目的 建立同时测定白头翁总皂苷碱水解产物中8种皂苷成分的HPLC-ELSD方法。方法 采用Kromasil C₁₈色谱柱（250 mm×4.6 mm，5 μm）；流动相为甲醇-0.2%甲酸水溶液；柱温35 ℃；梯度洗脱条件：0～30 min，70%～100%甲醇；体积流量1.0 mL/min；ELSD气化室温度40 ℃；气体压力350 kPa。结果 白头翁皂苷D、常春藤皂苷元3-O-β-D-吡喃葡萄糖基-(1→4)-α-L-吡喃阿拉伯糖、白头翁皂苷A、黑海常春藤苷A1、白头翁皂苷F、齐墩果酸3-O-β-D-吡喃葡萄糖基-(1→4)-β-D-吡喃葡萄糖基-(1→3)-α-L-吡喃鼠李糖基-(1→2)-α-L-吡喃阿拉伯糖苷、齐墩果酸3-O-β-D-吡喃葡萄糖基-(1→3)-α-L-吡喃鼠李糖基-(1→2)-α-L-吡喃阿拉伯糖苷、齐墩果酸3-O-α-L-吡喃鼠李糖基-(1→2)-α-L-吡喃阿拉伯糖苷进样量分别在0.799～4.568、0.563～6.756、0.431～2.683、0.894～7.826、0.643～7.504、1.351～7.822、0.629～2.515、0.698～2.794 μg，峰面积（A）的对数值lgA与相应质量（M，μg）的对数值lgM呈良好的线性关系（n＝5），8种成分的平均回收率在99.0%～101.0%，且RSD值均小于1.5%。结论 本检测方法简便、准确，为白头翁总皂苷碱水解产物的质量控制提供了依据。

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 ℃ 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 ℃; 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.

国家自然科学基金资助项目（81273402）；重大新药创制国家科技重大专项资助（2011ZX11102）；江苏省自然科学基金（BK2012620）