Macroscopical and microscopical analysis of selected Indian medicinal plants and HPTLC method for quantification of chemical markers in a polyherbal formulation

Context: Pharmacognostical standardization of crude drugs is essential before carrying out its scientific validation as it furnishes the identity and authenticity of the plant. The present work was undertaken to establish the macroscopic, microscopical evaluation and HPTLC densitometric analysis of selected Indian medicinal plants and polyherbal mixture. Aims: To explore the pharmacognostical and quantitative estimation of chemical markers lupeol, rutin and shatavarin IV in polyherbal formulation using HPTLC analysis. Methods: Pharmacognostical features were studied by the macroscopic and microscopical analysis were done. High-performance thin-layer chromatography was performed to evaluate the presence of lupeol, rutin and shatavarin IV in n-hexane:ethyl acetate (6:4 v/v), ethyl acetate:acetic acid:formic acid; methanol (10:1.1:1.1:2.6 v/v) and chloroform:acetic acid:water:methanol (5:3.5:1.5:1 v/v) for lupeol, rutin and shatavarin IV, respectively. Results: The Rf values for lupeol (0.90), rutin (0.36) and shatavarin IV (0.89) in samples and reference standard were found comparable under UV light at 550, 254 and 429 nm, respectively. The quantified values of lupeol, rutin and shatavarin IV (0.067, 0.053 and 0.048 ng/g, respectively) in the polyherbal formulation were detected using HPTLC analysis. The high-performance thin-layer chromatography method developed for quantization was simple, accurate and specific. Conclusions: The present study provides useful information about plant characters and standardization of specific and accurate tools to develop quantifications for identity, transparency and reproducibility of chemical markers in polyherbal formulation.