A validated high-performance thin-layer chromatographic method for the quantitation of protodioscin in Trigonella foenum-graecum and its herbal formulations

A simple, rapid, and sensitive high-performance thin-layer chromatographic (HPTLC) method was developed for the estimation of protodioscin from Trigonella foenum-graecum. Chromatographic separation was achieved on aluminum plates precoated with silica gel G60 F254 as the stationary phase and chloroform–methanol–glacial acetic acid (6.5:3.0:0.5 v/v) as the mobile phase. Densitometric evaluation was carried out at 515 nm after derivatizing the spots with p-dimethyl aminobenzaldehyde. The RF value of the protodioscin was found to be 0.31 ± 0.02. The response in terms of peak area was found to be linear over the concentration range from 400 to 1400 ng spot−1 with a regression coefficient of 0.9981 and mean recovery of 100.90 ± 3.00%. The developed method was validated as per the International Conference on Harmonization (ICH) guidelines. The limit of detection and limit of quantitation were found to be 100 ng spot−1 and 400 ng spot−1, respectively. Robustness of the developed method was assessed using experim...