UPLC-MS/MS法同时测定康艾注射液中11种成分

Simultaneous determination of 11 components in Kang'ai Injection by UPLC-MS/MS

目的建立UPLC-MS/MS法同时测定康艾注射液中11种成分(毛蕊异黄酮、毛蕊异黄酮苷、人参皂苷Rg1、人参皂苷Re、芒柄花素、黄芪甲苷、人参皂苷Rf、人参皂苷Rb2、人参皂苷Rc、人参皂苷Rd、人参皂苷Rb1)的方法。方法采用正负离子切换多反应监测模式(MRM),ESI离子源;Phenomenex C18色谱柱(100 mm×3.0 mm,2.6μm)进行分离,流动相为甲醇(A)、乙腈(B)、5 mmol/L乙酸铵-0.05 mmol/L乙酸钠水溶液(C),梯度洗脱,分析时间为7.0 min。结果所测11种主要有效成分在测定浓度范围内线性关系良好,r均大于0.99;精密度、重复性和稳定性良好;平均加样回收率为95.2%~104.4%,RSD≤4.64%。4个批次康艾注射液中毛蕊异黄酮、毛蕊异黄酮苷、人参皂苷Rg1、人参皂苷Re、芒柄花素、黄芪甲苷、人参皂苷Rf、人参皂苷Rb2、人参皂苷Rc、人参皂苷Rd、人参皂苷Rb1测得量分别为0.004~0.006μg/m L、0.002~0.003μg/m L、125.75~148.00μg/m L、51.75~77.00μg/m L、0.010~0.013μg/m L、51.50~87.75μg/m L、27.83~30.73μg/m L、4.23~5.15μg/m L、8.40~13.35μg/m L、17.33~27.68μg/m L和9.03~11.00μg/m L。结论首次建立可同时测定康艾注射液的11种主要成分的UPLC-MS/MS方法,该法操作简便,灵敏度高,专属性好,分析速度快,可用于康艾注射液的质量控制。

Objective To develop and validate an ultra-high performance liquid chromatography-tandem mass spectrometry（UPLCMS/MS） method for the simultaneous determination of 11 components（calycosin, calycosin-7-glucoside, ginsenoside Rg1, ginsenoside Re, formononetin, astragaloside IV, ginsenoside Rf, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, and ginsenoside Rb1） in Kang＇ai Injection（KI）. Methods Multiple reaction monitoring（MRM） with a polarity-switching electrospray ionization（ESI） source between positive and negative mode was used for the quantification. The 11 components were separated within 7.0 min on a Phenomenex Luna C18 column（100 mm × 3.0 mm, 2.6 μm） using a mobile phase consisted of methanol, acetonitrile, 5 mmol/L ammonium acetic-0.05 mmol/L sodium acetate water solution with gradient elution. Results The linear relationships showed well in the area of determination. The average recovery varied between 95.2% and 104.4% with RSD ≤ 4.64%. The precision, repeatability, and stability of the method were good for the determination of 11 components. The content ranges of calycosin, calycosin-7-glucoside, ginsenoside Rg1, ginsenoside Re, formononetin, astragaloside IV, ginsenoside Rf, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, and ginsenoside Rb1 in four batches of KI were 0.004—0.006 μg/m L, 0.002—0.003 μg/m L, 125.75—148.00 μg/m L, 51.75—77.00 μg/m L, 0.010—0.013 μg/m L, 51.50—87.75 μg/m L, 27.83—30.73 μg/m L, 4.23—5.15 μg/m L, 8.40—13.35 μg/m L, 17.33—27.68 μg/m L, and 9.03—11.00 μg/m L, respectively. Conclusion A new rapid and reliable UPLC-MS/MS method was developed and validated to evaluate the quality of KI for the first time.