TLC Determination of Betulinic Acid from Nymphodies macrospermum: A New Botanical Source for Tagara

The accepted botanical source of Tagara, an ayurvedic drug is Valeriana jatamansi Jones. In South India, a drug by the name of Granthika Tagara is used as Tagara in several therapeutic preparations. Currently, no analytical procedures appear to be available for quality control purposes. A sensitive, selective and precise thin-layer chromatographic method has been developed and validated for the analysis of betulinic acid in Nymphoides macrospermum. Separation and quantification was achieved by TLC using ternary mobile phase of hexane:ethyl acetate:acetic acid (7:3:0.03, v/v) (R-F 0.60) on precoated silica gel 60F(254) aluminium plates and densitometric determination was carried out after derivatization with anisaldehyde-sulphuric acid reagent in the reflection/absorption mode at 540 nm. The calibration curve was linear in the concentration range of 100-600 ng spot(-1). The method was validated for precision, repeatability and accuracy. The proposed method was found to be simple, precise, specific, sensitive and accurate for the quantification of betulinic acid. This is the first TLC report for the identification and quantification of betulinic acid in N. macrospermum and may be useful for the routine quality control of Granthika Tagara.