基于HPLC指纹图谱及化学计量学对温性经筋通贴膏的质量评价

Quality evaluation of Warming Jingjintong Paste based on HPLC fingerprint and stoichiometry

目的:建立温性经筋通贴膏HPLC指纹图谱及其7种成分含量测定方法,并结合化学计量学的方法对其进行分析,为其质量评价提供科学依据。方法:采用HPLC法,测定12批温性经筋通贴膏指纹图谱,确定共有峰并进行相似度评价;通过对照品确定盐酸小檗碱、补骨脂素、异补骨脂素、丹皮酚、毛蕊异黄酮苷、芒柄花素和大黄素成分,并进行含量测定;借助SPSS 16.0、SIMCA-P 14.1等软件对12批温性经筋通贴膏进行聚类分析、主成分分析、最小二乘判别分析(OPLS-DA)等化学计量学分析。结果:12批温性经筋通贴膏指纹图谱共确定了32个共有峰,相似度均>0.9,并指认了7个成分峰,7种成分的线性关系均比较良好(R2≥0.9990),重复性、精密度、稳定性考察中的RSD均<5%,回收率试验按照已知含量的100%分别加入各对照品,回收率RSD均<5%;12批次温性经筋通贴膏7种成分含量范围分别在17.69~52.48,24.07~50.86,22.73~58.39,74.76~130.30,7.71~10.25,1.07~5.10,16.18~46.89μg/g;化学计量学方法分析显示可将S5划分为Ⅰ类,其他批次为Ⅱ类,OPLS-DA共确定15个差异标志物。结论:本研究建立的指纹图谱和含量测定方法科学、可靠,并结合了化学计量学,为温性经筋通贴膏质量控制及后期剂型研究奠定了基础。

Objective:To establish the HPLC fingerprint of Warming Jingjintong Paste and its 7 components content determination method with stoichiometric method,so as to provide scientific basis for its quality evaluation.Methods:The fingerprint of 12 batches of Warming Jingjintong Paste were determined by high performance liquid chromatography(HPLC)and the common peak was determined and the similarity was evaluated.The components of berberine hydrochloride,psoralen,isopsoralen,paeonol,calycosin-7-O-β-D-glucoside,formononetin and emodin were determined by reference,and the contents of these components were determined.With the help of SPSS 16.0,SIMCA-P 14.1 and other software,cluster analysis,principal component analysis,least squares discriminant analysis(OPLS-DA)and other stoichiometric analysis were performed on 12 batches of Warming Jingjintong Paste.Results:A total of 32 common peaks were identified by 12 batches of Warming Jingjintong Paste fingerprint,and the similarity was greater than 0.9.At the same time,7 component peaks were identified,and the linear relationship of the 7 components was relatively good(R’≥0.9990).RSDs of repeatability,precision and stability were all less than 5%,and RSDs of recovery tests were all less than 5%when reference substances were added to 100%of known contents.The contents of 7 components in 12 batches of Warming Jingjintong Paste were 17.69~52.48,24.07~50.86,22.73~58.39,74.76~130.30,7.71~10.25,1.07~5.10,16.18~46.89μg/g,respectively.Stoichiometric analysis showed that S5 could be divided into class I and other batches into class II.A total of 15 difference markers were identified by OPLS-DA.Conclusion:The fingerprint and content determination methods established in this study are scientific,reliable and combined with stoichiometry,which lays a foundation for the quality control of Warming Jingjintong Paste and the research of late dosage form.