LC-MS/MS法同时测定复方附子口服液中10种成分含量

Simultaneous determination of ten components in compound Fuzi oral solution by LC-MS/MS

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,1.8μm),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 m L·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9.29μg·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、7.69μg·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 m L·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（r0.998 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.