Investigation on the spectrum-effect relationships of Da-Huang-Fu-Zi-Tang in rats by UHPLC-ESI-Q-TOF

来源 :2014年全国中药炮制学术年会暨中药饮片创新发展论坛及协同创新联盟会议 | 被引量 : 0次 | 上传用户:wmstudio
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  Ethnopharmacological relevance: Da-Huang-Fu-Zi-Tang (DHFZT) is a crucial TCM formula commonly used for the treatment of acute pancreatitis in clinical application.Our previous work found that DHFZT could act against pancreatic injury in rats with severe acut epancreatitis (SAP).Aim of the study: The goal of this paper was to study the underlying correlations between the chemical spectra and the protective effect of DHFZT on pancreatic acinar cell to reveal the real bioactive compounds in DHFZT.Materials and methods: The fingerprint chromatograms of rat serum after oral administration of DHFZT were established by UHPLC-ESI-Q-TOF-MS technique.At the same time, the model of anti-acute pancreatitis on cells was established by adding tree frog to AR42J cell line, and the protective effects of the serum on pancreatic acinar cell from injury was evaluated by detecting the efficacy of amylase.Then, the spectrum-effect relationships between UHPLC fingerprints and anti-acute pancreatitis activities were evaluated using canonical correlation analysia (CCA) statistical method.The chromatogram separation was performed on a C18 reversed phase UHPLC column (2.1 mm × 100 mm, 3.5 μm, Agilent), the column temperature was set at 35C.The mobile phase consisted of 0.1% formic acid and acetonitrile with gradient elution.The serum samples were analysised both in negative and positive ion mode.TOF MS and TOF MS/MS were scanned within the mass range ofm/z 100-1200 and 50-1200,respectively.A thorough analysis of a great deal of information of the constituents in the rat serum was undertaken.The structure identification of the detected compounds was confirmed by using high resolution MS values as well as the MS/MS fragments.Results: 18 peaks in serum after oral administration of DHFZT were detected within only 30 min recorded chromatograms.The structure of the 18 compounds were then given out, of which 11 were the original form of compounds absorbed from DHFZT, 7 were the metabolites of the compounds existed in rat serum.According to the CCA results, talatisamine, rhein glucoside,rhein isomer methylation, hypaconine, hydroxyl-chrysophanol, emodin glucuronide conjugation, and chrysophanol glucuronide conjugation were finally found to be the main anti-acute pancreatitis components in DHFZT.Conclusions: The model presented in this paper successfully discovered the spectrum-effect relationships of DHFZT, which showed a representative way to discover the primary active ingredients from the complicated herbal drugs.
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