In this research,a metabolomics method of ultra high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) was developed for the detection of bile acid metabolites in serum of type 2 diabetic patients,obtaining metabolic markers and exploring potential preventive and therapeutic targets that may predict diabetes.The samples were separated on a Waters ACQUITY UPLC BEH C18 column(150 mm×2.1 mm,1.7 μm) by gradient elution with 0.05% acetic acid aqueous solution and acetonitrile as mobile phases at a flow rate of 0.40 mL/min.And then this method was used to determine the bile acid metabolites in serum of 30 diabetic patients and 50 normal persons.41 bile acid standard solutions were well isolated within 30 min.There were good linear relationships for 41 bile acids in certain concentration ranges,with their correlation coefficients(r2) of 0.986 7-0.999 7.The limits of detection(LOD) and the limits of quantitation(LOQ) were in the ranges of 0.24-7.81 nmol/L and 0.49-15.63 nmol/L,respectively.Results showed that there was a correlation between the bile acid metabolic spectra of the serum and the two groups.Through the orthogonal projections to latent structures-discriminant analysis(OPLS-DA),there are 12 kinds of bile acids,including glycoursodeoxycholic acid,glycocholic acid,taurochenodeoxycholic acid,cholic acid,ursodeoxycholic acid,glycochenodeoxycholic acid,glycodeoxycholic acid,12 ketolithocholic acid,chenodeoxycholic acid,deoxycholic acid,isolithocholic acid and lithocholic acid carrying a group of important information.Combined with the receiver operating characteristic curve(ROC curve),six bile acid metabolic markers,including taurochenodeoxycholic acid(AUC=0.68),glycochenodeoxycholic acid(AUC=0.66),glycoursodeoxycholic acid(AUC=0.63),cholic acid(AUC=0.63),deoxycholic acid(AUC=0.63) and glycodeoxycholic acid(AUC=0.58) were finally obtained,which have a potential value for the early diagnosis and further research of diabetes.
关键词
超高效液相色谱-串联质谱(UPLC-MS/MS)胆汁酸代谢2型糖尿病血清
Keywords
ultra-high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)bile acid metabolismtype 2 diabetes mellitusserum
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College of Chemistry and Materials Science,Shanghai Normal University
University of Chinese Academy of Sciences
Chengdu Institute of Organic Chemistry,Chinese Academy of Sciences
Chengdu Institute of Biology,Chinese Academy of Sciences
Department of Medical Laboratory,Affiliated Minda Hospital of Hubei University for Nationalities