Preliminary Study Of Metabolomics In Rats After Oral Administration Of Si-Wu-Tang Based On UPLC-QTOF MS

Objective:Establish the endogenous metabolites database of SD rats serum and urine to distinguish the endogenous metabolites and exogenous metabolites. The database is the foundation of preliminary study of metabolomics in rats after oral administration of Si-Wu-Tang. Study the serum and urine metabolites in SD rats after oral administration of Si Wu Tang for once and for 7 days. Then investigating the changes in contents of the metabolites to illustrate the function of Si Wu Tang.Method:Using UPLC-QTOF MS to detect the serum and urine of normal SD rats and with the help of in-house software named "Searcher" and in-house localized Excel-format HMDB database to establish the endogenous metabolites database of SD rats serum and urine. Then using UPLC-QTOF MS to investigate the serum and urine metabolites in SD rats after oral administration of Si Wu Tang. The metabolites data were analyzed by principal component analysis (PCA) and orthogonal partial least-squares to latent structures discriminate analysis (OPLS-DA). The constituents identification of selected compounds includes comparing with reference compounds, accurate m/z value analysis of TOF-MS data and QTOF-MS/MS fragment analysis.Result:Establish the endogenous metabolite database of SD rats serum and urine. 950 significant signals were detected in serum by ES+,155 of them were given the possible chemical structures. There are 1280 signals were identified by ES".190 of them can be predicted by the possible chemical structures.15 of the signals detected in serum were verified by multiple-coincided HMDB-matched compounds.2260 significant signals were detected in urine by ES+,1060 of them were given the possible chemical structures.3000 signals were identified by ES",1270 of them can be predicted by the possible chemical structures.92 of the signals detected in serum were verified by multiple-coincided HMDB-matched compounds. There are 821 differences compounds were found in serum after oral administration of Si Wu Tang for once in 7 different times.182 compounds having differences were identified in serum after oral administration of Si Wu Tang for 7 days. The results demonstrate that compounds such as creatine, citric acid and cortexolone occur changes in contents by comparing with reference compounds, accurate m/z value analysis of TOF-MS data and QTOF-MS/MS fragment analysis.136 compounds were detected in urine after oral administration of Si Wu Tang for once and 77 compounds were identified in urine after oral administration of Si Wu Tang for 7 days. Compounds such as 4-heptanone, Sebacic acid, methylladenine,3-hydroxysebacic acid or 2-hydroxydecanedioic acid, 5-hydroxy-6-methoxuindole glucuronide or 6-hydroxy-5-methoxuindole glucuronide, n-acetyl-7-o-acetylneuraminic acid and formylanthranilic acid or noradrenochrome were demonstrated that change in content by QTOF-MS/MS fragment analysis.Conclusion:The research establishes the endogenous metabolite database of SD rats serum and urine. The work provides the preconditions to the study of metabolomics in rats after oral administration of Si-Wu-Tang and gives the chance to distinguish endogenous substances and exogenous substances. Discussing the physiological significance of compounds having changes in content after oral administration of Si Wu Tang and the role Si Wu Tang may play. The results provide references from endogenous metabolites content changing for discussing the pharmacological effects of Si Wu Tang.