HPLC-MS/MS法同时测定参乌健脑胶囊中8种成分

Simultaneous determination of eight components in Shenwu Jiannao Capsules by HPLC-MS/MS

目的建立高效液相色谱-串联质谱（HPLC-MS/MS）同时测定参乌健脑胶囊（SJC）中8种成分（葛根素、丹参酮IIA、二苯乙烯苷、芍药苷、人参皂苷Re、人参皂苷Rg1、毛蕊异黄酮苷、黄芩苷）的定量方法。方法选用YMC-Triart C18色谱柱,以甲醇-0.2%甲酸水溶液为流动相,梯度洗脱,体积流量为0.6 m L/min,柱温40℃。质谱条件：ESI源,采用多反应监测模式（MRM）,以定量离子对峰面积进行定量。结果 SJC中葛根素、丹参酮IIA、二苯乙烯苷、芍药苷、人参皂苷Re、人参皂苷Rg1、毛蕊异黄酮苷、黄芩苷分别在20.31～20 310、25.66～25 660、20.45～20 450、50.79～50 790、50.57～50 570、50.38～50 380、10.32～10 320、25.74～25 740 ng/m L内线性关系良好（r≥0.999 0）;精密度良好,重复性良好,RSD均小于2.0%;在室温条件下24 h内稳定;加样回收率为97.98%～102.48%,RSD小于1.6%。所测8种成分在6批样品中的质量分数依次为葛根素1.843～1.860 mg/g、丹参酮IIA 1.618～1.629 mg/g、二苯乙烯苷2.116～2.129 mg/g、芍药苷2.537～2.547mg/g、人参皂苷Re 0.034～0.041 mg/g、人参皂苷Rg1 0.048～0.055 mg/g、毛蕊异黄酮苷0.551～0.564 mg/g和黄芩苷2.333～2.346 mg/g。结论该方法快速、简便、重复性好,可用于同时测定SJC中多种成分。

Objective To develop a high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） method for the simultaneous determination of eight components（puerarin, tanshinone IIA, stibene glucoside, paeoniflorin, ginsenoside Re,ginsenoside Rg1, calycosin-7-glucoside, and baicalin） in Shenwu Jiannao Capsules（SJC）. Methods Chromatographic column：YMC-Triart C18, mobile phase： methanol-water（including 0.2% formic acid）, and gradient elution, flow rate： 0.6 m L/min, column temperature： 40 ℃. MS condition： electrospray ionization（ESI） and multiple reaction monitoring（MRM） were adopted, the detected peak areas of ion pairs were used for quantitative determination. Results There was a good linearity between the absorption peak area and the concentration for puerarin, tanshinone IIA, stibene glucoside, paeoniflorin, ginsenoside Re, ginsenoside Rg1, calycosin-7- glucoside, and baicalin in the ranges of 20.31—20 310, 25.66—25 660, 20.45—20 450, 50.79—50 790, 50.57—50570, 50.38—50 380, 10.32—10 320, and 25.74—25 740 ng/m L, respectively（r ≥ 0.999 0）. The precision was good and RSD was less than 2.0%. The repeatability was good and RSD was less than 2.0%. The stability was good in 24 h. The average recoveries were ranged from 97.98% to 102.48%（RSD ≤ 1.6%）. The contents of puerarin, tanshinone IIA, stibene glucoside, paeoniflorin,ginsenoside Re, ginsenoside Rg1, calycosin-7-glucoside, and baicalin in six batches of samples were in the ranges of 1.843 —1.860,1.618 — 1.629, 2.116 — 2.129, 2.537 — 2.547, 0.034 — 0.041, 0.048 — 0.055, 0.551 — 0.564, and 2.333 — 2.346 mg/g, respectively.Conclusion The method is rapid, simple, repeatable, and can be used to determine the multi components in different batches of SJC simultaneously.