目的 建立HPLC-MS法同时测定六神丸中9种蟾蜍二烯内酯类化合物（和蟾蜍他灵、沙蟾毒精、远华蟾毒精、去乙酰华蟾毒它灵、蟾毒它灵、华蟾毒它灵、蟾毒灵、华蟾酥毒基和脂蟾毒配基）的方法。方法 采用ODS-2（250 mm×4.6 mm，5 μm）色谱柱，以乙腈-0.15%磷酸水溶液为流动相，梯度洗脱，体积流量1 mL/min，柱温40℃，检测波长296 nm。HPLC-MS联用的色谱条件与HPLC分离基本条件一致，流动相为乙腈-0.2%甲酸水溶液。通过电喷雾离子源，正离子模式扫描，对六神丸中9种蟾蜍二烯内酯类化合物进行分析鉴别。结果 9种蟾蜍二烯内酯类化合物均得到良好分离，线性关系良好（r≥0.999 6）。加样回收率均在92%～105%，RSD<5%。应用建立的HPLC-MS方法测定了10批六神丸中9种蟾蜍二烯内酯类化合物的量，分别为和蟾蜍他灵0.40～0.88 mg/g，沙蟾毒精0.32～0.83 mg/g，远华蟾毒精0.61～2.11 mg/g，去乙酰华蟾毒它灵0.15～0.32 mg/g，蟾毒它灵0.84～1.85 mg/g，华蟾毒它灵25.76～62.14 mg/g，蟾毒灵0.96～2.06 mg/g，华蟾酥毒基2.30～4.75 mg/g，脂蟾毒配基1.20～2.61 mg/g。结论 所建立的定量测定方法简便、快速、准确可靠，专属性强，可用于六神丸的质量控制。

Objective To establish an HPLC-MS method for determining nine bufadienolides (gamabufotalin, arenobufagin, telocinobufagin, desacetylcinobufotalin, bufotalin, cinobufotalin, bufalin, cinobufagin, and resibufogenin) in Bufonis Venenum and Liushen Pill. Methods HPLC analysis was performed on ODS-2 column (250 mm×4.6 mm, 5 μm) with the mobile phase made up of acetonitrie and 0.15% phosphoric in water. Flow rate was set at 1 mL/min and column temperature was 40℃. The detection wavelength was 296 nm. Nine bufadienolides were determined by HPLC-ESI-TOF-MS with the mobile phase made up of acetonitrie and 0.2% formic acid in water. Eletrospray ionization source was applied and operated in the positive ion mode. Results Nine bufadienolides were separated at baseline with good linearity (r≥0.999 6). The recovery rates were 92%-105% (RSD<5%). Conclusion The method established is rapaid, simple, and accurate, which is helpful to control the quality of Liushen Pill.

国家中药标准化项目（ZYBZH-C-JS-30）；江苏省科技成果转化项目（BA2016104）；江苏高校优势学科建设工程资助项目（PAPD）