HPLC-MS/MS法同时测定参麦注射液7种主要有效成分

Simultaneous determination of seven components in Shenmai Injection by HPLC-MS/MS

目的建立HPLC-MS/MS方法同时测定参麦注射液中7种主要有效成分(人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、麦冬皂苷D、麦冬皂苷D′、甲基麦冬二氢黄酮A和甲基麦冬二氢黄酮B)。方法采用多反应监测(MRM)模式同时监测参麦注射液中5种皂苷类和2种黄酮类成分。色谱柱为Phenomenex Luna C18柱,流动相为乙腈-0.03%乙酸水溶液,梯度洗脱,分析时间15 min。采用所建立的方法对6批参麦注射液进行了测定。结果所测7种有效成分在测定质量浓度范围内线性关系良好,相关系数(r)均大于0.996 3,线性关系分别为人参皂苷Rg1 Y=15.6 X+1.63×104;人参皂苷Re Y=14X+5.36×103;人参皂苷Rb1 Y=2.46 X+4.74×103;麦冬皂苷D Y=11 X+9.73×103;麦冬皂苷D′Y=5.56 X+1.64×103;甲基麦冬二氢黄酮A Y=3.58×103 X+2.33×104;甲基麦冬二氢黄酮B Y=4.87×103 X+2.72×104。实验精密度、重复性和稳定性良好;平均加样回收率为95.3%～104.3%。结论本法简便、快速、灵敏度高、专属性好,可用于参麦注射液中人参皂苷Rg1、Re、Rb1及麦冬皂苷D、麦冬皂苷D′、甲基麦冬二氢黄酮A和甲基麦冬二氢黄酮B的定量测定。除人参皂苷外,其他4种成分均为参麦注射液中首次测定。

Objective To develop a liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method for simultaneously analyzing seven components （ginsenosides Rgl, ginsenosides Re, ginsenosides Rbl, ophiopogonin D, ophiopogonin D＇, methylophiopogonanone A, and methylophiopogonanone B） in Shenmai Injection. Methods Multiple reaction monitoring （MRM） scan mode was used for the quantification of five saponins and two flavones. The seven constituents were separated within 15 min on a Phenomenex Luna C18 column （150 mm ~ 2.1 mm, 5 ~tm） using a mobile phase consisted of acetonitrile and 0.03% acetic acid water solution with gradient elution. Results The linear relationships between the concentration and peak areas of the seven target components were ginsenoside Rgl Y= 15.6X＋ 1.63 104, ginsenoside Re Y： 14X＋ 5.36 x 103, ginsenoside Rbl Y=2.46X＋ 4.74 x 103, ophiopogonin D Y= 11 X ＋ 9.73 103, ophiopogonin D＇ Y： 5.56 X ＋ 1.64 ~ 103, methylophiopogonanone A Y： 3.58 x 103X ＋ 2.33 ~ 104, and methylophiopogonanone B Y--4.87 x 103X＋ 2.72 x 104, respectively. The precisions, repeatabilities, and stabilities of the method were good for the seven components. The average recovery ranged from 95.3%--104.3%, and the precision in terms of RSD was less than 2.4%. Conclusion The method is rapid and reliable for the determination of the seven constituents in Shenmai Injection. Among these constituents, ophiopogonin D, ophiopogonin D＇, methylophiopogonanone A, and methylophiopogonanone B are quantified in the Shenrnai Injection for the first time.