Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
com
ISSN 2249-8516
Original Article
ISOLATION AND SEPARATION OF PHENOLIC COMPOUND FROM
CORIANDER FLOWERS
Dr. P. Nazni* and R. Dharmaligam
Department of Food science and Nutrition, Periyar University, Salem, Tamilnadu, India.
raiseandrich@gmail.com
*
Corresponding author: naznifsn@gamil.com
Received 16 January 2014; accepted 21 January 2014
Abstract
The present study is carried out with coriander flowers; A simple and fast method was developed for simultaneous
quantitative determination of three biologically active phenolic compounds such as apigenin, quercetin, keamphferol in
flowers of Coriandrum sativum separation using by flash column chromatography then purification were conducted by
preparative chromatography and HPLC. The phenolic compound determine with different spectroscopic method such as
LC-MS, HC NMR, FTIR and Melting point. The present method is being reported first time and may be used for routine
quality control of the flowers of Coriandrum sativum.
© 2014 Universal Research Publications. All rights reserved
Key words: Apigenin, Quercetin, Keamphferol, Chromatography.
Introduction effects as well as their potential estrogenic and anticancer
Extracts of aromatic herbs, spices, and medicinal plants are activities (Springob and Saito, 2002). Quercetin belongs to
used in food and pharmaceutical processing to impart this group of plant pigments called flavonoids that are
flavor or other functional properties, as well as ingredients largely responsible for the colours of many fruits, flowers
in the pharmaceutical products. Plant based natural and vegetables. Quercetin works as anti-inflammatory,
constituents can be derived from any part of the plant like antioxidant, anticancer agents. (Lamson and Brignale
bark, leaves, flowers, roots, fruits, seeds, etc (Gordon and (2000) and Mahesh Chand Meena and Vidya Patni 2008).
David, 2001) i.e. any part of the plant may contain active Flavonoids possess a variety of biological activities that are
components. The beneficial medicinal effects of plant thought to contribute towards protecting humans against
materials typically result from the combinations of degenerative diseases. Quercetin, kaempferol and
secondary products present in the plant. The medicinal isorhamnetin have been shown to have an anti-
actions of plants are unique to particular plant species or inflammatory effect on activated macrophages. In addition,
groups and are consistent with this concept as the quercetin and kaempferol show chemopreventive properties
combination of secondary products in a particular plant is in brain tumours (medulloblastoma) and synergistically
taxonomically distinct (Wink, 1999). They are usually suppress cell proliferation in human gut cancer lines.
subdivided according to their substituents into flavanols Quercetin a main aglycone in human nutrition is a potent
(kaempferol, quercetin), anthocyanins, flavones, flavonones free radical scavenger and antioxidant, which is why it is
and chalcones. These flavonoids display a remarkable array thought to protect humans against several types of cancer
of biochemical and pharmacological actions viz.,anti - and cardiovascular diseases. Antioxidant effects of
inflammatory, antioxidant, anti -allergic, hepatoprotective, quercetin on the mucosa of the nasal turbinate were
antithrombotic, antiviral and anticarcinogenic activities demonstrated. Higher intakes of kaempferol resulted in a
(Middleton and Kandaswami, 1993). These compounds lower risk of coronary heart disease. In addition,
appear to play vital roles in defence against pathogens and isorhamnetin revealed distinct vasodilatory effects in
predators and contribute to physiological functions such as animal models, suggesting vascular protective effects in
seed maturation and dormancy (Winkel-Shirley, 2002). human cardiovascular diseases. (Susanne Schmidt1,
They are synthesized from phenyl propanoid and acetate Michaela Zietz).
derived precursors. Flavonoids are important for human Flavonoids are polyphenolic compounds that occur
beings due to their antioxidative and radical scavenging ubiquitously in plant foods. Flavonols and flavones are
International Journal of Agricultural and Food Science 2014, 4(1): 13-21
13
subclasses of flavonoids (Kühnau J 1976, Markham KR single spot using by iodine vapour (solvent system for
1989, Herrmann K 1988, and 1979). In 1992 the average t1and t2 chloroform: methanol; acetic acid; water 95:5;5:2
daily intake of the flavonols quercetin, kaempferol, and and t3).
myricetin from the Dutch diet was 16, 4, and 1 mg, Analysis of Crude Extract by HPLC
respectively; the average intake of the flavones apigenin The resultant flavonoid aglycons were qualified by using
and luteolin was 1 mg each (. Hertog MGL et al 1993). The reversed-phase HPLC series 6330 from Agilent
intake of these five dietary flavonoids was associated with technologies. Solvent A consisted of 100% water and 0.5%
a reduced risk for ischemic heart disease and stroke in acetic acid; solvent B was 100% acetonitrile. The following
several (Keli SO et al 1996, Knekt P et al 1996 Hertog gradient was used for eluent B: 5–7% (0–12 min), 7–9%
MGL, 1995-93,) although not all (Rimm ER, et al 1996, (12–25 min), 9– 12% (25–45 min), 12–15% (45–100 min),
Hertog MGL, et al 1997 ), epidemiologic studies. Two 15% isocratic (100– 150 min), 15–50% (150–155 min),
types of mechanisms have been proposed to explain this 50% isocratic (155–165 min), 50–5% (165–170 min), 5%
protective effect: inhibition of LDL oxidation and isocratic (170–175 min) ( Susanne Schmidt et.al 2010). The
inhibition of platelet aggregation (Hertog MGL and flow rate used was 0.8mL/min, and the measured detector
Hollman PCH et al 1996). Results obtained by incubation wavelengths were set at 280, 320, 330, 340 and 370 nm.
of human platelets or animal cells with isolated isolated Purification of phenolic compound by using Pre-HPLC
flavonoids suggest that flavonoids inhibit platelet The fraction sample collected from chloroform methonal
aggregation probably by inhibition of cyclooxygenase solvent system, collected sample purified through semi
activity (P Mensink1997). preparative HPLC (using a PU-2080 pump (Jasco, Japan)
In addition, many studies have demonstrated the potential equipped with a MD-2010 multi wave length detector
of plant products as antioxidants against various diseases (Jasco, Japan) and a 250 -10.0 mm i.d., 4- lm Synergi
induced by free radicals (Hou et al., 2003), and it has been Polar-RP column (Phenomenex, Torrance, CA). The
determined that the antioxidant effect of plants is mainly mobile phase was solvent A, 100% ACN; and solvent B,
attributed to phenolic compounds, such as flavonoids, ultrapure water. Elution condition was 0–80 min of 20–
phenolic acids, tannins, and phenolic diterpenes (Pietta, 100% A–B (linear gradient) at a flow rate of 4 ml/min
2000). The aim of the present study is to deals with the (Shang-Tse Ho et.al., 2010).
isolation and identification of flavonoid "quercetin" The structures of compounds were identified by LC-MS
keamphferol and apigenin from coriander flower. (Agilent 6330 Ion Trap ),FT-IR and NMR (Varian Unity
Materials and Methods Inova-600, USA), and all spectral data were consistent with
Plant materials the literature (Baderschneider & Winterhalter, 2001; Jiang
The plant materials of C. sativum were collected from the et al., 2001; Villegas & Kojima, 1985; Yoshida, Ahmed,
plants grown in and around Salem district, Tamilnadu, Memon, Okusa, 1991& Shang-Tse Ho et.al., 2010, Nedime
India. The plant materials were dried in shade separately. Durust et .al 1999 and).
Reagents Purified CF flower extract samples were subjected to LC-
Reagents and chemicals Methanol and acetic acid were of MS and NMR for the structural elucidation and
HPLC grade and were purchased from Merck Company. identification of constituents were performed, three
Deionized water was prepared by a Milli-Q Water compounds were separated at different retention time.
Purification system. Moreover the particular compounds were determined by
Extraction and separation of crude sample FTIR and DSC.
The dried samples (flower 250 g) were chopped into small Result and discussion
pieces and extracted with methanol: water (80:20) by The fresh flower harvested to under our study director
soaking for 48 Hrs at room temperature (25°C). The guidelines and dried with room temperature. The dried
methanolic extract was decanted, filtered under vacuum, material soaking with methanol: water (80:20) for 24hrs.
concentrated in a rotary evaporator (40°C.). The resulting After completion of soaking filter to whattmen paper and
crude extract from the flower of CF (25.0g) was concentrated by rotary vapor at 40°c. The crude sample
fractionated successively with ethyl acetate (EtOAc), n- subject to inject HPLC and separated to all peak using
butanol (BuOH), and the yield of soluble fractions of different gradient programmed based on that separation to
EtOAc (2.0g), BuOH (1.0g), and water (20.g). (Janmejai K developed for column chromatography method.
Srivastava et. al 2009, Tereschuk et.al. Naira Nayeem and The methanolic extract was subjected to column
Karvekar MD 2010). The ethyl acetate (methanol) soluble chromatography on silica gel using solvents of increasing
fraction from flower of CF (2.0g) was loaded into a flash polarities starting from petroleum ether, chloroform, ethyl
chromatography column (silica gel for mesh 260-400 acetate and methanol in different ratios to yield several
revelries grace column silisep) and eluted with different Sub-fractions. Fractions 10-20 (40% chloroform in
solvent system for gradient programme (like, EtOAc/n- methanol) were mixed due to their similar TLC pattern this
hexane and MeOH/chloroform solvent systems). For this fraction was coded as T1corander flower. The solvent
separation were separated by 5 sub fractions (EA5– EA10 system methanol: acetic acid: water in the ratio 95:5:5. This
and methanol: chloroform 7-10). Frequently each fraction fraction compound 1 was eluted with chloroform and
was checked with TLC. S. (ĐORĐEVIĆ, M. CAKIĆ, S. methanol in different ratios to get fractions of 20ml
AMR 2001, hui-ling cheng et al 2013 and jolanta nazaruk .Fraction 5 showed a single spot. This fraction was
and jan gudej 2001) for different mobile phase to getting collected dried and was recrystallized using methanol to get