Study On Effective Fractions In Chinese Herbal Prescription "ZHIZICHITANG"

"ZHIZICHITANG" was written in Treatise on Febrile Diseases by Zhongjing Zhang at Han dynasty, and it consisted of classical herb pairs of traditional Chinese medicinal compound, Gardenia jasminoides Ellis and Semen Sojae Praeparatum, which was used to regulate and improve sub-health status in clinical, such as anxiety, depression, insomnia, neurasthenia and so on.The complexity of constituents of Traditional Chinese Medicine (TCM) brought great challenges to its modern study, especially the compound of TCM. Although the study of chemical constituents and pharmacological activities of Gardenia jasminoides Ellis and Semen Sojae Praeparatum provided theorial basis for the treatment to sub-health status by "ZHIZICHITANG", less modern studies of compatibility and effective fractions of "ZHIZICHITANG" had been reported.Firstly, we studied the compatibility of "ZHIZICHITANG" and the effective fractions of Gardenia jasminoides Ellis and Semen Sojae Praeparatum. The solution rate of iridoid glycosides and isoflavones at different compatibility was measured by UV spectrophotometry, and that determined the best compatibility of "ZHIZICHITANG"’.Gardenia jasminoides Ellis: Semen Sojae Praeparatum (2:1). The qualitative and quantitative change of major index components were compared between the single decoctions and combined decoctions by HPLC. The results showed that the traditional Chinese medicines could be affected each other when they decocted together, which could affect the pharmacodynamics of TCM by changing the amount of decoction. Then the effective parts of "ZHIZICHITANG" were separated and prepared by D101 macroporous adsorptive resins, iridoid glycosides and isoflavones were obtained successfully through the optimization of separation and purification technology.Gardenia jasminoides Ellis from 17 different areas and Semen Sojae Praeparatum from 15 different areas were studied for building and evaluating fingerprints. The chemometrics methods including similarity evaluation, principal components analysis (PCA) and hierarchical clustering analysis (HCA) were introduced to evaluating Gardenia jasminoides Ellis and Semen Sojae Praeparatum.23 common peaks of Gardenia jasminoides Ellis and 12 common peaks of Semen Sojae Praeparatum were marked. Meanwhile, TCM from different areas were classified into cluters by HCA, and the PCA results showed that peak 5、6、7、12、15、17、19 of Gardenia jasminoides Ellis and peak 6、7、8、9、10、11 of Semen Sojae Praeparatum had larger influence on their quality. Gardenia jasminoides Ellis from Nanning, Guang Xi and Shanghai, Semen Sojae Praeparatum from Hangzhou Zhe Jiang and Chongqing were chosen for the better quality. The fingerprint chromatogram was established from "ZHIZICHITANG" combined by Gardenia jasminoides Ellis from Shanghai and Semen Sojae Praeparatum from Hangzhou, Zhe Jiang. The constituents affect the quality most have been qualitative analyzed by HPLC-ESI-MS. The ESI-MS fragmentation patterns of iridoid glycosides, organic acids and isoflavones were summarized and applied in rapid qualitative identification of active components from "ZHIZICHITANG".23 active components were identified(Deacetyl-asperulosidic acid, Gardoside, Shanzhiside, Geniposidic acid, Deacetyl-asperalosidic acid methyl este, Gardenoside, Scandoside methyl ester, Genipin-1-β-D-gentiobioside,5-O-caffeoylquinic acid, Geniposide, Daidzin, Glycitin, Quercetin-3-rutinoside, Genistin,3,4-Di-O-caffeoylquinic acid,3,5-Di-O-caffeoylquinic acid,6 "-O-trans-feruloylgenipin gentiobioside,4,5-Di-O-caffeoylquinic acid, crocin-, 3,4-Dicaffeoyl-5-(3-hydroxy-3-methylglutaroyl) quinic acid, 4-Sinapoyl-5-caffeoylquinic acid,6"-O-trans-ci nnamoylgenipn gentiobiosid, Daidzein, Glycitein, Genistein) and 17 active components were determined.