UPLC-MS/MS法同时测定灯盏生脉胶囊中的11个活性成分的含量

目的:建立超高效液相色谱-串联质谱法应用于灯盏生脉胶囊中11个活性成分的定量方法。方法: 采用Agilent ZORBAX RRHD Eclipse Plus C₁₈(2.1 mm×50 mm,1.8 μm)色谱柱;以0.15%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱,流速为0.35 mL·min^-1;离子源为电喷雾电离源,采用正/负离子模式检测,多重反应监测模式采集并定量。结果: 灯盏生脉胶囊中11个活性成分在30 min内达到基线分离,绿原酸、咖啡酸、灯盏花乙素、人参皂苷Rg₁、人参皂苷Re、人参皂苷Rb₁、五味子醇甲、麦冬皂苷D、五味子酯甲、五味子甲素、五味子乙素的线性范围分别为0.0060~40、0.040~50、1.00~625、0.020~50、0.020~60、0.0020~40、0.0040~40、0.010~25、0.0020~40、0.00080~40、0.0012~48 mg·L^-1各自质量浓度在相应的范围内与峰面积呈良好的线性关系,r>0.9990;检出限分别为1.5、10、2.0、5.0、5.0、0.50、1.0、2.5、0.50、0.20、0.30 μg·L^-1。11个成分的加样回收率为96.5%~105%,RSD(n=3)均小于3.0%。结论: 本方法准确,灵敏,简便快速,重复性好,可用于灯盏生脉胶囊的质量控制。

Objective: To develop a quantitative method for simultaneous determination of eleven effective components in Dengzhanshengmai capsules by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). Methods: The UPLC separation was performed on a ZORBAX RRHD Eclipse Plus C₁₈column (2.1 mm×50 mm,1.8 μm) by using water containing 0.15% formic acid and acetonitrile as the mobile phase with the gradient elution at a flow rate of 0.35 mL·min^-1.The negative/positive ion mode was utilized for collection and quantification in a multiple reaction monitoring mode with icon source as the electrospray ionization (ESI) source. Results: Eleven active components in Dengzhanshengmai capsules were baseline separated in 30 minutes.Under the optimized conditions,the calibration curves were linear (r>0.9990) in the range of 0.0060-40 mg·L^-1 for chlorogenic acid,0.040-50 mg·L^-1 for caffeic acid,1.00-625 mg·L^-1 for scutellarin,0.020-50 mg·L^-1 for ginsenoside Rg1,0.020-60 mg·L^-1 for ginsenoside Re,0.0020-40 mg·L^-1 for ginsenoside Rb1,0.0040-40 mg·L^-1 for schisandrin,0.010-25 mg·L^-1 for ophiopogonin D,0.0020-40 mg·L^-1 for schisantherin,0.00080-40 mg·L^-1 for deoxyschizandrin,0.0012-48 mg·L^-1 for γ-schisandrin with detection limits of 1.5,10,2.0,5.0,5.0,0.50,1.0,2.5,0.50,0.20,0.30 μg·L^-1,respectively.The recoveries ranged from 96.5% to 105% with all relative standard deviations no more than 3.0%. Conclusion: The developed method is accurate,sensitive,simple,rapid and highly repeatable which can be used for quality control of Dengzhanshengmai capsules.