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 (H-1 NMR and C-13 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 R10.51.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 tglg 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 add 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. (C) 2018 SAAB. Published by Elsevier B.V. All rights reserved.