HPLC-MS/MS法同时测定骨刺片中7个生物碱的含量

目的：建立HPLC-MS/MS同时测定骨刺片中延胡索乙素、苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、次乌头碱、乌头碱含量的方法，为骨刺片的质量评价提供依据。方法：采用Agilent Zorbax Eclipse Plus色谱柱（2.1 mm×50 mm，1.8 μm），以0.1%甲酸甲醇-0.1%甲酸溶液为流动相，梯度洗脱，流速0.3 mL·min^-1；电喷雾离子源正离子扫描，多反应监测模式（MRM）进行定量分析。结果：7个成分在一定浓度范围内与峰面积呈良好线性关系，线性相关系数r均大于0.999 0；平均回收率（n=6）在90.3%~104.8%之间，RSD在0.9%~2.8%之间。18批样品中双酯型生物碱（以乌头碱、次乌头碱、新乌头碱总量计）含量范围为0~10.11 μg·片^-1，安全性风险较小；单酯型生物碱（以苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱总量计）、延胡索乙素含量范围分别为1.99~141.58 μg·片^-1和0.02~16.43 μg·片^-1。结论：本方法可为骨刺片的质量控制提供依据。

Objective:To establish an HPLC-MS/MS method for the simultaneous determination of tetrahydropalmatine, benzoylmesaconine, benzoylaconine, benzoylhypaconine, mesaconitine, hypaconitine and aconitine in Guci tablets. Methods:HPLC separation was performed on an Agilent Zorbax Eclipse Plus column(2.1 mm×50 mm, 1.8 μm) with a mixture of 0.1% formic acid in methanol and 0.1% formic acid in water as the mobile phase in gradient elution at a flow rate of 0.3 mL·min^-1. For quantitation, electrospray ionization(ESI) source was applied and the mass spectrometer was operated in multiple reaction monitoring(MRM) modes. Results:All 7 alkaloids showed good linearity(r>0.999 0). The average recoveries(n=6) were in the range of 90.3%-104.8% with RSDs in the range of 0.9%-2.8%. In 18 batches of tested samples, the diester-alkaloids contents(the total contents of mesaconitine, hypaconitine and aconitine) ranged from 0 to 10.11 μg per tablet, which indicated low risk of drug safety;the monoester-alkaloids contents(the total contents of benzoylmesaconine, benzoylaconine and benzoylhypaconine) ranged from 1.99 to 141.58 μg per tablet, and the contents of tetrahydropalmatine ranged from 0.02 to 16.43 μg per tablet. Conclusion:The proposed method offered reference for quality control of Guci tablets.