乌头类制品中3个双酯型生物碱的RRLC-MS/MS法分析

RRLC-MS/MS simultaneous determination of 3 diester diterpenoid alkaloids in processed products of Aconitum

目的:进行乌头类制品制川乌、制草乌、白附片、黑顺片中新乌头碱、乌头碱、次乌头碱3个双酯型生物碱的超高效液相色谱-质谱(RRLC-MS/MS)联用法分析。方法:样品经乙酸乙酯-异丙醇(1∶1)超声处理(水温在25℃以下);采用Agilent Extend C18(2.1mm×50mm,1.8μm)色谱柱,以40mmol·L-1醋酸铵溶液(用氨水调pH至10.5)-乙腈为流动相进行梯度洗脱,流速0.2mL.min-1,柱温35℃。电喷雾离子化电离源(ESI),雾化气为N2,干燥气温度为350℃,干燥气流量为10L.min-1,雾化气压力为241.3Pa,扫描模式为多反应监测(MRM),定量离子对为m/z632→572、m/z646→586、m/z616→556,扫描范围为150～650aum。结果:新乌头碱、乌头碱、次乌头碱的线性范围分别为1.006～201.2ng·mL-1(r=0.9975),1.014～202.8ng·mL-1(r=0.9991)、1.072～214.4ng·mL-1(r=0.9994);检测限分别为0.2,0.1,0.2pg。结论:该方法具有专属性强、灵敏度高的特点,可用于乌头制品中的微量双酯型生物碱的测定。

Objective：To develop a specific and sensitive rapid resolution liquid chromatography-tandem mass spectrometry（RRLC-MS/MS）method for simultaneous determination of 3 diester diterpenoid alkaloids：mesaconitine,aconitine,hypaconitine.in processed products of Aconitum.Methods：The sample was extracted with ethyl acetate-isopropanol（1：1）by ultrasonication（water temperature under 25 ℃）.The separation was performed on an Agilent Extend C18（2.1 mm×50 mm,1.8 μm）column and a gradient elution of 40 mmol·L-1 ammonium acetate in water（pH 10.5,adjust the pH by ammonia water）and acetonitrile was used as mobile phase.The flow rate was 0.2 mL·min-1,and the column temperature was 35 ℃.RRLC-MS/MS system coupled with an electrospray ionization（ESI）source was performed in multiple reaction monitoring（MRM） mode.The acquisition parameters for positive mode were：drying N2 temperature,350 ℃,10 L·min-1;nebulizer pressure 241.3 Pa.The transitions of diester diterpenoid alkaloids executed as following：m/z 632→572 for mesaconitine,m/z 646→586 for aconitine,m/z 616→556 for hypaconitine.Scan ranges were 150-650 aum.Results：The linear ranges were 1.006-201.2 ng·mL-1（r=0.9975）for mesaconitine,1.014-202.8 ng·mL-1（r=0.9991）for aconitine,and 1.072-214.4 ng·mL-1（r=0.9994）for hypaconitine;The limits of detection for these 3 diester diterpenoid alkaloids were 0.2,0.1,0.2 pg.Conclusion：The validated method can be used for simultaneous determination of 3 trace components：mesaconitine,aconitine,hypaconitine in processed products of Aconitum.