LC-MS/MS法同时测定复方附子口服液中10种成分含量

目的:建立LC-MS/MS同时测定复方附子口服液中10个成分(毛蕊异黄酮葡萄糖苷、苯甲酰新乌头碱、苯甲酰次乌头碱、苯甲酰乌头碱、党参炔苷、新乌头碱、次乌头碱、乌头碱、黄芪甲苷和麦冬皂苷D)含量的分析方法。方法:采用Agilent SB-C₁₈色谱柱(2.1 mm×50 mm,1.8 μm),柱温为25 ℃,流动相A为0.05%甲酸水溶液,流动相B为0.05%甲酸甲醇溶液,梯度洗脱(0~3.5 min,40%B;3.5~4 min,40%B→70%B;4~5 min,70%B→90%B;5~6 min,90%B;6~6.1 min,90%B→40%B;6.1~7 min,40%B),流速0.3 mL·min^-1;电喷雾离子源(ESI),正、负离子扫描方式,多反应监测扫描模式进行定量分析。 结果:所测10种成分在一定范围内具有良好的线性关系,线性相关系数r均大于0.998 1,精密度、重复性、稳定性的RSD均小于3.6%;加样回收率在95.24%~104.1%,RSD均小于4.2%。3个批次双酯型生物碱(以新乌头碱、次乌头碱、乌头碱总量计)平均含量为9.29 μg·mL^-1,满足用药安全。单酯型生物碱(以苯甲酰新乌头碱、苯甲酰次乌头碱、苯甲酰乌头碱总量计)、毛蕊异黄酮葡萄糖苷、党参炔苷、黄芪甲苷、麦冬皂苷的平均含量分别为604.3、72.3、47.8、308、7.69 μg·mL^-1。结论:所建立的分析方法经方法学验证,可为复方附子口服液的质量控制提供依据。

Objective:To establish an LC-MS/MS method for simultaneous determination of ten components (calycosin-7-O-β-D-glucoside,benzoylmesaconine,benzoylhypaconine,benzoylaconine,lobetyolin,mesaconitine,hypaconitine,aconitine,astragoloside and ophiopogonin D) in compound Fuzi oral solution.Methods:The HPLC analysis was performed on an Agilent SB-C₁₈ column (2.1 mm×50 mm,1.8 μm).The mobile phase consisted of 0.05% formic acid in water (A) and 0.05% formic acid in methanol (B) with gradient elution (0- 3.5 min,40%B;3.5-4 min,40%B→70%B;4-5 min,70%B→90%B;5-6 min,90%B;6-6.1 min,90%B→40%B;6.1-7 min,40%B).The flow rate was 0.3 mL·min^-1,and the column temperature was 25℃.Electrospray ionization source was used.Positive and negative ions scanning and selected multiple reaction monitoring mode was applied for quantization.Results: The method has a good linearity in the given ranges (r >0.998 1).The RSDs of the precision,repeatability and stability tests were less than 3.6%.The average recoveries were in the range of 95.24%~104.1% with RSDs less than 4.2%.The average content of the diester aconitum alkaloids (with the total quantity of mesaconitine,hypaconitine,aconitine) was 9.29 μg·mL^-1,which met the drug safety.The average contents of the monoester aconitum alkaloids(with the total quantity of benzoylmesaconine,benzoylhypaconine,benzoylaconine),calycosin-7-O-β-D-glucoside,lobetyolin,astragoloside and ophiopogonin D were 604.3,72.3,47.8,308 and 7.69 μg·mL^-1,respectively.Conclusion:The method can provide a basis for the quality control of compound Fuzi oral solution.