ABSTRACT
Objective: To develop an HPLC method for the simultaneous determination of eight saponins in alkali hydrolysate of total saponins from Pulsatilla chinensis. Methods: HPLC was performed on a Kromasil C18 analytical column (250 mm × 4.6 mm, 5.0 μm) at 35°C with MeOH-0.2% HCOOH solution as the mobile phase by gradient elution and the step gradients were as follows: 0-30 min, 70%-100% MeOH; The flow rate was 1.0 mL/min; ELSD gasification chamber temperature was 40°C; Gas pressure of carrier gas N2 was 350 kPa. Results: The linear response (the log values of peak areas with corresponding log values of sample introducing amounts) ranges were 0.799-4.568 μg for pulsatilla saponin D, 0.563-6.756 μg for hederagenin 3-O-β-D-glucopyranosyl-(1→4)-α-L-arabinopyranoside, 0.431-2.683 μg for pulsatilla saponin A, 0.894-7.826 μg for hederacolchiside A1, 0.643-7.504 μg for pulsatilla saponin F, 1.351-7.822 μg for oleanolic acid 3-O-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl - (1→3)-α-L-rhamnopyranosyl-(1→2) - α-L-arabinopyranoside, 0.629-2.515 μg for oleanolic acid 3-O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl - (1→2)-α-L-arabinopyranoside, and 0.698-2.794 μg for oleanolic acid 3-O-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranoside, respectively (n = 5). The average recoveries of the eight saponins were between 99.0% and 101.0%, and RSD values were less than 1.5%. Conclusion: The results demonstrate that the established method has the adequate accuracy and selectivity for the quality control of alkali hydrolysate of total saponins from P. chinensis.