摘要
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,1.7μm）,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 m L/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,r^2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782.68～1 034.27μg/m L、绿原酸786.35～1 103.77μg/m L、隐绿原酸898.00～1 238.94μg/m L、咖啡酸82.85～115.17μg/m L、3,5-二咖啡酰奎宁酸226.02～461.63μg/m L、3,4-二咖啡酰奎宁酸191.59～404.21μg/m L、异连翘酯苷243.62～298.51μg/m L、连翘酯苷A 978.43～1 487.37μg/m L、连翘苷351.97～435.82μg/m L、芦丁41.19～75.65μg/m L、黄芩素40.28～73.73μg/m L、黄芩苷10 080.54～10 820.35μg/m L、野黄芩苷40.88～50.51μg/m L、汉黄芩苷187.73～204.85μg/m L、千层纸素A-7-O-β-D-葡萄糖苷144.92～335.08μg/m L、汉黄芩素9.30～25.58μg/m L、千层纸素A 7.51～16.58μg/m L。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 m L/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（r^2〉 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/m L for neochlorogenic acid, 786.35—1 103.77 μg/m L for chlorogenic acid, 898.00—1 238.94 μg/m L for cryptochlorogenic acid, 82.85 — 115.17 μg/m L for caffeic acid, 226.02 — 461.63 μg/m L for 3,5-dicaffeoylquinic acid, 191.59 — 404.21 μg/m L for 3,4-dicaffeoylquinic acid, 243.62—298.51 μg/m L for isoforsythiaside, 978.43—1487.37 μg/m L for forsythoside A, 351.97—435.82 μg/m L for forsythin, 41.19—75.65 μg/m L for rutin, 40.28—73.73 μg/m L for baicalein, 10 080.54—10 820.35 μg/m L for baicalin, 40.88—50.51 μg/m L for scutellarin, 187.73—204.85 μg/m L for wogonoside, 144.92—335.08 μg/m L for oroxylin A-7-O-β-D-glucuronide, 9.30—25.58 μg/m L for wogonin, and 7.51—16.58 μg/m L 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.
出处
《中草药》
CAS
CSCD
北大核心
2015年第22期3357-3363,共7页
Chinese Traditional and Herbal Drugs
基金
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)
