This study aimed to obtain information on bile acid profiles in diabetic (db/dh) mice and their wild type (wt) littermates for the understanding of pathogenesis and discovery of potential biomarkers of type 2 diabetes. Analytical methods based on protein precipitation or solid-phase extraction together with liquid chromatography-tandem mass spectrometry were developed for the determination of 25 bile adds in plasma, urine and feces samples collected from db/db and wt mice. GLP-1 concentration and hepatic genes related to bile acid synthesis were also investigated. The results showed that the concentrations of individual bile adds varied notably both interindividually and temporally. However, plasma, urine and feces samples displayed discriminating bile acid profiles between the db/db and wt groups, with the plasma profile showing the best differentiation capacity. In plasma and urine, the concentration variation of taurine-conjugated bile adds was more correlated with that of other taurine-conjugated bile acids, and vice versa for the unconjugated bile acids. Transcription of hepatic gene Cyp7b1 was downregulated, and Hsd3b7 upregulated in db/db mice. In conclusion, the bile add profile, particularly that in plasma, can distinguish the two animal groups and is a promising biomarker for type 2 diabetes. (C) 2016 Elsevier B.V. All rights reserved.