Isolation, cytotoxic evaluation, and simultaneous quantification of eight bioactive secondary metabolites from Cicer microphyllum by high-performance thin-layer chromatography

Chemical investigation of Cicer microphyllum resulted in the isolation and characterization of eight natural products viz. Stigmasterol, Oleanolic acid‐3‐acetate, Oleanolic acid, Biochanin A, Genistein, Pratensein, Chrysoeriol, and Luteolin. Herein, we report a novel, accurate, and cost‐effective high‐performance thin‐layer chromatography method for the simultaneous quantification of the isolated natural products on silica‐gel 60F254 plates using the solvent system n‐hexane/ethyl acetate/formic acid (9.0:6.5:0.8, v/v/v). Natural products were quantified after postchromatographic derivatization with ceric ammonium sulfate. The method was validated as per the International Conference on Harmonization guidelines. All calibration curves showed a good linear relationship (r > 0.9943) within the test range. Precision was assessed by intra‐ and interday tests with relative standard deviations