DETERMINATION AND QUANTIFICATION OF FLAVONOIDS AND PHENOLIC CONTENT OF WRIGHTIA TINCTORIA R.BR

Bhagwan M. Waghmare and Rahul K. Dhabale Botany Research Centre, Department of Botany, Maharashtra Mahavidyalaya, Nilanga. Dist. Latur (M.S.) India. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History

Wrightia tinctoria have been studies for their chemical constituents. The powder of fruit of W. tinctoria was subjected to hot extraction with methanol, petroleum ether and ethyl acetate and subjected to chromatography. The flavonoids in form of Rutin and Catechol were isolated from fruit extract of W. tinctoria. The results revealed that, methanol fruit extracts of W. tinctoria contains maximum amount of flavonoids as 90 mg/g of Rutin equivalent and the phenolic content as 3.0 mg/g of Catechol equivalents. While, fruit extract of ethanol and petroleum ether has been found minimum, whereas, fruit extract of W. tinctoria was found highest presence of catechol equivalents. The whole plant and its specific parts such as, bark, leaf, seed fruits, roots are known to have high medicinal properties have a back history of use indigeneous communities in India. (Nadkarni K. M., 1976).
The medicinal value of W. tinctoria for the treatment of a large number common ailments of human being which were reported in Ayurveda, Siddha, Unani and folk medicine (Kirtikar andBasu 1975, Warrier et.al. 1996) and screening of scientific data revealed that a large number of various drugs of indigeneous drug have been investigated (Khare C. P. 2007 and Niir board 2008).
Apart from that, W. tinctoria also provide comprehensive information on the traditional use of mankind. Such as, ethnopharmacology, phytochemistry and pharmacological study. Present study helpful to determination and quantity content of flavonoids and phenolic compounds such compounds is useful for producing safer drugs for the treatment of common various ailments of human beings (Bharat N. S. 2015).

Determination of flavonoids from fruits extracts of Wrightia tinctoria R.Br:-
Rutin was used as standard flavonoid. Different concentrations (20 to 100 µg/ml) of rutin were analyzed at 510 nm and a calibration curve was plotted as absorbance versus concentration. 10 µg/ml of each test substance (Extracts of Wrightia tinctoria R.Br. was analyzed by using the similar procedure and quantity of flavonoids in mg per gram of rutin equivalent was determined for each extract. 195 Procedure:- Known volume of samples was pipetted out in series of test tubes and volume was made up to 0.5 ml with distilled water.  Sodium nitrite (5%; 0.03 ml) was added to each tube and incubated for 5 minutes at room temperature.  Aluminum chloride solution (10%; 0.6 ml) was added and incubated for 5 minutes at room temperature.  Sodium hydroxide solution (1 M; 0.2 ml) was added and total volume was made up to 1 ml with distilled water.  Absorbance was measured at 510 nm against a reagent blank.  Standard curve using different concentrations of rutin was prepared.  From the standard curve, concentration of flavonoids in the test samples was determined and expressed as µg of rutin equivalent.

Determination of Phenolic compounds from fruit extracts of Wrightia tinctoria R.Br:-
Catechol was used as standard phenolic compound. Different concentrations (0.5 to 2.5 µg/ml) of catechol were analyzed at 650 nm and a calibration curve was plotted as absorbance versus concentration. 1 µg/ml of each test substance (extracts of Wrightia tinctoria R.Br. were analyzed by using the similar procedure and quantity of phenolic compounds in mg per gram of catechol equivalent was determined for each extract. Procedure:- Aliquot of each sample was pipette out in series of test tubes and volume was made up to 3 ml with distilled water.  Folin-Ciocalteu Reagent (0.5 ml) was added to each tube and incubated for 3 minutes at room temperature.  Sodium carbonate (20%; 2 ml) solution was added, mixed thoroughly and the tubes were incubated for 1minute in boiling water bath.  Absorbance was measured at 650 nm against a reagent blank.  Standard curve using different concentrations of standard phenolic catechol was prepared.  From the standard curve, concentration of phenols in the test samples was determined and expressed as µg of catechol equivalent.