[关键词]
[摘要]
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,2.6 μm)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0.004~0.006 μg/mL、0.002~0.003 μg/mL、125.75~148.00 μg/mL、51.75~77.00 μg/mL、0.010~0.013 μg/mL、51.50~87.75 μg/mL、27.83~30.73 μg/mL、4.23~5.15 μg/mL、8.40~13.35 μg/mL、17.33~27.68 μg/mL和9.03~11.00 μg/mL。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。
[Key word]
[Abstract]
Objective To develop and validate an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/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/mL, 0.002-0.003 μg/mL, 125.75-148.00 μg/mL, 51.75-77.00 μg/mL, 0.010-0.013 μg/mL, 51.50-87.75 μg/mL, 27.83-30.73 μg/mL, 4.23-5.15 μg/mL, 8.40-13.35 μg/mL, 17.33-27.68 μg/mL, and 9.03-11.00 μg/mL, 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.
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[基金项目]
河北医科大学大学生创新性实验计划项目(USIP2016137);河北省医学科学研究重点课题计划项目(20150560);河北省食品药品监督管理局(重点)科技项目(ZD2015023);国家自然科学基金资助项目(21402139)