ANALYSIS OF ANTIOXIDANT AND
ANTIMICROBIAL ACTIVITY IN ESSENTIALS OILS.
Daniela Salom Freire Zapata.
Departamento de Energa y Mecnica, Universidad de las Fuerzas Armadas ESPE, Latacunga, Cotopaxi, Ecuador.
Keywords: antioxidant properties, antimicrobial properties, Photochemiluminescence assay, DPPH assay, ABTS assay, agar disk
diffusion
1. Introduction:
Essential oils (EOs) are natural plant products
and they possess various biological properties.
EOs are complex mixtures of volatile compounds
produced by living organisms, from a whole
plant or plant part of known taxonomic origin,
and isolated by physical means only.
The composition of essential oils is mainly
mono- and sesquiterpene hydrocarbons and their
oxygenated, along with aliphatic aldehydes,
alcohols, and esters. In this respect, analytical
methods applied in the characterization of
essential oils have to account for a great number
of molecular species. Considering an essential oil
chemical profile is very important the choice of
an appropriate extraction method; "the following
extraction techniques can be applied: steam
distillation (SD), possibly followed by
rectification and fractionation, solvent extraction
(SE), fractionation of solvent extracts,
maceration, expression (cold pressing of citrus
peels), enfleurage, supercritical fluid extraction
(SFE), pressurized-fluid extraction,
simultaneous distillation extraction (SDE),
Soxhlet extraction, microwave-assisted
hydrodistillation (MAHD), dynamic (DHS) and
static (SHS) headspace (HS) techniques, solvent-
assisted flavor evaporation (SAFE), solid-phase
microextraction (SPME), and direct thermal
desorption (DTD), among others". (1).
Currently the bacterial infections have been
increased, thus, the search for effective and safe
medicines that could be used to treat particularly
persistent bacterial infections, is on. A possible
solution would be the essential oils, and this is
the razon for study their properties against certain
bacteria, fungi, viruses, and protozoa.
The essential oils of thyme, oregano, mint,
cinnamon, cumin, salvia, clove, and eucalyptus
have been found to possess the strongest
antimicrobial properties. The synergy of action,
as well their broad and complex activity of the
essential oils can make them a weapon against
multidrug resistant bacterial strains (2). In the
thyme essential oil the microbial activity was
determined by Agar diffusion, and it strongly
inhibited the growth of the clinical strains of
bacteria tested.
The potential activation of sunflower protein
films with antioxidant and antimicrobial
properties conferred by the phenolic compounds
of sunflower seeds was assessed. Like raw
material was used two sunflower protein
concentrates obtained from the residual pellet of
oil industry and the film-forming dispersions and
the films obtained were analyzed regarding their
antioxidant properties using ABTS, FRAP and
PCL assays, and their antimicrobians properties
by agar disk diffusion tests (3).
The antioxidant capacity of the filmogenic
dispersions and the supernatants obtained were
characterized by its cationic radical scavenging
ability (ABTS assay), its ferric ion reducing
capacity (FRAP assay), and its capacity to
quench superoxide anion radicals (PCL assay).
The ABTS and FRAP assays was described
previously in (4). The luminol
photochemiluminescence assay was carried out
in a PHOTOCHEM (Analytik Jena AG,
Germany) system with the kits of antioxidant
capacity of water-soluble substances (ACW) and
antioxidant capacity of lipid-soluble substances
(ACL), wherethe luminol plays a double role as
the photosensitizer and the radical detecting
agent. For assessed the antimicrobial activity 26
microbial strains was selected according the
importance in human health or for being
responsible for food spoilage, and with these the
antimicrobial activity of the filmogenic
dispersions and the resulting protein films were
determined by the agar disk diffusion method.
After the all essays the investigators concluded
that the phenolic compounds conferred important
antioxidant properties to both dispersions and
films, this activity being dependent on their
content and their free or protein bound nature.
However they no conferred their antimicrobial
properties probably due to their interaction with
proteins and the pH of the film dispersions.
The addition of sunflower protein films with
clove essential oil (CEO) allowed to prepare
biodegradable and edible films with increased
antioxidant properties and important in vitro
antimicrobial properties (5).
The film's antioxidative capacity was determined
for two diferrente antioxidative mechanisms: the
radical scavenging capacity and the reducing
capacity. The radical scavenging capacity was
measured using two different radicals: ABTS+
radical (ABTS assay) and Superoxide anion
radical (Photochemiluminescence assay). The
reducing capacity was measured following the
ferric ion reducing capacity (FRAP) assay.
Sunflower protein films incorporated or not with
clove essential oil were found flexible and
homogeneous.
The antioxidant properties assessed by ABTS,
FRAP and PCL and the both films exhibited
antioxidant capacity, due to the natural phenolic
compounds present in both sunflower and clove.
The increase of antioxidant capacity upon adding
CEO to the formulation was remarkable. And the
results acording the antimicrobial activity
showed that the CEO retains its antimicrobial
activity after its addition to sunflower protein
films.
In other investigation job the antioxidant and
antimicrobial activities of essential oil of leaves
Piper pubinervulum were studied using means of
the free radical inhibition methods DPPH,
ABTS, and photochemiluminescence (for the
antioxidant activity) and the agar diffusion
method (for the antimicrobial activity). (6) Also
the essential oil of T. vulgaris was studied and
compared with the previous results
corresponding to essential oil of leaves Piper
pubinervulum.
With the DPPH and ABTS methods the essential
oil showed low DPPH and ABTS radical
scavenging activity (20.399 g / mL) and (0.416
g / mL), respectively, compared to the essential
oil of T. vulgaris (0.414 g / mL) and (0.231 g
/ mL) respectively. The results of
photochemiluminescence showed that the P.
pubinervulum oil present a lower antioxidant
activity that T. vulgaris oil and the antibiotic
technique showed that the essential oil of Piper
pubinervulum possesses antimicrobial activity
against the study strains in concentrations from
24.13 mg / mL.
In other case, the antioxidant activity of oleoresin
from rosemary (Rosmarinus officinalis L.)
(ROSM) and sage (Salviaofficinalis L.) (SAG)
was evaluated using photochemiluminescent
method (PCL assay), spectrophotometricmethod
based on 1,1-diphenyl-2-picrylhydrazyl (DPPH)
assay and Rancimat test. (7)
Photocheminstrument (Ana-lytik Jena,
Leipzig, Germany) against the superoxide anion
radicalsgenerated from lumino, was used for
PCL assay. The radicals were eliminated in party
antioxidants present in the samples. The residual
radicals caused the detector substance luminol to
luminescence. For DPPH assay all the samples
were dissolvedin methanol and tested at different
concentrations in a reactionmixture ranging from
5 to 100 mg/L. The decrease in
solutionabsorbance was measured
spectrophotometrically at 517 nm. In the
Rancimat test an automated Metrohm Rancimat
model 743 (Herisau,Switzerland) was utilized to
determine the oxidative stability of thesunflower
oil (SO) blended with ROSM (SOROSM) and
SAG (SOSAG)at the concentration of 500 and
1000 mg/kg. In addition, the SOwere blended
with TBHQ (SOTBHQ) at 200 mg/kg to serve as
positive control.
Finally, the EC50 value obtained from PCL and
DPPH assays were significantly correlated(r =
0.997, p < 0.01). The SAG showed significantly
(p < 0.05) higher antioxidant activity than ROSM
inall the tested methods. The OSI and %AA of
ROSM and SAG blended oil was significantly
higher (p < 0.05)than the control sunflower oil
sample. In conclusion, the PCL and Rancimat
test were successfully usedfor direct and
dependable monitoring of antioxidant activity
and intended as the alternative to DPPHassay for
industrial application(7).
References:

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