HPLC-APCI-MS/MS法同时测定猪苓颗粒中麦角甾酮与麦角甾醇的含量

目的： 建立同时测定猪苓配方颗粒中麦角甾酮与麦角甾醇的液相色谱-串联质谱法。 方法： 采用超声提取方法制备供试品溶液，以Zorbax Eclipse XDB-C₁₈（2.1 mm×150 mm，3.5 μm）色谱柱进行分离，流动相为甲醇-10 mmol·L^-1醋酸铵缓冲液（98：2，V/V），流速0.4 mL·min^-1。试样在三重四极杆串联质谱仪中经过大气压化学离子化（APCI）离子源，在正离子方式下采用多反应离子监测（MRM）模式进行检测。 结果： 麦角甾酮与麦角甾醇浓度在1～3000 ng·mL^-1范围内与峰面积线性关系良好，麦角甾酮和麦角甾醇的平均加样回收率分别为100.8%和100.7%，日内和日间变异（RSD）均小于6.5%，色谱峰保留时间分别为4.2 min和4.6 min。 结论： 本实验建立的HPLC-APCI-MS/MS法用时短、专属性强，可以准确、灵敏、快速地检测猪苓配方颗粒中麦角甾酮与麦角甾醇的含量， 对猪苓颗粒质量控制和评价具有一定意义，为下一步深入研究其活性成分的药理机制奠定基础。

Objective: To develop a liquid chromatography-tandem mass spectrometry for the simultaneous determination of ergot sterone and ergosterol in Polyporus umbellatus particles. Methods: The test sample solution was prepared with methanol by ultrasonic extraction system.Ergot sterone and ergosterol were analyzed on a Zorbax Eclipse XDB-C₁₈ column(2.1 mm×150 mm,3.5 μm)with the mobile phase of methanol-10 mmol·L^-1 ammonium acetate buffer(98:2,V/V)at a flow rate of 0.4 mL·min^-1.The sample was injected into a triple quadrupole LC-MS with APCI probe and analyzed by the multi reaction monitoring(MRM)in the positive ion detection mode. Results: The assay was linear over the entire range of the calibration standard i.e.,a concentration range of 1 ng·mL^-1 to 3000 ng·mL^-1 which had good relationship with the peak area.The average recoveries of ergot sterone and ergosterol after ultrasonic extraction were 100.8% and 100.7%.The intra-day and inter-day variations(RSD)were less than 6.5%.And the chromatographic peak retention time of ergot sterone was 4.2 min.The retention time of ergosterol was 4.6 min. Conclusion: The method established in the experiment is characterized by short analysis time and strong specificity.It can be applied to simultaneous determination of the levels of ergot sterone and ergosterol in the same sample accurately,sensitively and rapidly.Therefore the developed method has significance for quality control and evaluation of Polyporus umbellatus particles,laying the foundation for further research on the pharmacological mechanism of the active components.