Content determination ，fingerprint establishment and differential markers screening of Rougui renshen granules / 中国药房

ABSTRACT

OBJECTIVE To determine the conte nts of 4 main components in Rougui renshen granules ，and to establish the fingerprint and to screen differential markers affecting its quality. METHODS HPLC method was employed to determine the contents of ammonium glycyrrhizinate ，glycyrrhizin，cinnamic acid and cinnamaldehyde. HPLC fingerprints of 10 batches of Rougui renshen granules were established simultaneously. Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition）was used to evaluate the similarity and determine the common peak ；SPSS 25.0 and SIMCA 14.1 software were applied for cluster analysis （CA），principal component analysis （PCA）and partial least square-discriminant analysis （OPLS-DA）. The differential markers affecting sample quality were screened by using the variable importance in projection （VIP）value＞ 1 as standard. RESULTS The methodology of content determination met the relevant requirements. The contents of ammonium glycyrrhizinate，glycyrrhizin，cinnamic acid and cinnamaldehyde were 1.808 4-2.770 0，1.137 2-1.481 4，0.076 5-0.091 8 and 0.130 9-0.478 4 mg/g，respectively. A total of 16 common peaks were found in the fingerprints of 10 batches of Rougui renshen granules. Four chromatographic peaks were identified ，i.e. glycyrrhizin （peak 6），cinnamic acid （peak 10），cinnamaldehyde（peak 11）and ammonium glycyrrhizinate （peak 15）. The similarities of samples were ＞0.95. Results of CA showed that 10 batches of samples could be classified into three categories ：S3 was grouped into one category ；S1-S2，S4-S5 and S 10 were grouped into one category；S6-S9 were grouped into one category. The results of PCA showed that the cumulative contribution rate of the first three principal components was 91.918％，and the classification results were consistent with CA. The results of OPLS-DA showed that the four peaks with VIP value ＞1 were peak 11（cinnamaldehyde），peak 15（ammonium glycyrrhizinate ），peak 6（glycyrrhizin） and peak 9. CONCLUSIONS Established methods of content determination and fingerprint are accurate and reproducible ，and can be used for the quality evaluation of Rougui renshen granules. The components as ammonium glycyrrhizinate ，cinnamaldehyde， glycyrrhizin may be differential markers affecting the quality of Rougui renshen granules.