Purification of the total steroidal saponins from fenugreek seeds (Trigonella foenum-graecum L.) using aqueous two-phase system and determination of diosgenin content using micellar electrokinetic chromatography method

The total steroidal saponins, particularly its major steroidal sapogenin (diosgenin), are the main active principles of fenugreek seed extract. In this study, an ethanol-salt aqueous two-phase system (ATPS) was explored for the purification of the total steroidal saponins, and the process conditions were optimized by response surface methodology (RSM). Under the optimized conditions, the RSM predicted recovery of the total steroidal saponins in the top phase of ATPS was 97.9%, which agreed with the average experimental recovery (98.3 ± 4.2% ( n = 6)). Moreover, a rapid micellar electrokinetic chromatography (MEKC) method was developed for the determination of diosgenin from extracts. The diosgenin content in the ATPS top phase extract was 3-fold higher than that in crude extract, suggesting this ATPS having a great potential for purification pharmacological active ingredients from fenugreek seeds.