Objective: To identify the bile acid components in rats by UHPLC-Q Exactive Orbitrap MS technology, and to study the distribution of bile acids in various tissues during bile acid cycling by utilizing the bile acid database established in this study. Methods: 0.1% formic acid-acetonitrile was used as mobile phase gradient elution at a flow rate of 0.2 mL · min-1. The mass spectrum data of SD rat serum, liver, bile, intestine, intestinal contents, urine, and feces were collected under the negative ion mode of electrospray ion source. The bile acid components were systematically screened by mass defect filtering (MDF), key ion filtering (KIF) and extracting ion chromatography (EIC) techniques. Finally, the bile acid composition of each tissue was comprehensively analyzed. Results: As a result, a total of 449 bile acids were identified or preliminarily identified, including 139 free bile acids, 112 taurine-conjugated bile acids, 44 glycine-conjugated bile acids, 56 sulfated bile acids, 36 glucuronidated bile acids, 3 N-acetylglucosamine-conjugated bile acids, 7 hexose-conjugated bile acids, 19 non-C24 bile acids, 2 acetylated bile acids, and 15 bile acids conjugated with other amino acids. Moreover, 12 novel bile acids conjugated with small organic acids were characterized for the first time. The distribution of bile acids in various tissues showed that taurine-conjugated bile acids were the most abundant in liver, bile, duodenum, jejunum tissues, and their intestinal contents. The content of free bile acids in ileal contents increased and was close to that of tauro-conjugated bile acids. The highest content of free bile acids was found in serum, colonic contents, urine, and feces. Conclusion: This study can provide basic data for further exploring the role of bile acid metabolism in bile acid-related diseases.
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