RESUMEN
OBJECTIVE To establish quantitative analysis of multi -components by single marker （QAMS） method to simultaneously detect the contents of cinnamic acid ，cinnamaldehyde，plantamajoside，verbascoside，isoacteoside，calceolarioside B ， psoralen，isopsoralen，neobavaisoflavone and bavachin in Gushen dingchuan pill ，and to perform quality evaluation of Gushen dingchuan pill by combining with chemical pattern recognition . METHODS High-performance liquid chromatography was adopted . Using psoralen as internal standard ，the relative correction factors of the other 9 components were established ，and the contents of each component were calculated and compared with those determined by external standard method . Cluster analysis ，principal component analysis and partial least squares discrimination analysis were performed by the results of QAMS method ，and the qualities of 15 batches of Gushen dingchuan pills were evaluated . RESULTS The above 10 components showed a good linear relationship in their respective ranges （r＞0.999 0）. RSDs of precision ，repeatability，stability and recovery tests were all lower than 2.00%. There was no significant difference between QAMS method and external standard method （P＞0.05）. The results of cluster analysis and principal component analysi showed that 15 batches of Gushen dingchuan pills could be clustered into 3 categories. The results of partial least squares discrimination analysis showed that psoralen ，verbascoside，cinnamaldehyde and isopsoralen were the main potential markers affecting the quality of Gushen dingchuan pills . CONCLUSIONS Established QAMS method for quantitative control of multi index components and chemical pattern recognition can be used for the quality evaluation of Gushen dingchuan pills .
RESUMEN
To solve the problems of the poor resolution of chromatographic separation,the weak durability of the relative correction factors,and the low accuracy of content determination results in the quantitative analysis of multi-components by single-marker( QAMS) method with andrographolide as the internal reference substance in the existing research of Andrographis Herba,a new QAMS method using dehydroandrographolide as the internal reference substance was established for the first time in this study. This new method can be used to simultaneously determine four diterpene lactones,including andrographolide( A),neoandrographolide( B),14-deoxyandrographolide( C),and dehydroandrographolide( S) through the optimization of chromatographic conditions and systematic investigation of methodology. At the present HPLC chromatographic conditions,four components could be well separated( R > 1. 5),and the methodology validations could satisfy the requirement of quantitative analysis. The relative correction factors( RCFs) of fA/S,fB/S,fC/S were determined as 0. 65,0. 54,0. 78,respectively. The relative standard deviations( RSDs) of their RCFs ranged between 1. 3%-5. 1%,0. 25%-0. 33%,0. 070%-0. 15%,0. 070%-0. 22%,respectively with three brands of HPLC instruments,five brands of C18 column,different flow rates( 0. 9,1. 0,1. 1 m L·min~(-1)),and different column temperatures( 25,30,35 ℃),indicating good durability of the RCFs. The relative retention value( RRV) method was used to locate the chromatographic peak of the components to be determined.The RRVs of rA/S,rB/S,and rC/Swere 0. 44,0. 86,0. 97,respectively. The RSDs of the RRVs ranged between 0. 030%-1. 6% with different HPLC instruments and columns,showing accurate peak location. The present QAMS method and the external standard method( ESM)were both used to determine the contents of four diterpene lactones from Andrographis Herba( 6 batches of medicinal materials and 18 batches of cut crude drugs). The relative errors of the determined content results between two methods were less than 2. 0%. It demonstrated that there was no significant difference in content results between these two methods,indicating good accuracy of the present QAMS method. Therefore,in this study,an accurate and highly durable QAMS method using dehydroandrographolide as the internal reference substance was established for simultaneous determination of four diterpene lactones. This method could be used to effectively control the quality of Andrographis Herba and provide technical basis for the formulation of traditional Chinese medicine industry standard and improvement of the Chinese Pharmacopoeia standard of Andrographis Herba.