Isolation, characterization and quantification of a bioactive compound from Cleome viscosa L. seeds by HPTLC-Densitometric method

Aim The current study presents the first report of isolation of bioactive compound (salicylic acid) from seeds of Cleome viscosa L. confirmed by its characterization by infrared (IR) spectroscopy, nuclear magnetic resonance (1H NMR and 13C NMR) spectroscopy and mass spectroscopy. The present work is the first report of HPTLC densitometric method, which has been developed and validated for quantification of a marker compound (salicylic acid) from ethyl acetate extract using the solvent system of ethyl acetate: acetonitrile (1.1:0.9, v/v). Material and methods TLC glass plates pre-coated with silica gel F366 (stationary phase) were used. Densitometric analysis of salicylic acid was performed in reflectance mode at 366 nm. Result The developed compact spot for salicylic acid at Rf 0.51. ICH guidelines were used to validate this method in terms of precision, repeatability and accuracy. Linearity range for salicylic acid was 10–50 ng/spot and the content of salicylic acid was found to be 0.9 ± 0.04 μg/g of seed. The limit of detection (LOD) and limit of quanitification (LOQ) values for salicylic acid were 4 ng and 8 ng, respectively. Better resolution from other present constituents present in the extract were obtained by this developed simple, precise and accurate method. Conclusion The study concluded the presence of Salicylic acid as active compound present in seeds of C viscosa. The present HPTLC-densitometric method may be used for routine quality control analysis for salicylic acid quantification from Cleome viscosa L.