乌头类制品中3个双酯型生物碱的RRLC-MS/MS法分析

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(2.1 mm×50 mm,1.8 μm)色谱柱,以40 mmol·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0.2 mL·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10 L·min^-1,雾化气压力为241.3 Pa,扫描模式为多反应监测(MRM),定量离子对为m/z 632→572、m/z 646→586、m/z 616→556,扫描范围为150~650 aum。 结果: 新乌头碱、乌头碱、次乌头碱的线性范围分别为1.006~201.2 ng·mL^-1(r=0.9975),1.014~202.8 ng·mL^-1(r=0.9991)、1.072~214.4 ng·mL^-1(r=0.9994);检测限分别为0.2,0.1,0.2 pg。 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.