HPLC-MS法同时测定六神丸中9种蟾蜍二烯内酯类化合物  

Simultaneous determination of nine bufadienolides in Liushen Pill by HPLC-MS

作　　者：李娟[1,2,3] 狄留庆[1,2,3] 李俊松[1,2,3] 钱静[1,2,3] 王恒斌 李全 LI Juan DI Liu-qing LI Jun-song QIAN Jing WANG Heng-bin LI Quan(College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China Jiangsu Engineering Research Center for Efficient Delivery System ofTCM, Nanjing 210023, China Nanjing Engineering and Technology Research Center for TCM Pellets, Nanjing 210023, China Suzhou Leiyunshang Pharmaceutical Co. Limited, Suzhou 215009, China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]江苏省中药高效给药系统工程技术研究中心,江苏南京210023 [3]南京市中药微丸产业化工程技术研究中心,江苏南京210023 [4]雷允上药业有限公司,江苏苏州215009

出　　处：《中草药》2017年第4期700-705,共6页Chinese Traditional and Herbal Drugs

基　　金：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)

摘　　要：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×4.6mm,5μm)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.

关 键 词：六神丸 蟾酥 蟾蜍二烯内酯 HPLC 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 脂蟾毒配基 

分 类 号：R286.02[医药卫生—中药学]