液质联用技术同时测定金钱草颗粒中4种成分的含量

Simultaneous determination of four components in Jinqiancao Granules by HPLC-MS/MS

目的 建立液相色谱-串联质谱(HPLC-MS/MS)方法同时测定金钱草颗粒中没食子酸、槲皮苷、杨梅苷、芦丁共4种成分的含量,用于不同厂家或同一厂家不同批次样品的质量分析。方法 采用液相色谱-串联质谱(HPLC-MS/MS)法。色谱柱为Agilent ZORBAX SB-C_(18)(150 mm×4.6 mm, 5μm)柱,柱温为40℃,流动相为甲醇(A)-体积分数0.1‰的甲酸水溶液(B)进行梯度洗脱,流速为1.0 mL·min~(-1)。质谱采用电喷雾离子源(ESI),进行负离子模式检测,在多反应监测模式下,没食子酸、槲皮苷、杨梅苷、芦丁的监测离子对分别为168.7/124.9、447.4/299.9、463.4/316.1、609.5/299.9;出峰时间分别为2.76、8.09、7.17、7.85 min。结果 没食子酸、槲皮苷、杨梅苷、芦丁的线性关系良好,相关系数分别为0.997 0、0.998 4、0.996 8、0.998 4。方法的重复性试验中4种成分含量的RSD值分别为1.0%、5.0%、6.1%和6.9%,平均加样回收率均在92.7%~107.9%内。样品测定结果表明,不同批次各成分含量有明显差异。结论 本方法可为金钱草颗粒的质量标准研究提供依据。

Objective A liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for simultaneous determination of gallic acid,quercitrin,myricitrin and rutin in Jinqiancao granules was established for the quality analysis of different batches of samples from different manufacturers or from the same factory.Methods The chromatographic column was Agilent ZORBAX SB-C 18 column(150 mm×4.6 mm,5μm)with mobile phase consisted of methanol(A)-0.1‰formic acid water(B).The column temperature was 40℃and the flow rate was 1.0 mL·min-1.The determination was performed using the electrospray ionization source in negative ion mode.In the multi-reaction monitoring mode,the monitoring ion pairs of gallic acid,quercitrin,myricitrin and rutin were 168.7/124.9,447.4/299.9,463.4/316.1 and 609.5/299.9,respectively;the retention time was 2.76,8.09,7.17 and 7.85 min,respectively.Results The results of methodological verification showed that the linearity of analytical response was good.The correlation coefficients of gallic acid,quercitrin,myricitrin and rutin were 0.9970,0.9984,0.9968 and 0.9984,respectively.The RSD values of the four components in the repeatability test were 1.0%,5.0%,6.1%and 6.9%,respectively.The average recoveries were in the range of 92.7%-107.9%.The results of sample determination showed that there were significant differences between different batches samples.Conclusion The method can provide a basis for the study of the quality standard of Jinqiancao Granules.