目的 建立超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定双黄连口服液中17种成分(新绿原酸、绿原酸、隐绿原酸、咖啡酸、3,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、异连翘酯苷、连翘酯苷A、连翘苷、芦丁、黄芩素、黄芩苷、野黄芩苷、汉黄芩苷、千层纸素A-7-O-β-D-葡萄糖苷、汉黄芩素、千层纸素A)的分析方法。方法 采用Acquity UPLC BEH C₁₈色谱柱(100 mm×2.1 mm,1.7 μm),以乙腈-甲醇(4:1)-0.4%甲酸水为流动相,梯度洗脱,体积流量为0.4 mL/min,柱温40 ℃,采用电喷雾电离源(ESI),多反应离子监测(MRM)扫描方式进行检测,分析时间为7 min。结果 所测17种有效成分在测定质量浓度范围内线性关系良好,r²均大于0.990 1,实验精密度、准确度和稳定性良好,平均加样回收率为95.81%～102.42%,所测5批样品中17种成分定量范围依次为新绿原酸782.68～1 034.27 μg/mL、绿原酸786.35～1 103.77 μg/mL、隐绿原酸898.00～1 238.94 μg/mL、咖啡酸82.85～115.17 μg/mL、3,5-二咖啡酰奎宁酸226.02～461.63 μg/mL、3,4-二咖啡酰奎宁酸191.59～404.21 μg/mL、异连翘酯苷243.62～298.51 μg/mL、连翘酯苷A 978.43～1 487.37 μg/mL、连翘苷351.97～435.82 μg/mL、芦丁41.19～75.65 μg/mL、黄芩素40.28～73.73 μg/mL、黄芩苷10 080.54～10 820.35 μg/mL、野黄芩苷40.88～50.51 μg/mL、汉黄芩苷187.73～204.85 μg/mL、千层纸素A-7-O-β-D-葡萄糖苷144.92～335.08 μg/mL、汉黄芩素9.30～25.58 μg/mL、千层纸素A 7.51～16.58 μg/mL。结论 方法简单、快速、准确度及灵敏度高、专属性好,可用于双黄连口服液中3大类17个主要成分的快速定量测定。

Objective To establish a UPLC-MS/MS method for simultaneous determination of 17 bioactive compounds in Shuang-Huang-Lian Oral Liquid. Methods The separation was performed on an Acquity UPLC HSS T3 C₁₈ column (100 mm ×2.1 mm, 1.7 μm) and the flow rate was set at 0.4 mL/min with acetonitrile/methanol (4:1)-0.4% formic acid as the mobile phase. The column temperature was 40 ℃ and only 7 min was needed for separating the five pairs of isomers. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source with positive and negative modes. Results All calibration curves had good linearity (r² >0.990 1). The precisions and accuracies of the 17 analytes were all satisfied and the recoveries were 95.81%—102.42%. The content ranges of the 17 compounds were 782.68—1 034.27 μg/mL for neochlorogenic acid, 786.35—1 103.77 μg/mL for chlorogenic acid, 898.00—1 238.94 μg/mL for cryptochlorogenic acid, 82.85—115.17 μg/mL for caffeic acid, 226.02—461.63 μg/mL for 3,5-dicaffeoylquinic acid, 191.59—404.21 μg/mL for 3,4-dicaffeoylquinic acid, 243.62—298.51 μg/mL for isoforsythiaside, 978.43—1487.37 μg/mL for forsythoside A, 351.97—435.82 μg/mL for forsythin, 41.19—75.65 μg/mL for rutin, 40.28—73.73 μg/mL for baicalein, 10 080.54—10 820.35 μg/mL for baicalin, 40.88—50.51 μg/mL for scutellarin, 187.73—204.85 μg/mL for wogonoside, 144.92—335.08 μg/mL for oroxylin A-7-O-β-D-glucuronide, 9.30—25.58 μg/mL for wogonin, and 7.51—16.58 μg/mL for oroxylin A, respectively. Conclusion The developed UPLC-MS/MS method is simple, sensitive, and accurate, and could be applied for simultaneous determination of the 17 components in Shuang-Huang-Lian Oral Liquid.

江苏省“333高层次人才培养工程”资助项目(BRA2013201);江苏高校优势学科建设工程资助项目(ysxk-2010)