HPTLC METHOD FOR QUANTIFICATION OF ELLAGIC ACID IN IN-VIVO AND IN-VITRO SAMPLES OF WOODFORDIA FRUTICOSA (LINN.) KURZAbstract
Realising the immense medicinal values of secondary metabolites in drug formulations, the present study was conducted on an important sec metabolite i.e., ellagic acid (tannin) in in-vivo (leaf) and in-vitro callus tissues of Woodfordia fruticosa a much used medicinally important plant. The present method provided a quick and easy approach for the detection and quantitation of biomarker ellagic acid. Qualitative analysis showed the presence of ellagic acid in both in-vivo and in-vitro (callus) samples of Woodfordia fruticosa. Analytical method high-performance thin-layer chromatography (HPTLC) was developed for the quantification of ellagic acid in in-vivo and in-vitro samples of the various solvent systems tried, the system containing toluene-ethyl acetate-formic acid-methanol (3:3:0.8:0.2 v/v) gave the best resolution of ellagic acid (Rf 0.65) from the other components of the sample extracts, and enabled their simultaneous quantification. Quantitative evaluation of the plate was performed in the remission/absorption mode at 280 nm by the use of a TLC Scanner III CAMAG. The amount of ellagic acid was estimated to be about 0.21% w/w for leaf powder and 1.75% w/w for callus powder. The presence of isolated ellagic acid was confirmed by the superimposable IR spectra of isolated and authentic samples of ellagic acid. The estimated ellagic acid values for the two extracts indicate that Woodfordia fruticosa is the richer source of ellagic acid and that the plant is a promising model for the extraction of ellagic acid as it is present in appreciable quantities in both plant extracts.