Metabolic Analysis Of Acute Liver Failure Based On GC-MS And UPLC-MS Technology

AimThe study was to discover the dynamic change of serum metabolites during acute liver failure(ALF),identify potential biomarkers and gain new insight into the pathophysiology of ALF.MethodsGalN were administrated at a dose of 1.3 g/kg body weight to ten Bama experimental miniature pigs.Blood samples(5 ml whole blood)were collected before the administration of GalN(0 h group as the normal control)and 12 h(12 h group),24 h(24 h group),36 h(36 h group),and 48 h(48 h group)after GalN administration.Hepatic injury was quantified by determining liver function detect and histopathological assessment of the liver.After sample preparation,they were analyzed by GC-MS and UPLC-MS respectively.The data were analyzed and exported to SIMCA-P+12.0(Umetrics)for subsequent multivariate analyses.Then the potential biomarkers were selected based on VIP and Splot.ResultsThe mean survival duration of animals was 53.4�3.41 h.Dynamic progression of ALF was demonstrated by histopathological comparison of hepatic tissues obtained at the indicated time points.Three overlap TIC chromatograms from GC-MS show stable retention times,and the main peaks did not exhibit any retention time drift.The QC samples in a PCA scatter plot were tightly clustered,suggesting the UPLC-MS system to be highly reproducible.After data normalisation,OPLS-DA was carried out using the SIMCA-P+12.0 software.The OPLS-DA scatter plot of three-dimensional diagram demonstrated the process of liver failure,showing marked separation among the groups and tight clustering within each group;this suggested that the deteriorating pathogenesis of liver injury may be accompanied by changes in metabolism.Based on VIP and Splot,we identified four group metabolites that have changed dramatically during the process of ALF:amino acids(AA),bile acids,lysophosphatidylcholines(LPC)and phosphatidylcholines(PC).These metabolites were related to the whole pathological process of liver failure and could be used as a potential marker.ConclusionAn integrated global GC-MS and UPLC-MS-based metabolomics approach were applied to evaluate dynamic metabolic changes and identify candidate biomarkers of ALF.We identified four groups of serum metabolites�amino acids,conjugated bile acids,LPCs,and PCs.Amino acids could be good indicator to evaluate severity of ALF;Conjugated bile acids could be good biomarkers of ALF during the early stages;LPCs,and PCs contribute to not only the early diagnosis of ALF but also its severity and prognosis evaluation.