Estimation of Rutin and Quercetin in Terminalia chebula By HPLC

 

Ashok Kumar BS1, Lakshman K2, Jayaveera KN3, Vamshi Krishna N1, Manjunath M4, Suresh MV4, Shivatej5, Hanumantha Reddy5 and Sudheer Naik5

1Department of Pharmacognosy, Sri K.V.College of Pharmacy, Chickballapur, Karnataka (INDIA).

2Department of Pharmacognosy, PES College of Pharmacy, Bangaluru, Karnataka (INDIA).

3Department of Chemistry, Jawaharlal Nehru Technological University, Anantapur, Andhra Pradesh (INDIA).

4Department of Pharmacognosy, Rural College of Pharmacy, Devanahalli, Karnataka (INDIA).

5Department of Pharmacognosy, Sri Krishna Chaithanya College of Pharmacy, Madanapalle, Andhra Pradesh, (INDIA).

*Corresponding Author E-mail: ashok4vani@gmail.com

 

ABSTRACT

Tannins and Flavonoids present in the Terminalia chebula, Flavonoids like Rutin and quercetin possess many biochemical effects like inhibition of enzymes, regulatory role on different hormones and pharmacological activities like antimicrobial, antioxidant, anticancer, antihepatotoxic, protection of cardio vascular system. An HPLC method was developed for the estimation of rutin and quercetin from methanol methanolic extract of Terminalia chebula

 

KEYWORDS:  Terminalia chebula; Rutin; Quercetin; HPTLC; Standardization.

 

 

INTRODUCTION:

Terminalia chebula is one of the ingredients present in the many ayurvedic and other traditional medicine system. Terminalia chebula is traditionally used in formulation for anti-diabetic, anti-inflammatory, laxative, antibacterial, antifungal, cardiotonic, diuretic, hyperlipidemic activity, jaundice1-6 . Flavonoids are a group of polyphenolic compounds, which are widely distributed through out the plant kingdom. To date about 300 varieties of flavonoids are known7. Many have low toxicity in mammals and some of them are widely used in medicine for maintenance of capillary integrity8.  Rutin, 5,7,3’, 4’,tetrahydroxy flavonol -3-rhamnoglucoside and quercetin 5,7,3’,4’,- tetrahydroxy flavonol  are exhibits anti-inflammatory, antihepatotoxic9, antiulcer10, antiallergic, antiviral actions and some of them provides protection against cardiovascular mortality11,12. Both possess antioxidant activity and reduce low density lipoproteins (LDL) oxidation13, quercetin in combination with other flavonoids, are inhibiting a number of enzymes like bradykinin14, tyrosine kinase15, and 5’-nucleotidase activity16. Rutin and quercetin have shown regulatory activity of hormones like affect the transport, metabolism and action of thyroid hormones. High performance layer chromatography17 (HPLC) method is the suitable method for estimation of chemical constituents present in plant materials. Hence Terminalia chebula contains rutin and quercetin are important active constituents and is estimated by HPLC method.

 

MATERIALS AND METHODS:

Instrumentation:

The Shimadzu class LC-10AT HPLC, Hichrom C18 and a Rheodyne 7725i injector fitted with a 20 ml loop, column oven, and a photodiode array detector. The output signal was monitored and processed using chromquest version3.0 software on Pentium computer (Hewlett Packard).

 

Solvent and Chemicals:

Rutin and quercetin obtained from natural remedies (Bangalore) chromatographic grade methanol, formic acid and acetonitrile (AR), were obtained from Merck (Mumbai, India).

 

Plant material:

Terminalia chebula fruits were extracted with distilled water by Soxhlet apparatus. The pooled aqueous extract was evaporated under vacuum to dryness, yielding was noted. Aqueous extract was subjected for estimation of rutin and quercetin.

 

Preparation of Standard and Sample Solutions:

Rutin and quercetin 10 mg were accurately weighed into a 10 mL volumetric flask, dissolved in 5 mL methanol and the solution was made up to 10 mL with the same solvent (1 mg/mL). T. Chebula fruit extract was accurately weighed (10 mg) into a 10 mL of volumetric flask and dissolved in methanol the solution was filtered through Whatman filter paper No. 42 and the filtrate was made up to the mark with methanol.

 

Chromatography:

Flow rate                : 0.9 ml/min

Detection               : 340 nm

Injection quantity                : 50 ml

Column used   : Hichrom C18 (150 mmx4.6 mm i.d., 5m)

Column temperature     : 35°C

Mobile phase ration      :70:30 % v/v

Mobile phase                : 0.5% Formic acid: Acetonitrile

 

Graph 1: STANDARDS (Rutin and Quercetin)

 

Graph-2. Terminalia chebula (Extract)

 

RESULTS AND DISCUSSION:

The retention time of standards rutin and quercetin were found to be 4.072 and 19.104 (Graphs1). The retention time of rutin and quercetin in Terminalia chebula were found to be 4.802 and 19.040 (Graphs2), which are matching with standard Rt values respectively. Then the amount of rutin and quercetin in Terminalia chebula was found to be 59.52 and 9.06 % w/v respectively graphs

 

ACKNOWLEDGEMENT:

The authors are thankful to K.V.Naveen Kiran, Chairman, Sri K.V.College of Pharmacy, Chickballapur, for providing required facilities.

 

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Received on 31.01.2009        Modified on 22.03.2009

Accepted on 28.04.2009        © AJRC All right reserved

Asian J. Research Chem. 2(4):Oct.-Dec. 2009 page 388-389