UPLC-MS/MS法同时测定银黄制剂中26种成分含量

Simultaneous determination of twenty-six components in Yinghuang preparations by UPLC-MS/MS

目的建立超高效液相色谱串联三重四极杆质谱法(UPLC-MS/MS)同时测定银黄制剂中26种化学成分的含量。方法采用UPLC-MS/MS法,色谱柱为Waters CORTECS C18色谱柱(2.1 mm×100 mm,1.6μm),流动相为乙腈-0. 1%甲酸水,梯度洗脱,流速为0. 25 m L/min,质谱检测采用多反应监测(MRM)模式。结果 26种分析物的线性关系分别在各自考察的浓度范围内呈良好的线性关系,各相关系数均大于0.990 0;加样回收率在93.68%~100.53%,RSD为2.02%~3.82%。12批样品中新绿原酸、绿原酸、隐绿原酸、咖啡酸、异绿原酸A、异绿原酸B、异绿原酸C、马钱苷酸、马钱苷、獐牙菜苷、断氧化马钱苷、芦丁、异槲皮苷、野黄芩苷、黄芩苷、汉黄芩苷、木犀草苷、千层纸素A-7-O-葡萄糖醛酸苷、黄芩素、汉黄芩素、木犀草素、白杨素、千层纸素A、灰毡毛忍冬皂苷甲、灰毡毛忍冬皂苷乙、川续断皂苷乙的质量分数分别为0.587~4.285、3.321~15.286、0.241~4.497、0.046~0.280、0.277~2.524、0.093~1.498、0.422~3.875、0.023~0.405、0.008~0.477、0.010~0.503、1.315~19.792、0.003~0.021、0.023~0.174、0.117~0.906、20.373~110.206、0.062~2.936、0.006~0.665、0.975~5.658、1.145~5.975、0.056~0.803、0.005~0.068、0.011~0.094、0.375~3.817、0.009~3.574、0.039~8.547、0.088~1.339 mg/g。结论本文所建立的UPLC-MS/MS法快速、准确、重复性好,可作为同时测定银黄制剂中26种成分的方法,并且首次报道了银黄制剂中同时测定有机酸、黄酮、环烯醚萜苷、三萜皂苷四大类成分的UPLC-MS/MS液质分析方法。

Objective An ultra high performance liquid chromatography coupled to triple quadrupole mass spectrometry （UPLC-MS/MS） method has been established to simultaneously determine the contents of twenty-six components（neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,loganic acid, loganin, sweroside, secoxyloganin, rutin,isoquercitrin,scutellarin,baicalin,wogonoside,luteoloside,oroxylin A-7-O -glucuronide, baicalein, wogonin,luteolin, chrysin, oroxylin A,macranthoidin A,macranthoidin B and dipsacoside B） in Yinghuang preparations. Methods This chromatographic separation was performed on a Waters CORTECS UPLC C 18 （2.1 mm×100 mm,1.6 μm） column with the mobile phase consisting of acetonitrile and 0.1% formic acid water in a gradient elution at a flow rate of 0.25 mL/min. Mass spectrometer with ESI source was using MRM mode. Results All calibration curves of the twenty-six components showed good linear regressions （ r ≥0.990 0） within the test range. The average recoveries were 93.68%-100.53% with the RSD ranging from 2.02% to 3.82%. The content ranges of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,loganic acid,loganin,sweroside,secoxyloganin,rutin,isoquercitrin,scutellarin,baicalin,wogonoside,luteoloside,oroxylin A 7-O -glucuronide,baicalein,wogonin,luteolin,chrysin,oroxylin A,macranthoidin A,macranthoidin B and dipsacoside B in Yinghuang preparations from 12 batches of samples were 0.587-4.285,3.321-15.286,0.241-4.497,0.046-0.280,0.277-2.524,0.093-1.498,0.422-3.875,0.023-0.405,0.008-0.477,0.010-0.503,1.315-19.792,0.003-0.021,0.023-0.174,0.117- 0.906, 20.373-110.206,0.062-2.936,0.006-0.665,0.975-5.658,1.145-5.975,0.056-0.803,0.005- 0.068, 0.011-0.094,0.375-3.817,0.009-3.574,0.039-8.547,0.088-1.339 mg/g respectively. Conclusion The established UPLC-MS/MS method is rapid,accurate,and has high repeatability,which could provide scientific evidences for the quality control of Yinghuang preparations.