三黄片一测多评的方法学研究

目的:探索三黄片多组分色谱峰定位方法,同时建立其多组分一测多评的含量测定方法。方法:以三黄片为研究对象,在不同色谱柱和色谱仪器上,研究保留时间法、双标法以及对照提取物法对盐酸小檗碱、黄芩苷、大黄素、大黄酚4种待测成分色谱峰定位的准确性。同时,建立一测多评法测定三黄片中4种主要有效成分含量的分析方法。结果:用保留时间法或双标法峰定位时,其待测成分峰相对保留时间与理论相对保留时间差在不同色谱柱上RSD均大于5%,而对照提取物法峰定位时,在不同条件下均可快速准确定位样品中待测成分色谱峰。进而采用一测多评法测定三黄片中4种主要有效成分的含量,结果3批不同厂家生产的三黄片中4种主要有效成分一测多评法测定值与外标法测定值间无显著差异,相对误差均小于5%。结论:对照提取物法在不同色谱仪器和色谱柱上均能快速准确定位样品中待测成分色谱峰,准确性好。故本试验采用对照提取物法进行色谱峰定位,一测多评法测定三黄片中盐酸小檗碱、黄芩苷、大黄素、大黄酚4种有效成分含量,此方法准确、可行。

Objective:To explore a peak positioning method for multicomponent analysis in Sanhuang tablets,and establish a quality evaluation method for quantitative study of multi-component with a single-marker(QAMS).Methods:The accuracy of positioning chromatographic peaks of four kinds of analyte,including berberine hydrochloride,baicalin,emodin and chrysophanol,by the methods of relative retention time,two reference substances for determination of multiple components(TRSDMC),and standard reference extract(SRE)were studied using different columns and chromatography instruments.Meanwhile,a quality evaluation method,quantitative analysis of multi-component with a single-marker(QAMS),was established to determine the multi-component in Sanhuang tablets.Results:When chromatographic peaks were positioned by relative retention time or the TRSDMC in different columns,the RSD values of the relative retention time of the remaining ingredients were all above 5%.When chromatographic peak was positioned by the SRE,position chromatographic peaks of analyte can be quickly and accurately identified on different conditions by comparison analyte retention time of the reference extraction.Furthermore,QAMS was applied to determine the contents of these four kinds of active ingredients in Sanhuang tablets.No significant difference was found in the quantitative results of the four main active ingredients in 3 batches of three different manufacturers of Sanhuang tablets between the measured value and the calculated value,and its relative error were all below 5%.Conclusion:The positioning chromatographic peaks identified by the standard reference extract method were more accurate than relative retention time and TRSDMC in different columns and chromatographs.Taken together,in this study,chromatographic peak is positioned by the standard reference extract method.The QAMS is accurate and feasible in evaluating the contents of the four main active ingredients in Sanhuang tablets.