Development and Validation of an HPLC Method for Diosgenin, A Bioactive Compound in the Rhizomes of Chamaecostus cuspidatus

Chamaecostus cuspidatus is an important medicinal herb, with rhizomes which are rich in diosgenin content and are mainly used as an antidiabetic and anticancer. Diosgenin is a steroidal sapogenin (5α-Spirosten-3β-ol) and belongs to the spirostanol group. A validated High-Performance Liquid Chromatography (HPLC) method has been developed to estimate diosgenin. C. cuspidatus is native to India and Sri Lanka. In this study, diosgenin was separated by HPLC using a welchrom 5 μm-C₁₈ column, at a flow rate of 0.5 mL /min, by keeping the injection volume 10 μL. The column temperature was maintained at 25ºC and detection was carried out at wavelength 232 nm. The retention time for diosgenin was found to be 7.086 min. Data analysis was performed using lab solution software. The Limit of Detection (LOD) was found to be 0.753 μg/mL for diosgenin. The Limit of Quantification (LOQ) was found to be 2.341 μg/mL for diosgenin. Recovery of diosgenin was found to be 91.90%. The proposed HPLC method was validated as per ICH guidelines and was found to be selective, sensitive, and accurate for the estimation of diosgenin from the rhizome of C. cuspidatus. To the best of our knowledge, there are limited studies on the HPLC quantification of diosgenin from C. cuspidatus this is the novel study on HPLC of diosgenin from the rhizomes of C. cuspidatus.