液相色谱-质谱联用法测定参蛇软膏中雷公藤甲素的含量

Determination of Triptolide in Shenshe Ointment by LC-MS/MS

目的:建立一种液相色谱-质谱联用法测定参蛇软膏中雷公藤甲素的含量。方法:色谱条件:色谱柱为Agilent TCC_(18)柱(250 mm×4.6 mm,5μm),流动相为甲醇-0.1%甲酸水溶液(73∶27,v/v),流速为0.5 ml·min^(-1),柱温为35℃,进样量为2μl;质谱条件:采用电喷雾离子源(ESI),多反应离子监测(MRM),正离子检测模式,以甲硝唑为内标,检测雷公藤甲素;监测离子:雷公藤甲素m/z 361.4→43.2,甲硝唑m/z 172.1→128.0。结果:雷公藤甲素在10~500 ng·ml^(-1)范围内呈良好的线性关系,r=0.999 9;定量限(LOQ)为9.5 ng·ml^(-1);日内和日间精密度(RSD)均小于3%,平均回收率为96.87%(RSD=2.79,n=6)。结论:本方法简单、快速、灵敏、准确,可用于参蛇软膏中雷公藤甲素的含量测定。

Objective： To establish an LC-MS/MS method for the content determination of triptolide in Shenshe ointment. Methods： The chromatographic separation was performed on an Agilent TC-C1s （250 mm×4.6 mm, 5 μm）column with the mobile phase consisting of methanol -0.1% formic acid solution （73：27,v/v）. The flow rate was 0.5 ml·min^-1 and the column temperature was maintained at 35℃. An electrospray ionization （ESI） source was applied with multiple reaction monitoring （MRM） combined with a positive mode. Metronidazole was used as the internal standard. The mass transitions were 361.4→43.2 for triptolide and 172.1→ 128.0 for metronidazole, respectively. Results： The linear range was 10-500 ng·ml^-1 with good correlation coefficient （r = 0.999 9）. The limit of quantification （LOQ） was 9.5 ng·ml^-1. The intra- and inter-day RSDs of peak areas were all less than 3% and the average recovery was 96.87% （RSD = 2.79, n = 6）. Conclusion： The established LC-MS/MS method is simple, efficient, sensitive and accurate in the quality control of Shenshe ointment.