Abstract
The research work was carried out to develop and validate new, rapid, precise and robust high performance thin layer chromatographic method for concurrent quantitative determination of gymnemagenin and gallic acid in selected herbal formulation with densitometric detection. Separation was attained on Merck aluminium HPTLC plates precoated with silica gel 60 F254. The solvent system which is optimaized contained toluene: ethyl acetate: methanol: acetic acid: formic acid (10.4: 4: 4: 0.4: 0.3, v/v/v/v). Developed plates were derivatized by 5% sulphuric acid reagent followed by heating at 110°C for 4 min in a preheated oven followed by scanning at 456 nm in reflectance-absorbance mode. The Rf (Retention factor) was found to be 0.58±0.02, for gymnemagenin and 0.41±0.02, for gallic acid. Results were found to be linear over a range of 200-1000 ng band-1 and 80-240 ng band-1 for gymnemagenin and gallic acid respectively. The proposed HPTLC method was validated according to ICH Q2 (R1) guideline. The proposed HPTLC method can be applied for quality-control testings to quantitative analysis of gymnemagenin and gallic acid simultaneously in selected marketed formulation.
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