You are on page 1of 4

International Journal of Research in Pharmaceutical and Biomedical Sciences

______________________________________________________________________Research Paper

Quantitative Analysis of Quercetin in Natural Sources by


RP-HPLC
Ch. R. S. Phani*, Ch. Vinaykumar, K. Umamaheswara rao and G. Sindhuja
R. V. Labs, Guntur, India
__________________________________________________________________________________________
ABSTRACT
A Rapid and specific reversed phase high pressure liquid chromatography (RP-HPLC) method for the
quantitative analysis of flavonoid quercetin in the extract of solanum Trilobatum and various dietary sources. The
flavonoid is analyzed on a kromasil Rp c18 column. Using a mobile phase consisted of methanol-acetonitrile-water
(60:20:20 v/v/v) under the following conditions. Detecting the wavelength at 262 nm and flow rate at 1.1 ml /min.
The sensitivity 0.05 AUFS and the volume of injecting sample is 10l. The HPLC system was operated at ambient
0
temperature (+ 28 C). The stop time was setted at 6 mins. The standard Quercetin was diluted using the mobile
phase at a known concentration of 0.5 mg/ml; the sample was filtered through sample filter contain 0.45 m prous
nylon membrane filter paper. The filterate was introduced on to a Reverse phase analytical column. The content of
Quercetin which is present in solanum Trilobatum and in various dietary sources (Red onion, green apple, lemon,
green tea) was between 2.03 to 2.30 min. Recovery of the flavonoid Quercetin is between 21.1% to 98.6%. The
method was applied to the quantitative analysis of flavonoid in solanum Trilobatum & various dietary sources and
was found to be simple rapid and efficient. The HPLC method showed on excellent performance in separating the
flavonoid quercetin in dietary sources and in medicinal plants.
Key Words: RP-HPLC, Flavonoids, quercetin, solanum Trilobatum, Quantitative Analysis
INTRODUCTION
Quercetin is a flavonol, it is plant derived flavonoid
used as a nutritional supplement found in fruits and
vegetables. Flavonoids also collectively known as
Vitamin-P and citrin are a class of Secondary
Metabolites. Flavonoids are almost universal pigments
of plants1. They are important parts of Human diet and
considered as active principles of many medicinal
plants. Quercetin is thought to have potent antioxidant,
Antidiabetic and anti tumour, and antiviral, anti
inflammatory benefits2. Quercetin is mainly found in
many often consumed foods include green apple, onion,
green tea, lemon as well as many seeds, flowers, barks,
and leaves3. It is also found in medicinal botanicals
those are Solanum Trilobatum, Ginkgobiloba,
Hypercerium perforatum and in many others. There has
been increasing interest in Quercetin in the sports,
science and athletic communities due to scientific and
clinical research results that show Quercetin
antioxidant, anti-inflamatory and other properties as
likely to improve mental and physical performance4. An
American, cancer society says that while Quercetin
has been promoted as being at effective against wide
variety of diseases including cancer5. Adequate dietary
intake of fruits and vegetables and may reduce the risk
of cancer (e.g. onion, lemon, green apple etc)6. In this
present
study
_________________________________________
*Address for correspondence
E-mail: phani.r.s.ch@gmail.com

Vol. 1 (1) Jul Sep 2010

HPLC used for the quantitative analysis of flavonoid


quercetin from medicinal plant Solanum Trilobatum
and from dietary sources. The results indicate that this
method is fast, sensitive and suitable for quantitative
analysis.
EXPERIMENTAL
Equipments
Electronic Balance ELB 300, DiGISUN pH meter,
PEAK 7000 isocratic HPLC following configurations.
PEAK 7000 delivery system. Rheodyne manual sample
injector with switch (7725). Analytical column
Kromasil 100-5 C18 (250 x 4.6 mm)
Chemicals, Reagents and Materials
Methanol (Chromatographic grade, Jiangsu
Hanbon Sci &Tech. Co., Ltd.,) Acetonitrile
(Chromatographic grade, Hanbon), H2O (HPLC grade,
sigma chemicals), Quercetin (Sigma, St. Louis) is used
as an external standard. The whole plant of Solanum
Trilobatum was collected from the local market of
Chennai, Tamilnadu. And the dietary sources collected
from local market, those are lemon, green apple, green
tea, red onion.
Chromatographic Conditions
Chromatographic analysis was carried out by
Kromasil 100 C18v reversed phase column (250 x 4.6
mm) packed with 5m diameter particles. The mobile
phase was methanol:acetonitrile:water (60:20:20 v/v/v).

www.ijrpbsonline.com

19

International Journal of Research in Pharmaceutical and Biomedical Sciences


The mobile phase was filtered through a 0.45 m
membrane filter. Then deaerated ultrasonically prior to
use. RP-HPLC separation of standard Quercetin at 262
nm. Flow rate and injection volume were 1.1 ml /min
and 10 l. The Chromatographic peaks of the analytes
were confirmed by comparing their Retention time and
UV Spectra with those of the reference standards.
Quercetin was carried out by the integration of the peak
using external standard method. All chromatographic
operations were carried out at ambient temperature.
Preparation of Standard Solution
About 5 mg of given Quercetin sample was
accurately weighed and taken into 10 ml volumetric
flasks and dissolved in 10 ml mobile phase. To prepare
0.5 mg/ml sample solution. The volumetric flask
contains standard solution kept for sonication for 10
mins. Then standard solution was filtered with 0.45 m
membrane filter paper with sample filter

Preparation of Sample Solution


In medicinal Plant: The whole plant after being wash
with water and dried in the shade for several days were
powdered about 5 mg of given solanum Trilobatum
plant sample was accurately weighed and taken into 10
ml volumetric flask and dissolved in 10 ml mobile
phase. To prepare 0.5 mg/ml sample solutions. The
volumetric flask contains standard solution kept for
sonication for 10 mins. Then sample solution was
filtered with 0.45 m membrane filter paper and with
sample filter.
In dietary sources: 5 mg of given green tea powdered
weighed and transferred into 10 ml volumetric flask and
dissolved in 10 ml mobile phase. And kept for
sonification for 10 mins and filtered by using sample
filter. Like that only 10 gms of green apple, red onion,
lemon pieces are taken from that 10 ml of sample was
extracted. The extract sample was filtered with 0.45 m
membrane filter paper with sample filter.

Structure of Flavonoid Quercetin


IUPAC Name: 2-(3, 4- dihydroxyphenyl)- 3,5,7- trihydroxy- 4H- chromen- 4-one
Molecular Formula: C15H10O7
RESULTS AND DISCUSSION
Optimization of Standard sample preparation
5 mg of the standard sample was weighed and
taken into 10 ml volumetric flask and dissolved in 10
ml mobile phase. To prepare 0.5 mg /ml sample
solution and filtered. The filtrate was injected directly.
The obtained results shows Retention time at 1.915
and area is 903880.9
Optimization of Dietary Sources sample
Preparation
5 gms of green tea powder was weight and taken
into 10 ml volumetric flask. And dissolved in 10 ml
mobile phase to prepare 0.5 mg/ml sample solution
and filtered. Like that 10 gms of green apple, red

Vol. 1 (1) Jul Sep 2010

onion, lemon pieces are taken. From that 10 ml of


sample extracted and filtered and these samples are
injected directly. The results obtained shows that
retention time at 2.03 to 2.30 and areas from 18080.9
to 161192.6. By choosing of mobile phase a mixture
of methanol acetonitril water (60:20:20:v/v/v) as
well as other chromatographic conditions showed
high performance in the separation of the flavonoid
quercetin. This quercetin mainly found in many often
consumed foods. So, adequate dietary intake of fruits
and vegetables may reduce the risk of various
diseases (e.g. Allergies, cardio vascular diseases,
inflammation, antiulcer, viral infections, anticancer,
antidiabetic).

www.ijrpbsonline.com

20

International Journal of Research in Pharmaceutical and Biomedical Sciences


Table 1: Percentage of Quercetin which is present in various natural sources
S. No.
1.
2.
3.

Sample
Standard
Solanum Trilabatum
Green Apple

Percentage of Quercetin
Present in sample
100%
98.66%
48.08%

4.

Lemon

23.55%

5.

Onion

21.19%

6.

Green Tea

6.51%

Fig 1: Standard flavonoid Quercetin, Rt at 2.27

Fig 2: Extract yield of Flavonoid Quercitin from Solanum Trilobatum, RT at 2.05 (a)

Fig 3: Extract yield of flavonoid Quercetin from Green tea, RT at 2.03 (b)

Vol. 1 (1) Jul Sep 2010

www.ijrpbsonline.com

21

International Journal of Research in Pharmaceutical and Biomedical Sciences

Fig 4: Extract yield of flavonoid Quercetin from Lemon, RT at 2.2 (c)

Fig 5: Extract yield of flavonoid Quercetin from Onion, RT at 2.30 (d)

Fig 6: Extract yield of flavonoid Quercetin from Apple, RT at 2.17 (e)

CONCLUSION
The content of Quercetin which is present in
solanum Trilobatum and various dietary sources were
found between 2.03 to 2.30 min. and the concentration
of the Quercetin found between 21.1% to 98.6%. The
method was applied to the Quantitative analysis of
Quercetin and various dietary sources was found to be
simple, rapid & efficient.
REFERENCES
1. Galeotti, F., Barile, E., Curir, P., Dolci, M.
and Lanzotti, V., Flavonoids from carnation
(Dianthus
caryophyllus)
and
their
antifungal. Phytochemistry Letters. 2008;
1: 44-50.

Vol. 1 (1) Jul Sep 2010

2. Spencer et al., Flavonoids: Modulators of


brain function. British Journal of Nutrition.
2008; 99: 6077.
3. http://www.jci.org/cgi/content/full/107/2/13
5?ijkey=a1e09ce2dbca283cec170598f2410
b15d5f4304f&keytype2=tf_ipsecsha
4. Cushnie, T. P. T. and Lamb, A. J.,
Antimicrobial activity of flavonoids.
International Journal of Antimicrobial
Agents. 2005; 26 (5): 343356.
5. Sousa et al., Phosphoprotein levels,
MAPK activities and NF kappa B
expression are affected by fisetin. J.
Enzyme Inhib. Med. Chem. 2007; .22
(4): 439444.
6. http://www.frenchscout.com/polyphenols
#procyanidins.

www.ijrpbsonline.com

22

You might also like