基于UHPLC-MS/MS技术的导赤丸中24个成分定量测定联合化学计量学质量评价

Quantitative determination of 24 components in Daochi Pills based on UHPLC-MS/MS technology combined with chemometrics quality evaluation

目的建立UHPLC-MS/MS法同时测定导赤丸中黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、千层纸素A、盐酸小檗碱、表小檗碱、盐酸黄连碱、盐酸药根碱、盐酸巴马汀、大黄素、大黄酸、大黄素甲醚、芦荟大黄素、大黄酚、栀子苷、西红花苷I、西红花苷II、哈巴苷、哈巴俄苷、木通苯乙醇苷B、芍药苷、连翘酯苷A和连翘苷的含量。方法采用Phenomenon C18柱(100 mm×2.1 mm,2.6μm)色谱柱;以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱;体积流量0.3 mL/min;柱温40℃;进样量为1μL;采用电喷雾离子源(ESI),以多反应监测(MRM)模式正、负离子同时扫描。结果24种成分在各自的质量浓度范围内线性关系良好(r≥0.9977);精密度、稳定性、重复性等均符合方法学要求,平均加样回收率为92.7%~107.9%,RSD均小于6.5%。15批样品中24种成分的平均质量分数在6.834~7667.200μg/g,聚类分析、主成分分析结果一致,均可将3个厂家的样品准确划分为3类。结论所建立的UPLC-MS/MS法24个成分含量测定方法重现性好,简便、准确、稳定,能客观、全面、有效地确定不同生产企业样品中主要成分的差异,可为导赤丸质量评价体系的完善提供依据。

Objective To establish an UHPLC-MS/MS method for the simultaneous determination of baicalin,wogonoside,baicalein,wogonin,melaeuca A,berberine hydrochloride,epiberberine,coptisine chloride,jatrorrhizine,palmatine hydrochloride,emodin,rhein,physcion,aloe emodin,chrysophanol,geniposide,crocetin Ⅰ,crocetin Ⅱ,harpagide,harpagoside,calceolarioside B,paeoniflorin,forsythoside A,and forsythin in Daochi Pills.Methods The components were separated on phenomenon C18 column(100 mm×2.1 mm,2.6μm)with acetonitrile and 0.1%formic acid aqueous solution as the mobile phase by gradient elution at a flow rate of 0.3 mL/min.The column temperature was set at 40℃.The injection volume was 1μL.The detection was carried out by electrospray ionization(ESI),positive and negative electrospray ionization was performed in multiple reaction monitoring(MRM)mode to scan.Results A total of 24 constituents showed good linear relationships within their own ranges(r≥0.9977),and the precision,stability and repeatability complied with the requirements of methodology.The average recoveries were between 92.7%—107.9% respectively,and RSD were below 6.5%.The average mass fractions of 24 constituents in 15 batches of samples were 6.834—7667.200μg/g.The results of cluster analysis and principal component analysis were consistent,and could accurately classify three manufacturers of samples into three categories.Conclusion The established UHPLC-MS/MS method for determining the content of 24 components has good reproducibility,which is simple,accurate and stable,and can objectively,comprehensively,and effectively determine the differences in the main components in samples from different production enterprises.It can provide a basis for improving the quality evaluation system of Daochi Pills.