基于UPLC-MS/MS技术与化学计量学相结合的风湿安泰片中18个成分含量测定及质量评价研究

Content determination and quality evaluation of 18 components in Fengshiantai tablets based on UPLC-MS/MS technology and chemometrics

目的:建立UPLC-MS/MS法,同时测定风湿安泰片中18个特征性成分含量,并对测定结果进行化学计量学分析,为其质量控制提供依据。方法:采用Waters BEH C_(18)(100 mm×2.1 mm,1.7μm)色谱柱,以甲醇-0.1%甲酸水溶液为流动相,梯度洗脱,流速0.2 mL·min^(-1),柱温40℃,进样量1μL;质谱采用电喷雾离子源(ESI),正离子检测,多反应监测(MRM)模式。结果:所建方法在一定范围内线性关系良好(r≥0.9986),平均加样回收率为90.2%~101.7%,RSD为1.1%~4.9%。39批次样品中麻黄碱(以盐酸麻黄碱计)、伪麻黄碱(以盐酸伪麻黄碱计)、绿原酸、紫丁香苷、士的宁、马钱子碱、小檗碱(以盐酸小檗碱计)、柚皮苷、桂皮醛、人参皂苷Re、人参皂苷Rg1、人参皂苷Rb1、朝藿定C、淫羊藿苷、川续断皂苷Ⅵ、甘草酸(以甘草酸铵计)、羌活醇、异欧前胡素的含量范围分别为20.23~141.75、10.50~121.69、215.24~697.34、8.46~42.42、135.24~363.32、91.29~242.87、19.65~80.31、22.57~105.05、4.69~86.88、10.32~241.15、3.36~65.07、10.79~390.69、6.51~86.29、4.10~89.75、195.55~566.37、96.98~448.95、0~132.41、7.09~232.71μg·片^(-1)。化学计量学结果显示,5个生产企业39批样品聚为3类,不同生产企业样品质量存在一定差异,筛选出影响质量的8个差异性标志物(柚皮苷、甘草酸、异欧前胡素、小檗碱、紫丁香苷、麻黄碱、伪麻黄碱、士的宁)。结论:建立的方法简单、可行,结合化学计量学方法,可用于风湿安泰片的质量评价。

Objective:To establish an UPLC-MS/MS method to measure the content of 18 characteristic components in Fengshiantai tablets,and conduct chemometric analysis on the measurement results to provide a basis for quality control.Methods:The components were separated on Waters BEH C_(18)(100 mm×2.1 mm,1.7μm)column with methanol and 0.1%formic acid aqueous solution as the mobile phase by gradient elution at a flow rate of 0.2 mL·min^(-1).The column temperature was set at 40℃.The injection volume was 1μL.The detection was carried out by electrospray ionization(ESI),and electron spray ionization source was adopted in positive ion detection with multiple reaction monitoring mode(MRM).Results:The established methods showed a good linear relationship in a certain range(r≥0.9983),whose average recovery rates of samples were 90.2%-101.7%with the RSD of 1.1%-4.9%.The content of ephedrine(calculated as ephedrine hydrochloride),chlorogenic acid,syringin,strychnine,brucine,berberine(calculated as berberine hydrochloride),naringin,cinnamaldehyde,ginsenoside Re,ginsenoside Rg_1,epimedin C,icariin,asperosaponinⅥ,glycyrrhizic acid(calculated as glycyrrhizic acid ammonium salt),ginsenoside Rb_1,notopterol,isoimperatorin in 39 batches of samples were 20.23-141.75μg per tablet,10.50-121.69μg per tablet,215.24-697.34μg per tablet,8.46-42.42μg per tablet,135.24-363.32μg per tablet,91.29-242.87μg per tablet,19.65-80.31μg per tablet,22.57-105.05μg per tablet,4.69-86.88μg per tablet,10.32-241.15μg per tablet,3.36-65.07μg per tablet,10.79-390.69μg per tablet,6.51-86.29μg per tablet,4.10-89.75μg per tablet,195.55-566.37μg per tablet,96.98-448.95μg per tablet,0-132.41μg per tablet,7.09-232.71μg per tablet.The results of chemometrics showed that 39 batches of samples from 5 production enterprises were classify into three categories,and there were certain differeces in sample quality among different production enterprises,eight differential markers(naringin,glycyrrhizic acid,isoimperatorin,berberine,syringin,ephedrine,pseudoephedrine,strychnine)that caused quality differences were found.Conclusion:The established method is simple and feasible,combined with chemometric methods,and can be used for quality evaluation of Fengshiantai tablets.