Ultra performance liquid chromatography-mass spectrometry profiling of bile acid metabolites in biofluids: application to experimental toxicology studies.

TitleUltra performance liquid chromatography-mass spectrometry profiling of bile acid metabolites in biofluids: application to experimental toxicology studies.
Publication TypeJournal Article
Year of Publication2010
AuthorsWant EJ, Coen M, Masson P, Keun HC, Pearce JTM, Reily MD, Robertson DG, Rohde CM, Holmes E, Lindon JC, Plumb RS, Nicholson JK
JournalAnal Chem
Volume82
Issue12
Pagination5282-9
Date Published2010 Jun 15
ISSN1520-6882
KeywordsAnimals, Bile Acids and Salts, Chromatography, High Pressure Liquid, Galactosamine, Glycine, Linear Models, Liver, Male, Mass Spectrometry, Rats, Rats, Sprague-Dawley, Reproducibility of Results
Abstract

We have developed an ultra performance liquid chromatography-mass spectrometry (UPLC-MS(E)) method to measure bile acids (BAs) reproducibly and reliably in biological fluids and have applied this approach for indications of hepatic damage in experimental toxicity studies. BAs were extracted from serum using methanol, and an Acquity HSS column coupled to a Q-ToF mass spectrometer was used to separate and identify 25 individual BAs within 5 min. Employing a gradient elution of water and acetonitrile over 21 min enabled the detection of a wide range of endogenous metabolites, including the BAs. The utilization of MS(E) allowed for characteristic fragmentation information to be obtained in a single analytical run, easily distinguishing glycine and taurine BA conjugates. The proportions of these conjugates were altered markedly in an experimental toxic state induced by galactosamine exposure in rats. Principally, taurine-conjugated BAs were greatly elevated ( approximately 50-fold from control levels), and were highly correlated to liver damage severity as assessed by histopathological scoring (r = 0.83), indicating their potential as a sensitive measure of hepatic damage. The UPLC-MS approach to BA analysis offers a sensitive and reproducible tool that will be of great value in exploring both markers and mechanisms of hepatotoxicity and can readily be extended to clinical studies of liver damage.

DOI10.1021/ac1007078
Alternate JournalAnal. Chem.
PubMed ID20469835
Grant List / / Medical Research Council / United Kingdom