HPLC-MS法同时测定白蒲黄片中7种五环三萜皂苷类化学成分

目的: 建立同时测定白蒲黄片中7种五环三萜皂苷类成分白头翁皂苷A3、白头翁皂苷B4、23-羟基白桦脂酸、白头翁皂苷B、白头翁皂苷C、刺人参苷S和齐墩果酸的HPLC-MS法。方法: 采用Diamonsil ODS C₁₈(250 mm×4.6 mm,5 μm)色谱柱,流动相为含0.1%甲酸的水(A)-含0.1%甲酸的甲醇(B),梯度洗脱程序为25%A(0 min),5%A(0~6 min),5%A(6~14 min),25%A(保持6 min),流速0.8 mL·min^-1,柱温30 ℃,进样量10 μL;采用正离子和负离子模式同时监测,正离子和负离子监测模式下源喷射电压分别为5.5 kV和-4.5 kV,离子源温度为650 ℃,雾化气(Gas 1)344 kPa,加热气(Gas 2)412 kPa,接口持续加热,帘气172 kPa,全程氮气通入状态。结果: 在14 min内白蒲黄片中7种有效成分白头翁皂苷A3、白头翁皂苷B4、23-羟基白桦脂酸、白头翁皂苷B、白头翁皂苷C、刺人参苷S和齐墩果酸被完全分离;峰面积与其浓度呈良好的线性;平均回收率范围为95.11%~107.3%,RSD为0.86%~3.26%。结论: 本文建立的方法经验证简便、重现性好、专属性高,可为白蒲黄片质量控制提供参考。

Objective: To establish a method based on high performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry(HPLC-ESI-MS/MS)for the simultaneous determination of anemoside A3,anemoside B4,23-hydroxybetulinic acid,pulsatilloside B,pulsatilloside C,cirenshenoside S and oleanolic acid in Baipuhuang tablets. Methods: The samples were separated on a Diamonsil ODS C₁₈ column(250 mm×4.6 mm,5 μm)by a mobile phase of 0.1% formic acid aqueous solution(A)-methanol containing 0.1% formic acid(B)at the flow rate of 0.8 mL·min^-1.The gradient elution program was 25%A(0 min),5%A(0-6 min),5%A(6-14 min),and then back to the original 25%A held for 6 min.The column temperature was 30 ℃,and the injection volume was 10 μL.Multiple-reaction monitoring scanning was employed for quantification with switching electrospray ion source polarity between positive and negative modes in a single run.The ion spray voltage for the two modes was set to 5.5 kV and-4.5 kV,respectively.The turbo spray temperature was maintained at 650 ℃.Nebulizer gas(Gas 1)and heater gas(Gas 2)was set at 344 and 412 kPa,respectively.The curtain gas was kept at 172 kPa and the interface heater were kept operating.Nitrogen was used throughout the course. Results: The complete separation was obtained within 14 min for the seven compounds(anemoside A3,anemoside B4,23-hydroxybetulinic acid,pulsatilloside B,pulsatilloside C,cirenshenoside S and oleanolic acid).Seven regression equations showing linear relationships between peak area and content of each compound were obtained.The recoveries(n=6)of the compounds were 95.11%-107.3% and RSDs were 0.86%-3.26%,respectively. Conclusion: The method is proved to be simple,accurate and with good reproducibility by the method validation,and can be used for the determination of seven triterpenoid saponins in Baipuhuang tablets.