Abstract

SIMULTENOUS ESTIMATION OF AMINO ACID AND STEROIDAL SAPOGENIN IN BULK AND HERBAL FORMULATION

Sonal More*, Dr. Varsha M. Jadhav and Dr. Vilasrao J. Kadam

ABSTRACT

The present work deals with development of HPTLC method for simultaneous estimation of l-dopa and diosgenin in Mucuna pruriens and Tribulus terristris respectively and also in a marketed herbal formulation containing Mucuna pruriens seeds powder and Tribulus terristis fruits powder as the major ingredients. Chromatographic separation of the drugs was performed on Merck TLC aluminium plates pre-coated with silica gel 60F254 as the stationary phase. The mobile phase selected was toluene: ethyl acetate: methanol: formic acid: glacial acetic acid(4: 2.5: 2:1.20:0.30 v/v/v/v/v). The sample solutions were prepared in methanol and linear ascending development was carried out in twin trough glass chamber. The two markers were satisfactorily resolved with Rf values 0.24±0.02 and 0.73±0.02 for l-dopa and diosgenin, respectively. The linear regression analysis data for the calibration plots for l-dopa and diosgenin showed good linear relationship with regression coefficient (r2) 0.9991 and 0.998 in the concentration range of 200-1400 ng/spot and 10-70 ng/spot, respectively. The method was validated for linearity, specificity, recovery, precision, robustness as per ICH guidelines. The developed method was found to be precise, specific and reliable for the determination of l-dopa and diosgenin from these individual herbs as well as from marketed herbal formulations.

Keywords: L-dopa, Diosgenin, HPTLC, Validation, Vigomax Forte tablets, Analytical method development.