Pan et al.,
2007 reported that the cytotoxicity of modified gold nanoparticles depended

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110 Brajesh Kumar, Kumari Smita and Brajendra Kumar

primarily on their size and not on ligand chemistry. Particles 1–2 nm in size
were highly toxic and both smaller gold compounds (Tauredon) and larger 15-
nm.
Iron oxide nanoparticles are generally considered as safe, biocompatible
and non-toxic materials (Arami et al., 2015). LD-50 (the median lethal dose or
the dose required to kill half of the tested animals during a specified time) of
the uncoated iron oxide nanoparticles was reported to be 300–600 mg Fe kg-1
body weight. This value was increased to 2000–6000 mg Fe kg-1 when the
IONPs were coated with stabilizing and biocompatible dextran molecules
(Wada et al., 2001). Fe3O4 nanomaterials can induce microglial activation and
subsequent increased levels of pro-inflammatory cytokines release, resulting in
cytotoxicity effects in a broad range of neuronal cells (Xue et al., 2012). In
another study by Geppert et al., 2009 showed that the incubation of astrocyte
primary cultures with magnetic iron oxide nano-particles has also
demonstrated that the particles do not induce any acute damage to these brain
cells. Lower doses of the iron nanoparticles can cause mild side effects such as
nausea, vomiting or flatulence whereas acute doses can cause severe side
effects such as inhibition of acetylcholinestrase in red blood cells, inhibition of
Na+–K+, Mg2+, and Ca2+-ATPases activities in brain and activation of the
hepatotoxicity marker enzymes in serum and liver (Arami et al., 2015).
The toxic effect of ZnO nanoparticles is due to their solubility in the
extracellular region, which in turn increases the intracellular Zn2+ level. The
exposure of ZnO nanoparticles induces oxidative stress and cytotoxicity,
mitochondrial dysfunction in RKO colon carcinoma cells, increased oxidative
stress, increased intracellular Ca2+ level, decreased mitochondrial membrane
potential, and interleukin-8 productions in the BEAS-2B bronchial epithelial
cells and A549 alveolar adenocarcinoma cells (Pandurangan and Kim, 2015).
Owing to its biological inertness, metallic Pd(0), forming the Pd
nanostructures, shows a priori the safest toxicity profile among palladium
species. Nevertheless, it progressively accumulates in the environment,
especially in aquatic ecosystem. Chen et al., 2015 studied the dose-dependent
toxic effect of Pd on zebrafish development. It indicated that acute Pd
exposure significantly decreased both the survival rate (LC50: 292.6 μg/L, viz.
2.75 μM) and hatching rate (IC50: 181.5 μg/L, viz. 1.71 μM) of zebrafish
during embryonic development. The heartbeat rate of zebrafish embryos was
also decreased after Pd exposure. In other study, Peric et al., 2012 compared
the toxicity of inorganic and organic palladium compounds on cardiovascular
system of rat. It seems that palladium, when bound in an organic compound
(linked to TEA in Pd complex), does not contribute significantly to cardio-

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toxicity. Besides this, it was also reported that the phytochemicals protect
metal nanoparticles from degradation and their functionalization with silver
and gold nanoparticles suppresses the toxicity against various cance cell lines
(Kumar et al., 2016f, g).

CONCLUSION
In conclusion, we have reviewed the current progress in the environmental
application of metal nanoparticles mediated remediation of organic dyes.
Phytochemical functionalized metal nanoparticles, and its beneficial effects,
offer the solar light harvesting potential for the degradation of organic dyes,
shorter reaction time, low-cost reagents, and possible environment friendly
alternative to chemical methods. The obtained results declared that the
phytochemical functionalized nanocatalysts (Ag, Au, Fe, Pd and Zn) showed
markedly high removal efficiency of the organic dye pollutants and promising
catalyst. The mechanism of catalysis in the presence of metal nanoparticles
can be described as an electron transfer process from donor NaBH4 to an
acceptor dye. The photocatalytic degradation effect of silver nanoparticles was
higher as compared to gold and iron oxide nanoparticles. The photocatalytic
effect of smaller and spherical nanoparticles was also higher as compared to
larger and triangular nanoparticles. Organic moiety and phytochemical
functionalization on the surface of metal nanoparticles not only reduces the
toxicity but also increase the stability of nanoparticles and lower the
manufacturing cost of catalyst. It supports the use of renewable energy for
environmental protection and this technology to be adequately attractive
compared to other techniques.

ACKNOWLEDGMENT
This scientific work has been funded by the TATA College, Kolhan
University, Chaibasa, Jharkhand, INDIA and Prometeo Project of the National
Secretariat of Higher Education, Science, Technology and Innovation
(SENESCYT), Ecuador.

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112 Brajesh Kumar, Kumari Smita and Brajendra Kumar

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BIOGRAPHICAL SKETCH
Dr. Brajesh Kumar, PhD
Department of Chemistry, TATA College, Kolhan University,
Chaibasa, 833202, Jharkhand, India
Centro de Nanociencia y Nanotecnologia,
Universidad de las Fuerzas Armadas ESPE,
Sangolqui, Ecuador

Education: MSc and PhD (University of Delhi, Delhi, India)

Honors: NET, GATE, JPSC, Prometeo Fellow

Publications Last 3 Years: 50 research articles

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120 Brajesh Kumar, Kumari Smita and Brajendra Kumar

Dr. Brajesh Kumar is currently working as an Assistant Professor in the


Department of Chemistry, TATA College, Chaibasa, INDIA. He received his
M.Sc and Ph.D in Chemistry from the University of Delhi, India. His research
interest is the development of sustainable and ecofriendly technique for natural
product extraction, purification and analysis, natural polymers, peptide
chemistry, microwave and ultrasound assisted organic synthesis, nanoparticles
synthesis and their applications for environmental remediation, active films of
organic solar cells, nanomedicine, sensors and organic synthesis. He is
credited different national and international fellowship and worked as a faculty
member in different universities of India, Ecuador and South Korea. He
published more than 90 research articles, patents, book chapters and
conference paper in the international and national level. He is an active
member of the different societies like ACS, ISCB, WASET, IICBEE. He is an
editorial member of several international journal and also an active reviewer of
more than 50 international journals (Elsevier, Wiley, Springer, Hindwai, De
Gruyter, IEEE, etc).

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 4

IONIC LIQUID-PROMOTED
SYNTHESIS OF PHOSPHINATES AND
BISPHOSPHONIC ACID DERIVATIVES

Nóra Zsuzsa Kiss, Dávid Illés Nagy


and György Keglevich†
Department of Organic Chemistry and Technology,
Budapest University of Technology and Economics, Budapest, Hungary

ABSTRACT
These days, the ionic liquids (ILs) are in the focus as green and
tunable solvents in organic syntheses. However, another application of
ILs has also emerged based on their potential as catalysts or additives in
different reactions. IL additives may enhance the rate of the reactions,
and promote complete conversions. Literature examples are shortly
overviewed in this paper. We found that the microwave (MW)-assisted
esterification of phosphinic acids may also be facilitated by the presence
of 10% of a suitable IL. The presence of e.g., [bmim][PF6] allowed a
lower temperature of ca. 160–180°C (instead of ca. 220°C), shorter
reaction times, and higher conversions/yields. Hence, a novel method was
developed for the syntheses of cyclic and acyclic phosphinates. In another
field, the synthesis of dronic acid derivatives, such as pamidronic acid


Corresponding author E-mail address: zsnkiss@mail.bme.hu.

Corresponding author E-mail address: gkeglevich@mail.bme.hu.

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122 Nóra Zsuzsa Kiss, Dávid Illés Nagy and György Keglevich

and alendronate useful in the treatment of osteoporosis was influenced


positively by the presence of 30% of [bmim][BF4]. Both in the case of
using sulfolane as a solvent, and in the solvent-free variation, significant
increase in the yields up to 80% could be observed that means a record in
the dronic acid discipline.

1. INTRODUCTION
Ionic liquids (ILs) became widely used reaction media due to advantages
meant by the fine tunability of their properties (e.g., polarity and lipophilicity)
and recirculability, not speaking about the fact that they are regarded green
solvents [1–6].
ILs are especially suitable for homogeneous catalytic reactions, such as
hydrogenations [7] and hydroformylations [8], as the transition metal catalyst
remains in the IL phase, and can be recycled [9, 10]. The work-up may
comprise a simple phase separation [11].
As a typical industrial application, the synthesis of dodecylbenzene by
Friedel–Crafts alkylation involves the use of imidazolium [AlCl4] ILs as a
solvent [12].
The second generation of ILs include task-specific species with side
chains promoting special reactions [13, 14]. Thus, the ILs may serve as
solvents, and also as catalysts in a number of cases giving rise to twin
advantages of non-volatility and unnecessity for an additional catalyst.
Examples among others, include Friedel–Crafts acylations [15], hydrolyses
[16], multicomponent reactions [17, 18], and syntheses of heterocycles
[19–21].
Recently, ILs have started finding applications as catalysts, or as
additives, meaning that they are used only in smaller amounts, and often
together with microwave (MW) irradiation. This protocol seems to be a green
approach
[3, 22, 23].
FeCl3-based ionic liquids (e.g., [bmim]Cl∙FeCl3) applied in a catalytic
amount can promote MW-assisted Friedel–Crafts acylations and alkylations
[24, 25].
The Fischer esterification of carboxylic acids was carried out in the
presence of 3 equivalents of an acidic IL ([bmim][HSO4]) under MW
irradiation, resulting in the corresponding esters in almost quantitative yields
[26]. Later on, other Brønsted acidic ILs with a [HSO4]– anion applied in

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Ionic Liquid-Promoted Syntheses 123

quantities of 5–50% were also found to be suitable catalysts for direct


esterifications [27]. Both basic and acidic ILs were found to be efficient
catalysts in transesterifications during biodiesel production, when the ionic
species were applied in an amount of 1–10% [28]. Moreover, solid material-
supported, functionalized ILs were also applied for a better recyclability and
easier handling, thus, reducing the amount of the catalyst to <4% [29].
There are numerous examples for condensation-type reactions catalyzed
by ILs. In the three-component Mannich condensation of acetylenes,
aldehydes and secondary amines, it was pointed out that the use of 20% of
[bmim][PF6] in dioxane as the solvent resulted in a significantly improved
yield, as compared to the cases, when dioxane, or the IL was used alone [30].
The benzoin condensation of benzaldehyde was found to be more efficient in
the presence of a small amount (2–5%) of an IL, as compared to the instances,
when 40–90% of the additive was measured in [31]. Task-specific ILs were
applied in an amount of 10–40% in a three-component solvent-free Biginelli
reaction to give pyrimidine derivatives in high yields [32, 33]. Similarly, the
Pechmann condensation of phenols and β–keto esters benefited from the IL
catalysis. Moreover, the typically used corrosive catalysts (H2SO4 or HClO4)
could be replaced by a catalytic amount of an acidic IL, such as [bmim][HSO4]
in the synthesis of cumarine derivatives [34]. IL catalysis was also applied in
the syntheses of indole [35].
Other examples, such as halogenation reactions [36–38] and
dehalogenations [39] were also described, and Lewis acidic ILs have also been
applied as catalysts in the MW-promoted solvent-free reaction of alcohols with
tetrahydropyrane in order to protect the alcohols [40, 41].
It is worth mentioning that the use of a small amount of an IL as an
additive may also function as a polar “heating aid” enhancing the absorption of
MWs [42].

2. THE USE OF IL ADDITIVES IN THE


SYNTHESIS OF PHOSPHINATES
Within heterocyclic chemistry, the five- and six-membered P-heterocycles
represent a prominent class [43–45]. The P-cycles may bear a phosphinic or
phosphine oxide function. 1-Alkoxy- or 1-aryloxy-phospholene 1-oxides are
usually prepared from the corresponding phosphinic chlorides by reaction with
an alcohol or phenol as the nucleophiles [45–47]. This method is not atom

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124 Nóra Zsuzsa Kiss, Dávid Illés Nagy and György Keglevich

efficient, and the use of acid chlorides means cost. A “greener” approach could
start from phosphinic acids, however, these species do not undergo direct
esterification with alcohols [45]. Interestingly, we found that the direct
esterifications may take place under microwave (MW) conditions [48, 49].
Other possibilities for the synthesis of phosphinates, involve alkylating
esterification of the phosphinic acids by alkyl halides [50], or the use of
activating agents, such as the T3P® reagent [51]. The Arbuzov reaction
represents another approach to phosphinates [52]. Among these variations, the
most environmentally-friendly protocol is the direct esterification [53]. Under
MW irradiation, the direct esterification of phosphinic acids with alcohols
used in a 15-fold molar excess afforded the phosphinates in variable yields.
The esterifications were especially efficient using longer carbon atom chain
(n ≥ 4) alcohols at or above 200°C. Phenols were not found to be suitable
reaction partners in MW-assisted direct esterifications. The desired products
could be identified in the reaction mixtures, but their quantity remained below
3% [54, 55].
The energetics of the esterifications under discussion were explored by
quantum chemical calculations. The direct esterification was found to be
thermoneutral with a rather high (≥130 kJ mol–1) enthalpy of activation. Such
reactions may be promoted by MW irradiation via the beneficial effect of the
statistically occurring local overheatings [56, 57].
The obvious disadvantage of the MW-assisted esterification is the high
reaction temperature (200–235°C), the longer reaction times (up to 6 hours)
required, and the limitation in terms of the nucleophile partner: when alcohols
with low boiling point were used, the yields of the phosphinates remained low,
and the attempts to perform esterifications with phenol derivatives remained
unsuccessful.
As demonstrated in the Introduction, certain organic chemical
transformations became more efficient in the presence of ILs, especially under
MW irradiation. We wished to try this possibility utilizing ILs as additives for
the synthesis of cyclic phosphinates [58].
First, we wished to test different ILs as potential catalysts in the
esterification of 1-hydroxy-3-phospholene oxide 1 with n-pentanol at 180°C
(Scheme 1). The esterifications were carried out as described earlier [49], but
in the presence of a catalytic amount of the imidazolium salts. As can be seen
in Table 1, the blind probe experiment led to a conversion of 82% after a 2 h’s
irradiation time (Table 1/Entry 1). The use of 10% of [bmim][BF4] led to
phosphinate 2g in a conversion of 41% (Table 1/Entry 2), while the
[bmim][PF6] additive had a more remarkable effect, the conversion became

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Ionic Liquid-Promoted Syntheses 125

complete after 30 min (Table 1/Entry 3). Applying the same reaction
conditions, [emim][HSO4], [emim][MeSO4], [emim][EtSO4] and [emim][Cl]
additives were less efficient (Table 1/Entries 4–7). In the presence of these
additives, the conversions remained under 30%. The addition of [emim][Ac] to
the reaction mixture led to a similar outcome obtained with [bmim][BF4]
(Table 1/Entry 8). It is worth mentioning that in a few cases, the conversions
could be improved allowing longer irradiation times (Table 1/Entries 2 and 5–
7).

Scheme 1.

Table 1. Direct esterification of hydroxyphospholene oxide 1 by


pentyl alcohol at 180°C in the presence of 10% of an ionic liquid

Entry Catalyst t (min) Conversiona (%)


1 – 120 82
2 [bmim][BF4] 30 41c
3 [bmim][PF6] 30 100
4 [emim][HSO4] 30 30b
5 [emim][MeSO4] 30 27c
6 [emim][EtSO4] 30 21c
7 [emim][Cl] 30 29c
8 [emim][Ac] 30 40b
a
On the basis of relative 31P NMR intensities
b
No change upon further irradiation
c
Prolonged heatings resulted in higher conversions

In the next step, we wished to extend the scope of the esterifications


applying [bmim][PF6] as an additive in the reaction of 1-hydroxy-3-
phospholene oxides 1 and 3 with a series of simple alcohols (Scheme 2).

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126 Nóra Zsuzsa Kiss, Dávid Illés Nagy and György Keglevich

Scheme 2.

Table 2. Direct esterification of 1-hydroxy-3-phospholene 1-oxides


(1 and 3) in the absence or presence of 10% of [bmim][PF6] as an additive

Entry R1 R2 IL T p t Conversion (%)a Yield Ref. Product


(°C) (bar) (h) MW (Δb) (%)
1 H Me – 160 18 4 38 32 [58] 2a
2 H Me 10% 160 18 3 62 38 [58] 2a
3 H Et – 160 17 4 38 30 [58] 2b
4 H Et 10% 160 17 3 86 60 [58] 2b
5 H nBu – 200 16 2 62 (11) 58 [48] 2c
6 H nBu 10% 180 14 0.5 90 (19) 83 [58] 2c
7 H nPent – 220 9 2.5 100 82 [58] 2e
8 H nPent 10% 180 5 0.5 100 (52) 94 [58] 2e
9 H iPent – 235 14 3 100 76 [49] 2f
10 H iPent 10% 180 5 0.5 100 95 [58] 2f
11 H nOct – 220 2 2 100 71 [56] 2g
12 H nOct 10% 180 1.5 0.33 100 85 [58] 2g
13 H nDodecyl – 230 2 2 100 95 [56] 2h
14 H nDodecyl 10% 180 1 0.33 100 94 [58] 2h
15 Me nBu – 220 18 3 65 [48] 4c
16 Me nBu 10% 190 15 2 100 93 4c
17 Me iBu – 200 17 2 34 (4) 30 4d
18 Me iBu 10% 180 15 2 100 (61) 67 4d
19 Me nPent – 235 11 3 90 67 4e
20 Me nPent 10% 200 8 1 95 72 4e
21 Me nOct – 230 2.5 2 100 90 4g
22 Me nOct 10% 180 1.5 1 100 83 4g
a On the basis of relative 31P NMR intensities
b Comparative thermal experiment

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Ionic Liquid-Promoted Syntheses 127

The esterifications were carried out in the temperature range of 160–


210°C under MW irradiation, in the presence of 10% of [bmim][PF6]. The
results can be found in Table 2 together with the data obtained without the use
of any additive. The esterifications with the most volatile alcohols (MeOH and
EtOH) were not too efficient under MW irradiation (Table 2/Entries 1 and 3).
However, when the reactions were performed in the presence of [bmim][PF6]
at 160°C for 3 h, conversions of 62% and 86% were obtained (Table 2/Entries
2 and 4). The uncatalyzed method required longer reaction times, and resulted
in lower conversions. The esterification of phosphinic acid 1 took place readily
with longer carbon-atom chain alcohols at 180°C in the presence of 10% of the
IL selected to provide complete conversions and excellent yields after 20–30
min (Table 2/Entries 6, 8, 10, 12 and 14).
Direct esterification of the dimethyl derivative (3) required somewhat
higher temperatures and longer reaction times due to its lower reactivity.
However, the same tendencies were observed as those reported above (Table
2/Entries 15–22).
It can be concluded that the MW-assisted direct esterifications of 1-
hydroxy-3-phospholene oxides 1 and 3 became much more efficient in the
presence of [bmim][PF6] as an additive in respect of temperature, reaction time
and outcome.
The IL catalysis was effective also for the derivatization of other cyclic
phosphinic acids, such as monomethyl- and dimethyl-hydroxyphospholane
oxides (5A and 5B), and a hydroxy-hexahydrophosphinine oxide (5C) as well
(Scheme 3). As can be seen in Table 3, the IL-promoted experiments could be
performed under milder conditions (lower temperature and shorter reaction
times) resulting in higher conversions and yields.

Scheme 3.

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128 Nóra Zsuzsa Kiss, Dávid Illés Nagy and György Keglevich

Table 3. Direct esterification of cyclic phosphinic acids 5A–C with n-


pentanol in the absence or presence of 10% of [bmim][PF6] as an additive

Entry Starting material IL T p t Conversion (%)a Yield Ref. Product


(°C) (bar) (h) (%)
1 5A – 235 11 3 85 79 [58] 6A
2 5A 10% 220 9 1 100 89 [58] 6A
3 5B – 235 11 5 72 60 [58] 6B
4 5B 10% 220 9 2 95 84 [58] 6B
5 5C – 220 9 4 45 31 [58] 6C
6 5C 10% 220 9 2 60 42 [58] 6C
a On the basis of relative 31P NMR intensities.

Encouraged by the above results, and realizing the potential in the use of
ILs as additives, we attempted to extend the MW-assisted and IL-promoted
direct esterification to the reaction of phenol derivatives as well. As was
mentioned above, previous attempts to obtain the aryl phosphinates via MW-
assisted direct esterification remained unsuccessful [55].
Along the lines of the above, we tested several ILs in the reaction of
hydroxy-3-phospholene oxide 1 with phenol (Scheme 4). To our surprise,
while there was practically no reaction in the absence of an IL, all IL tested
promoted the esterification to a smaller or greater extent (Figure 1). In
accordance with our previous experience, the application of [bmim][PF6] had
the most beneficial effect.
Thus, in the next stage of our work, we wished to synthesize new
aryloxyphospholene oxides (7) applying a catalytic amount of [bmim][PF6]
under MW irradiation (Scheme 5). The direct esterifications were carried out
in a closed vial, irradiating the reaction mixture under conditions given in
Table 4.

Scheme 4.

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Ionic Liquid-Promoted Syntheses 129

Figure 1. The effect of different ILs in the synthesis of 1-phenoxy-3-methyl-3-


phospholene 1-oxide.

Scheme 5.

Table 4. Direct esterification of 1-hydroxy-3-phospholene 1-oxide (1)


with phenol derivatives in the absence or
presence of 10% of [bmim][PF6] as an additive

Entry Ar IL T p t Conversion (%)a MW (Δb) Ref. Product


(°C) (bar) (h)
1 Ph – 200 2 4 <3 [55] 7i
2 Ph 10% 160 1 0.5 100 (0) 7i
3 4-Me-Ph – 200 2 4 <3 [55] 7j
4 4-Me-Ph 10% 180 1.5 0.5 100 7j
5 4-MeO-Ph – 200 2 4 <3 [55] 7k
6 4-MeO-Ph 10% 180 1.5 0.5 >95 7k
7 4-Cl-Ph – 200 2 4 <3 [55] 7l
8 4-Cl-Ph 10% 160 1 0.5 100 7l
a On the basis of relative 31P NMR intensities.
b Comparative thermal experiment.

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130 Nóra Zsuzsa Kiss, Dávid Illés Nagy and György Keglevich

The reaction of phosphinic acid 1 with phenol derivatives took place


readily in the presence of the IL additive to afford aryloxyphospholene oxides
(7i–l) in moderate yields of 30–50%. Further optimizations, especially in
respect of the work-up procedure are to be performed.
We were successful in demonstrating the advantageous effect of the
catalytic amounts of ILs in the direct esterification of phosphinic acids not
only with alcohols, but also with phenols.

3. THE ROLE OF THE ILS IN THE SYNTHESES


OF DRONIC ACID DERIVATIVES

The hydroxy-bisphosphonic acid (dronic acid) derivatives have two


pentavalent, tetracoordinated phosphorus functions (two phosphonate groups)
at the same methylene unit, and hence they incorporate a phosphorus-carbon-
phosphorus triad. They are capable to create complex with calcium ions, thus,
they bind rapidly and strongly to the hydroxyapatite crystals in the bones.
Several members of this family are used as drugs in the treatment of
osteoporosis, the Paget-disease, and tumor-induced hypercalcaemia.
According to the recent researches, they also show direct (breast and prostate)
antitumor-, as well as antiparasitic activity [59–63]. The biological activity and
effect of hydroxy-bisphosphonic acid derivatives depend significantly on their
side chain.
In general, during the preparation of dronic acid derivatives, the
corresponding carboxylic acid is reacted with different P-reagents in various
solvents. In most cases, phosphorus trichloride and phosphorous acid were
applied as the P-reactants in different ratios, but phosphorus acid, phosphoryl
chloride or phosphorus pentoxide also emerged in the syntheses. Although, the
cheap and easily available reagents make attractive the preparation outlined,
the reaction mixtures are so heterogeneous that they cannot be stirred.
Moreover, the hydrolysis is too vigorous and exothermic, and a large amount
of hydrochloric acid is released. In most cases, methanesulfonic acid (MSA)
was preferred as the solvent, as it assists the safe implementation of the
reactions, and ensures a certain extent of homogeneity. In addition to MSA,
many other solvents, such as sulfolane, chlorobenzene, toluene, p-cresol,
acetonitrile, n-octane etc. [64] have been applied in the syntheses of hydroxy-
bisphosphonic acid derivatives. The synthesis of dronic acid derivatives may
also be performed in the absence of any solvent [64]. The preparation in IL, or

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Ionic Liquid-Promoted Syntheses 131

in the presence of IL has been less studied [64]. Pamidronic acid (12),
alendronic acid (13), risedronic acid (14) and zoledronic acid (15) were
prepared in tributylammonium chloride as the solvent, starting from the
appropriate acetic acid (8-11), and using 2 equivalents of phosphorus
trichloride, and 1 equivalent of phosphorous acid as the P-reagents (Scheme
6). Low yields of 12–31% were reported, and the purities were not provided
[65].

Scheme 6.

γ-Aminobutyric acid (9) was reacted with 2 equivalents of phosphorus


trichloride and 1,5 equivalents of phosphorous acid in the presence of only 0.5
equivalents of different ILs as the additives. Depending on the ILs used,
sodium alendronate trihydrate (13-Na) was claimed to had been obtained in
yields of 92–94%, in a pure a form based on HPLC [66]. Keglevich, Grün and
Nagy reproduced the synthesis described for alendronate (13-Na) in the
presence of [bmim][BF4], and the dronate under discussion (13-Na) could be
obtained in a yield of only 19%, while the purity was 95% (Scheme 7) [67].
Hence, it was proved that the high yields (e.g., 92%) described are incorrect
and misleading, that were probably related on crude products.

Scheme 7.

In the case of zoledronic acid (15), imidazol-1-yl-acetic acid (11) was the
starting material, and phosphorus trichloride and phosphorus acid were applied
in a quantity of 2.5 and 1.7 equivalents, respectively, in 1.6 equivalents of
various ILs. Again excessive yields of 90-92% were reported due to neglect of
the criterions of purity [68]. In one instance, sodium zoledronate (15-Na) was
prepared starting from crude zoledronic acid (15) obtained using 1.6

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132 Nóra Zsuzsa Kiss, Dávid Illés Nagy and György Keglevich

equivalents of [bmim][BF4]. After pH adjustment and crystallization, a more


realistic yield of 60% was reported with a purity of 99.8% (Scheme 8) [68].

Scheme 8.

The role of the ILs in the syntheses of hydroxy-bisphosphonic acid


derivatives was studied by Keglevich, Grün and Nagy in detail. So far, the
effect of ILs has been investigated in the synthesis of pamidronic acid (12) and
alendronate (13-Na) [67, 69]. The corresponding carboxylic acid [β-alanine (8)
or γ-aminobutyric acid (9)], 2 or 3 equivalents of phosphorus trichloride, 2
equivalents of phosphorous acid, along with different amounts of an IL
[(bmim)(PF6) or (bmim)(BF4)] were measured in. The reactions were carried
out at 75°C for 3 h that was followed by hydrolysis, pH-adjustment [in the
case of alendronate (13-Na)] and crystallization (Scheme 9) (Table 5) [67, 69].

Scheme 9.

In contrast to alendronate (13-Na), it is possible to synthetize pamidronic


acid (12) in the absence of any solvent in a moderate yield (44%) (Table
5/Entry 1). Increasing the amount of the IL, the yields also increased. For
pamidronic acid (12) using 0.3 equivalents of [bmim][PF6], the yield was 71%,
while for alendronate (13-Na) measuring in 0.3 equivalents of [bmim][BF4],
the outcome was 66% (Table 5/Entry 4). Surprisingly, a further increase in the
quantity of the ILs, led to a decrease in the yields. Measuring in more than one

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Ionic Liquid-Promoted Syntheses 133

equivalent of the IL, the yields were almost identical with those of the solvent-
free variations, for pamidronic acid (12) 42% vs. 44% (Table 5/Entries 7 and
1), and for alendronate (13-Na) 5% vs. 0% (Table 5/Entries 6 and 1). It can be
concluded that it is enough to use ILs as an additive in a quantity of 0.3 or 0.6
equivalents [67, 69]. Hence, we demonstrated the catalytic effect of ILs in the
syntheses of hydroxy-bisphosphonic acid derivatives. The catalytic effect of
ILs has not yet been clarified. According to one explanation, they may
enhance the electrophilic character of the carbonyl group in the starting
carboxylic acid.

Table 5. Synthesis of pamidronic acid (12) and alendronate (13-Na)


from β-alanine (8) and γ-aminobutyric acid (9), respectively,
using 2 or 3 equivalents of phosphorus trichloride,
and 2 equivalents of phosphorous acid in different amounts of an IL

Entry Amount of ILa Pamidronic acida,b Alendronatea,c


(equiv.) Purityd,e (%) Yielde (%) Purityd,e (%) Yielde (%)
1 0 >99 44 – 0
2 0.1 >99 64 90 46
3 0.2 >99 65 – –
4 0.3 >99 71 98 66
5 0.6 >99 70 98 63
6 1.1 – – 90 5
7 2 >99 42 – –
a The IL was [bmim][PF6] (pamidronic acid) or [bmim][BF4] (alendronate)
b 2 equivalents of PCl3 and 2 equivalents of H3PO3
c 3 equivalents of PCl and 2 equivalents of H PO
3 3 3
d On the basis of acid-base titration
e From at least two parallel experiments

Alendronate (13-Na) was also prepared in sulfolane as the solvent, or in


the mixture of sulfolane and an IL additive. Applying phosphorus trichloride
and phosphorous acid in a molar ratio of 2:2 or 3:2 in sulfolane, the yields
were 46% and 52%, respectively. Interestingly, when the same amounts of the
P-reactants were used in sulfolane together with 0.3 equivalents of the
[bmim][BF4] additive, the target dronate was obtained in outstanding yields of
72% and 80%, respectively, in a pure form [67]. It can be seen that sulfolane
and [bmim][BF4] have a synergistic effect (Figure 2).

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134 Nóra Zsuzsa Kiss, Dávid Illés Nagy and György Keglevich

Figure 2. The effect of the solvent or/and additive in the preparation of alendronate
(13-Na).

SUMMARY
It was shown that both in the MW-assisted direct esterification of
phosphonic acids, and in the synthesis of certain dronic acid derivatives from
the corresponding carboxylic acids and phosphorus trichloride/phosphorous
acid, catalytic amounts of suitable imidazolium ILs promoted the reactions
regarding reaction time and yield. It may be a general trend that the use of less
IL is more advantageous, than its use as a solvent.

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 5

LOW MOLECULAR MASS REACTIVE


ACRYLAMIDE COPOLYMERS AS CARRIERS
OF BIOLOGICALLY ACTIVE COMPOUNDS

M. V. Solovskiy, PhD, M. Yu. Smirnova*, PhD,


E. B. Tarabukina, PhD, A. I. Amirova, PhD,
and N. V. Zakharova, PhD
Institute of Macromolecular Compounds
of Russian Academy of Sciences, Saint-Petersburg, Russia

ABSTRACT
Low molecular mass linear statistic copolymers of acrylamide (AA)
are promising as carriers for drugs and biologically active compounds
(BAC). Minor toxicity of polyacrylamide (PAA) for warm-blooded
animals makes it possible to use AA copolymers with reactive
comonomers for the purpose of drug delivery. Since they are not
biodegradable, their molecular mass should be restricted to (2–5) × 104
for their complete removal from the organism.
The aim of the current work was to synthesize low molar mass ionic
AA copolymers for drug delivery and to study their behavior in water
solutions.

*
Corresponding Author Email: smirnova_mariann@list.ru.

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142 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

As anionic systems, non-toxic statistical AA copolymers with acrylic


(AAc), methacrylic (MAAc) and 2-acrylamido-2-methylpropansulfonic
(AAMPSAc) acids having different comonomer composition were
synthesized by means of radical copolymerization using AIBN as
initiator. AA with AAc copolymers were also obtained by means of
alkaline hydrolysis. The structure of the obtained copolymers was
confirmed using IR and NMR spectroscopy.
It was shown that AA is more active when copolymerizing with
MAAc than with AAc. Different pattern of functional group distribution
along the chains of AA with MAAc and AA with AAc copolymers
results in their different complexing ability with respect to cationic BAC.
The synthesized anionic copolymers were used as drug carriers in
complex with aminoglycoside antibiotics to reduce the toxicity of the
latter. The complexation was confirmed using the methods of equilibrium
dialysis, potentiometric titration and molecular hydrodynamics. It was
found that the toxicity and antimicrobial activity of the resulting
aminoglycoside complexes depend on their stability. The toxicity of
aminoglycoside antibiotics in polymeric form proved to be reduced
3-4 times on average as compared with corresponding intact
aminoglycosides, while completely retaining their antimicrobial activity.
The antiviral activity of AA with AAMPSAc copolymers was also
discovered. These copolymers were successively used to obtain water-
soluble nontoxic polymer complexes of antiviral arbidol drug.
As cationic carriers, AA copolymers with 2-aminoethylmethacrylate
hydrochloride (2-AEMH) were synthesized. A method of ion exchange
conversion of AA with 2-AEMH copolymers into AA with 2-
aminoethylmethacrylate (2-AEM) copolymers containing highly reactive
primary amino groups was developed. Polymeric ketimin derivatives of
doxycycline antibiotic were obtained. They proved to be nontoxic and
showed a marked antimicrobial activity. They influenced the antibody
response at intraperitoneal injection with antigen, demonstrating
immunosuppressant properties depending on the dose. The obtained
polymeric conjugates of doxycycline can be used as antibacterial
formulations after tissue and organ transplantations.

Keywords: anionic and kationic copolymers of acrylamide, polymer drugs,


antibiotics-aminoglycoside, arbidol, doxycycline

1. INTRODUCTION
An important problem of modern polymer chemistry is the use of
macromolecular compounds for controlled modification of properties of

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Low Molecular Mass Reactive Acrylamide Copolymers … 143

biologically active compounds (BAC), i.e., drugs, haptens and biocydes [1, 2].
Currently, it is possible to obtain composites between low molecular weight
BAC and polymers and, thus, to achieve increased solubility in water,
prolonged action, increase in biological activity, to avoid side effects, and
provide controlled delivery of a BAC into target cell. Among other methods of
biologically active polymer synthesis is formation of ionic or covalent bonds
between BAC and water-soluble non-toxic polymers (carriers). Ready
medicinal preparations can be used as BAC, and thus the main problem is a
development of suitable polymer carriers. Synthetic carbon-chain copolymers
of N-vinylpyrrolydone [3] and N-(2-hydroxypropyl)methacrylamide [4] have
proven effectiveness as carriers for BAC.
Non-toxic hydrophilic acrylamide (AA) copolymers are also promising as
BAC carriers [3]. However, they are still not thoroughly studied in this
respect; only the use of acrylamide/acrylic acid copolymers for immobilizing
protein BAC has been reported [5, 6]. There are no data published on
modification of low molecular weight BAC with AA copolymers. Carbon-
chain polymers, including acrylamide copolymers, do not undergo noticeable
metabolism and are removed intact from the organism via kidney filtration (if
their molecular mass (MM) does not exceed 40000). Therefore, in the present
work we focused on developing methods for synthesizing low molecular mass
(ММ = (20-40)·103) ionogenic AA copolymers (both anionic and cationic).
The goal of the present work was synthesizing reactive ionogenic AA
copolymers, determining their composition and molecular mass and using
these products as BAC carriers.

2. ABOUT OUR RESEARCH


2.1. Synthesis and Properties of Anionic Acrylamide Copolymers

The copolymers were obtained by radical heterophase copolymerization of


monomers in 2-propanol, ethanol, or 2-propanol/ethanol mixture. We have
synthesized the following anionic copolymers: copolymers I (AA/acrylic acid,
AAc), copolymers II (AA/methacrylic acid, MAAc) and copolymers III
(AA/2-acrylamido-2-methylpropane sulfonic acid, AAMPSAc).
The interest to AA/AAc and AA/MAAc copolymers is caused by the
differences in the distribution of COOH groups along copolymer chains [7].
In the case of AA/MAAc copolymers, the microblock structure consisting of
MAAc units is formed in the polymer backbone. On the contrary, AA/AAc

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144 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

copolymer chains consist of AA microblocks, which may affect the ability of


copolymers I and II to form complexes with BAC cations. The interest to
copolymers III is caused by the fact that these copolymers, like other
sulfocontaining polymers, can possess their own antiviral activity.

CH 2 CH CH 2 CH
m2
C O COOH

NH2

CH 2 CH CH 2 CH
m2
C O C O

NH2 NH

H3C C CH3

CH 2

SO3H
I (R = H); II (R = CH3-) III

Taking into account the high reactivity of AA and its tendency towards
formation of crosslinked structures, copolymerization of AA with AAc,
MAAc and AAMPSAc was carried out at rather low temperature (50°C). The
results of copolymerization experiments are summarized in Table 1. It can be
seen that MM of anionic AA copolymers can be controlled by varying
comonomer concentration in the initial mixture (Experiments 2-3, 11-12, 13-
14), varying initiator concentration (Experiments 1-2) and introducing 2-
propanol, which is used as a chain transfer agent in production of
poly(acrylamide) (Experiments 3-5, 7-8). The values of viscometric average
molecular masses (Мη) of copolymers I-III (with m2 = 18.2-22.8 mol.%) were
equal to (10 – 39)·103 (Table 1). In the case of copolymers I-7, I-8, MM
determined by sedimentation and diffusion analysis (MSD) turned out to be
41·103 and 50·103, respectively.
Table 2 shows that with increasing MSD, translational diffusion
coefficients D0 decrease, and, correspondingly, the values of efficient
hydrodynamic radii of macromolecules RhD (calculated from the D0 values)
increase. The values of hydrodynamic invariant A0 are close to the average

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Low Molecular Mass Reactive Acrylamide Copolymers … 145

experimentally obtained value (A0 = 3.2·10-10 erg/grad/mol1/3) for flexible-


chain polymers in good solvents [8]. This indicates the agreement between
hydrodynamic parameters (sedimentation and diffusion coefficients (S0 and
D0) and intrinsic viscosity [η]) determined by hydrodynamic methods and
reliability of the MSD values calculated from these parameters. The values of
specific partial volumes of the copolymers (υ) are close to that of
poly(acrylamide) (0.73).
The structure of copolymers I-III was confirmed by IR and NMR
spectroscopy [9, 10].
The copolymer obtained in Experiment III-10 demonstrated antiviral
activity against both herpes virus and H1N1 influenza virus.

Table 1. Heterophase copolymerization of acrylamide [М1]


with unsaturated carboxylic acids [М2] at 50°С

№ The initial reaction mixture АА-АAc (I), АА-MAAc (II) and AA-
AAMPSAc (III) copolymers
[М2], [М1+М2], Solvent Yield,% m2, [η], cm3/g Мη·10-3,
mol. % wt. % mol. % (MSD)·10-3
Copolymerization of АА with АAc (I)
1 20 20 2-propanol 99.5 20.4 17 10
2 20 20 2-propanol 98.8 20.1 25 19
3 20 30 2-propanol 99.5 19.8 31 27
4 20 30 2-propanol + 99.8 20.1 40 39
ethanol
5 32 30 2-propanol + 99.7 31.1 20 (41)
ethanol
6 35 30 ethanol 96.9 34.7 32 (50)
Copolymerization of АА with MAAc(II)
7 20 30 2-propanol 99.9 19.0 25 19
8 20 30 2-propanol + 99.8 19.2 29 25
ethanol
9 20 30 ethanol 99.6 19.0 33 29
Copolymerization of АА with AAMPSAc (III)
10 25 25 ethanol 99.2 22.8 32 28
11 20 15 ethanol 99.6 19.3 21 16
12 20 40 ethanol 99.8 18.5 38 36
13 20 40 2-propanol 99.8 19.1 25 19
14 20 50 2-propanol 98.8 18.2 28 23
Note: AIBN concentration is 3 wt. %; in Experiments 1, 7, 8, 12, 13, 14 it is 4.5 wt. %

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146 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

Table 2. Molecular masses and hydrodynamic characteristics of AA-AAc


copolymers determined in 0.5 M NaCl solution

№ m2, [η], ν, So, Do·107, MSD·10-3 RhD,nm Ao·1010,


mol. % cm3/g cm3/g Sv cm2/s erg/grad/mol1/3
I-5 31.1 20 0.69 2.0 4.3 40 4.8 3.3
I-6 34.7 32 0.69 2.1 3.5 50 5.5 3.1

2.2. Alkaline Hydrolysis of Poly(Acrylamide)

Copolymers of AA with AAc were also obtained by alkaline hydrolysis of


low molecular weight polyacrylamide (IV). Alkaline hydrolysis of PAA
results in copolymers of AA with AAc salts (V) according to Scheme 1:

(1)
IV V

In order to obtain AA-AAc copolymers (I), the solution of copolymer V


was acidified to pH = 2.0 with 0.1 N solution of HCl (Scheme 2):

(2)
I

Sodium chloride liberated in reaction (2) was removed by dialysis. PAA


hydrolysis was carried out in 0.6 M NaOH solution (the initial polymer
concentration was 0.3 mol·L-1). The hydrolysis temperature and time were
varied. Table 3 presents results of Experiments 1-7 (PAA alkaline hydrolysis).
The data given in Table 3 imply that at the temperature of 100°С and low
concentration of alkali, hydrolysis proceeds at a high rate (Table 3,
Experiments 1-3). Already in 15 min, the degree of PAA hydrolysis (q)
reaches 50.7%, and in 1 h this value increases up to 60.5%. Under similar
hydrolysis conditions, with decreasing temperature, the q value also decreases.
After dialysis, the yield turned out to be 58-76%.

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Low Molecular Mass Reactive Acrylamide Copolymers … 147

Table 3. Alkaline hydrolysis of low molecular weight poly(acrylamide) IV

№ The initial PAA Conditions of Product of hydrolysis (АА-АAc


hydrolysis copolymer I)
[η]·102, Mη·10-3 T, °C τ, min Yield, % m2*, mol. % [η]·102,cm3·g-1
cm3·g-1
1 0.40 36 100 60 64.0 60.5 0.16
2 0.40 36 100 30 59.2 52.0 0.17
3 0.40 36 100 15 58.0 50.7 0.19
4 0.40 36 60 60 76.3 44.6 0.24
5 0.40 36 37 60 70.6 17.0 0.27
6 0.40 36 30 60 68.3 13.1 0.37
7** 0.40 36 60 60 71.7 32.3 0.21
*
) m2 = q is the degree of hydrolysis
**
) C(NaOH) = 0.3 mol·L-1

It should be noted that intrinsic viscosity of the hydrolysis products


decreases (from 37 to 16 cm3·g-1) with increasing q (from 13.1 to 60.5%).
Therefore, we can conclude that efficient hydrodynamic radius of hydrolyzed
PAA molecules decreases, since the value of intrinsic viscosity is sensitive to
the size of macromolecules. This decrease in hydrodynamic dimensions of
AA-AAc copolymer molecules may be caused, first, by the formation of
intrachain hydrogen bonds (i.e., by intramolecular contacts between carboxyl
groups of AAc units and carbonyl groups of AA units in the polymer chain).
Second, decrease in the [η] value may result from polymer destruction. In
order to establish the real cause of this process, we used sedimentation and
diffusion, which can be considered ultimate methods of determining MM
(Table 4).

Table 4. Hydrodynamic characteristics of PAA and


AA-AAc copolymers obtained by alkaline hydrolysis

Polymer m2, mol. % [η]·102, cm3·g-1 So, Do·107, cm2/s MSD·10-3 RhD, nm
Sv
PAA 0 0.40 1.7 3.3 50 6.3
I-7 32.3 0.21 1.8 4.5 36 4.6
I-3 50.7 0.19 1.6 5.6 23 3.7

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148 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

H2C NH2

H O H
H
OH H
CH3
HO
OH O H
H NH2 H
OH H
O
H3C

H HN CH3 OH
H
2
H R NH
H H2N 1
OH R CH2
O
H2N O
H
H
O

H2C NH2 H
HO NH2 O
H O CH2 O
H
H H H H OH
HO O H2N NH2

H NH2 NH2
NB GB
H2C OH

H O H H2 C OH
H O
NH2 H
H
HO NH2
H OH HO
OH

H2C NH2 H2C NH2


O O
H O
H H2N
OH OH
HO HO
H NH2 O OH O

OH
OH OH

NH2 HN CO HC CH2 CH2 NH2


NH2 NH2

КB АB

The increase in the degree of hydrolysis is accompanied not only by the


decrease in the [η] value, but also by the rise in translational diffusion
coefficient and, correspondingly, by decrease in the values of hydrodynamic

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Low Molecular Mass Reactive Acrylamide Copolymers … 149

radius RhD calculated from these parameters. Table 4 shows that with
increasing content of AAc units in the copolymer, its molecular mass drops
from MSD = 5.0·104 (for the initial PAA) to MSD = 2.3·104 (for the copolymer
with carboxyl group content equal to 50.7 mol. %); i.e., molecular mass of
PAA decreases by more than a factor of 2. These data indicate that in the
selected conditions, alkaline hydrolysis of PAA is accompanied by destruction
of polymer backbone, and decrease in macromolecule size is mainly related to
the decrease in MM of hydrolyzed PAA (AA-AAc copolymer) as compared to
that of the initial PAA. It should be noted that PAA hydrolysis in the selected
conditions and the accompanying thermodestruction of low molecular weight
AA-AAc copolymer result in simultaneous changes in MM and m2. The higher
degree of hydrolysis, the lower the molecular mass of the hydrolyzed PAA.
The relationship between the MM of AA-AAc copolymer and the degree of
hydrolysis in the studied interval of copolymer compositions can be described
by the following equation: M/M1 = 1-1.17·10-2·m2, where M and M1 are the
molecular masses (MSD) of the initial and hydrolyzed PAA, respectively. This
relationship can be used for predicting the corresponding characteristics in the
synthesis of low molecular weight AA-AAc copolymers by alkaline
hydrolysis.

2.3. Polymeric Complexes of Aminoglycoside Antibiotics Based


on Anionic Acrylamide Copolymers

Anionic copolymers (AA-AAc, AA-MAAc, AA-AAMPSAc) were used


as carriers in the formation of complexes with the following basic
aminoglycoside antibiotics: neomycin (NB), kanamycin (KB), gentamycin
(GB), amikacin (AB).
Aminoglycoside antibiotics are compounds with related structures which
show high activity against a wide spectrum of bacteria. At the same time, these
drugs demonstrate high toxicity when used for medicinal purposes. In this
connection, there is a problem of lowering the toxicity of aminoglycoside
antibiotics. One of the efficient methods of decreasing toxicity of ionogenic
BAC [10] which allows retaining their biospecific activity is complexation
with non-toxic polyelectrolytes. In this work, we used synthesized non-toxic
copolymers (AA-AAc, AA-MAAc, and AA-AAMPSAc) as polyelectrolytes
for decreasing toxicity of aminoglycoside antibiotics. Complexation between
these copolymers and antibiotics was carried out in aqueous solutions at the
antibiotic/copolymer mass ratio equal to 1-5÷7. Viscosimetric studies of water

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150 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

and 3 M urea solutions of these complexes (where hydrogen bonds are


destroyed) demonstrated that the obtained polymeric ionic complexes of
aminoglycosides are stabilized by hydrogen bonds [11]. Weighed portions of
copolymers were titrated with 0.3 N solution of an antibiotic; it was
established that the ability of different antibiotics to form complexes with the
same AA-AAc copolymer decreases in the order: amikacin> kanamycin >
gentamycin > neomycin. Amikacin forms the most stable complex.Irrespective
of the antibiotic structure, increase in the amount of charged groups in the
copolymer leads to an increase in the maximum binding capacity (Qmax.) of the
copolymer towards the antibiotic, which confirms that complexation occurs
due to electrostatic interactions. The Qmax. values for carboxyl-containing
polymers are higher by a factor of 2.5-3 than those for sulfo-containing
copolymers of similar composition. This is due to the steric hindrances created
by two methyl groups located in the vicinity of the sulfonic groups; these
groups prevent the antibiotic from approaching the reactive site.
Equilibrium dialysis was used in the studies of complexation between
gentamycin and AA-AAc and AA-MAAc copolymers of close composition
and similar molecular masses (Mη = 19000). It was found that more stable
complex based on AA-MAAc copolymer (with increased density of charged
groups on polymer backbone) demonstrate the highest binding degree in water
(q = 75.5%) as compared to that for AA-AAc (54.2%). Increase in ionic
strength of the solution led to the drop in the q value down to 50-37% (in 0.15
М NaCl solution); thus, electrostatic forces prevail in this interaction.
The data obtained by hydrodynamic and optical methods in 1 M aqueous
solution of NaNO3 at high ionic strength confirmed high stability of the
complex of AA-MAAc copolymer (Mη = 19000) with kanamycin antibiotic
(Table 5).

Table 5. Hydrodynamic characteristics of AA-MAAc copolymer


and its complex in 1 M NaNO3 solution

Sample [η], ν, S0, D0·10-7, MSD·10-3 MW·10-3 RhD,


cm3/g cm3/g Sv cm2/s nm
Copolymer (II-1) 26 0.72 3.3 4.9 66 65 4.5
Complex between 27 0.73 2.1 2.8 80 87 7.4
Copolymer (II-1) + 15.0
wt. % of kanamycin

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Low Molecular Mass Reactive Acrylamide Copolymers … 151

Table 5 shows that the MSD and MW for AA-MAAc copolymer are
virtually similar, and in the case of polymeric complex, these parameters are
higher by a factor of 1.2-1.3. The hydrodynamic radius of the macromolecules
in the complex (RhD) obtained from the values of diffusion coefficient (which
were determined by isothermal translational diffusion) is significantly higher
than that of AA-MAAc copolymer (by a factor of 1.6). Increase in the values
of the MSD, MW and RhD parameters after interaction between the copolymer
and the antibiotic confirms formation of a complex.
In Vitro toxicity of all studied polymeric complexes (in cell cultures) was
lower by a factor of 4-4.5 than that of non-modified antibiotics [11, 12].
Polymeric complexes retain high antimicrobial activity inherent in the non-
modified antibiotic, despite rather low content of the drug in complexes (15-20
wt. %). The stable complex AA-MAAc +amikacin demonstrates the lowest
antibacterial activity and, at the same time, the lowest in vitro toxicity. Direct
relationship between biospecific activity and toxicity of polymer-BAC
complex and the stability of this complex allows controlling antibacterial
activity of polymeric complexes of cationic antimicrobial drugs and their
toxicity via the changing content of COOH groups in the macromolecule
and their distribution along the copolymer backbone.
In addition to complexation with aminoglycoside antibiotics, copolymers
III were used for modification of antiviral preparation Arbidol (VI).

CH3
H2C N
CH3
HO O
C * HCl * H2O
OC2H5

Br N CH 2 S
CH3
VI

Arbidol is an efficient antiviral preparation, an immunostimulant and


interferon inducer. However, it has several disadvantages, such as insolubility
in water and significant toxicity after parenteral introduction. These
disadvantages can be avoided after its complexation with low molecular
weight copolymers AA-AAMPSAc.

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152 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

CH 2 CH CH 2 CH CH 2 CH
m1 m2 m3
C O C O C O
HN HN
NH2
H3C C CH3 H3C C CH3

CH 2 CH 2 R
- +
SO3H SO3 N XVI

H R
VII

Water-soluble polymeric complexes VII had the following composition:


m1 = (80.9-76.6) mol.%; m2 = (7.6-9.8) mol.%; m3 = (11.5-13.6) mol.%, and,
according to the UV spectroscopy data, contained 26.4-32.1 wt.% of Arbidol.
Antiviral activity of different variations of Arbidol polymeric complexes
turned out to be approximately similar and comparable to the activity of non-
modified preparation against the model influenza strain A; however, their
toxicity in vitro was 10 times lower [13].

2.4. Cationic Copolymers of Acrylamide

Cationic copolymers of AA with 2-aminoethylmethacrylate IX intended


for use as BAC modifiers were synthesized by passing aqueous solutions of
copolymers AA-2-aminoethylmethacrylate hydrochloride VIII through the
column containing EDE-10 P anionite in the OH- form:

VIII IX

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Low Molecular Mass Reactive Acrylamide Copolymers … 153

Copolymers IX were obtained by heterophase radical copolymerization of


AA (M1) with 2-aminoethylmethacrylate hydrochloride (M2) (in ethanol or
propanol-2) at 50°С in the presence of AIBN initiator for 24 hrs. The
characteristics of the obtained copolymers are given in Table 6. The table
shows that copolymers VIII in all cases were synthesized with high yield (90-
99%). Water-soluble copolymers VIII of various compositions were obtained
(m2 = 18.7-34.6 mol. %); their composition was determined from the data of
elemental analysis for chlorine. The Mη values of copolymers VIII-1 – VIII-7
were determined by viscometry using the conventional Mark-Kuhn-Houwink
equation for poly(acrylamide) and were equal to 6000-39000.

Table 6. Copolymerization of АА with


2-aminoethylmethacrylate (AEM) hydrochloride

№ The initial reaction mixture Copolymer VIII


[M2], [M1+M2], Solvent Yield, [Cl], m2, [η], Мη·10-3,
mol. % wt. % % wt. % mol. % cm3/g MSD*·10-3
1 20 10 ethanol 89.2 7.49 18.7 24 18
2 20 15 -//- 91.0 8.25 21.2 34 30.5
3 25 20 -//- 99.8 7.95 20.2 40 39
4 25 10 2-propanol 96.2 7.93 20.1 12 6
5 20 20 -//- 99.7 8.08 20.7 15 9
6 15 20 -//- 87.3 7.01 17.3 30 25
7 25 20 -//- 90.9 8.27 21.3 24 18
8 30 20 -//- 98.0 9.01 23.7 34 33*
9 35 20 -//- 93.8 11.81 34.6 36 39*
Note: In all initial reaction mixtures, AIBN content was 4.5 wt. %.

A series of hydrodynamic parameters and molecular characteristics was


obtained for copolymers VIII-8 – VIII-9 using sedimentation, diffusion,
densitometry and viscometry, as well as dynamic light scattering in 0.5 M
aqueous solution of NaCl (Table 7). The values of partial specific volume (υ)
for the samples containing markedly different amounts of m2 units are virtually
similar to that of poly(acrylamide) (υ = 0.73 cm3/g) [14]. The S0, D0 and [η]
values of the studied samples are in agreement with each other.
The values of hydrodynamic invariant A0 are close to the average
experimental value known for flexible-chain polymers (A0 = 3.2·10-10
erg/grad/mol1/3). Insignificant difference between this average value and our
data on A0 is possibly due to polydispersity of the studied samples (which were
not fractionated). The studies of copolymer solutions by dynamic light

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154 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

scattering demonstrated that in solutions of both samples, two kinds of


scattering species are present, i.e., macromolecules with hydrodynamic radius
(Rh1DLS) of 4.4 and 5.4 nm (samples VIII-8 and VIII-9, respectively) and large
scattering particles with a hydrodynamic radius (Rh2DLS) of approximately 60-
70 nm. The Rh1DLS values are in accordance with the values of hydrodynamic
radius RhD (4.8 and 5.1 nm) calculated from the data on translational diffusion
with the use of the Stokes-Einstein formula (Table 7).

Table 7. Hydrodynamic and molecular characteristics of acrylamide


copolymers VIII obtained in 0.5 M NaCl solution

№ m2, [η], S0, D0×107, MSD RhD, Rh1DLS, Rh2DLS, А0×1010,


mol.% cm3/g Sv cm2/s nm nm nm erg/grad/mol/3
VIII-8 23.7 34 1.7 4.5 33 4.8 4.4 60 3.4
VIII-9 34.6 36 1.9 4.3 39 5.1 5.4 70 3.7

The particles of smaller size are apparently individual copolymer


molecules. Large particles are associates of macromolecules, since strong
intermolecular interactions between molecules in solutions are revealed.
Concentration dependence of the calculated radius (see Figure 1) obtained for
large particles is indicative of the formation of associates.

Figure 1. Concentration dependence of hydrodynamic radius of the slow mode Rh2DLS


of copolymer VIII-8 in solution.

Acrylamide-2-aminoethylmethacrylate hydrochloride copolymers VIII


were quantitatively transformed into AA-2-AEM (IX) by ion exchange [15].

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Low Molecular Mass Reactive Acrylamide Copolymers … 155

2.5. Polymeric Ketimine Derivative of Doxycycline Antibiotic

Copolymers of AA and 2-aminoethylmethacrylate were used as carriers


for doxycycline hydrochloride antibiotic (X) in the synthesis of its polymeric
ketimine derivative.

OH O OH O NH2
OH
O
* HCl
OH
H OH
CH3 OH N
H3C CH3
X

Currently, doxycycline is the most widely used tetracycline antibiotic; it


displays bacteriostatic effect due to inhibition of synthesis of bacterial protein.
Besides, doxycycline contains phenolic fragments; thus, it is expected that its
polymeric ketimine (like polymeric derivatives of other phenol-containing
compounds) will demonstrate immunomodulating activity. At the same time,
doxycycline has some side effects, of which the most important is
hepatotoxicity. Thus, this antibiotic was modified with a polymer to reduce
toxicity.

Figure 2. Outlet elution curves.

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156 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

In the synthesis of polymeric ketimine XI, we used AA-2-AEM


copolymer (MSD = 33000) containing 23.7 mol. % of NH2-groups in side
chains distant from the copolymer backbone. According to the data of gel
filtration experiments (using the column containing Acrylex P-150 gel, and
H2O as an eluent), the obtained product was rather monodisperse. The
elutioncurve obtained at the outlet included only one maximum (Figure 2,
curve 1).
In the reference experiment, free doxycycline hydrochloride was passed
through the column (Figure 2, curve 2); it was found that the retention volume
of the antibiotic is significantly higher (approximately by a factor of 2.3) than
that of polymeric ketimine XI.

H3C CH3
N OH CH3
OH H
HO

O
OH
NH2 O OH N OH

(CH 2)2

NH2 O

C O C O

CH 2 CH CH 2 C
m1 m2
CH3
XI

The formation of polymeric ketimine of doxycycline (XI) was confirmed


by IR and UV spectroscopy. In the UV spectrum of reaction product, the
absorption band with λmax = 274 nm is observed, in common with the UV-
spectrum of the initial doxycycline (λmax = 271 nm). Since the obtained
conjugate does not contain non-reacted antibiotic, appearance of this band
(λmax = 274 nm) in the UV spectrum of reaction product and its small (3 nm)
bathochromic shift are indicative of the presence of bound doxycycline.
According to the UV spectroscopy data, the content of the attached antibiotic
is 32.7 wt. %. In the IR spectrum (Figure 3), the absorption band at 1583 cm-1
is present which is attributed to valence vibrations of C=N ketimine bond.

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Low Molecular Mass Reactive Acrylamide Copolymers … 157

Figure 3. The IR spectrum obtained by subtraction of IR spectrum of the polymer


carrier from the IR spectrum of the product.

The LD50 value found for polymeric ketimine of doxycycline was equal to
1200 mg/kg, which is approximately 2 times higher than that for pure
antibiotic; thus, the derivative is not toxic (Table 8). In case of intraperitoneal
introduction with an antigen (ram erythrocytes), polymeric ketimine of
doxycycline exerts an influence on antibody response and demonstrates
immunosuppressive properties (depending on the administered dose); these
properties are more pronounced than in the case of pure antibiotic, particularly
when the dose is equal to 10 mg/kg.

Table 8. Biological properties of


polymeric ketimine derivative of doxycycline

Sample Acute Immunomodulating Antimicrobial


toxicity** activity activity
IRC* MIC, μg/mL
LD50, 25 mg/kg 10 mg/kg Staph. E.
mg/kg aur. coli
Polymeric ketimine (XI) 1200 0.6 0.4 1.8 31.0
Doxycycline hydrochloride 512 0.7 0.6 0.2 7.5
(reference sample)
* IRC is the immune response coefficient.
** In experiments with mice after intraperitoneal introduction.

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158 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

Besides, polymeric ketimine derivative of doxycycline demonstrated


noticeable antimicrobial activity which was almost similar to that of the initial
doxycycline hydrochloride (Table 8). In order to reveal the mechanism of
antimicrobial action of polymeric ketimine derivative of doxycycline, we
estimated the rate of antibiotic elimination from the polymer carrier in buffer
solution (pH = 7.4, T = 37°C). As seen from Figure 4, in the conditions under
study, polymeric ketimine of doxycycline undergoes slow hydrolysis (degree
of hydrolysis achieves 16% in 96 hrs).

Figure 4. Hydrolysis of polymeric ketimine derivative of doxycycline.

The low rate of hydrolysis is apparently related to steric hindrances which


are created by tetracyclic system of doxycycline and shielding of C=N bond.
At the same time, polymeric ketimine demonstrates rather high antimicrobial
activity, despite the low degree of hydrolysis. The obtained results are in
agreement with the reported data [16] on high antibacterial activity of polymer
derivatives of tetracycline based on N-vinylpyrrolidone/vinylamine
copolymers. These copolymers were synthesized by the Mannich reaction and
contain strong covalent bond (–NH–CH2–NH–). Thus, it can be assumed that
the synthesized ketimine of doxycycline affects bacterial cell in polymeric
form.
Thus, in this work, we have for the first time obtained polymeric ketimine
derivatives of a drug using doxycycline antibiotic as an example. The obtained
polymeric conjugates are non-toxic, demonstrate a new type of polyfunctional
biological activity (combination of antimicrobial and immunosuppressive
properties). They are promising preparations for use in surgical procedures
involving transplantation of organs and tissues complicated by bacterial
infections.

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Low Molecular Mass Reactive Acrylamide Copolymers … 159

3. EXPERIMENTAL
3.1. Monomers and Reagents

Acrylamide was twice recrystallized from ethyl acetate (m.p. 84.5°С).


Calculated: С 50.65%; H 7.03%. Found %: С 50.48; H 7.10.
Acrylic acid was purified by distillation under vacuum in the presence of
an inhibitor (bis-naphthoquinonyl-p-phenylenediamine). The fraction with
b.p. =32°С (5 mm Hg) was taken, nD20 = 1.4200; d420 = 1/0507 g/cm3,
R = 17.34 cm3/g, R(calc.) = 17.34 cm3/g.
Methacrylic acid was distilled in the presence of bis-naphthoquinonyl-p-
phenylenediamine; the fraction with b.p. = 48°С (5 mm Hg) was taken,
nd20 = 1.4310 (literature data: nd20 = 1.4314).
2-Acrylamido-2-methylpropanesulfoacid was purified according to the
technique described elsewhere [9].
2-Aminoethylmethacrylate hydrochloride was synthesized using the
method described elsewhere [15].
DMFA was prepared using the technique described in [17].

3.2. Polymeric Derivatives of Biologically Active Compounds


Based on Reactive Acrylamide Copolymers

Synthesis of aminoglycoside antibiotics and anionic acrylamide


copolymer complexes was described in [11, 18].
Preparation of Arbidol complexes was described in [13].
Synthesis of polymeric ketimine of doxycycline.
Solution of 0.15 g of doxycycline hydrochloride in 4 mL of H2O
was added to the solution of 0.309 g of copolymer acrylamide-2-
aminoethylmethacrylate (IX-9) in 40 mL of H2O. The obtained solution (pH =
8.0) was stirred for 4 h at 25°С and dried by sublimation of water under
vacuum; the product was dialyzed against water for 15 hrs and isolated by
lyophilization. The yield was 77.8%, doxycycline content was 32.6 wt. %.
Kinetics of hydrolysis of polymeric derivative of doxycycline.
The studies were performed in the model medium (buffer with pH = 7.4, T
= 25°С) by dialysis through semipermeable cellophane membrane in static
conditions; the amount of the antibiotic passed through membrane over a
certain time period was registered.

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160 M. V. Solovskiy, M. Yu. Smirnova, E. B. Tarabukina et al.

3.3. Methods of Investigation of Ionogenic AA Copolymers and


BAC Derivatives Based on These Copolymers

3.3.1. The methods of determination of molecular mass of the synthesized


AA copolymers by sedimentation and diffusion analysis (MSD) and light
scattering (MW) were described elsewhere [15].
3.3.2. Physico-chemical methods used in the studies of complexation
between anionic AA copolymers and aminoglycoside antibiotics are described
elsewhere [12, 18].
3.3.3. The following analytical instruments were used: an «Avance II
400» NMR spectrometer (Bruker); «IFS 88», Bruker and «Midac» Fourier IR
spectrometers; “SF-256” UV spectrometer (LOMO Fotonika); and a «Vario»
CHNS elemental analyzer. Refractive index (nD20) was determined with the
use of an Abbe “IRF-24” refractometer. Potentiometric titration was conducted
with the use of an HI 2210 pH-meter (Hanna Instruments). The polymers were
isolated from aqueous solutions with the use of a «FreeZone 6» sublimation
dryer (Labconco).
3.3.4. The methods used in biological studies of the obtained polymers
and BAC derivatives based on these polymers are described elsewhere [9, 11,
12, 19].

CONCLUSION
Some methods were proposed for the synthesis of novel polymer carriers
of BAC based on low molecular weight acrylamide copolymers which contain
–COOH, -SO3H and –NH2 groups. The composition, molecular masses and
other parameters, as well as in vitro toxicity of the polymers, were studied.
Complexation between acrylamide anionic copolymers and aminoglycoside
antibiotics in water and water-salt solutions was studied by physico-chemical
methods. The factors which affect the stability of the formed polymer
complexes were revealed. It was established that toxicity and antimicrobial
activity of the polymer complexes depend on their stability. Copolymers of
acrylamide with 2-acrylamido-2-methylpropanesulfonic acid were used to
obtain water-soluble non-toxic complexes of Arbidol; these complexes possess
broad-spectrum antiviral activity.
It was demonstrated that the obtained low molecular weight anionic and
cationic copolymers of acrylamide are promising carriers for various drugs.

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Low Molecular Mass Reactive Acrylamide Copolymers … 161

The authors are grateful to M. Yu. Eropkin, E. M. Eropkina and E. N.


Vlasova for their help in performing the experiments and characterizing the
samples.

REFERENCES
[1] Plate, N. A., &Vasil’ev A. E. (1986). Physiologically Active Polymers.
Moscow: Chemistry.
[2] Thatte, S., Datar, K. and Ottenbrite R. M. (2005). Perspectives on:
polymeric drugs and drug delivery systems. J. Bioact. Compat. Polym.,
20: 585-601.
[3] Panarin E. F., Lavrov N. A., Solovskiy M. V., Schal’nova L. I. (2014).
Polymers – Carriers of Biologically Active Substantives. Saint-
Petersburg: Profession.
[4] Kopecek J. &Kopeckova P. (2010). HPMA copolymers: origins, early
developments, present and future.Adv. Drug Deliv. Rev., 62: 122-149.
[5] Torchilin, V. P., Reyzez, I. L., Tischenko, E. F. et al. (1976).
Immobilization of enzymes on biocompatible carriers. IV. Modification
α-chymotrypsin of water-soluble copolymers of vinyl series. Evaluation
of accessibility in various ways immobilized α-chymotrypsin for protein
inhibitor. Bioorg. Chem., 2 (12): 1687-1694.
[6] Martensson, K. &Mosbach, K. (1972). Covalent coupling of pullulanase
to an acrylic copolymer using a water-soluble carbodiimide. Biotechnol.
Bioeng. 14: 715-724.
[7] Solovskiy, M. V., Gavrilova, I. I., Smirnova, M. Yu., Schul’tseva, E. L.
(2008). Synthesis of low molecular weight water soluble anionic
copolymers of acrylamide.IzvestijaVUZov. Series Chem. and Chem.
Technol., 51: 72-74.
[8] Tsvetkov, V. N. (1989). Rigid-chain polymers. New-York: Plenum.
[9] Solovskiy, M. V., Eropkin, M. Yu., Eropkina, E. M. et al. (2007).
Synthesis and properties of low-molecular-weight copolymers of
acrylamide with 2-acrylamido-2-methylpropanesulfonic acid, as
potential drug carriers. Russ. J. Appl. Chem., 80 (10): 1703-1707.
[10] Afinogenov, G. E. and Panarin, E. F. (1993). Antimicrobial polymers.
Saint-Petersburg: Hyppocrat.

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[11] Solovskiy, M. V., Eropkin, M. Yu., Eropkina, E. M. et al. (2010).


Aminoglycoside antibiotic complexes with copolymers of acrylamide
with acrylic and methacrylic acids. Pharm. Chem. J., 44 (6): 28-32 (in
Russian).
[12] Tarabukina, E. B., Solovskii, M. V., Pautov, V. D. et al. (2015).
Physicoshemical, molecular, and biological properties of complexes
formed between aminoglycoside antibiotics and some anionic
copolymers of acrylic series: Part II. J. Bioact. Compat. Polym., 30 (6):
571-583.
[13] Eropkin, M. Yu., Solovskiy, M. V., Smirnova, M. Yu. et al. (2009).
Synthesis and biological activity of water-soluble polymer complexes of
arbidol. Pharm. Chem. J., 43 (10): 27-31 (in Russian).
[14] Tarabukina, E. B., Amirova, A. I., Schul’tseva, E. L., Solovskiy, M. V.
(2009). Effect of synthesis conditions on molecular characteristics of
acrylamide copolymers with acrylic acid, carriers of cationic
biologically active substance. Rus. J. Appl. Chem., 82 (9): 1606-1614.
[15] Solovskiy, M. V., Smirnova, M. Yu., Tarabukina, E. B., Zakharova, N.
V. (2012). Synthesis of copolymers acrylamide with 2-hydrochloride
aminoethylmethacrylate - carriers of biologically active compounds.
Russ. J. Gen. Chem., 82 (10): 1650-1655.
[16] Schukovskaya, L. L., Dumova, A. P., Pal’chik, R. I. et al. (1970). Some
physicochemical and biological properties of polymer derivatives of
tetracycline. Antibiotics., 5: 775-779 (in Russian).
[17] Keil, B., Herout, V., Hudlicky, M. et al. (1966). Laboratory technique of
organic chemistry. Edited by B. Keil. Мoscow: Mir.
[18] Solovskii, M. V., Tarabukina, E. B., Amirova A. I. et al. (2014).
Complexation of anionic copolymers of acrylamide and N-(2-
hydroxypropyl)methacrylamide with aminoglycoside antibiotics. Russ.
J. Phys. Chem. A., 88 (3): 428-432.
[19] Solovskiy, M. V., Yeropkin, M. Yu., Yeropkina, Ye. M. et. al. (2012). In
vitro and in vivo studies of toxicity of polyacrylamide and certain
acrylamide anionic copolymers. ToksikologicheskiiVestnik., 2: 24-26.

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 6

THE PHENOMENON OF
MASS CONCENTRATION PERIODICITY
IN AMORPHOUS MATTER:
A NEW KIND OF PERIODICITY IN NATURE

K. Zubow2, A. Zubow1 and V. A. Zubow2,


1
Department of Telecommunication Networks Group, TU Berlin, Germany
2
R&D, Dr. Zubow Consulting, Germany

ABSTRACT
Periodic structures in some amorphous substances (water, starch,
heart biomatrix, styrene-maleic anhydride copolymer (1:1) esterified by
bis-tri-n-butyltinoxide and gelatin) were investigated by the gravitational
mass spectroscopy method (GMS). There are periodic sub ensembles
(PSE) of masses in the interval from 30 mio. to 4 billion Daltons. PSE
were characterized to have 5 mass peaks, at log m = 7.62; 8.57; 9.02; 9.31
and 9.54 (m in Daltons), they were represented as sub micelle and micelle
structures in the expanded form. In white gravitational noises, PSE were
observed to be stable, in colored gravitational noises, however, these
periodic structures were destroyed with the emergence of new sub
ensembles both in the expanded and the collapsed forms. It was tried to
find is there any correlation between the principles of periodic structures
in the amorphous molecular matter and those in the universe.


Corresponding author Email: aist@zubow.de.

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164 K. Zubow, A. Zubow and V. A. Zubow

Keywords: amorphous materials, long-range order, periodic structures,


gravitational mass spectroscopy.

1. INTRODUCTION
It is known that molecular mass concentrations (clusters, domains) are
formed under the influence of gravitational noises (GN) [1]. GN themselves
are generated by a huge number of galaxies, stars, nebulae, planets and black
holes. GN in the universe cause clusters of galaxies (https://en.wikipedia.
org/wiki/Millennium_Run) and giant voids (https://en.wikipedia.org/wiki/
Void_(astronomy)) resembling on molecular level some porous materials [2],
Figure 1.

Figure 1. Heterogeneity of baryonic mass distribution in the universe (A, 1500 Mpc/h,
http://bigclosetr.us/topshelf/blog/13640/universal-brain-neuron-network) and
molecular matter (B, C, ~ 10- 6m) (bone: http://www.umich.edu/~bme332/ch9bone/
bme332bone.htm).

For a long time in the colloid chemistry, the phenomenon of periodic


colloidal structures at the level of some μm [3] has been well-known, but there
is not a clear understanding how they are formed. It is believed that the
structure of the baryonic universe and the formation of the amorphous
molecular mater in it are realized under GN influence. At nano level,
amorphous matter is forced by GN to a certain periodicity in the cluster mass
distribution (domains, sub micelles and micelles, and their agglomerates).
Let’s prove this hypothesis and find some periodicities in amorphous
substances.
Thus, the aim of this work was to look for periodic structures in
amorphous molecular matter.

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The Phenomenon of Mass Concentration Periodicity … 165

2. EXPERIMENTAL
As objects of the long-range order (LRO) investigation served distilled
water, starch, proteins (biomatrix of duck heart muscle and gelatin, Dr. Oetker,
FRG), and an organotin polymer [4]. LRO was analyzed using the GMS
method [1].

3. RESULTS AND DISCUSSION


Figure 2 showed the GMS spectra of potato starch.

Figure 2. GMS spectra of potato starch powders: A – in white noises [1], B - heated at
523 K for 30 s (color noise [1]). The signal of the ring oscillation (α-D-glycopyranose)
was indicated by arrow.

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166 K. Zubow, A. Zubow and V. A. Zubow

As shown in Figure 2 at colored noise influence LRO was destroyed


especially in the range of giant micelle and sub micelle structures. Here should
emphasized the periodic structures of sub ensembles (S1...S8) with expanded
and collapsed structures in the original starch being destroyed in colored
noises.

Figure 3. GMS spectra of two water samples: A - storage for 2 years under quasi-white
noise conditions (iron, grounded, heat and sound isolated box, 295 ± 3 K), B -
exposition to colored noises (mixing and UV radiation). The water cluster models were
kindly provided by the professors Lenz [5] and Chaplin [6]. Amorphous structure of
water at 225K was given the right (http://phys.org/news/2011-11-supercool-doesnt-
.html).

Figure 3 showed the GMS spectra of two water samples, the first sample
was stored for a long time in white noises and the second one shortly in
colored noises [1]. In the range of sub micelle and micelle structures, there
were periodic mass accumulations at log m = 7.62; 8.57; 9.02; 9.31 and 9.54

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The Phenomenon of Mass Concentration Periodicity … 167

Daltons, which were also present in some synthetic polymers, for example, in
polyethylene, polystyrene, polypropylene etc. The form of the mass
distribution curves in the sub ensembles was almost the same, i.e., there is a
certain analogy in the periodicity of recurring events. On the other hand, the
spectrum showed any periodic voids between the sub ensembles of masses.
These voids in the spectra were not an indication for a balance between the
signals from collapsed and expanded micelle forms that mutually extinguish
each other.
In these mass/frequency ranges [1], the formation of micelles is simply
forbidden by the gravitational white noises, voids therefore periodically arise.
Similar effects can be observed when sound is directed on powders
(https://en.wikipedia.org/wiki/Cymatics).
External energy influences on water caused by colored noises destroyed
the LRO organization in the range of sub micelle and micelle structures while
in the area of small clusters the equilibrium expanded-collapsed clusters was
shifted toward collapsed structures (-f).
Similar regularities we observed in solutions of salts, acids, organic
liquids, plants’ biomatrices, animal proteins and synthetic polymers, too.
Figure 4 showed photographs of some supra molecular structures in
amorphous polymers and colloidal systems. One can notice some kind of a
diffuse periodicity in the voids of the nano material further, stationary waves
in the solid mineral agate. However, observe a periodicity in the mass
distribution on the nano level in amorphous matter using the electron
microscopy is impossible.

Figure 4. Supramolecular structures in organic (A, http://akvilon-sakh.ru/images1/


57649dcacc117.png) and inorganic amorphous polymers (B, http://domapestovo.
ru/images1/576042462e576.png). For comparison - periodic colloidal structure in
minerals (C, [3]).

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168 K. Zubow, A. Zubow and V. A. Zubow

In amorphous bio matrices, the visual observation of periodic structures is


also impossible, Figure 5. For example, the structure of the neuronal network
in the brain can only suggest the existence of a certain regularity, but at the
micrometer level. Using electron microscopy, it cannot be explored too.

Figure 5. Network of neurons in brain (A, http://en.academic.ru/dic.nsf/enwiki/


838519) and micro structure of a biomatrix (B, http: //rsfs.royalsocietypublishing. org /
content / 4/2 / 20,130,058). Given for a better understanding, only.

Figure 6. GMS spectrum of fresh heart duck biomatrix (10 min. After slaughter). Weak
shock waves.

Figure 6 showed the GMS spectrum of a duck heart biomatrix. In the


range of large sub micelle and micelle structures there were signals at: log m =
7.62; 8.57; 9.02; 9.31 and 9.55, which agree with the periodic structures of sub
ensembles in water (Figure 3). Here a periodical distribution of voids (market
with arrows) was also found. After removing water these periodic structures
disappeared (Figure 7) hence at drying the colored noises destroyed the
periodicity of sub ensembles in the amorphous biomatrix.

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The Phenomenon of Mass Concentration Periodicity … 169

Figure 7. GMS spectrum of dried duck heart biomatrix (Figure 6). A – signals from
myosin heads, B – signals from actin heads, C - range of sub ensembles (sub micelles
and micelles). Weak shock waves.

Figure 8. GMS spectrum of TOP. Weak shock waves, Zubow constant 9.4E-15 N/m. C
- range of large periodic mass concentrations (sub ensembles). The first cluster signal
belongs to -SnBu3, the second one to the domain consisting of 3 repeating polymer
units.

Figure 8 showed the GMS spectrum of a TOP film. As visible in the


synthetic polymer, there are periodic oscillations of sub ensembles of masses
(interval C), too however, because of the unfinished LRO formation process
the interval C is insufficiently expressed.

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170 K. Zubow, A. Zubow and V. A. Zubow

As we know this class of copolymers is characterized to be a porous


material, which pore structure is periodically [7]. Some periodic mass
concentrations in the walls were further found [4].
This periodicity is not based on the spatial principle. It does not matter
how far the mass concentrations are from each other. In GMS (around GMS
sensor), the distances between similar masses lies at a few millimeters,
nevertheless far distant mass analogues may influence the signal intensity as it,
for example, was found for water, polystyrene, starch [1] etc.

Figure 9. GMS spectra of fish collagen (bladder, FB) and gelatin (film, G). RASC -
domain consisting of: Gly 22 –Pro % 13% - Hyp 10% - Glu 9.8% - Ala 8% - Arg 7.6%
(weight %). NC1-hexamer model (168,931 Da) is taken from [9]. Weak shock waves.

Figure 9 showed the GMS spectra of collagen (fish bubble) and gelatin.
We remember that gelatin is a protein, which was extracted from various
animal organs at a hard impact of color noises from chemical technologies.
Both spectra were very different especially in the interval C. The exposure to
colored noises (boiling, extraction, filtration, drying and pressing a film) led
not only to an easier spectrum of the sub ensembles, but also to an increased
number of periodic structures from 4 to 7, at the same time in the gelatin
appeared some voids. Moreover, it was visible that the harmony of small- and
medium-sized domains in the expanded form was disturbed, in gelatin, besides

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The Phenomenon of Mass Concentration Periodicity … 171

the expanded forms collapsed domains appeared. So, the RASC signal (base
domain of 11 average units [8]) strongly pronounced in the native collagen
disappeared in gelatin. From this it follows that the investigation of high
structures of biopolymers (tertiary, quaternary and higher structures) should be
carried out under conditions being adequate to their existence in the body (in
vivo).
The analogy between the structure of the molecular amorphous matter and
the visible structure in the universe suggested that there should also be applied
the periodicity of mass and void distribution in the latter. In the universe, such
periodic mass concentrations (sub ensembles) are known, for example, the
galaxy clusters [10] with a period of 500 ... 650 million l.y. however, no data
yet on the periodicity of clusters in the dark matter (https://en.wikipedia.
org/wiki/Dark_matter#/media/File:COSMOS_3D_dark_matter_map.png),
such as of neutrino clouds [11]. If it will be found so it should be caused by a
certain generator of super-gravitational noises being outside the baryonic
universe.
To find this generator could be possible using a gravitational compass [1].
On the other hand, if the analogy exists, so the properties of the universe being
similar to those of colloidal systems, can be described applying the well-
known rules of the colloidal chemistry. Remember the process of coalescence
in colloidal chemistry is the basis for the gravitation [12].
In Figure 10, the GMS spectrum of water under microgravity conditions
(free fall, first 0.4 s, state maximally approximated to the real white noises,
[1]) was given.

Figure 10. GMS spectrum of water under microgravity conditions. Weak shock waves.

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172 K. Zubow, A. Zubow and V. A. Zubow

From the spectrum it was evident that the microgravity weakly influenced
the periodicity of the mass concentrations in the range C (Figure 3), that means
the periodicity of sub ensembles and voids was preserved, generally. However,
all water clusters were transformed into the collapsed forms, the number of
sub ensembles in the range C slightly increased and the signal from the base
water cluster (H2O)12 completely disappeared. The transition of water clusters
into the collapsed form is due to reduced lengths of the hydrogen bonds -
H...O-. This is possible when the influence of external gravitational fields on
the balance between protons and neutrinos becomes smaller (see Figure 11).
The balance really determines the bond length [12] and is characterized by
canonical structures (below in Figure 11). The weakening of the external
gravitational field increased the potential energy of the canonical structures
with strengthening the hydrogen bonds and the densifying all water clusters.

Figure 11. Modeled proton oscillations between physical vacuum (PV) and baryonic
state, and proton (p) interaction with neutrinos (ν) in a chemical bond (1-2). The
number of canonical structures characterizing the chemical bonds was given below
[12].

It should be mentioned that the time of the free fall (state of acceleration,
first 0.4 s) could not have been sufficient for the formation of a new
thermodynamic state of the entire water cluster ensemble under microgravity.
Here the signal of the rare cluster (H2O)178 appeared it emerged under strong
color noise influences (boiling, stirring, etc.), normally.
The new type of periodicity in amorphous matter can be illustrated as a
family of pendulums each with its own mass/frequency (see first Zubow
equation in [1].), Figure 12, but with one oscillation vector for the whole
pendula family agreeing with the direction of the gravitational compass [1]. In
turn this vector of all pendula, oscillating as protons’ ensembles [13], is
directed to large mass-analogues in our universe. Here each oscillator type is
characterized by its own mass concentration (domains, domains’ associates or

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The Phenomenon of Mass Concentration Periodicity … 173

micelle formations). Creating an individual resonance field the mass clusters


interacted distantly with their analogous around them [1, 8], where the
resonance field is typically for a given oscillator, only. The interaction with
other mass concentrations is limited by the Newtonian forces, and for the
information interaction with analogues resonance fields are applied.

Figure 12. Model of a new type of periodic mass concentration in amorphous matter.
For understanding, three different mass ensembles and their hypothetical GMS
spectrum were given, only. The arrow showed the direction of the dominant mass in
the universe.

CONCLUSION
The amorphous molecular matter is characterized by periodic structures of
mass sub ensembles appearing mostly when external energy influences (white
noises) are excluded, and they are less pronounced in the colored noises.
Periodic structures in the amorphous matter occur in the mass range from
30 million to 4 billion Dalton.

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174 K. Zubow, A. Zubow and V. A. Zubow

Periodic structures in the amorphous matter are the result of gravitational


influences from our universe.
Periodic structures in the amorphous matter are formed according to the
similarity principle in the nature.
Similar mass ensembles have their own oscillation fields and interact
remotely with each other.
Besides to the periodic masses periodical voids are generated forcibly by
gravitational noises.

REFERENCES
[1] Zubow K., Zubow A. V., Zubow V. A. The Way to the ETIs. Applied
gravitational mass spectroscopy. Nova Sci. Publ. NY, 2014.
https://www.novapublishers.com/catalog/product_info.php?products_id
=42668&osCsid=5bd85d42dc273360fd48126de7be9daf.
[2] https://arxiv.org/pdf/astro-ph/0504097v2.pdf.
[3] Efremov J. F. Periodic colloidal structures. - L.: Chemistry, 1971. - 192
p (in Russian). http://jes.ecsdl.org/content/120/6/185C.abstract.
[4] Zubow K., Zubow A. V., Zubow V. A. Long-Range Order in Organotin
Monomers and Polymers. Physical Chemistry-An Indian Journal. 2013,
v.8, no.5, pp.198-206. ID: Phy3482676.
[5] Annika Lenz, Lars Ojamäe. On the stability of dence versus cage-shaped
water clusters: Quantum-chemical investigations of zero-point energies,
free energies, basis-set effects and IR spectra of (H2O)12 and (H2O)20.
Chemical Physics Letters. 2006, v. 418, pp. 361-367.
[6] Chaplin M., SBU London, http://www.lsbu.ac.uk/water/index.html.
[7] Ke Zhang, Liwei Zhang, Yongming Chen. Robust Organic/Inorganic
Hybrid Porous Thin Films via Breath-Figure Method and Gelation
Process. Macromolecular Rapid Communications. 2007, vol. 28, pp.
2024-2028. DOI: 10.1002/marc.200700384.
[8] Zubow K., Zubow A., Zubow V. A. Phenomenal properties of the
domain ensembles in proteins. Biochemistry and Molecular Biology
Journal. Delaware. USA. 2016, vo. 2, no., 1, pp. 1-10.
http://biochem-molbio.imedpub.com/phenomenal-properties-of-the-
domain-ensembles-in-proteins.pdf.
[9] Manuel E. Than, Stefan Henrich, Robert Huber, Albert Ries. The 1.9- α
crystal structure of the noncollagenous (NC 1) domain of human
placenta collagen IV shows stabilization via a novel type of covalent

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The Phenomenon of Mass Concentration Periodicity … 175

Met-Lys cross-link. Proc Natl Acad Sci U S A. 2002 May 14, vol. 99,
no.10, pp. 6607–6612. doi: 10.1073/pnas.062183499..
[10] http://www.wissenschaft.de/archiv/-/journal_content/56/12054/1542
367/Kosmischer-Schaum/.
[11] Zubow A., Zubow K., Zubow V. A. Radiation of Axions and Neutrinos
from the Center of the Milky Way: Structure of the Center. Horizons in
World Physics. Ed. Albert Reimer. 2015, v. 283, pp. 1-21. Nova Sci.
Publ. NY. ID_30510. https://www.novapublishers.com/catalog/pro
duct_info.php?products_id=54346.
[12] Zubow K., Zubow A., Zubow V.A. Revision of the Nature of
Gravitation and the Nature of the Chemical Bond, Ed. Advances in
Chemistry Research. Ed. J. C. Taylor. NY, NovaPublish. Inc. 2016, vol.
32, pp. 141-164.
[13] Zubow A., Zubow K., Zubow V. A. Nature of Energy. Phenomenon of
Electric Neutral Particles’ Emission in Chemical and Mechano-
Chemical Reactions. Horizons in World Physics. Nova Sci. Publish. NY,
2014, vol. 22, pp. 37-52. https://www.novapublishers.com/catalog/
product_info.php?products_id=50340&osCsid=994455a7afd05deb7075
94aaf444ed06.

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 7

USE OF FURAN DERIVATIVES


ACTING AS ELECTROPHILIC DIENOPHILES:
AN EXPERIMENTAL AND
THEORETICAL ANALYSIS OF
POLAR CYCLOADDITION REACTIONS 

In memory of Ernest Wenkert

Pedro M. E. Mancini* , Mauro Cainelli, †

Carla M. Ormachea and María N. Kneeteman


Laboratorio Fester, Química Orgánica, IQAL (UNL-CONICET),
Facultad de Ingeniería Química, Universidad Nacional del Litoral,
Santa Fe, Argentina

ABSTRACT
Furans have, fundamentally, biological and chemical uses. For
several years, we have been working with substituted furans as


Author Contributions: M. Cainelli realized joint to C. Ormachea the theoretical study. Authors
M. Kneeteman and P.M.E. Mancini wrote the protocol, and wrote the first draft of the
manuscript. All authors read and approved the final manuscript.
*
Corresponding author Email: pmancini@fiq.unl.edu.ar.

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178 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

eletrophilic dienophiles joint to different nucleophilic dienes in Polar


Diels-Alder Reactions (P-DA). The principal objective of this analysis
was the preparation of benzofurans and dibenzofurans. To develop these
reactions, we used conventional thermal conditions and microwave
irradiation. The solvents employed were organic ones and protic ionic
liquids (PILs). Also, we worked in free solvent conditions. We
demonstrated that substituted furans result good electrophiles in
cycloaddition processes. The best experimental condition was the
combination of microwave irradiation in presence of PILs like
ethylamonium nitrate (NEA) and N-methylimidazolium tetrafluoroborate
([HMIM][BF4]). When the nitro group is used as substituent in the
electrophile, the process is irreversible due to the loss of nitrous acid.
Then this substituent is convenient for this transformation. Moreover, a
computational theoretical study of furan reactivity as dienophile in Polar
Diels-Alder reactions (P-DA) was performed. For this purpose, the
Density Functional Theory (DFT) method was employed. The principal
aim of this theoretical study was to analyze the reactivity,
regioselectivity, solvent effect (organics and ionic liquids) and reaction
mechanisms of these reactions. Gaussian 09 is the software used to
develop the theoretical calculations. The functional applied was B3LYP
and the basis set 6-31G(d). The structures of reactants, products and
transition states were optimized and validated through the calculation of
the vibrational frequencies. For the analysis of reactivity and
regioselectivity, the nucleophilic and electrophilic global and local
indexes were used, respectively. The solvent effect was considered
employing two different solvatation models: the Polarizable Continuum
Model (PCM) and the Supermolecular Approach. For the mechanistic
analysis, the stationary points were located in the Potential Energy
Surface (PES) and then optimized and validated. The activation energy
values and reactions paths were analyzed for each reaction.

Keywords: polar cycloaddition reaction, carbocyclic, heterocyclic,


electrophile, DFT

ABBREVIATIONS
[BMIM][BF4]: Tetrafluoroborate n-butylimidazolium
[BMIM][PF6]: Hexafluorophosphate n-butylimidazolium
CA: Cycloadduct
DA: Diels-Alder
DFT: Density Functional Theory
FMO: Frontier Molecular Orbitals

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 179

HBD: Hydrogen Bonding Donor


[HMIM][BF4]: Tetrafluoroborate 1-butyl-3-methylimidazolium
[HMIM][PF6]: Hexafluorophosphate 1-butyl-3-methylimidazolium
HOMO: Highest Occupied Molecular Orbital
ILs: Ionic Liquids
IRC: Intrinsic Reaction Coordinates
LUMO: Lowest Unoccupied Molecular Orbital
MW: Microwave
NEA: Nitrate ethylamonium
P-DA: Polar Diels-Alder
PCM: Polarizable Continuum Model
PES: Potential Energy Surface
PILs: Protic Ionic Liquids
RTILs: Room Temperature Ionic Liquids
SEAr: Aromatic Electrophilic Substitution
TCE: Tetracyanoethylene
TS: Transition State

INTRODUCTION
Furan is an aromatic heterocyclic compound with, fundamentally,
biological and chemical uses. Two important derivatives are 2-furoic acid and
furfural. Nowadays, several synthesis processes are very well known. Among
them, it is convenient to cite the following ones: synthesis of Paal-Knorr,
synthesis of Feist-Berani, and those that use acyloins or alkyniloxiranes as
starting material. Industrially, the furan is obtained by decarbonylation of
furfural.
Although the principal reaction that the furan ring suffers is the
electrophilic aromatic Substitution (SEAr), it has been known that aromatic
heterocycles such as furan, thiophene, and pyrrole undergo Diels-Alder
reactions despite their aromaticity and hence its inertness expected. In view of
their electron-rich constitution and electron-donor properties, they have been
involved mostly as the diene component in the cycloaddition process. Thus,
furans have been used efficiently in this capacity since the early days of the
Diels-Alder reaction [1]. Also there are a limited number of examples of five-
membered aromatic heterocycles acting as dienophiles in Diels-Alder
reactions. A special driving force permits expression of such unusual
heterocycle behavior.

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180 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

In 1988 E. Wenkert demonstrated that β-acylfurans can act as electrophilic


dienophiles in its reaction with isoprene, showing to undergo high-yielding
cycloaddition products. This observation constituted the first indication of the
feasibility of normal Diels-Alder reaction with five-membered aromatic
heterocycles, holding electron-withdrawing groups, as electrophilic
dienophiles. The Diels-Alder reaction is considered to be a pericyclic process.
However, depending on the structure of the diene and the dienophile, the
nature of this reaction can change. For many years the Diels-Alder (DA)
reaction (Scheme 1) has remained as one of the most powerful transformation
methods in chemical synthesis, particularly for obtaining polycyclic rings.
With its potential to form carbon-carbon, carbon-heteroatom, and heteroatom-
heteroatom bonds, the reaction underlies the synthesis of diverse carbon and
heterocyclic compounds. It is a key step in the synthesis of many natural
products and pharmaceutical compounds [2].

Scheme 1. The Diels-Alder reaction.

In the last years, we have worked with aromatic nitro-substituted


compounds as electrophiles in cycloaddition reactions with different dienes.
They were able to show that a new aromatic system can act as electrophilic
dienophile in cycloaddition reactions. In the last cases the nitro group is lost
during the process as nitrous acid. This transforms it in an irreversible reaction
with the aromatization of the primary cycloadduct. In this context, mono and
disubstituted nitropyrroles, nitrofuranes, nitrothiophenes, nitroselenophenes,
nitroindoles, nitrobenzofuranes and nitrobenzothiophenes, were studied [3].
More recently a series of processes which facilitate the discussion of this
type of reactivity of furans derivatives were developed. One objective of this

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 181

analysis was the preparation of benzofurans and dibenzofurans. Then the


influence of the reactions variables of them, specially the solvent effect on this
polar Diels-Alder (P-DA) cycloaddition process, was studied [4].
For understanding the polar character trend in a series of P-DA reactions,
a theoretical study was developed using a known method.
Then the aims of this work are related to the electrophilic behavior of
furans, substituted with different electron withdrawing groups, when they
participate in polar reactions with nucleophilic dienes. For the development of
these reactions, we used conventional thermal conditions and microwave
irradiation (MW). The solvents employed were organic ones and protic ionic
liquids (PILs). Also, it was possible to work in free solvent conditions under
MW. Moreover, a theoretical study of the reactivity and the mechanism of
these reactions was done.

RESULTS AND DISCUSSION


Monosubstituted Furans as Electrophile

Cycloadditions reactions of isoprene, 1,3-butadiene and a 1,3-dioxy


derivative, respectively, with furfural, methyl 2-furoate, 3-furaldehyde, methyl
3-furoate, methyl 2-methyl-3-furoate and 3-formylbenzofuran were described
by Wenkert et al. The determination of the reaction parameters has led to the
conclusion that the 3-acylfuran/1,3-diene reaction is an efficient process,
useful for organic synthesis [5]. The cycloadditions in which the dienophile is
a 2-furan derivative have two problems. The first one is the low yields
observed. The second one is the presence of the cycloadduct in a relation 1:2
as principal product. In contrast to the 2-acylfurans, their 3 isomers, 3-
furaldehyde and methyl 3-furoate, showed to be excellent dienophiles, e.g.,
yielding bicycles derivatives (74 and 64% yields, respectively) interacting with
isoprene [6] (Scheme 2).

Scheme 2. Adducts of the reaction of 3-carbonyl substituted furans with isoprene.

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182 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

In an earlier work [7], the 2-nitrofuran was studied in DA reactions under


thermal conditions. In contrast to 2-acylfuran, 2-nitrofuran act as an efficient
dienophile when reacts with isoprene, 1-N-acetyl-N-propyl-1,3-butadiene, 1-
diethyl-amino-3-tert-butyldimethyl-siloxy-1,3- butadiene (Rawal’s diene) and
1-methoxy-3- trimethylsilyloxy-1,3-butadiene (Danishefsky’s diene) in
different reaction conditions and using organic solvents as reaction media. In
that direction, the efficacy of 2-nitrofuran as electrophilic dienophile in P-DA
reactions was tested. This study was completed testing the behavior of 3-
nitrofuran [8]. 2- and 3-Nitrofuran react efficiently with the mentioned dienes
since the nitro group induce side selectivity. Treatment of 2-nitrofuran and 3-
nitrofuran with isoprene gave a mixture of isomeric dihydrobenzofurans and
isomeric benzofurans as principal products in relation 1:1:3:3, with moderated
yield (with thermal extrusion of the nitro group in the form of nitrous acid
from the initial DA adduct) (Scheme 3). In general, the yields increase with
the increment of the temperature. It should be noted that isolation of the
primary nitro-adduct has not been achieved under these conditions.

Scheme 3. Reaction of 2- and 3-nitrofuran with isoprene.

The reaction of nitrofuran isomers with 1-N-acetyl-N-propyl-1,3-


butadiene afforded benzofuran with the loss of N-acetyl-N-propylamino and
nitro groups. In the same way, in the reactions with Rawal´s and
Danishefsky’s dienes, the aromatic cycloadducts, 5- and 6-
hydroxylbenzofurans, were obtained with moderate to high yields and
complete regioselectivity. It is necessary to take into account that, when the
Rawal’s diene was used, the first aromatic product obtained was the
benzofuran derivative with a trimethylsylil substituent in the positions 5- and
6-, which is then hydrolyzed during the purification process (Scheme 4).

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 183

Scheme 4. Reaction of 2- and 3-nitrofuran with Rawal’s diene.

The reaction of 3-formylbenzofuran and isoprene gave a mixture (67%) of


cycloadducts (see below) in a relation 2:1. Once again the presence of the
substituent in position 3- facilitates the process (Scheme 5).

Scheme 5. 3-carbonylbenzofuran and the adducts of its cycloaddition reaction with


isoprene

Moreover, the presence of a methyl group in position 2- joint to a carbonyl


group in position 3- of the furan ring, proved to be a poor electrophilic
dienophile. The blockage of cycloaddition on the acyl side of the furan by the
methyl group makes this 3-acylfuran behave like a 2-acylylfuran.
On the other hand, 2- and 3-nitrobenzofurans were studied in polar
thermal Diels–Alder reactions with normal electron demand using several
structurally different dienes -isoprene, 1-trimethylsilyloxy-1,3-butadiene and
Danishesfky’s diene-, under different thermal reaction conditions [9]. The
Rawal’s diene was used only with 2-nitrobenzofuran as a counterpart,
considering the class of silyloxy substituent this compound has. A very strong
electron-acceptor group, such as the nitro group, pushes the dienophilic
character of these heterocyclic compounds. The presence of the nitro group
enhanced the cycloaddition by favoring aromatization, thus preventing
retrocycloaddition (irreversibility). Since this substituent is easily extruded
under thermal conditions, this reaction sequence becomes a simple method for
the preparation of families of organic compounds with heteroatomic rings like
dibenzofurans derivatives. They are important heteroatomic compounds which
display a wide variety of biological activities. Firstly, we explored 2-
nitrobenzofuran as substrate, with the cited dienes. In this way, there is a
difference with other electron withdrawing groups, the acyl- substituent in the
heteroatomic ring do not promote the cycloaddition reaction when they are in
position 2-. The thermal reactions of 2-nitrobenzofuran with isoprene

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184 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

afforded the mixture of aromatic regioisomeric cycloadducts 3- and 4-


methyldibenzofuranes as principal products and 3- and 4-
methyldihydrodibenzofurans, with reasonable yield (total yield 75%). On the
other hand, reactions with 1-trimethylsilyloxy-1,3-butadiene, yielded
dibenzofuran (ca. of 70%) with the loss of trimethylsilyloxy and nitro groups.
In the same way, in the thermal reaction of 2-nitrobenzofuran with
Danishesfky’s diene, an aromatic cycloadduct (4-hydroxybenzofurane) was
obtained with very good yield and complete regioselectivity. Finally, the
reaction between this electrophile and Rawal’s diene was explored with
similar results to those of the cycloaddition using Danishesfky’s diene. In the
last two processes we observed the expected aromatization of the initial nitro
adduct, promoted by the loss of nitro, the metoxyl and the dimethylamine
groups, respectively (Scheme 6).

Scheme 6. Cycloaddition reaction of 2- and 3-nitrobenzofuranes with different dienes.

Disubstituted Furans as Electrophile

One efficient Diels-Alder reaction was that of dimethyl 2,4-


furandicarboxylate and isoprene. It afforded a 1:1 mixture (73%) of bicycles
derivatives and a ca. 1:1 mixture (25%) of tricycles derivatives [5] (Scheme 7).

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 185

Scheme 7. 2,4-furandicarboxylate and the cycloadducts of its reaction with isoprene.

Methyl 5-nitrofuran-3-carboxylate and methyl 5-nitrofuran-2-carboxylate-


demonstrated to be good electrophilic dienophiles. Exposure of methyl 5-
nitrofuran-3-carboxylate to isoprene yielded the mixture of isomeric
benzofuranes as principal product with good yield, and a mixture of double
addition adducts in low yield. On the other hand, reactions of methyl 5-
nitrofuran-2-carboxylate with isoprene gave a mixture of aromatic
cycloadducts of simple addition, with moderate yields. In general, the yield
increases with the temperature in every experiment [7] (Scheme 8).

Scheme 8. Cycloaddition reaction of 5-nitrofuran-2- and 3-carboxylate with isoprene.

When methyl 5-nitrofuran-3-carboxylate or methyl 5-nitrofuran-2-


carboxylate react, in thermal conditions, with 1-N-acetyl-N-propyl-1,3-
butadiene, the Rawal‘s diene and the Danishefsky’s diene, respectively, the
aromatic cycloadducts were obtained with excellent yield. All products
showed extrusion of the nitro group in the form of nitrous acid. These
reactions proceed by stereoselective addition of the diene to the nitro-
substituted double bond of the furan. Only 1:1 adducts whose structure reveals
site and regioselectivity were obtained (Scheme 9).

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186 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

Scheme 9. Cycloaddition reaction of 5-nitrofuran-2- and 3-carboxylate with different


dienes.

2-nitrofurans, in contrast to the 2-acylated furans, reacts efficiently with


dienes of different nucleophilicity in normal electron demand P-DA reactions,
with the nitro group inducing side selectivity. This substituent is easily
extruded under thermal conditions, giving cycloadducts of high interest as
intermediaries in the synthesis of some alkaloid families as morphine,
kreysiginine and codeine. Selecting the appropriate reagents (diene-
dienophile), the P-DA adducts for the total synthesis of these natural products
could be obtained in a simple way. Similarly, 3-nitrofuran derivatives showed
their dienophilic character taking part in a normal demand P-DA cycloaddition
reaction. These reactions could be considered domino processes that are
initiated by a polar DA reaction, and the subsequent concerted elimination of
nitrous acid from the [4 + 2] cycloadduct to yield the corresponding products.
A very strong electron-acceptor group, such as a nitro group, induces a
similar reactivity at 2- and 3-positions in the furan ring. The ease of thermal
extrusion of nitrous acid accompanying the DA reaction of 2- and 3-
nitrofurans and 2- and 3-nitrobenzofurans, followed by the further
aromatization, makes this reaction sequence a simple, economical and efficient
method of benzofurans and dibenzofurans preparation. They are direct
intermediates in the synthesis of important heteroatomic compounds that
display a wide variety of biological activities.

Solvent Effects in this Type on P-DA Reactions


The mechanisms of the analyzed reactions can be considered as concerted
asynchronous processes, that provides a polar character to the reaction (P-DA)
being able to experiment solvent effects [10]. Ionic liquids (ILs) have shown

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 187

that they can be a good alternative compared to conventional solvents due to


their physical properties such as low vapor pressure, reutilization capacity, and
less environmental aggressive preparation and degradation ways [11].
An IL is a salt-substance exclusively composed of cations and anions.
This fact differentiates them from simple ionic solutions, in which ions are
dissolved in the molecular medium. They are also different from inorganic
molten salts because their melting points are lower than 100 ºC. Room
temperature ionic liquids (RTILs) have been the subject of considerable
research efforts. RTILs have attracted considerable attention because they are
expected to be ideal solvents that could aid in novel reactions in green
chemistry [12]. In particular, the possibility of fine-tuning chemical and
physical properties by an appropriate choice of cations and anions as
stimulated much of the current excitement with respect to these compounds
and has led to the term “design solvents.” As a consequence, the
characterization of the properties of different classes of ILs used as solvent for
specific applications in chemical reactions and catalysis, has been intensively
investigated.
Recently, the reactions between 2- and 3-nitrofurans and dienes of
different nucleophilicity using protic ionic liquids (PILs) as solvents, were
analyzed. In these cases, furan acts as an electrophilic dienophile. The
experimental data showed that the presence of an IL as reaction media results
in higher reaction yields and in lower reaction conditions (temperature and
time) than those in which organic solvents are employed. It has been
demonstrated that, in this conditions, 2- and 3-nitrofuran react efficiently with
the selected dienes in normal electron demand P-DA reactions, with the nitro
group inducing the formation of a selective product. In that direction, the
results are similar in a structural form than those obtained with conventional
solvents. The PILs used were 1-methylimidazolium tetrafluoroborate
[HMIM][BF4], 1-methylimidazolium hexafluorophosphate [HMIM][PF6] and
ethylamonium nitrate [NEA]. The dienes employed were isoprene, 1-
trimethylsyliloxi, 1,3-butadiene and the Danishefsky´s diene. In thermal
conventional conditions, the experimental results showed that the reactions
developed using neoteric solvents were faster that those developed in a
traditional organic solvent [13]. In these cases, the yields for the reactions
developed in presence of PIL were good. Probably the influence of the PILs on
the electrophilic character of the dienophiles is related to the hydrogen-
bonding character of the nitro group.
When 2- and 3-nitrofuran were allowed to react with the less nucleophilic
diene (isoprene) in a sealed ampoule, using NEA as solvent, the reactions

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188 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

produced a mixture of isomeric benzofurans (1:1) as primary products in


moderate yield, and dihydrobenzofurans derivatives (1:1) in lower yield. The
reaction time was 24 h [14]. In the other way, the best yield for the reactions of
2- and 3-nitrofuran with 1-trimetylsylyloxi-1,3-butadiene was obtained with
NEA as solvent and a reaction time of 24 h. [HMIM][BF4] is a better solvent
than [BMIM][BF4], probably due to its HBD (Hydrogen Bond Donor)
character. Finally, the reaction of Danishefsky’s diene with 2- and 3-nitrofuran
(60ºC, 24 h) using the three PIL’s cited above, produced 5-
hydroxyibenzofuran in a reasonable yield. Similar results were obtained for
the reactions developed in these neoteric solvents with 2- and 3-
nitrobenzofuranes.

P-DA Reactions under Microwave Irradiation


The microwave-assisted controlled heating has become a powerful tool in
organic synthesis. This condition is being used to accelerate organic reactions
and, generally, increase the yield. The region of the microwave (MW)
irradiation is located between the infrared and the radio waves. In the MW
region, the electromagnetic energy affects the molecular rotation without
changing the molecular structure. The application of MW in organic synthesis
has been improved over the last years. This methodology results in better
reaction rates and yields. Using MW irradiation, the reaction mixture
undergoes heating by a combination of thermal effect, dipolar polarization and
ionic conduction. Then, the polar compound can absorb energy very efficiently
[15]. Although the MW effect is a topic of discussion yet, this proceeding has
been employed successfully, observing even modifications of the selectivity
[16].
Considering the influence of MW irradiation to improve organic reaction,
some experiences on P-DA process using this methodology were carried out.
Furan derivatives as electrophilic dienophiles, and dienes of different
nucleophilicity, were employed. At this point, it is necessary to take into
account that the combination of MW radiation and a protic ionic liquid (PIL)
as solvent (1-methylimidazolium tetrafluorborate) produces, on the reaction
between 2-nitro-N-tosylpyrrole and isoprene, a remarkable improvement on
the yield in a short time of reaction [17].
To explore the electrophilicity of 2-nitrobenzofuran, 3-nitrobenzofuran
and 5-nitrobenzofuran in the normal electron demand P-DA reaction, we chose
isoprene, 1-trimethylsyliloxy-1,3-butadiene, and Danishefsky’diene as dienes.
The reactions of 2- and 3-nitrobenzofurans with isoprene under MW
irradiation, using PILs [HMIM][BF4], [BMIM][BF4] and NEA as reaction

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 189

media, offer the isomeric dibenzofurans derivatives (1:1) in reasonable yield


(ca. 60%). In these reactions, it is not observed the dihydrobenzofurans
derivatives. When these electrophiles react with 1-trimethyl-1,3-butadiene,
using MW irradiation and the cited PILs as solvent, the only product was the
dibenzofuran. All reactions show a good yield (ca. 75%). On the other hand,
the reactions of 2- and 3-nitrobenfuran with Danishefsky’s diene in the
conditions indicated before, offer the isomeric hydroxilated dibenzofurans,
respectively, with good yield (ca. 75%). The reactions are regioselective as a
consequence of the diene substitution combined with its nucleophilicity.
When 5-nitrobenzofuran react with isoprene under MW irradiation in
presence of the cited PILs, in all cases the isomeric mixture (1:1) of methyl
benzo[e]benzofurans was obtained with reasonable yield (ca. 60%). On the
other hand, the reactions of this electrophilic dienophile with 1-trimethyl-1,3-
butadiene, in all the reaction conditions cited, produce the benzo[e]furan with
good yield (ca. 70%). Finally, the reactions of 5-nitrobenzofuran with
Danishefsky’s diene yielded in regioselective form the hydroxylic
benzo[e]indole with good yield (ca. 75%). Once again, the regioselectivity
observed is consequence of the type of diene employed and the nitro group
induce the electrophilic behavior in the ring in which it is present.
Reactions developed under MW irradiation and in free solvent conditions
offer similar results than when PILs are the reaction media.
Although the reactions with different pars diene/dienophile offer similar
behavior under conventional heating or with microwave irradiation, in the last
experimental condition we observe shorter times of reaction and better yields,
especially when the solvents are PILs or in free solvent conditions. Comparing
these two situations, the yields obtained are similar. The free solvent condition
is environmentally friendly allows a more simple manipulation of the system
and guarantee a good separation of the products at a lower cost. These results
explain why the free solvent condition is preferred.

Theoretical Calculations
Electronic properties of reactions involving 2- and 3- nitrofuran and
different dienes were studied theoretically in order to explain the reactivity and
mechanism of these processes.

Global Properties of dienophiles and dienes


The global electrophilicity (ω) of 2-nitrofuran and 3-nitrofuran is 2.51 eV
and 2.35 eV respectively (Table 1).

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Table 1. Dienophile electronic properties

Dienophile εHOMO(eV) εLUMO(eV) μ(eV) Η(eV) ω(eV) N(eV)


2-nitrofuran -0.26971 -0.09222 -0.180965 0.17749 2.51 1.78
3-nitrofuran -0.26712 -0.08632 -0.176700 0.1808 2.35 1.85

The dienes used were 2-methyl-1,3-butadiene (Isoprene), 1-methoxy-1,3-


butadiene and 1-methoxy-3-trimethylsilyloxy-1,3-butadiene (Danishefsky’s
diene). The differences of global electrophilicity (Δω) between the dienophile
and the different dienes (Table 2), allows analyzing the reactivity of the
reactions.

Table 2. Diene electronic properties

Diene εHOMO(eV) εLUMO(eV) μ(eV) η(eV) ω(eV) N(eV)


Isoprene -6.18 -0.41 -3.30 5.77 0.94 2.93
1-methoxy-1,3-butadiene -5.57 -0.14 -2.85 5.43 0.75 3.55
Danishefsky’s diene -5.56 0.04 -2.76 5.60 0.68 3.56

The dienophile has the highest value of electronic chemical potential (μ),
which indicates that the charge transference process derives from the dienes to
the first one. In these cases, the diene is going to act as nucleophile and the
dienophile as electrophile.
The diene that presents the highest difference of global electrophilicity
(Δω), respect to the dienophile, is the Danishefsky’s diene. Then, the polarity
of the reactions involving this diene would be higher and the results more
favorable than those in which 1-methoxy-1,3-butadiene and isoprene are
employed.

Local properties of dienophiles and dienes


The regioselectivity could be analyzed using the local electronic properties
of the dienophiles (Table 3) and the dienes (Table 4).

Table 3. Dienophiles’ local electrophilicity

Dienophile ωk(eV)
C2 0.10
2-nitrofuran
C3 0.38
C2 0.43
3-nitrofuran
C3 0.06

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 191

Table 4. Dienes’ local nucleophilicity

Diene Nk(eV)
C1 1.20
Isoprene
C4 0.92
C1 0.74
1-metoxy-1,3-butadiene
C4 0.94
C1 0.56
Danishefsky’s diene
C4 1.46

In case of 2-nitrofuran, the local electrophilicities of C2 and C3 are 0.10 eV


and 0.38 eV respectively (Δωk = 0.28 eV), while for 3-nitrofuran are 0.43 eV
and 0.06 eV respectively (Δωk = 0.37 eV). The most electrophilic center is, in
both cases, the carbon atom next to the carbon atom substituted with the nitro
group.
On the other hand, the dienes have a local nucleophilicity in C1 and C4 of
1.20 eV and 0.92 eV (ΔNk = 0.28 eV) respectively for isoprene, 0.74 eV and
0.94 eV (ΔNk = 0.20 eV) respectively for 1-metoxi-1,3-butadiene, 0.56 eV and
1.46 eV (ΔNk = 0.90 eV) respectively for the Danishefsky’s diene. The
regioselectivity is expected to be higher for the processes that involve
Danishefsky’s diene due to the higher difference in the local nucleophilicity
between C1 and C4, which is a consequence of the type of electron donor
groups (-OMe y –OSiMe3) and its relative positions.
It’s expected that atoms of the dienophile with higher values of local
electrophilicity will react with the atoms with higher values of local
nucleophilicity of the diene (C4 in the case of Danishefsky’s diene).

Mechanism of Reaction
In cycloaddition reactions, such as DA with nitro-dienophiles, primary
adducts retaining the nitro group are not observed. Therefore, there is an
elimination stage of the substituent group as nitrous acid that needs to be
consider. This is a domino process that involves consecutive reactions. If
asymmetric dienes such as Danishefsky’s diene are used, an extra stage,
corresponding to the elimination of -OMe group and hydrolysis of –OSiMe3
group, is also present. Here, only the process until the formation of the primary
adducts is consider in the mechanism study. This is because it is the
determinant step of the reaction. It is necessary taken into account that the
irreversible character of these reactions is due to the elimination of the nitrous
acid and the subsequent aromatization of the products.

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2-Nitrofuran and 3-Nitrofuran with Isoprene


The ΔNk between C1 and C4 of the diene is not enough to observe
regioselectivity and both isomers appear as products (Scheme 10). The
elimination of the nitro group allows obtaining a mixture of dihydro and
aromatic isomers as final products.

Scheme 10. Cycloaddition process of nitrofurans with isoprene.

The Δω of the reaction is 1.57 eV and 1.41 eV when 2-nitrofuran and 3-


nitrofuran are employed respectively. This reactions present one transition
state (TS, Figure 1) for each, para and meta products.

2-nitrofuran

3-nitrofuran
Figure 1. Transition states for para (left) and meta (right) cycloadducts.

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 193

This process is concerted and asynchronous because both formatting


bonds vibrate at the same time and in an asymmetric form [(Δrpara= 0.12 Å),
(Δrmeta= 0.30 Å), (Δr = (r1 – r2)] for 2-nitrofuran and [(Δrpara= 1.23 Å),
(Δrmeta= 1.37 Å), (Δr = (r1 – r2)] for 3-nitrofuran, where Δr is the difference in
length of the forming bonds at TS.
The energy barriers of both isomers are similar. This validates the
obtention of the two products. The energy barrier of the reaction of 2-
nitrofuran is higher (24 kcal/mol) than those in which 3-nitrofuran is employed
as dienophile (17.5 kcal/mol) (Figure 2a and 2b).

30 TS
25
20
15
24 kcal/mol
10
ΔE (kcal/mol)

5
(a)
0
-5
-10
-15
-20
-25
-30
CA
25
TS
20
15
10
17.5 kcal/mol
5
ΔE (kcal/mol)

(b) 0
-5
-10
-15
-20
-25
-30 CA
Figure 2. Reaction path of 2-nitrofuran (a) and 3-nitrofuran (b) with isoprene.

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194 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

2-Nitrofuran and 3-Nitrofuran with 1-Methoxy-1,3-Butadiene


In this case, the methoxy group is eliminated as methanol after the
cycloaddition reaction, together with the nitro group to reach the same
aromatic product (scheme 11).

Scheme 11. Cycloaddition process of nitrofurans with 1-methoxy-1,3-butadiene.

The Δω of the reaction for this system is 1.76 eV and 1.60 eV for 2-
nitrofuran and 3-nitrofuran respectively. This reactions present one transition
state (TS, Figure 3) for each, ortho and meta products.

2-nitrofuran

3-nitrofuran

Figure 3. Transition states for ortho (left) and para (right) cycloadducts.

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 195

This process is concerted and asynchronous because both formatting


bonds vibrate at the same time and in an asymmetric form [(Δrpara= 0.95 Å),
(Δrmeta= 0.76 Å)] for 2-nitrofuran and [(Δrortho=1,24 Å), (Δrmeta=0,90 Å)] for 3-
nitrofuran.
The energy barrier of the 2-nitrofuran ortho isomer is 22.5 kcal/mol and
the energy of the para isomer is slightly higher (ΔEa = 2.52 kcal/mol) (Figure
4a). The energy barrier of the ortho cycloadduct is higher than those in which
3-nitrofuran is employed as dienophile (13.37 kcal/mol). Moreover, the energy
difference between both isomers in this case is higher (ΔEa = 14.81 kcal/mol)
(Figure 4b).

ΔEa=2.52 kcal/mol
30 TS
25
20
15
22.5 kcal/mol
ΔE (kcal/mol)

10
5
(a) 0
-5
-10
-15
-20
CA

35 TS
30 ΔEa=14.81 kcal/mol
25
20
15
ΔE (kcal/mol)

10
13.37 kcal/mol
5
(b) 0
-5
-10
-15
-20 CA

Figure 4. Reaction path of 2-nitrofuran (a) and 3-nitrofuran (b) with 1-methoxy-1,3-
butadiene.

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196 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

2-Nitrofuran and 3-Nitrofuran with Danishefsky’s Diene


The ΔNk between C1 and C4 in this diene is high enough to observe the
formation of only one isomer (Scheme 12). In this case, the methoxy group is
also eliminated as methanol and the trimethylsilyloxy group is hydrolyzated
together with the elimination of the nitro group to reach the final hydroxylated
aromatic product.

Scheme 12. Cycloaddition process of nitrofurans with Danishefsky’s diene.

The Δω of the reaction for this system is 1.83 eV for 2-nitrofuran and 1.67
eV for 3-nitrofuran. The only product is the para isomer, which derives from
the bond between the most electrophilic atom of the dienophile (C3 for 2-
nitrofuran and C2 for 3-nitrofuran) and the most nucleophilic atom of the diene
(C4). This reaction is highly asynchronous and presents two transition states
(TS1 and TS2, Figure 5). The TS1 is associated with the bonding formation
between these two reactive atoms and the TS2 corresponds to the second
bonding formation between the dienophile and the C1 of the diene. The
difference in the bonding distances are [(ΔrTS1=1,24 Å), (ΔrTS2=0,90 Å)] for 2-
nitrofuran and [(ΔrTS1=1,24 Å), (ΔrTS2=0,90 Å)] for 3-nitrofuran.
We are in presence of a two-step mechanism. This fact does not agree
with the pericyclic reaction concept. In a similar way, we detected, in a
theoretical study, that the reaction of 3-nitropyridine as electrophilic
dienophile with Danishefsky’s diene, presents a mechanism with two step and
a detectable intermediate state [25].
The primary cycloadduct is not observed in all of these cases because of
the relative stability of the nitrate adduct respect to the final product with
elimination of nitric acid. The irreversible step of the DA reaction (impulsive
force) is the extrusion of nitrous acid and the stability is related to the
aromaticity of the final product.

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 197

The energy barrier of the TS1 is 18.95 kcal/mol and the energy of the TS2
is 16.83 kcal/mol for 2-nitrofuran (Figure 6a). The energy barrier of the para
cycloadduct is higher than those in which 3-nitrofuran is employed as
dienophile (11.31 kcal/mol and 6.55 kcal/mol for TS1 and TS2 respectively,
Figure 6b). Moreover, the energy of the TS1 is the determinant step of the
reaction because it presents higher values of energy than the TS2.

2-nitrofuran

3-nitrofuran

Figure 5. Transition states for the para cycloadduct.

Influence of Neoteric Solvents

For this analysis, the dienophile used was the 3-nitrofuran and the IL
employed for the solvent effect was tetrafluoroborate of 1-methyllimidazolium
-[HMIM][BF4]- (Figure 7). This selection is related to the possibility of
hydrogen bonding formation between this molecules.
If the classical interaction between the electrophile and the IL is
considered, which means that the IL cation interact via hydrogen bonding with
the nitro group of the electrophile, the global electrophilicity index is too high
(ca. tetrafluoroborate of 1-methyllimidazolium and ethyl ammonium nitrate).
These results are not compatible with the experimental data, probably because

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198 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

the anion is not considered formally (although the rate of the reaction is higher
than those when a molecular solvent is used, the yields and the temperature of
the reactions do not change enough).
25
TS1
TS2
20

15

10 18.95 kcal/mol 16.83 kcal/mol


ΔE (kcal/mol)

5
(a)
0

-5

-10

-15

-20 CA
15 TS1
10 TS2

5 11.31 kcal/mol
6.55 kcal/mol
ΔE (kcal/mol)

0
(b)
-5

-10

-15
CA
-20

Figure 6. Reaction path of 2-nitrofuran (a) and 3-nitrofuran (b) with Danishefsky’s
diene.

Figure 7. [HMIM][BF4].

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 199

For this reason, the supermolecular approach was explored. In this case
the anion is considered. When one par anion-cation of IL is taken into account,
the anion interacts only with the cation (electrostatic interactions) without
affecting the 3-nitrofuran, enabling a planar arrangement of hydrogen bond
between the cation and the dienophile (Figure 8).

Figure 8. One and two pairs anion-cation of IL interacting with the dienophile

When a second IL par anion-cation is added, an additional electrostatic


interaction with the dienophile appears and, therefore, the value of decreases
from 5.01 to 3.78 eV. This trend continues when the 3rd and 4th IL molecules
are added, with ω values of 3.11 and 2.84 eV respectively (Figure 9).
Based on these results, it can be considered that when a specific
interaction of hydrogen bonding on the dienophile predominates, the value of
the global electrophilicity increases, while when we are also in the presence of
electrostatic interactions, the effect is counteracted.
It is noted that the global electrophilicity of the dienophiles increases
when the solvent effect is considered respect to the gas phase. Furthermore,
the highest values correspond to the influence of the IL.

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200 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

Figure 9. Optimized geometries observed when the 3rd and 4th IL molecules are
added.
There is a second ω value that is related to the structure where a same
anion is interacting with both, the dienophile and the cation, which reduces the
effect of the hydrogen bonding interaction due to the rotation in the plane
(Figure 10).

Figure 10. Structures observed when a same anion is interacting with both, the
dienophile and the cation.

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 201

In this case, ω values are lower, 3-nitrofuran+[HMIM][BF4]x1 (2.91 eV);


3-nitrofuran+[HMIM][BF4]x2 (2.92 eV); 3-nitrofuran+[HMIM][BF4]x3 (2.75
eV); 3-nitrofuran+[HMIM][BF4]x4 (2.74 eV). This seems to be due to the
diminution of the interaction between the orbital of both, the dienophile and
the solvent, which is a consequence of the non-planar arrangement.
Moreover, incorporating other IL molecules, the number of electrostatic
interactions increases, including new solvent-solvent interactions. The ω
values fluctuate around 2.80 eV due to the different relation between
electrostatic and hydrogen bonding effects.
Additionally, the global electrophilicity value of 3-nitrofuran in the
presence of IL using the PCM method, is lower than the one obtained using
the explicit method, its value is in fact similar to water, because it is a polar
solvent and, with this method, those solvents having similar dielectric constant
values will provide similar results regardless of their structures.
Global electrophilicity values obtained by the second situation provide
more consistent results with the experiences, where the reaction yields are
slightly higher and the reaction conditions, time and temperature, decrease.

Computational Methods
The theoretical DFT calculations were carried out using the Gaussian 09
[18] program. The hybrid functional [19] employed was B3LYP [20], together
with the standard 6-31G(d) basis set [21].
Initially, the geometric structures of reactants and products were
optimized using the Berny analytical gradient optimization method [22]. The
mechanistic study was realized through the construction of the Potential
Energy Surface (PES) for every system. The structures of transition states
(TSs) were located, optimized and then verified through IRC (Intrinsic
Reaction Coordinates) calculations [23]. The frequency calculations were
realized in order to validate the optimized structures. Reactants and
cycloadduct (CA) structures were verified by the absence of negative
frequencies and the TSs by the presence of only one imaginary frequency.
The reactivity of the systems was studied using some indexes defined in
terms of the electronic chemical potential (μ) and the chemical hardness (η).
The chemical potential is the charge transfer capacity of the system in basal
state and the hardness is the resistance to change the chemical potential when
the number of electrons variates. Both quantities may be approached in terms
of the one electron energies of the frontier molecular orbital HOMO and
LUMO, εH and εL [24].

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202 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

εH +εL
μ= 2
(1)

η = εL − εH (2)

The global electrophilicity index, ω, represents the capability of a


molecule to accept an electron considering the environment satured by them.
This index it’s given by the following simple expression.

μ2
ω = 2η (3)
Recently an empirical (relative) nucleophilicity index, N, has been
introduced based on the HOMO energies obtained within the Kohn-Sham
scheme.

𝑁 = εHOMO(Nu) − εHOMO(TCE) (4)

The nucleophilicity is referred to tetracyanoethylene (TCE), which


presents the lowest HOMO energy in relation to large series of molecules
already investigated in the context of polar cycloadditions [25].
Fukui function is a measure of the sensibility of the chemical potential in a
particular point when an external perturbation is present and the number of
electron remain constant. It’s also defined as the variation of the electronic
density in a point when the number of electron changes and the external
potential remains constant [26].

∂ρ(r) ∂μ
f(r) = ( ∂n
) = (∂v(r)) (5)
v(r) n

The resolution of this function can be obtained in terms of the FMO.

fkα = ∑μϵk fμα (6)

2
fμα = |cμα | + cμα ∑v≠μ cvα Sμv (7)

fk+ and fk- are the Fukui functions for a nucleophilic and electrophilic attack,
respectively. Local electrophilicity and nucleophilicity indexes, ωk and Nk, can
be obtained using the following expressions.

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 203

ωk = ωfk+ (8)

𝑁k = 𝑁fk− (9)

The optimization structures provide the values of HOMO and LUMO


orbitals that are used to quantify the reactivity indexes [27].
The solvent effect was studied using an explicit model of solvation, the
supermolecular method, which consist on adding molecules of ILs together
with the dienophile and then optimize the system. The molecules of ILs were
added progressively until a total amount of 4. The effect is analyzed only with
the dienophile due to the capacity of this one to form hydrogen bonds. In order
to compare the results of this model, we analyzed the solvation effect using the
polarizable continuum model (PCM), where the solute (dienophile) is placed
into a cavity and the interaction with the solvents are considered only through
their dielectric constants, independently of their structure [28].

CONCLUSION
In this work the use of furan derivatives acting as electrophilic dienophiles
in P-DA cycloaddition was demonstrated. These reactions are useful for the
preparation of benzofurans and dibenzofurans in one step. The discussion as
extended considering solvent effects influence, specially the employment of
PIL´s. In this conditions it could be demonstrated that the TS of the reactions
have charge separation and then, the solvent effect is important. Moreover, the
influence of different forms to develop the process was analyzed. In this sense,
the use of microwave irradiation gave the best results.
On the other hand, a computational theoretical study of the reactivity of
furans 2- and 3- nitro substituted as dienophiles in P-DA reactions involving
several nucleophilic dienes was realized. For this purpose, the Density
Functional Theory (DFT) method was employed. When isoprene and 1-
methoxy-1,3-butadiene are employed, only one TS is observed. However
when Danishefsky’s diene is employed, two TSs are noted corresponding with
each bonding formation. The similar activation energies of the reactions
employing isoprene as diene, explain that the obtention of both isomers is
equally possible. Moreover, when 1-metoxy-1,3-butadiene is employed, the
mechanism suggests a preference for the ortho cycloadduct. However, the final
product is the same and the experimental determination is not possible.
Finally, the reactions involving the Danishefsky’s diene are completely

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204 Pedro M. E. Mancini, Mauro Cainelli, Carla M. Ormachea et al.

regioselective and the product is the result of the bonding formation of the
most nucleophilic atom of the diene with the most electrophilic atom of the
dienophile. The TS1 is the determinant step of these reactions due to it is
higher value of activation energy.
The global electrophilicity of the dienophile increase when IL is employed
as a reaction media. The PCM is not adequate to explain this effect because it
does not consider the solute-solvent specific interactions. On the other hand,
the supermolecular approach considers the hydrogen bonding interaction and
is more consistent with the experimental results obtained when 2- and 3-
nitrofuran are used as electrophiles in DA reactions using ILs as solvents.
In general the experimental results were coincident with the theoretical
analysis.

ACKNOWLEDGMENT
This research was supported by the Agencia Nacional de Ciencia y
Tecnología (ANCyT) of Argentina -PICT 2014 No.1587-, and by CAI+D
2011 -No 66, 501 201101 00478 LI-at the Universidad Nacional del Litoral,
Santa Fe, Argentina. C. Ormachea thanks to the Consejo Nacional de
Investigaciones Científicas y Técnicas (CONICET) of Argentina for the
scholarship.

Funding Sources

ANCyT (Agencia Nacional de Ciencia y Tecnología de la República


Argentina). MINCyT (Ministerio de Ciencia y Tecnología de la República
Argentina). Universidad Nacional del Litoral, Santa Fe, República Argentina.
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET),
Argentina.

REFERENCES
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(b) Diels O; Alder K. Justus Liebigs Ann. Chem. 1931, 490, 243.

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The Use of Furan Derivatives Acting as Electrophilic Dienophiles 205

[2] Carruthers W. Cycloaddition Reaction in Organic Synthesis. Pergamon


Press, Oxford, UK, 1990.
[3] (a) Biolatto B; Kneeteman M; Paredes E; Mancini PME. J. Org. Chem.
2001, 66, 3906. (b) Paredes E; Brasca R; Kneeteman M; Mancini PME.
Tetrahedron, 2007, 63, 3790.
[4] Hayes BL. Microwave Synthesis. Chemistry at the Speed of Light. CEM
Publishing, USA, 2002.
[5] Wenkert E; Piettre SR. J. Org. Chem. 1988, 53, 5850.
[6] Wenkert E; Moeller PDR; Piettre SR. J. Am. Chem. Soc. 1988, 110,
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[7] Della Rosa C; Kneeteman MN; Mancini PME. Tetrahedron Lett. 2005,
46, 8711.
[8] Brasca R; Della Rosa C; Kneeteman M; Mancini PME. Letters in
Organic Chemistry, 2011, 8, 82.
[9] Della Rosa C; Sanchez JP; Kneeteman M; Mancini PME. Tetrahedron
Lett. 2011, 52, 2316.
[10] Bini R; Chiappe C; Mestre VL; Pomelli CS; Welton T. Theor. Chem.
Acc. 2009, 123, 347.
[11] Mancini PME; Fortunato G; Bravo MV; Adam C. Ionic Liquids: Binary
Mixtures with Selected Molecular Solvents. Caracterization of its
Molecular-Microscopic Properties. Reactivity. Chapter 13 in Green
Solvent Book 2, Springer, UK, 2012, 335.
[12] (a) Hichcock PE; Mohammed TJ; Seddon JA; Hussey CL; Ward EH.
Inorg. Chim. Acta. 1986, 19. (b) Welton T. Chem. Rev. 1999, 99, 2072.
[13] Kneeteman MN; Della Rosa CD; Lopez Baena F; Mancini PME.
International Journal of Pure and Apllied Chemistry, 2015, 8, 229.
[14] Della Rosa CD; Ormachea CM; Kneeteman MN; Adam CG; Mancini
PME. Tetrahedron Lett. 2011, 52, 6754.
[15] Deb S; Wähälä K. Steroids, 2010, 75, 740.
[16] (a) Tsuji M; Nishizawa Y; Kubokawa M; Tsuji T. Chemistry-A
European Journal, 2005, 11, 440. (b) Topsett GA; Conner W C;
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Cruz P. Combinatorial Chemistry and High Throughput Screening,
2007, 10, 766.
[17] Mancini PME; Kneeteman MN; Della Rosa CD; Ormachea CM; Suarez
A; Domingo LR. Tetrahedron Lett. 2012, 53, 6508.
[18] Frisch MJ, et al. Gaussian 09, revision C.01, gaussian, Inc., Wallingford
CT; 2010.

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[19] (a) Parr RG; von Szentpaly L; Liu S. J. Am. Chem. Soc. 1999, 121,
1922. (b) Parr RG; Pearson RG. J. Am. Chem. Soc. 1983, 105, 7512. (c)
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Phys. 1988, 37, 785. (b) Becke AD. J. Chem. Phys. 1993, 98, 5648.
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Sheikhet I. Quantum chemical and statistical theory of solutions a
computational approach. Ellis Horwod, London, 1995.

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In: Advances in Chemistry Research. Volume 37 ISBN: 978-1-53611-041-8
Editor: James C. Taylor © 2017 Nova Science Publishers, Inc.

Chapter 8

THE USE OF SURFACTANTS IN THE


CHEMICAL TREATMENT OF A CRUDE CASE
ORIENTAL ZONE IN VENEZUELA FOR
ASPHALTENE MITIGATION

Juan Pereira1, Henry Labrador1, Víctor Perez1,


Ivan Villanueva1, José Bustamante1, Sofia Guevara2,
Maria Mannello2 and Miguel Parra2
1
Laboratorio de Petróleo, Hidrocarburos y Derivados,
Facultad Experimental de Ciencias y Tecnologia Departamento
de Química, Universidad de Carabobo, Venezuela
2
PDVSA Oriente Division Furrial, Venezuela

ABSTRACT
Petroleum asphaltenes are interesting molecules that have been
studied extensively due to various problems caused by the petroleum
industry. The high tendency of self association of asphaltene behavior is
the main strength of this complex behavior of crude oil. The phenomenon
of aggregation is the association of asphaltene molecules in particles of
different shape and size. This chapter of book discusses the use of
surfactants in oil chemical treatment to remedy the problems caused by
asphaltenes during production of crude oil. In this particular and specific
case it will be presented of Furrial area in eastern Venezuela, which is
one of the most important operational areas of the oil industry at
Venezuela. Different changes in pressure, temperature, and secondary

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208 Juan Pereira, Henry Labrador, Víctor Perez et al.

recovery methods may induce aggregation asphaltenes. These changes


affect their interfacial properties because they can alter the
physicochemical properties in the medium (crude oil or solvent) where
they are present. The adsorption process of asphaltenes in liquid-liquid
aggregates interfaces, it is slower than the adsorption of asphaltene
molecules. The aggregation state of asphaltene also depend on the nature
of crude oil. Furrial crude oil has always been characterized by the high
tendency to precipitation of asphaltenes.
Asphaltenes aggregation significantly affects the stability of
emulsions, oil viscosity and asphaltene precipitation. Recently, Acevedo
showed the colloidal nature of asphaltenes focused on the point of view
of its fractions A1 and A2, the first insoluble in toluene. The A1 fraction
is responsible for the aggregation behavior due to its solubility behavior.
This model can explain in detail their aggregation in crude oil and other
organic solvents. Surfactants are commonly used in the solution to the
problem of asphaltenes in crude oil production, and stability of water in
oil and asphaltene precipitation.
Surfactants are the main asset of the formulation components (a
chemical cocktail) used in the chemical treatment. Also the other
formulation components and additives such as polar solvents, play a key
role. A good formulation should increase surfactant´s properties such as,
surface tension, adsorption, interfacial activity, solubility, etc. The
appropriate combination between surfactants in solvent is important to
achieve high dispersion thereof. Besides the proper structure and
conformation they are key in the work of chemical treatment. For this
reason the addition of molecules that help disperse the surfactant is very
important. There are still many things that explain the action of
surfactants in the chemical treatment. However, this chapter aims to
provide an efficient contribution to this important issue related to the oil
industry.

Keywords: surfactants, asphaltenes, emulsions, asphaltene precipitation

1. SURFACTANTS: STRUCTURE AND PROPERTIES


Surfactants are chemical species amphiphilic, ie, molecules that exhibit
areas with opposite affinities typically identified by a polar zone (head) and a
nonpolar area (tail), see Figure 1. The polar area (hydrophilic region), favors
dissolution in polar solvents, this type interactions can occur by hydrogen
bonding or electrostatic interactions. The area nonpolar (hydrophobic region),
interacts weakly with the water, so it is responsible for activating the
hydrophobic effect. This interaction is based on the London dispersion force
[1].

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The Use of Surfactants in the Chemical Treatment … 209

Polar Zone

Non Polar Zone

Figure 1. Typical Conformation of a surfactant.

In the surfactants there is a close relationship between the structure and


properties have exhibited in solvents, ie, the molecular structure thereof
determines its behavior at the interfaces and the physicochemical properties of
their solutions.
The structure of a surfactant is quite diverse and even not all obey to that
shown in Figure 1. They have also been synthesized a wide range of
surfactants, which do not comply with this model, the case of polymeric
surfactants. In all cases, the existence of distinct zones with opposite affinities
met, which gives them the amphiphilic character and the corresponding
interfacial activity.
One of the properties of aqueous systems is affected by the presence of a
surfactant is the surface tension. This decreases as the concentration of
surfactant in an aqueous system increases to a limit value reached in the
critical micelle concentration (cmc). From this point the surface tension be
maintained constant. This surface tension drop is due to the adsorption of the
surfactant in the water-air interface. When the cmc is reached the interface is
saturated with surfactant, so a further increase forces it to be located within the
body of liquid. The area per molecule is a measure of the ability to form

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210 Juan Pereira, Henry Labrador, Víctor Perez et al.

packaged in the interface layers. It is generally accepted that lower molecular


interfacial areas indicate an increase in packaging capacity at the interface [2].
The viscosity of aqueous system is another property that can be influenced
by the presence of surfactants. The rheology of systems is modified by the
formation of wormlike micelles (WLM), for example. One of the most
extensively studied WLM systems, is based on the combination of the cationic
surfactant cetyltrimethylammonium bromide (CTBA) with the aromatic
cosolute or sodium salicylate. It has been shown, in the presence of the
cationic surfactant CTAB, small variations in the structure of aromatic co-
solutes significantly affect this type of training and thus the rheology of the
system [3].
There are different parameters that allow surfactants relate their structure
with the behavior exhibited at the interface. This provides a tool that allows
the appropriate selection depending on the application for which will be
employed. Useful parameters for characterizing surfactants are, among others,
hydrophilic-Lipophilic Balance (HLB), the phase inversion temperature (PIT),
the R Winsor and surfactant affinity difference (SAD).

1.1. Surfactant: The Balance Hydrophilic-Lipophilic (HLB)

The HLB indicates a balance between hydrophilic and lipophilic


tendencies of the surfactant. This is based on an experimental method of
attributing a number HLB surfactants from data concerning the stability of an
emulsion. This parameter, which is a scale between 0 and 20, was introduced
by Griffin in 1949. He found a relationship between the nature of the
surfactant and its properties as a surfactant and emulsifier and used it to make
emulsions of O/W. This number, which is invariable, can be estimated
according to the molecular structure or by reference, two known HLB
surfactants [4].
The HLB of a surfactant is related to its solubility. That is, a surfactant
having an HLB number low (below 9.0) tend to be oil soluble (lipophilic).
This shows that the lipophilic part is dominant in the type of interaction
established with the solvent. One having a high HLB number (above 11.0)
tend to be water soluble (hydrophilic). In this case, the dominant region is the
hydrophilic part. Those in the range of 9 to 11 are intermediate [5].
Two surfactants can have the same HLB and still exhibit different
solubility characteristics. This shows some of the limitations of this value as
the only parameter to be considered in the formulations.

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The Use of Surfactants in the Chemical Treatment … 211

The HLB of a surfactant, based on their structure, can be determined by


two methods: Griffin method or the method of Davies.
The HLB by Griffin’s method can be obtained using equation 1:

HLB = 20 * MH/M (1)

Where MH is the mass of the hydrophilic area and M the molecular mass
of the whole molecule.
The HLB Davies method assigns a specific value to different groups
which are present in the surfactant and the contribution of each to the HLB is
determined by equation 2:

HLB = 7 + hydrophilic groups - hydrophobic groups (2)

The HLB of a mixture of surfactants responds to a linear behavior. When


two or more surfactants are mixed, the resulting mixture will have an HLB
intermediate. It could be calculated from a linear average mixing rule based on
weight composition. That is obtained by Equation 3:

HLBMixture = x1* HLB1 + x2*HLB2 (3)

where HLB1 and are HLB2 HLB surfactants numbers 1 and 2, x1 and x2 their
weight fractions in the mixture and HLBMixture, the HLB of the surfactant
mixture. For example, suppose you want to determine the HLB of a mixture of
70% of TWEN 80 (HLB = 15) with 30% SPAN 80 (HLB = 4.3). The
calculation proceed as follows:

HLBMixture = 0,7 * (15) + 0,3 * (4,3) = 11,8

the mixture of these surfactants generates an HLB of hydrophilic nature. One


can see that, by varying the proportions of the surfactants, can obtain a range
of intermediate HLB. Also, when it is desired to obtain a required HLB, it can
be used to find the proportions in which should be mixed surfactants that are
available. HLB values for a wide variety of surfactants have been calculated
[6].
Salager found that these methods are consistent with each other, ie, both
the rule of linear mixing as the stability of the required HLB of the oil, are
obtained with an accuracy of plus or minus one unit in most cases for HLB
between 4 and 14. However for HLB values greater than 14, the mixing rule
exhibits nonlinear trend [7].

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212 Juan Pereira, Henry Labrador, Víctor Perez et al.

The HLB is a valuable tool to identify the nature of the surfactant,


however, in preparation of formulations use is limited, because it is based only
on the structural aspects of the surfactants. The physico-chemical environment
of the solution and temperature, it’s not considered. All these aspects affect the
interfacial activity of the surfactant in the future applications.

1.2. Polymeric Surfactants: Case Triblock Copolymers and


Ethoxylated Nonylphenolics Resins

The simplest model of a polymeric surfactant is represented by a


homopolymer, formed from the same repeating units (same monomers).
Examples of this type is poly (ethylene oxide) (PEO) and poly
(vinylpyrrolidone) (PVP). Homopolymers have low surface activity at the oil/
water (O/W) which does not make the most suitable emulsifiers [9].
Same polymers may exhibit interfacial activity with changes in its
structure. Such is the case of silicones. Silicones consist of a backbone with
pendant organic groups siloxanes, usually methyl groups. Both the nature of
the backbone and the pendant organic groups of contributing to surface
activity of the polymer. Polydimethylsiloxanes are the most common and have
the most interesting interfacial properties. Its overall structure is (CH3)3SiO
[(CH3)2SiO]nSi(CH3)3 wherein n is about 0 to 2500. This type of polymer has
low intermolecular forces, resulting from the intrinsic surface activity together
with the methyl group the unique flexibility of the siloxane backbone,
allowing the polymer adopt a variety of configurations leaving exposed
surface methyl groups [10].
Siloxane surfactants are different from conventional hydrocarbon
surfactants in being surface active in media and lowering nonaqueous surface
tension to 20 dynes/cm [11].
The polymers exhibit higher interfacial properties are copolymers,
macromolecules formed by the repetition of different monomers. The best
polymeric surfactants are the block and graft copolymer type. Polymeric
surfactants block (A-B or A-B-A) or graft type (BAn) are essential materials
for the preparation of many emulsion systems. Most block copolymers and
graft have low critical micelle concentrations (cmc), and in many cases is not
easy to measure the cmc for these blocks and graft copolymers.
These include the Pluronics and the ethoxylated alkylphenolic resins.
These are nonionic surfactants. Usually, they have been used in the chemical

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The Use of Surfactants in the Chemical Treatment … 213

treatment of crude oil. These have important interactions with asphaltenes,


responsible for different effects on interfacial petroleum.
The Pluronic Block Copolymers or poloxamers, are widely used in
various drug delivery systems and in tissue engineering. “Pluronics” are
triblock copolymers of “poly” (ethylene oxide) (PEO) and “poly” (propylene
oxide) (PPO), arranged in the PEO-PPO-PEO structure. Depending on the
length of the blocks of hydrophilic-lipophilic balance (HLB) of the
copolymers changes. In the “Pluronics” micelles form spontaneously solution
above the CMC.
In the Pluronics solution spontaneously form micelles above the critical
micelle concentration (CMC). The core of the micelles containing PPO blocks
and allows the incorporation of hydrophobic molecules [12].
Each group Pluronics series are identified by a letter and two digits. The
letter denotes the presentation of the substance: (L) liquid (P) to form a paste,
(F) solid, enough to form flakes. The first digit indicates the relative molecular
weight of the hydrophobic unit and the second, the relative hydrophilic-
hydrophobic relationship. Thus, the Pluronics L62, L64 and L68 have the
same molecular weight of the hydrophobic unit and its difference is increased
hydrophilic block. Instead, L44 and L64, has hydrophobic unit with different
molecular mass and equal hydrophilic-hydrophobic balance [13].
The alkyl phenolic resins are copolymers of an alkyl phenol and
formaldehyde. These are synthesized in a wide range of proportions. The
effect of various “amphiphiles” in the asphaltene stability dissolved in toluene
was reported and it was concluded that the solutions in toluene, containment of
alkylphenols as nonylphenol were less sensitive to precipitation by addition of
low molecular weight alkanes. This effect was attributed to acid-base
interactions between asphaltenes and “amphiphile” as well as steric
stabilization provided by the hydrocarbon tail.
Simulations were performed which revealed that alkylphenols tend to
form hydrogen bonds with -OH and -N periphery of asphaltenes. It has been
confirmed by high-resolution transmission electron microscopy (HRTEM)
octylphenol that can reduce the size of the aggregates of asphaltenes.
Octylphenol achieved saturating the hydrogen bonds asphaltene inhibiting
aggregation among themselves [15].
This is of particular interest, considering that hydrogen bonds play an
important role in the phenomenon of aggregation of asphaltenes. An
amphiphilic dispersant generally contains “anchor” polar group, which adheres
to the surface of asphaltenes, and an alkyl group “blocking” blocks other
asphaltene molecules and allows the dispersant is soluble in aliphatic solvents.

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214 Juan Pereira, Henry Labrador, Víctor Perez et al.

The heteroatom-containing polar group while the aliphatic tail, typically, have
less than 16 carbon atoms to prevent crystallization

1.3. Interactions of Surfactants in the Mixtures

When nonionic and anionic surfactants are mixed often exhibit strong
interactions that affect surface activity. These interactions give rise to
significant deviations from ideal mixing laws and adsorption behavior [16].
For example, AOT adsorption at the oil/water precoated consider Tween
80. Tween 80 layers adsorbs irreversibly after the interface is rinsed with
deionized water. The presence of Tween 80 at the oil water inhibits adsorption
of AOT [17].
May interface is acting to fractionate species in more persistent surfactant
commercially. However, the irreversible adsorption is consistent with recent
molecular simulations, due to interactions between poly (ethylene oxide) head
groups and the interface [18].

2. ASPHALTENES: INTRODUCTION
Asphaltenes are the fraction of petroleum compounds defined based on its
solubility, and insoluble in n-heptane and soluble in toluene. Here the
problems of asphaltenes begins in its very definition. Also are compounds of
higher polarity oil. Among its main features is that they are a complex mixture
of hydrocarbons. But undoubtedly are molecules with fascinating properties.
They have represented the scientific and technical challenge because of its
complex phase behavior in the oil industry. In exploration and production, the
adsorption of asphaltenes on mineral produces wettability changes and
blockage of pores in the reservoir rock. Adsorption at interfaces also raw water
produces very stable emulsions. Aggregation leads flocculation and
precipitation in different production facilities and even in the same reservoir
rock. During refining asphaltenes produce poisoning of catalysts and are
responsible for the formation of coke in the process of improvement. In most
of these cases are surfactants which can counteract the negative effect of
asphaltenes in oil. This chapter discusses this and some examples of
operational areas in Venezuela.

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The Use of Surfactants in the Chemical Treatment … 215

From the structural standpoint asphaltenes oil molecules are characterized


by a polyaromatic core which is substituted in the peripheries by alkyl chains.
This petroleum fraction as much heteroatoms (sulfur, nitrogen and oxygen)
and metals (vanadium and nickel) is concentrated.
The main intermolecular interactions in the asphaltenes are: acid-base
hydrogen bonds, pi-pi interactions, among others. It found an interesting
interaction of alkylphenols with asphaltenes.

2.1. Asphaltenes: Solubility

Asphaltenes are solid dark color, its chemical structure has been the
subject of many investigations [19, 20, 21] are pericondensaded polyaromatic,
with the presence of heteroatoms (N, S, O) trace metals (V, Ni, Fe) with and
naphthenic alkyl chains attached to the aromatic centers. In the oil it is in form
of colloid being in equilibrium pseudo which may rupture thermodynamic
changes as pressure and temperature, and this may cause precipitation
generating multiple disadvantages in the oil industry [22, 23] and operationally
defined as present in the fossil fuel as soluble and insoluble aromatic solvent
on a paraffin low molecular mass fraction.
The problems caused by this fraction during oil production, are connected
to the tendency of separation of the asphaltenes from the liquid phase,
increasing costs in the oil industry, flocculation can be induced by changes in
pressure, temperature, composition that generates a reduced stability of
colloids of asphaltenes in oil. Crude Oil is a complex mixture and describe
each fraction present in detail is impractical. The asphaltenes found in
petroleum bordered by the other fractions keeping in suspension, which can
use the Flory-Huggins theory, applied in polymers, which are mixtures of
large molecules with many small solvent, which is you can be used to
determine the solubility of asphaltenes in mixtures of thermodynamic manner.
The first model solubility for asphaltenes was proposed by Hirschberg [24],
which has been used in many applications and have made many adaptations.
Applying the theory Flory-Huggins of the Gibbs energy change by mixing
asphaltene with maltenes at constant pressure and can be evaluated by the
following equation:

(4)

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216 Juan Pereira, Henry Labrador, Víctor Perez et al.

Where R is the gas constant, T absolute temperature, n the number of


moles, is Ф volume fraction, Vm is the molar volume and δ is the solubility
parameter. The first term represents the Gibbs free energy which changes by
the entropic changes in the mix. This would be a thermodynamic path explain
their low solubility of asphaltenes and applicable to diluted solutions,
assuming the asphaltene behavior are similar to polymers.
Other thermodynamic model proposed for the study of the solubility of the
asphaltenes, was presented by Cimino et al. [25], they assume that the
asphaltenes are not pure and the solvent phase pure, the disadvantage that
occurs can not be calculated the amount of asphaltenes separated in the pure
solvent and the asphaltene has a breakpoint or threshold separating the solvent.
Thermodynamic models have been derived from the theory of polymers
Flory Huggins, in early these models asphaltene solubility is completely
predictable a parameter which are stabilized by the properties of the
asphaltenes and the dissolution of the model results of solubility asphaltenes,
is given by three points with respect to the stability of asphaltenes in the
solvent: the pseudo stability which is associated with the ability of flocculation
of the asphaltenes, the part where the asphaltenes can be displayed with
miscroscopy technique and instability when asphaltenes separated from the
solvent [26]
The solubility parameter is also used in models solubility of the
asphaltenes [26-29], Mitchell and Speight [26] in the last century obtained
between the solubility of the asphaltenes and the Hildebrand solubility
parameter, an excellent correlation and it was probably because they
considered the polar and nonpolar part in their interactions, although many
solvents used in the study are the presence of polar and hydrogen bonding
forces are present. Wiehe [27, 28] developed two component in the solubility
parameter approach while other research has three components as used in the
model developed by Hansen [30]. The solubility parameter was originally
developed to describe the existing interactions between a solute and solvent.

2.2. Asphaltenes: Aggregation

One of the properties of the asphaltenes is the formation of nanoclusters in


low concentrations, nanoclusters can be regarded as the germ or origin of
colloids, it is widely accepted that the aggregates begin to form at
concentrations well diluted in good solvents, experimental describe the nature

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The Use of Surfactants in the Chemical Treatment … 217

of the nanoclusters and the aggregation process is subject to many


disagreements among researchers, Mullis [31] and Mullins et al. [32], indicate
that the molecules of asphaltenes begin to form aggregates in solutions very
diluted concentrations below 100 mg/L, but some research suggests that
asphaltenes can be added in small oligomeric at lower concentrations of 5 mg/
L species [33, 34], Evdokimon et al. [35], by the technique of fluorescence
emission indicate that the first aggregates are formed asphaltenes below 0.7
mg/L and typically fluorophores rings 2 and 3, but this is currently under
discussion which would be the minimum concentration where asphaltenes
begin their interactions between them. These nanoclusters formed on fossil
fuels are considered, which have a size limit in response to steric effects
caused by the alkyl chains are unit to aromatic centers asphaltenes, may
indicate that there may be a minimum concentration that nanoclusters stop
growing [36-39] and at very high concentrations and under certain conditions
form the cluster with cluster known as the minimum concentration (CCC).
Aggregation of asphaltenes is important to clearly define what type of
aggregate can begin to consider, you can take into account the size of the
aggregates and values have been reported between 5 and 15 nm, which can be
referred to as nanoclusters [40-42] in investigations of sweeps [43-45] make a
clear distinction between an initial state as molecule and an association of
these asphaltene molecules to form nanoclusters, are the union of several
molecules of asphaltenes and this partnership gives its behavior in crude oil, to
which a separate produces its many drawbacks.
As indicated above asphaltenes have a high tendency to aggregate in a
good solvent, they would be as monomers at very low concentrations and so
indicated by Evdokimon et al. [36], the concentrations are below 1 mg/L,
where some investigations the concentration of nanoclusters criticism is about
50 to 100 mg/L [46], and these nanoclusters are composed of approximately 4
to 10 macromolecules [46, 47] (29 and 30), this indicates that the nanoclusters
are forming at very low concentrations and they are formed by few molecules.
At concentrations above 1 g/L over the nanoclusters to the cluster or critical
concentration cluster, under unfavorable conditions separation of the liquid
phase occurs, but if that high concentration is stabilized by natural dispersants
affect the viscosity of augmenting crude oil and making transport difficult, so
crude oil viscosity is strongly dependent on the percentage of asphaltene
present, its shape and interactions between them.

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2.3. Asphaltenes: Viscosity

Many studies [48-51] have shown that oil viscosity are greatly influenced
by the colloidal structure of the asphaltenes present in fossil fuels. The
presence of these associated macromolecules have a profound influence on the
viscosity. Lin et al. [52], showed that the particles are associated giving an
increased viscosity, they used the model Pal-Rhodes [53] and changing two
parameters described the relationship between viscosity asphalt and asphaltene
content. The Pal-Rhode model [54] was originally to describe the
viscosity/concentration emulsions given in the following equation relationship.

Ƞr=ƞ/ƞ0= (1-Kɸ)-2.5 (5)

Where ƞ0 are ƞ and the viscosity of the emulsion and solvent respectively,
ƞr represents the relative viscosity and ɸ is the volume fraction of particles. K
is the constant of solvation which is affected by the associations and the
exponent is the result of -2.5 assume associations spherically. Sheu et al. [55],
using equation viscosity raised by Pal-Rhodes describes the viscosity of
solutions of asphaltenes in function of the concentration of asphaltenes in
different aromatic solvents, various investigations have determined that the
concentration of asphaltenes is determinant in viscosity.
Pal [56], leads to a new viscosity for solutions of asphaltenes, taking into
account the cluster and nanoclusters asphaltenes, at low concentration
asphaltenes are as nanoclusters disc-shaped and are separated not interacting,
and high concentrations nanoclusters interact to form cluster, resulting in an
increase in the fraction of effective volume due to a continuous phase
immobilized by cluster, this indicates that the viscosity is dependent on the
interactions of the cluster creating a kind of network throughout the fossil fuel
that its viscosity increases, making it difficult to transport.
Aggregation processes asphaltenes affect their properties producing
negative consequences, since their separation from the liquid phase and an
increase in viscosity, which can indicate that the problem of scale when
aggregates change from nanometers to microns sizes.

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3. MIXING ASPHALTENES AND SURFACTANTS:


ASPHALTENES-SURFACTANT COMPLEX INTERACTIONS,
ACTIVE SITES FOR AGGREGATION OF ASPHALTENE
BEHAVIOR “FILM” ASPHALTENE-SURFACTANT
3.1. Asphaltene-Asphaltene Interactions

Two types of asphaltenes are suggested: Asphaltenes type A1 and A2,


where A1 represents the insoluble fraction of aromatic solvents and A2 which
is soluble in aromatic solvents [57, 58]. The composition of these types of
asphaltenes in crude defines aggregation of asphaltenes in a solvent medium
and therefore their solubility. For example, asphaltene in toluene solution with
high A1 fraction concentration can therefore be present in insoluble as
colloidal aggregates (Figure 2) A1 fraction which is covered by the A2
fraction moiety can interact with the solvent.
Due to the variety of sizes and shapes of asphaltenes, the aggregates can
leave spaces and there complex may occur with other compounds such as the
solvent. A study of the influence of diethyl ether on the physicochemical
properties of Ayacucho crude oil evidenced by the inclusion of diethyl ether
molecules in the aggregates of asphaltenes through studies of
thermogravimetry (TGA) and NMR-H [59]. Insertion diethylether interpreted
in a broad peak observed in the temperature range 150-250°C which is not
observed in the thermogram of crude oil without using solvent. Since the
boiling point of the ether is about 34.5°C [59], if no insertion, all of the solvent
should be removed from the solid. It is also checked through the qualitative
study of the solvent in the crude oil by results from Pereira at al. [59], where a
peak was observed in the region close to 4 ppm protons belonging to the
solvent in insertion via homogeneous and heterogeneous compared to
untreated crude oil.

Figure 2. Form of colloidal asphaltene in an aromatic solvent: A1 representing the


insoluble fraction covered by the A2 fraction interacts with the solvent.

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Another case is shown in complexation with p-nitrophenol (PNP), which


is generated by charge transfer [60, 61] with amino groups present in the
asphaltene.

3.2. Asphaltenes-Surfactants Interactions

As has been mentioned, asphaltenes are primarily responsible for


generating the emulsions during the process of crude oil production and
implementation or use of demulsifying agents for the maximum use of crude
oil has been necessary and this requires reducing the interfacial area generated
by the contact between the asphaltenes and the aqueous phase. The addition of
a demulsifying agent must be an agent that allows the exchange of matter
between itself and the asphaltene, displacing these generating coalescence and
thus the recovery of crude oil. In Figure 3 model adsorption [62] is shown in
the interface both asphaltene and the demulsifier which shows that there is an
equilibrium adsorption desorption both the demulsifier and asphaltene in their
respective phases and there is a constant of proportionality between the
concentration of CA asphaltene and concentration of demulsifier CD:

(6)

A good demulsifier must break all interactions between asphaltene


molecules, providing the highest possible Kint and for that the mixing model
HLB is used:

(7)

Where XA and XD are the mole fractions of asphaltene and demulsifier


respectively. HLBA is the measure of asphaltene hydrophilicity, HLBD the
hydrophilicity of the demulsifier and HLBm of the mixture, the latter two
being experimentally determined through studies of optimal formulation.
Disregarding the pressure and temperature variables that can affect the
stability of emulsions are the concentration and nature of crude oil, demulsifier
and water salinity.

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Figure 3. Model asphaltene adsorption and demulsifier at the interface.

There is evidence of the influence of the concentration of asphaltene. In


nonylphenol which is employed as demulsifier EON = 5.5 and asphaltene
concentration is varied 100 to 10000 ppm in cyclohexane [62]. Regardless of
the concentration of asphaltene stability it leads to a minimum where the
concentration of demulsifier in this minimum is expressed as CD*. From 1000
to 10000 ppm is note that CD* is constant and is about 200 ppm. This is
assumed in the fact that at concentrations lower than 1000 or 350 ppm
asphaltenes, there is insufficient interaction thereof with the interface to
equation 1 is fulfilled and, beyond 1000 ppm, is asphaltene at a concentration
fixed due to saturation at the interface, and action can occur demulsifier
fulfilling equation 6.
The influence of the HLB of demulsifier to different concentrations of
asphaltene. By repeating the above procedure but with a family of ethoxylated
nonylphenols with EON 4.75 to 10, the stability of the emulsions follows the
same pattern independently of EON But as the EON increases, CD* it
decreases due to increased hydrophilicity of demulsifier offers lower
interaction energy with the aqueous phase asphaltene decreasing XA [62].
The nature of the demulsifier also influences the stability of emulsions.
Besides the already mentioned ethoxylated nonylphenol, it has also been
carried out the breaking of emulsions of crude oil in water using “extended
surfactants” whose structures [63, 64] have a polar group mixed with a
nonionic ionic fraction and a fraction such as sodium dodecyl polyethylene
sulphate.

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CH3 CH3
CH3

+ +
NH3 NH3 +
NH3
- -
O O -
O
O O CH3 CH3
CH3
O O O O O O O
O O O
- -
O H N+ O OH- N+ +
OH OH OH OH OH OH
3 3H N
3
N N N
N N N
O O O
O O H3 C H3 C H C NH2 NH2NH
O 3 2

- -
O O O O- OO
HO OHO HOO O + + +
NH3 NHNH
3 3

H3C H3CH3C

Figure 4. Changing the structure of asphaltene by interacting with hexylamine. The


asphaltene with carboxylic groups allow placed at the interface. By forming ion pairs
with hexylamine carboxylic groups become carbon chains asphaltene generating a
more hydrophilic character.

3.3. Asphaltenes-Interactions Base-Acid

Another aspect of interactions involving asphaltene based acid


interactions. Emulsion breaking tests were carried out with the use of the
surfactants mentioned in the previous section, but with the addition of organic
acids and amines. Among them hydrophilic character (ethylamine and acetic
acid) and hydrophobicity (hexylamine and ethylamine), in order to study the
behavior of these additives in improving demulsifier activity.
Test emulsion stability was performed by varying, as in the previous
section, the asphaltene concentration and thus finding the optimum
concentration of demulsifier [65]. This result was compared by performing the
same test but adding the acid and alkaline additives. It was observed that
hexylamine improves the demulsifier activity significantly compared to the
other additives. Silva et al. [65], shows the influence of demulsifier as blank
and the influence of each demulsifier additive through the variation of the CD*
to the variation of the concentration of asphaltene. Hexylamine reduce the
optimal concentration of demulsifier in 200 ppm of asphaltene, 20 to 3 ppm,
and 1000 ppm of asphaltene, 100 to 30 ppm.

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The Use of Surfactants in the Chemical Treatment … 223

The influence it may have on the demulsifier activity hexylamine


compared with other additives can explain two aspects, first, that there is
interaction of the amino group with donor groups of protons present in the
asphaltenes, however, if the interaction ionizes the asphaltene, generate
charged groups rather favor the emulsion, but to achieve this, the system pH
must be greater than 12 and the actual pH of the system is 8, at this pH, the
interaction of asphaltene with hexylamine may be of ion pair (R-COO- H +
NR3 ···) which modifies the structure of asphaltene (Figure 4) making it a
more hydrophilic species and facilitate the breaking of the emulsion. Unlike
ethylamine, possessing a short chain does not allow generating a lipophilic
structure significantly. The second aspect is that the asphaltene studied does
not have strong electron donating groups that allow form ion pairs with acidic
additives. These tests were conducted with the surfactant EO13-PO30-EO13 type
with HLB of 15, compared with other tests carried out with a surfactant type
EO8-PO30-EO8, HLB of 10.5; obtaining similar results [65].

4. EXAMPLES OF ASPHALTENE PROBLEMS


IN THE PRODUCTION OF FURRIAL CRUDE OIL
IN EASTERN VENEZUELA

4.1. Case 1: Viscosity Increased for W/O Emulsions

The Furrial is almost the most important oil field in Venezuela, located
north of Monagas state with more than 120 active wells. This is a mature oil
field; accordingly program Lift Gas (LPG) was implemented at the end of
2012. Following the implementation of LPG, was detected in most wells with
this production method, the formation of W/O emulsions stable high viscosity
because the high shear generated in the gas inlet to the production line. Figure
5 shows the evidence for this effect.
The tests were performed by applying a chemical treatment (improved
flow) in these wells showed high emulsion viscosity and thus increase
productivity in the “Furrial” field.
Testing in the laboratory to control parameters set (API, water and
sediments (W&S), bottles of test, measurement threshold flocculation and
viscosity).

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Figure 5. Crude oil from Well 1. Viscosity: 10.000 cP/210°F.

Once taken to the field for testing under dynamic conditions by online
reviews with multiphase flow meter AGAR, it was monitored in parallel other
parameters: API, A&S and viscosity. It was obtained by application of the
chemical, a decrease in viscosity with 96% and increase the production rate
tested in a range of 6% to 11% wells.
Applying a flow best, it allowed to observe an increase in the mobility of
well fluid collection system, optimizing production. In a gas lift system,
sufficient agitation to allow the water dispersed in oil in a form “W/O” high
viscosity emulsion, stabilized by different species of interfacial activity present
in crude occurs. This phenomenon directly affects the rheological
characteristics, and the fluid runs through the entire production system.
In this step, efficiency tests were performed with different asphaltene
dispersants intermediates and emulsion breakers for the selection of the most
favorable combination, to determine the proportions and the best concentration
of the final product. The analyses of control used in this phase were
asphaltenes flocculation threshold, bottle and viscosity tests. These activities
were carried out with samples of Well 1, which presented a significant
increase in viscosity with an approximate magnitude of 10,000 cP @ 210°F
when the change of method of production to gas lift occurs.
Laboratory tests allowed to determine that the viscosity increase is related
to emulsion present into the sample; dramatic reductions in viscosity levels
were obtained by applying increasing concentrations of product flow
improver, which contains an emulsion breaker. As can be seen in Figure 6.

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The Use of Surfactants in the Chemical Treatment … 225

Figure 6. Viscosity vs. product concentration.

Figure 7. Well 1: Bottle Test. Effect of chemical additive on emulsion stability.

Then, the hydraulic conditions of the well/tubing system were simulated


with the commercial simulator v.9.6 PipephaseTM to determine changes in the
friction coefficient, resulting in a considerable reduction from 88.65 psig @
10.000 cP to 360 cP @ 62.18 psig. This parameter is directly related to fluid
mobility.
In Figure 7, the test bottle is observed, to determine the best concentration;
the emulsion must be destabilized, but not totally because phase separation not
have to be reached. This condition affect well productivity by generating a
column of water with considerable weight inside the production tubing
downhole, which will restrict the fluid flow at borehole.
PipephaseTM v.9.6 simulator was used for hydraulic simulation and get to
know the behavior of the well, the evaluation was conducted considering
viscosities between 10.000cP and 360 cP at 210°F, keeping constant the other

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226 Juan Pereira, Henry Labrador, Víctor Perez et al.

parameters of the well. The assessment indicates that there is significant


improvement in the mobility of fluids, when the viscosity is decreased,
achieving a reduction in friction losses from 88.65 to 62.18 psig @ 10.000cP
psig @ 360 cP.
The initial conditions of the well 1 and the results of viscosity variation
performed in laboratory which was made from an initial viscosity of 10,000 cP
@ 210°F and using a product concentration of 300 ppm approaching
maximum viscosity of 360 cP @ 210°F.
The test with the selected chemical was made in well 1 GLP from El
Furrial oilfield (see Figure 8) to determine the efficiency of the technique in
operational dynamic conditions and measured by AGAR multiphase meter by
changes in behavior fluid, with constant monitoring to perform on real-time
control and analyzes (A&S, °API, viscosity, pressure and temperature on
wellhead) Figure 8 shows a diagram of the production process comes from the
well to the processing station fluids.
After the results obtained in well 1, test on wells 2 and 3 with the same
conditions and maintaining test criteria were made.
The activities were performed previous adjustment of surface in each
wellhead, providing the installation of an enhancer mechanical configuration
for injection of chemical through gas lift facilities, pneumatic pump (3700psi)
tank with capacity for 4 drums and chemical flow. Note that the values of
BS% W must be kept constant during the test to ensure that water does not
accumulate in the tubing downhole to.

Figure 8. Schematic of the production process.

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The Use of Surfactants in the Chemical Treatment … 227

4.2. Case 2: Influence of Asphaltene Precipitation on


Emulsion Stability

In the last 20 years there has been a special interest in the study of the
phenomenon of asphaltene, which is due, on the one hand, their presence in
most field applications of injection of miscible gases, and other, and
exploitation of increasingly deep deposits containing crudes with unstable
asphaltenes.
The problems generated by these colloidal agents at the level of lines of
transfers and processing stations crude oil are diverse, and is problematic field
is not exempt under review, equipped with a crude oil asphalt nature and after
high production by at least 30, also wet.
The crude oil Furrial, has a high tendency to destabilization and
precipitation of asphaltenes and this component an emulsifying agent,
influenced by high water cuts accompanying the oil produced, generated
during the various stages of the production process, emulsions stable affecting
the viscosity and hence the fluid velocity.
Based on this a project that monitored the effects of unstable asphaltenes
from the crude stream that comes from Furrial field selecting a chemical
treatment that stabilizes the system dewatering constant output of crude
outside specifications began one water and sediment percentage well above the
established by the manufacturer dehydrators station.
The constant output of dehydration OOS (Outside Of Specification),
coinciding with the migration of certain wells in the form of gas lift, has
generated a call from acercs care possible destabilization of asphaltenes by this
method of artificial lift. This control measure applied through a chemical
dispersant treament of asphaltenes can lead to stabilization of the system and
maintain constant dehydration and low current oil specifications handled by
the main station Jusepín 2.
Asphaltenes do not have a definite melting point so they remain in solid
form, massing and pluging the dehydration equipment. Such is the case of
electrostatic dehydrators that make up the main station Jusepín 2 wherein these
colloidal agents generate severe problems being deposited on the walls and
electrodes of the apparatus, serving as natural insulation to prevent the passage
of alternating current, destabilizing and to system.
The knowledge of the system is to be applied where the treatment is key,
equipment and processes them covering dewatering plant crude oil Jusepín
main station 2 is described in detail.

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228 Juan Pereira, Henry Labrador, Víctor Perez et al.

Figure 9. Plant scheme dewatering main station Jusepín 2.

Plant consists of the following equipment: washing tanks, pumps raw


water pumps and electrostatic dehydrators, it can be seen in Figure 9.
Washing tanks or 55004 and 55005 settlement dynamic (TQ-55004/5) are
used to separate free water associated with the wet crude from the production
modules. These tanks of 55,000 barrels of rated capacity and 42 '10 5/8 “feet
tall at the level of capacity, are based on the principle of differential gravity.
That is, being heavier water than crude oil it sits on the bottom of the tanks
containing the mixture of the two.
Notably stable emulsions by gravity do not allow rupture of the
emulsifying agent and prevent water droplets come together in a reasonable
time. The emulsified crude oil passes into the eccentric internal overflow tank
19 feet high, and from there to the electrostatic dehydrators through the P-01
pumps where it is finally dehydrated.
On the other hand, dehydration main function is to separate oil emulsions
- water by applying electric fields and consists of seven electrostatic
dehydrators, of which five were obtained in 1999 to the manufacturer NATCO
technology dual polarity AC/DC (polarity dual - alternating current and direct
current), two of them have the capacity to handle 60 MBD of crude (DE-
101/201/301), two 80 MBD (DE-401/501), and two electrostatic dehydrators
(DE - 601/701) with a capacity of handling 60 MBD were obtained in 2007 to
the manufacturer Industries Vander - Rohe CA, due to differences in the
design of the input and output of oil and differences in design internal; which

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The Use of Surfactants in the Chemical Treatment … 229

corresponds to an installed to treat crude MBD 460 and handle 182 MBDP
water capacity.
Each team is 120 inches in diameter and 45 feet long; They are also
designed to operate at a maximum and 125 psia and 210°F operate with a
voltage in the primary circuit (power) of 480 volts, and convert it by a
transformer secondary voltage of 23000 volts. Typically, the voltage of the
secondary circuit can be selected between 16000, 19500 and 23000 volts. This
selection is made by the manufacturer and basically depends on the
conductivity of crude oil and water content. Flow distribution dehydrators is
done by control valve located in the discharge of dry crude oil. The maximum
level of interface on computers is 51 inches or 43% filling of the container to
prevent short circuits.
The LAG, a production method that uses compressed gas at high pressure
and external energy source, the gas is injected at a point in the column of fluid
in the tubing, it is intended to lighten or displacing the fluid column within the
production tubing, reducing its weight. Thus, the energy reservoir is sufficient
to carry the fluids from the bottom to the surface.
However, as reflected in Figure 10, the migration of new wells of Furrial
to be produced by this method of artificial lift generated instability asphaltenes
present in the crude stream handled by the process dewatering the EPJ-2.
The injection of gases rich in this survey method promotes asphaltene
precipitation in training and at the level of the well respectively. This effect is
because these injections cause changes in the composition of the reservoir
fluids and pH changes of the medium, which destabilizes the asphaltenes
present in crude, causing them to flocculate and precipitate, causing thus
changes wettability and well plugging level of training and subsequently
transfer lines and processing equipment.

Unstable
asphaltenes. Dewatering
System OOS (% WyS>
0.7)

Figure 10. Behaviour percentage of water and sediment vs. time.

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230 Juan Pereira, Henry Labrador, Víctor Perez et al.

It is noteworthy that this migration field wells The Furrial that produced
by natural flow, and presetaban the problem of unstable asphaltenes and in
relation to the above the gas lift, increased this instability generating stronger
emulsions, reaching system dehydration Jusepín main station 2, a crude oil
emulsions with many problems and asphaltene.
Clearly, (see Figure 10), it is evident as following the incorporation of
new wells (taken as example the well FN-23) asphaltenes are unstable as a
result of this injection of paraffins (gas injection) that causes that the
protective resin layer is broken, leading to asphaltene particles aglomerarce
including flocculating and possibly precipitating on the walls and electrodes of
the electrostatic dehydrator operating in the EPJ-2, destabilized dewatering
system.
As indicated before, the issue of asphaltenes in the oil field is fairly new,
so it can be said that experimental laboratory methods are extremely important
when drawing conclusions and recommendations to decide the real options to
correct or prevent problems associated with asphaltenes.
Before any field test first involved a complete diagnosis of the system
(wells that had already migrated to LAG, transfer lines and the main station
Jusepín 2) it was performed. In the particular case of this work it was taken as
reference the Well FN - 23 were established, then the variables that influence
the problem, knowing the experiences they have with other chemical
treatments in order to define the parameters to be evaluated (before during and
after the test), a dispersing chemical aromatic base was selected to repair the
electrodes affected by unstable asphaltenes and the evaluation program was
established, preparing formats for data collection and selection of systems and
equipment chemical injection.
It was shown that by performing laboratory tests such as threshold
determination flocculating asphaltenes to be unstable, they are deposited at the
interface in the dehydration process, stabilizing the emulsion present in the
stream of crude oil from Furrial Field.
This method, called threshold determination flocculation is based on
determining absorbance change oil by adding a nonsolvent continuously (n-
heptane). At the beginning of the titration, the addition of n-heptane produces
a dilution effect leading to a decrease in absorbance. Later, when the process
of asphaltene, the particles scatter radiation and an increase in absorbance is
observed. Curves like those shown in Figure 11. The curve minimum
corresponds to the lower volume of n-heptane required to begin to form
particles and asphaltenes called flocculation threshold are obtained.

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The Use of Surfactants in the Chemical Treatment … 231

Figure 11. Examples threshold determining asphaltene flocculation.

Activity inhibitors asphaltene products is determined through flocculation


threshold measures oil at a concentration of 1% inhibitory products (10.000
ppm) relative to oil. According to this method, if the threshold of flocculating
a crude oil solution containing the inhibitor product is greater than that of the
solution without the inhibitor product (white), the product has inhibitory
activity asphaltene.
According to tests the optimal threshold Flocculation is 28 milliliters of
heptane, showing a value of this test heptane 19 milliliters of sample selected
for the crude oil in the lapsus study problem.
Another observation that emerged from the tests was the relationship
operational and asphaltenes temperature. When these unstable colloidal agents
found in crude, the temperature range under which the electrostatic dehydrator
operating in the EPJ-2, is reduced, pulling, faster, out of specification
processing crude.
The success was obtained by injecting the chemical treatment was based
on an amp knowledge of system dewatering affected by the problem, product
selection and program most appropriate for the case study treatment, special
attention was maintained in the continuity of treatment chemical for these
colloidal dispersion agents, evaluating, monitoring and controlling the system
so dehydration.

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INDEX

acrylic acid, 143, 162


# activated carbon, 96, 106, 114, 118, 119
activation energy, xi, 103, 178, 204
1,8-cineole, 18, 22, 23, 24, 25, 27, 28, 29,
active site, 64, 65, 69, 71, 101
30, 31, 32, 35, 36, 37, 38, 39, 41, 42, 45,
acylation, 135
46, 51, 52, 59, 63, 64, 65, 66, 70
additives, ix, xii, 88, 108, 121, 122, 123,
124, 125, 128, 131, 208, 222, 223
A adenocarcinoma, 110
adenosine, 74
AA copolymers, ix, x, 141, 142, 143, 144, adjustment, 132, 226
160 adsorption, xii, 13, 88, 96, 103, 116, 208,
absorption spectra, 100 209, 214, 220, 221
accessibility, 161 aesthetics, 92
acetic acid, 131, 222 aflatoxin, 85
acetone, 33, 34 Africa, 55
acetonitrile, 10, 130 age, viii, 16
acetylcholinesterase, viii, 16, 53, 54, 71, 72, aggregation, xi, xii, 100, 207, 208, 213, 217,
73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 219
85, 86 aggregation process, 217
acetylcholinesterase inhibitor, 72, 76, 78, AIBN, x, 142, 145, 153
81, 85 alanine, 132, 133
acid, vii, ix, xi, 10, 12, 23, 24, 26, 27, 28, alcohols, 16, 68, 123, 124, 125, 127, 130,
33, 34, 37, 39, 52, 59, 83, 84, 91, 95, 96, 137, 138, 139
100, 101, 102, 108, 121, 124, 127, 130, aldehydes, 2, 16, 123
131, 132, 133, 134, 136, 137, 138, 139, alendronate, ix, 122, 131, 132, 133, 134,
140, 143, 159, 160, 161, 178, 179, 180, 140
182, 185, 186, 191, 196, 213, 215, 222 alfalfa, 61
acidic, 11, 98, 122, 123, 223 algae, 108, 117
acidity, 2, 3 Algeria, 22, 23
Acoraceae, 54, 69, 70 alkaline hydrolysis, x, 142, 146, 147, 149
alkaloids, 89

Complimentary Contributor Copy


236 Index

alkylation, 122 Barbados, 60


almonds, 68 barriers, 193
aloe, 104 base, 7, 8, 133, 171, 172, 213, 215, 230
Altingiaceae, 54 beneficial effect, 111, 124, 128
amines, 4, 9, 108, 123, 222 benign, 101
amino groups, x, 142, 220 benzene, 24, 103, 135
aminoglycosides, x, 142, 150 benzodiazepine, 136
amorphous materials, 164 beverages, viii, 15, 108
amorphous polymers, 167 biodegradation, 88, 108, 114
Anacardiaceae, 55 biodiesel, vii, 2, 12, 123, 137
angiosperm, 57, 61 biofuel, v, vii, 1, 2, 4
Annonaceae, 56, 70 biological activities, 77, 79, 82, 84, 183, 186
anthocyanin, 98 biological activity, 130, 143, 158, 162
Anthraquinone, 91, 113, 117 biologically active compounds, vii, viii, ix,
antibiotic, x, 142, 149, 150, 151, 155, 156, 15, 141, 143, 162
157, 158, 159, 162 biomass, 95, 97, 114
antibody, x, 142, 157 biomolecules, 103, 104
antigen, x, 142, 157 biopolymers, 171
antioxidant, viii, 6, 11, 12, 13, 15, 16, 72, biosynthesis, 98, 101, 116
73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, bismuth, 13
84, 85, 99, 104, 105, 115, 116, 118 black hole, 164
antioxidation, 9 black tea, 93, 102
antitumor, viii, 15, 16, 130 blood, viii, 15, 109
Apiaceae, 56, 81 blood circulation, 109
aquatic life, 108 blood-brain barrier, viii, 15
aquatic systems, 97 body weight, 108, 110
aqueous solutions, viii, 2, 9, 13, 119, 149, Bolivia, 101
152, 160 bonding, 92, 187, 196, 197, 199, 200, 201,
Arbidol polymeric complexes, 152 203, 204, 208, 216
Argentina, 21, 25, 101, 177, 204 bonds, 150, 172, 180, 193, 195, 213
aromatics, 136 bone, 164
Artemia, 75 bones, 130
Asia, 65, 66 Boraginaceae, 58, 74
Asteraceae, 16, 57, 61, 69, 70, 82 borneol, 18, 23, 24, 25, 26, 30, 34, 35, 36,
astrocytes, 109, 113 37, 38, 39, 41, 42, 43, 47, 52, 63, 64, 66
atoms, 105, 191, 196 bornyl acetate, 18, 22, 24, 25, 26, 35, 42, 57
brain, viii, 15, 108, 110, 113, 118, 119, 164,
168
B Brassicaceae, 71
Brazil, 27, 28, 45, 46, 47, 49, 53, 101
BAC, ix, x, 141, 142, 143, 144, 149, 151,
bronchial epithelial cells, 110
152, 160
Buddlejaceae, 59, 70
bacteria, 88, 108, 117, 149
Bulgaria, 39
bacterial infection, 158
Burkina Faso, 32, 45, 76
bacteriostatic, 155
Burseraceae, 59
Bangladesh, 74

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Index 237

butadiene, 181, 182, 183, 185, 187, 188, catalytic properties, 105
189, 190, 191, 194, 195, 203 cell culture, 109, 113, 151
butyrylcholinesterase, viii, 16, 53, 54, 74, cell death, 109, 115
77, 79, 84 cell line, 111, 115
central nervous system, 108, 116
Ceramics, 116
C chain transfer, 144
chemical, vii, viii, xi, xii, 13, 15, 79, 88, 89,
Ca2+, 109, 110
90, 92, 109, 111, 124, 135, 139, 160,
cabbage, 71
170, 172, 174, 177, 179, 180, 187, 190,
caffeine, 103
201, 202, 206, 207, 208, 212, 215, 223,
calcium, 113, 130
224, 225, 226, 227, 230, 231
cancer, 115
chemical bonds, 172
capillene, 18, 23, 58
chemical reactions, 187
capillin, 18, 23, 58
chemical stability, 88
carbon, 13, 86, 117, 124, 127, 130, 143,
Chile, 101
180, 191, 214, 222
China, 22, 23, 33, 35, 44, 45, 46
carbon atoms, 214
chlorine, 153
carbon dioxide (CO2), 86, 107
chlorobenzene, 130
carbonyl groups, 147
cholinesterase, 53, 72, 73, 74, 75, 78, 79,
carboxyl, 147, 149, 150
81, 85
carboxylic acid, 2, 61, 122, 130, 132, 133,
chromatography, 103
134, 145
chymotrypsin, 161
carboxylic acids, 2, 61, 122, 134, 145
Cistaceae, 60
carboxylic groups, 222
classes, 90, 187
carcinoma, 110
classification, 74, 80, 108
cardiovascular system, 110
cleavage, 103
carotenoids, 89
cluster model, 166
carvacrol, 18, 21, 26, 32, 33, 39, 40, 41, 42,
clusters, 105, 164, 167, 171, 172, 173
43, 63, 64, 85
CMC, 213
carvone, 18, 29, 30, 31, 37, 62
coenzyme, 136
caryophyllene oxide, 18, 22, 23, 24, 25, 26,
cognitive performance, 75
28, 34, 35, 37, 38, 39, 42, 43, 44, 45, 49,
collagen, 170, 174
53, 58, 63, 70
combustion, 104, 105
case studies, 139
commercial, 66, 225
case study, 231
complexity, viii, 15
catalysis, 101, 106, 111, 123, 127, 134, 135,
composites, 143
136, 187
composition, vii, viii, x, 4, 15, 58, 62, 63,
catalyst, ix, 87, 100, 101, 107, 111, 122,
64, 70, 71, 72, 73, 75, 76, 77, 78, 79, 80,
123, 135, 137, 138
81, 82, 83, 84, 86, 89, 103, 105, 142,
catalyst deactivation, 135
143, 150, 152, 153, 160, 211, 215, 219,
catalysts, ix, 2, 99, 102, 106, 107, 121, 122,
229
123, 124, 135, 136, 137, 214
compounds, viii, 2, 54, 55, 56, 58, 59, 61,
catalytic activity, 98, 99, 101, 113
62, 63, 64, 68, 82, 88, 91, 98, 108, 109,
catalytic effect, 100, 133
catalytic hydrogenation, 135

Complimentary Contributor Copy


238 Index

110, 117, 142, 149, 155, 180, 183, 186, Cyprus, 21, 36, 57
187, 214, 219 cytokines, 110
condensation, 2, 123, 137 cytotoxicity, 82, 83, 109, 110, 115, 117
conduction, 104, 107 Czech Republic, 1
conductivity, 3, 229
conference, 12, 120
configuration, 9, 226 D
constituents, viii, 11, 15, 16, 19, 20, 21, 22,
decomposition, 7, 97, 100, 106
24, 25, 26, 28, 29, 30, 31, 32, 33, 34, 35,
degradation, viii, ix, 2, 11, 87, 88, 89, 92,
36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,
93, 96, 98, 99, 100, 101, 102, 103, 104,
47, 48, 49, 50, 52, 56, 57, 59, 61, 62, 67,
105, 107, 111, 112, 113, 114, 116, 117,
68, 69, 70, 71, 72, 79, 82, 84
118, 119, 187
construction, 201
degradation mechanism, 103
consumption, 55
degradation process, 89, 107
contact dermatitis, 108
degradation rate, 96, 102
contaminant, 92
dehydration, 227, 228, 230, 231
contradiction, 69
deposits, 227
controversial, 57, 58
derivatives, vii, ix, x, 12, 58, 85, 105, 121,
COOH, 90, 143, 151, 160
123, 124, 128, 129, 130, 132, 133, 134,
copolymerization, 143, 144, 145
136, 138, 140, 142, 155, 158, 160, 162,
copolymers, vii, ix, x, 141, 142, 143, 144,
179, 180, 181, 183, 184, 186, 188, 189,
145, 146, 147, 149, 150, 151, 152, 153,
203
154, 158, 160, 161, 162, 170, 212, 213
desorption, 220
correlation, xi, 76, 163, 216
destruction, 92, 149
cosmetics, 88, 91
detectable, 196
cost, vii, ix, 87, 89, 98, 100, 102, 103, 111,
detection, viii, 2, 3
113, 124, 189
detergents, 91
Costa Rica, 101
detoxification, 92, 108, 114
cotton, 91
DFT, xi, 178, 201, 203
coumarins, 137
diabetes, 84, 86
covalent bond, 91, 143, 158
dialysis, x, 142, 146, 150, 159
covering, 3, 227
dielectric constant, 201, 203
criticism, 217
dienes, xi, 178, 180, 181, 182, 183, 184,
Croatia, 15, 21, 42, 48, 57
186, 187, 188, 189, 190, 191, 203
crop, 96, 101
diet, 71, 101
crude oil, xi, xii, 11, 13, 207, 208, 213, 217,
differential scanning, 3, 12
219, 220, 221, 227, 228, 229, 230, 231
differential scanning calorimetry, 3, 12
crystal structure, 174
diffusion, 103, 144, 147, 148, 151, 153, 154,
crystalline, 97, 98, 99, 106, 118
160
crystallization, 132, 214
diseases, viii, 16, 70, 71, 76, 79, 80, 86, 109,
crystals, 130
130
CTAB, 210
distillation, 16, 113, 159
cultivation, 55
distilled water, 3, 165
Cupressaceae, 60
distribution, x, 142, 143, 151, 164, 167, 168,
cyclic and acyclic phosphinates, ix, 121
171, 229

Complimentary Contributor Copy


Index 239

DMFA, 159 environmental protection, ix, 88, 111


DNA damage, 108, 109 enzymes, viii, 16, 54, 62, 63, 65, 66, 68, 71,
domestication, 73 75, 77, 83, 85, 110, 161
double bonds, 90 equilibrium, x, 142, 167, 215, 220
drawing, 230 erythrocytes, 157
dronic acid derivatives, ix, 121, 130, 134 essential oils, vii, viii, 15, 16, 18, 19, 62, 70,
drug carriers, x, 142, 161 72, 73, 74, 75, 76, 77, 78, 79, 82, 83, 84,
drug delivery, ix, 100, 116, 141, 161, 213 86, 89, 105
drugs, ix, 130, 141, 142, 143, 149, 151, 160, esterification, ix, 121, 122, 124, 125, 126,
161 127, 128, 129, 130, 134, 137, 138, 139
dyeing, 88, 91, 92 estragole, 18, 23, 58
dyes, ix, 61, 87, 88, 89, 90, 91, 92, 93, 94, ethanol, viii, 2, 6, 10, 26, 84, 143, 145, 153
95, 96, 99, 101, 102, 103, 106, 107, 108, ethers, 2, 16
109, 111, 112, 114, 115, 116, 117, 118 ethyl acetate, 159
ethylene, 212, 213, 214
ethylene oxide, 212, 213, 214
E eucalyptus, 16, 65
eugenol, 18, 21, 27, 32, 43, 44, 45, 47, 56,
E. coli, 157
61, 63, 64, 65, 70
East Asia, 54, 62, 69
Euphorbiaceae, 60
ecosystem, 88, 110
Eurasia, 60
Ecuador, 87, 101, 111, 119, 120
European Union, 108
effluents, 92, 103, 108
experimental condition, xi, 178, 189
Egypt, 23, 27, 32, 33, 34, 45, 51, 73
exposure, 98, 100, 110, 118, 170
electric field, 228
extraction, viii, 2, 11, 86, 120, 170
electrochemical, v, vii, 1, 4, 5, 11, 13
extracts, 63, 79, 82, 85, 96, 99, 102, 104,
electrochemistry, 2, 13
113, 115, 116, 118
electrode surface, 4
extrusion, 182, 185, 186, 196
electrodes, viii, 2, 4, 11, 13, 227, 230
electrolyte, viii, 2, 5, 7, 8, 9, 10
electromagnetic, 188 F
electron, 90, 97, 101, 102, 104, 106, 111,
113, 167, 168, 179, 180, 181, 183, 186, Fabaceae, 61, 75
187, 188, 191, 201, 202, 223 fabrication, vii, ix, 87, 96, 98, 104, 105,
electron microscopy, 97, 102, 167, 168 114, 115
electrons, 89, 96, 107, 201 fatty acids, 3, 5, 137
electrophilicity, 188, 189, 190, 191, 197, fertilizers, 61
199, 201, 202, 204 filtration, 143, 156, 170
emulsions, xii, 208, 210, 214, 218, 220, 221, fish, 170
223, 227, 228, 230 flavonoids, 89, 102, 104
energy, 89, 90, 96, 100, 105, 106, 167, 172, flexible-chain polymers, 145, 153
173, 188, 193, 195, 197, 202, 221, 229 flocculation, 214, 215, 216, 223, 224, 230,
engineering, ix, 88, 96, 109 231
enthalpy of activation, 124 flowers, 68, 80, 82
environment, 88, 89, 92, 110, 111, 116, 202, fluid, 82, 85, 224, 225, 226, 227, 229
212 fluid extract, 82, 85

Complimentary Contributor Copy


240 Index

fluorescence, 217
fluorophores, 217
H
food, viii, 15, 57, 60, 61, 81, 85, 108
halogenation, 123
food additive, 60, 108
hardness, 201
food products, 108
harvesting, 111
formaldehyde, 213
Hawaii, 101
formation, 98, 99, 101, 102, 104, 105, 143,
HBD, 179, 188
144, 147, 149, 151, 154, 156, 164, 167,
hemisphere, 60, 68, 69
169, 172, 187, 191, 196, 197, 203, 210,
hepatotoxicity, 110, 155
214, 216, 223
heptane, 214, 230, 231
France, 23, 24, 28
herpes virus, 145
free energy, 216
hexane, 83
free radicals, 104, 105
hippocampus, 109
friction, 225, 226
homogeneity, 130
fruits, 16, 55, 65, 68, 75
HRTEM, 213
FTIR, 3, 99, 102, 103, 105
Hungary, 23, 121
fuel, viii, 2, 3, 11, 12, 104, 215, 218, 232,
hybrid, 98, 106, 201
233, 234
hydrocarbons, 16, 55, 56, 58, 214
functionalization, 111
hydrogen, 105, 147, 150, 172, 187, 197,
furan, vii, xi, 178, 179, 181, 183, 185, 186,
199, 200, 201, 203, 204, 208, 213, 215,
187, 189, 203
216
hydrogen bonds, 147, 150, 172, 203, 213,
G 215
hydrolysis, 92, 130, 132, 146, 147, 148,
gene expression, 109 158, 159, 191
genus, 54, 59, 62, 66, 68 hydrophilicity, 220, 221
germacrene D, 18, 19, 20, 21, 23, 26, 29, hydrophobicity, 222
30, 31, 33, 34, 35, 37, 39, 43, 44, 48, 49, hydrothermal synthesis, 98
53, 58, 63 hydroxyapatite, 117, 130
Germany, 19, 52, 53, 113, 163 hydroxyl, 107
Gibbs energy, 215 hyperthermia, 119
glucosinolates, 89
gold compound, 110
gold nanoparticles, 100, 101, 109, 111, 114,
I
115, 116, 117
Illiciaceae, 62
gravitation, 171
illumination, 102, 104, 106
gravitational field, 172
immune response, 157
gravitational mass spectroscopy, x, 163,
immunostimulant, 151
164, 174
impulsive, 196
Greater Antilles, 62
in vitro, 72, 74, 76, 79, 81, 83, 85, 86, 115,
groundwater, 92
117, 119, 136, 151, 152, 160, 162
in vivo, 76, 112, 162, 171
India, 19, 23, 24, 28, 32, 45, 47, 87, 119,
120

Complimentary Contributor Copy


Index 241

industry(ies), xi, xii, 57, 60, 61, 65, 88, 91,


92, 207, 208, 214, 215
K
inflammation, 109
K+, 110
infrared spectroscopy, 102
ketones, 2, 16
inhibition, 54, 55, 56, 57, 58, 59, 60, 61, 62,
kinetics, 102, 103
63, 64, 68, 69, 70, 75, 78, 79, 83, 101,
Korea, 23
110, 155
inhibitor, 159, 161, 231
initial state, 217 L
injections, 229
insecticide, 58 Lamiaceae, 16, 62, 69, 70
insects, viii, 15, 70, 80 Lauraceae, 16, 64, 70, 77
interface, 209, 210, 214, 220, 221, 222, 229, Lebanon, 32, 33
230 legislation, 108
interface layers, 210 ligand, 102, 110
interferon, 151 light, 88, 89, 96, 99, 100, 101, 102, 103,
interleukin-8, 110 105, 108, 109, 111, 113, 118, 119, 153,
intermolecular interactions, 154, 215 160
intrinsic viscosity, 145, 147 light scattering, 99, 102, 153, 154, 160
ionic conduction, 188 limonene, 18, 19, 20, 21, 22, 23, 25, 27, 29,
ionic liquids, ix, xi, 121, 122, 134, 135, 136, 30, 31, 33, 37, 39, 44, 45, 46, 47, 48, 49,
137, 139, 178, 181, 186, 187 50, 51, 52, 53, 56, 57, 60, 67, 68, 69, 70
ionic solutions, 187 linalool, 18, 24, 28, 29, 31, 32, 33, 34, 35,
ionogenic, 143, 149, 160 41, 42, 43, 47, 51, 58, 62, 70
ions, 96, 99, 109, 130, 187 linalyl acetate, 18, 24, 28, 29, 31, 37, 41, 51,
IR spectra, 174 62
Iran, 22, 32, 37, 43, 73 liquid phase, 215, 217, 218
IRC, 157, 179, 201 liquids, ix, xi, 121, 122, 134, 135, 136, 137,
iron, vii, ix, 87, 89, 96, 102, 103, 110, 111, 139, 167, 178, 181, 186, 187
112, 113, 116, 166 liver, 108, 110, 139
irradiation, xi, 89, 96, 98, 99, 101, 102, 104, long-range order, 164, 165
109, 119, 122, 124, 125, 127, 128, 136, low molecular mass, vii, ix, 141, 143, 215
137, 138, 178, 181, 188, 189, 203 lubricants, 3
isomers, vii, 2, 38, 39, 181, 182, 192, 193, lubricating oil, 2, 3, 4
195, 203
isoprene, 180, 181, 182, 183, 184, 185, 187,
188, 189, 190, 191, 192, 193, 203 M
Italy, 23, 26, 36, 48, 51, 52, 84
macromolecules, 89, 144, 147, 151, 154,
212, 217, 218
J Malaysia, 43, 44, 45, 50, 54, 62, 69
Mark-Kuhn-Houwink equation, 153
Japan, 24, 46 mass spectrometry, 79, 103
materials, 11, 88, 89, 99, 109, 110, 112,
164, 212

Complimentary Contributor Copy


242 Index

matrix, 6, 7, 11 molecular weight, viii, 15, 16, 143, 146,


mechanism, 101, 103, 106, 107, 111, 116, 147, 149, 151, 160, 161, 213
139, 158, 181, 189, 191, 196, 203 molecules, xi, xii, 89, 92, 101, 107, 110,
Mediterranean, 54, 60 147, 154, 197, 199, 200, 201, 202, 203,
melting, 187, 227 207, 208, 213, 214, 215, 217, 219, 220
menthol, 18, 29, 30, 31, 62 monomers, 143, 212, 217
menthone, 18, 28, 29, 30, 31, 33, 62, 64 monoterpenes, 16, 18, 55, 57, 63, 66, 68, 70
mercury, 13 monoterpenoids, 16
metabolic, 82 monoterpenoids, 16, 18
metabolism, 143 Montenegro, 79
metabolites, viii, 15, 108 Morocco, 22, 27, 42, 45, 52
metal nanoparticles, vii, ix, 87, 88, 89, 96, morphology, 99, 102, 103, 105, 106
100, 107, 111, 113 Moscow, 161
metal salts, 89 mustard oil, 71
metals, 2, 11, 89, 215 myosin, 169
meter, 160, 224, 226 Myrtaceae, 16, 65, 69, 70, 78
methacrylic acid, 143, 162
methanol, 194, 196
methodology, 11, 100, 188 N
methyl group, 150, 183, 212
Na+, 110
methyl groups, 150, 212
Na2SO4, 6
methylene blue, 96, 99, 101, 105, 106, 116,
NaCl, 146, 150, 153, 154
117, 118
nanochemistry, 112
Mexico, 62
nanocrystals, 105
Mg2+, 110
nanomaterials, 110, 114, 116
mice, 157
nanomedicine, 120
microgravity, 171, 172
nanoparticles, 88, 92, 96, 97, 98, 99, 100,
micrometer, 168
101, 102, 103, 104, 105, 106, 109, 110,
microscopy, 98
111, 112, 113, 114, 115, 116, 117, 118,
microwave, ix, xi, 11, 98, 120, 121, 122,
119, 120
124, 136, 137, 138, 139, 178, 179, 181,
nanostructures, 110
188, 189, 203, 205
nanotechnology, 88, 109
microwave heating, 138
naphthalene, 24
migration, 227, 229, 230
natural polymers, 120
Milky Way, 175
negative consequences, 218
mixing, 166, 211, 214, 215, 220
negative effects, 108
models, xi, 103, 178, 216
Nelumbonaceae, 66
modifications, 109, 188
Netherlands, 232
modules, 228
neurodegenerative disorders, 74
molar volume, 216
neurological disease, viii, 16, 70, 71
molecular mass, vii, ix, 141, 143, 144, 149,
neuronal cells, 110
150, 160, 164, 211, 213, 215
neurons, 119, 168
molecular orbital, 201
neutrinos, 172
molecular structure, 167, 188, 209, 210
NH2, 156, 160
Nigeria, 21, 24, 32, 47, 52, 53

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Index 243

nitrite, 10 oxide nanoparticles, 104, 110, 111, 112, 113


nitrofuran, 182, 183, 185, 186, 187, 189, oxygen, 2, 3, 92, 107, 108, 215
190, 191, 192, 193, 194, 195, 196, 197, oxygen consumption, 3
198, 199, 201, 204
nitrogen, 61, 108, 215
nitroso compounds, 91 P
NMR, x, 79, 125, 126, 128, 129, 142, 145,
PAA, ix, 141, 146, 147, 149
160, 219
Pacific, 69
noble metals, 89
Pakistan, 23, 26, 47, 50, 51, 77, 79
nonionic surfactants, 212
palladium, vii, ix, 87, 89, 96, 106, 110, 112,
North Africa, 60
114, 117, 118
North America, 60, 66
pamidronic acid, ix, 121, 131, 132, 133, 140
nucleophiles, 123
parallel, 133, 224
nucleophilicity, 186, 187, 188, 189, 191,
PCM, xi, 178, 179, 201, 203, 204
202
periodic structures, x, 163, 164, 166, 168,
170, 173, 174
O periodicity, vii, 164, 167, 168, 170, 171,
172
octane, 130 Peru, 101
oil, xii, 3, 4, 6, 7, 11, 12, 13, 19, 20, 21, 22, PES, xi, 178, 179, 201
24, 25, 26, 28, 29, 30, 31, 32, 33, 34, 35, petroleum, xi, 13, 207, 213, 214, 215
36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, Petroleum, xi, 207
47, 48, 49, 50, 51, 52, 53, 54, 55, 60, 71, pH, 96, 97, 98, 102, 105, 132, 146, 158,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 159, 160, 223, 229
83, 84, 85, 86, 94, 96, 100, 114,115, 207, phase inversion, 210
208, 210, 211, 212, 213, 214, 215, 217, phenol, vii, 2, 4, 45, 64, 123, 124, 128, 129,
218, 219, 220, 221, 223, 224, 227, 228, 130, 155, 213
229, 230, 231, 232 phenolic compounds, vii, 1
oil production, xii, 208, 215 phenolic resins, 213
operating costs, 88 phenylpropanoids, viii, 15, 16, 18, 70, 71
optical properties, 117 phosphinic acids, ix, 121, 124, 127, 128,
optimization, 201, 203 130, 138, 139
optimization method, 201 phosphorous, 130, 131, 132, 133, 134
oral cavity, 119 phosphorus, 130, 131, 132, 133, 134
organ, x, 142 photocatalysis, 96, 106, 113
organic compounds, 90, 183 photochemical degradation, 88
organic matter, 117 photodegradation, 104, 105
organic solvents, xii, 138, 182, 187, 208 photons, 107
organism, ix, 141, 143 photosynthesis, 88, 108
organs, 16, 81, 158, 170 physical properties, 187
osteoporosis, ix, 122, 130 physicochemical properties, xii, 208, 209,
oxidation, vii, 1, 2, 3, 4, 7, 9, 10, 11, 12, 219
103, 116 Pinaceae, 66
oxidation products, 2, 3, 12 Pinus halepensis, 48
oxidative stress, 109, 110, 113 Piperaceae, 66, 70

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244 Index

PIT, 210
planets, 164
R
plants, viii, 15, 16, 55, 60, 72, 74, 75, 76,
radiation, 96, 175, 188, 230
80, 81, 89, 112, 113, 167
radical copolymerization, x, 142, 153
plastics, 91
radicals, 79, 107
platinum, 100
radius, 147, 149, 151, 154
PNA, 4, 10
rainforest, 68
Poaceae, 67
reactants, xi, 101, 130, 133, 137, 178, 201
polar, xii, 82, 123, 178, 181, 183, 186, 188,
reaction mechanism, xi, 178
201, 202, 208, 213, 216, 221
reaction rate, 188
polarity, 122, 190, 214, 228
reaction temperature, 124
polarization, 188
reaction time, ix, 111, 121, 124, 127, 134,
pollinators, viii, 15
188
pollutant, 88, 116
reactions, ix, xi, 2, 4, 101, 106, 121, 122,
pollutants, 88, 92, 105, 107, 111, 112, 116
123, 124, 127, 130, 132, 134, 135, 136,
polyacrylamide, ix, 141, 146, 162
137, 178, 179, 180, 181, 182, 183, 185,
polydispersity, 153
186, 187, 188, 189, 190, 191, 192, 194,
polyesters, 91
198, 203, 204
polygonaceae, 67
reactive oxygen, 109
polymer chain, 147
reactivity, xi, 103, 113, 127, 144, 178, 180,
polymer destruction, 147
181, 186, 189, 190, 201, 203
polymer synthesis, 143
reagents, 93, 94, 95, 111, 130, 137, 186
polymers, 143, 144, 145, 150, 160, 161,
recovery, xii, 82, 92, 208, 220
212, 215, 216
red blood cells, 110
polyphenols, 103, 104
regeneration, 5, 9
polypropylene, 167
regioselectivity, xi, 178, 182, 184, 185, 189,
polystyrene, 167, 170
190, 191, 192
population, 19, 20
remediation, 88, 89, 92, 97, 100, 102, 103,
porous materials, 164
111, 120
Portugal, 22, 24, 28, 31, 33, 34, 38, 40, 41,
renewable energy, 111
42, 43, 53
resins, 26, 59, 212
potato starch, 165
resistance, 201
proteins, 96, 165, 167, 174
retention volume, 156
protons, 172, 219, 223
rhizome, 54, 69
pulegone, 18, 28, 29, 30, 31, 33, 62, 64
rings, 90, 108, 180, 183, 217
pumps, 228
Romania, 38
purification, 120, 182
room temperature, 98, 99, 100, 101, 105,
pyrimidine, 123, 137
116, 135, 137
ROS, 109
Q Rosaceae, 68
roses, 68
quantification, 3, 6, 7 Russia, 141
quantum chemical calculations, 124 Rutaceae, 68, 70
quantum dots, 112
quaternary ammonium, 136

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Index 245

110, 122, 123, 124, 154, 208, 214, 217,


S 223, 224
spectroscopy, x, 97, 99, 142, 145, 152, 156,
safrole, 18, 45, 64
163, 164, 174
salinity, 220
stability, x, xii, 3, 9, 12, 98, 99, 111, 142,
salts, 92, 124, 135, 140, 146, 167, 187
150, 151, 160, 174, 196, 208, 210, 211,
Saudi Arabia, 28, 30, 52
213, 215, 216, 220, 221, 222, 225
sediment, 227, 229
stabilization, 99, 105, 135, 174, 213, 227
sedimentation, 144, 145, 147, 153, 160
stabilizers, vii, ix, 87
sediments, 223
stable complex, 150, 151
seed, 56, 63, 66, 69, 96
starch, x, 163, 165, 166, 170
selectivity, 69, 182, 186, 188
states, xi, 135, 178, 192, 194, 196, 197, 201
semiconductor, 89, 114, 118
storage, viii, 15, 166
sensors, 120, 170
strong interaction, 214
Sesquiterpenes, 16, 18, 55, 56, 63, 66, 67,
structure, x, xii, 71, 90, 99, 102, 104, 105,
70
106, 117, 142, 143, 145, 150, 164, 166,
Sesquiterpenoids, 16, 18, 75
167, 168, 170, 171, 180, 185, 200, 203,
shock waves, 168, 169, 170, 171
208, 209, 210, 211, 212, 213, 215, 218,
shrubs, 66, 67, 68
222, 223
side chain, 122, 130, 156
styrene, x, 163
side effects, 110, 143, 155
sulfur, 108, 215
signals, viii, 2, 6, 7, 10, 167, 168, 169
surface properties, 89
silicones, 212
surface tension, xii, 208, 209, 212
silver, vii, ix, 87, 89, 96, 98, 99, 100, 109,
surfactant, xii, 98, 208, 209, 210, 211, 212,
111, 112, 113, 114, 115, 116, 118, 119
214, 223
skin, 109, 115
surfactants, vii, xii, 207, 208, 209, 210, 211,
sodium, 100, 103, 131, 140, 210, 221
212, 214, 221, 222
solar cells, 120
survival rate, 110
solubility, viii, xii, 2, 15, 108, 110, 143,
susceptibility, 115
208, 210, 214, 215, 216, 219
synergistic effect, 70, 133
solution, xii, 10, 98, 101, 104, 105, 109,
syntheses, ix, 96, 121, 122, 123, 130, 132,
146, 150, 153, 154, 158, 159, 208, 212,
133, 135
213, 219, 231
synthesis, vii, ix, 85, 88, 96, 97, 98, 100,
solvation, 203, 218
101, 102, 103, 104, 105, 106, 112, 113,
solvent effects, 186, 203
114, 115, 116, 117, 118, 119, 120, 121,
solvents, ix, xi, xii, 98, 121, 122, 130, 137,
122, 123, 124, 129, 130, 131, 132, 134,
140, 145, 178, 181, 187, 188, 189, 201,
135, 136, 137, 138, 139, 140, 149, 155,
203, 204, 208, 209, 213, 216, 218, 219
156, 160, 162, 179, 180, 181, 186, 188
South America, 60, 65
synthetic polymers, 167
South Korea, 120
Spain, 33, 37, 40, 83
spathulenol, 18, 21, 23, 24, 25, 33, 34, 35, T
39, 45, 46, 47, 49, 55, 58, 63
species, 5, 16, 54, 55, 56, 57, 58, 59, 60, 61, tannins, 99, 102
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, target, viii, 2, 16, 70, 133, 143
73, 74, 75, 77, 79, 82, 83, 95, 107, 109, techniques, ix, 5, 88, 97, 98, 99, 111

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246 Index

technologies, 88, 106, 109, 170 universe, xi, 163, 164, 171, 172, 173, 174
technology, 96, 111, 228 UV absorption spectra, 104
TEM, 97, 99, 102, 104, 105, 106 UV light, 93, 98, 104
temperature, ix, xii, 3, 12, 93, 96, 101, 103, UV radiation, 166
121, 127, 136, 144, 146, 182, 185, 187, UV spectrum, 156
198, 201, 207, 210, 212, 215, 216, 219,
220, 226, 231
terpenes, viii, 15, 16, 17, 72, 84 V
terpenoids, viii, 15, 16, 71, 72, 77, 84, 105
Valerianaceae, 69
terpinen-4-ol, 18, 19, 20, 21, 23, 27, 32, 41,
vanadium, 215
42, 60, 65
vegetables, 56, 67
testing, 3, 4, 12, 70, 182, 224
Venezuela, vi, vii, xii, 207, 214, 223
textiles, 88, 112
Verbenaceae, 69, 80
Thailand, 28, 33, 35, 44, 45, 46, 49, 51
viscosity, xii, 2, 147, 208, 210, 217, 218,
thermodestruction, 149
223, 224, 226, 227, 234
thermogravimetry, 219
vitamins, 98
thin films, 118
voltammetric, viii, 2, 4, 5, 6, 9, 11, 12, 13
thymol, 18, 24, 32, 33, 39, 40, 41, 42, 43,
voltammetry, 2, 5, 9, 11, 12
58, 63, 64, 85
voltammogram, 6, 7
titanium, 99, 115
toluene, xii, 130, 208, 213, 214, 219
toxic effect, 110, 117 W
toxic products, 92
toxic substances, 89 waste, 99, 100, 104, 116, 117
toxicity, ix, x, 72, 75, 77, 108, 109, 110, wastewater, 88, 92, 96, 101, 108, 112, 116
111, 112, 116, 117, 118, 141, 142, 149, water, ix, x, xii, 9, 88, 91, 92, 98, 102, 105,
151, 152, 155, 157, 160, 162 108, 141, 142, 143, 149, 150, 151, 159,
transesterification, 137 160, 161, 162, 163, 166, 167, 168, 170,
transformation, xi, 8, 124, 178, 180 171, 172, 174, 201, 208, 209, 210, 212,
transition metal, 122 214, 220, 221, 223, 224, 225, 226, 227,
transition state, xi, 135, 178, 192, 194, 196, 228, 229
197, 201 water clusters, 172, 174
transmission electron microscopy, 99, 213 water purification, 98
transport, 217, 218 wells, 223, 224, 226, 227, 229, 230
treatment, vii, viii, ix, xii, 10, 16, 70, 80, 86, West Africa, 61
88, 90, 92, 96, 122, 130, 207, 208, 213,
223, 227, 231
Turkey, 24, 25, 28, 33, 34, 35, 36, 37, 38, X
39, 42, 48, 49, 51, 84
XPS, 106
X-ray diffraction (XRD), XRD, 98, 99, 101,
U 102, 105, 106

ultrasound, 118, 120


United States (USA), 12, 13, 23, 62, 112,
174, 205

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Index 247

β-caryophyllene, 18, 19, 20, 21, 23, 24, 25,


Y 28, 29, 30, 31, 33, 34, 35, 36, 39, 40, 42,
43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 57,
Yemen, 26
58, 63, 67, 68, 70
β-cubebene, 18, 57
Z β-eudesmol, 18, 22
β-pinene, 18, 19, 20, 21, 22, 23, 24, 25, 26,
zinc, vii, ix, 87, 89, 96, 104, 115, 117, 118 29, 31, 32, 33, 35, 36, 37, 38, 39, 42, 46,
zinc oxide, 104, 115, 117, 118 47, 48, 49, 51, 52, 66, 67, 68, 70
Zingiberaceae, 69
ZnO, 93, 95, 103, 104, 105, 110, 116, 118
γ

α γ-terpinene, 18, 19, 21, 23, 32, 33, 40, 41,


42, 43, 46, 51, 52, 56, 57, 63, 65, 68
α-humulene (α- caryophyllene), 18
α-pinene, 18, 19, 20, 21, 22, 24, 25, 26, 32,
Δ
33, 34, 35, 36, 37, 38, 39, 41, 42, 44, 45,
46, 47, 48, 49, 55, 56, 57, 58, 60, 63, 64,
δ-3-carene, 18, 48, 49, 52, 56, 60, 67
65, 66, 67, 70
δ-cadinene, 18, 21, 24, 43, 45, 46, 57

β-asarone, 18, 54, 69, 70

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