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Papers In Press, published online ahead of print January 1, 2009
J. Lipid Res., doi:10.1194/jlr.D800041-JLR200

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Journal of Lipid Research, Vol. 50, 173-180, January 2009
Copyright © 2009 by American Society for Biochemistry and Molecular Biology

Methods

Improved analysis of bile acids in tissues and intestinal contents of rats using LC/ESI-MS

Masahito Hagio^*, Megumi Matsumoto, Michihiro Fukushima, Hiroshi Hara^* and Satoshi Ishizuka^1,*

^* Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo, Hokkaido, Japan
Meiji Dairies Research Chair, Creative Research Initiative Sousei, Hokkaido University, Sapporo, Hokkaido, Japan
Department of Agriculture and Life Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of a figure.

Published, JLR Papers in Press, September 4, 2008.

¹ To whom correspondence should be addressed. e-mail: zuka{at}chem.agr.hokudai.ac.jp

To evaluate bile acid (BA) metabolism in detail, we established a method for analyzing BA composition in various tissues and intestinal contents using ultra performance liquid chromatography/electrospray ionization mass spectrometry (UPLC/ESI-MS). Twenty-two individual BAs were determined simultaneously from extracts. We applied this method to define the differences in BA metabolism between two rat strains, WKAH and DA. The amount of total bile acids (TBAs) in the liver was significantly higher in WKAH than in DA rats. In contrast, TBA concentration in jejunal content, cecal content, colorectal content, and feces was higher in DA rats than in WKAH rats. Nearly all BAs in the liver were in the taurine- or glycine-conjugated form in DA rats, and the proportion of conjugated liver BAs was up to 75% in WKAH rats. Similar trends were observed for the conjugation rates in bile. The most abundant secondary BA in cecal content, colorectal content, and feces was hyodeoxycholic acid in WKAH rats and -muricholic acid in DA rats. Analyzing detailed BA profiles, including conjugation status, in a single run is possible using UPLC/ESI-MS. This method will be useful for investigating the roles of BA metabolism under physiological and pathological conditions.

Supplementary key words HPLC • ultra performance liquid chromatography • electrospray ionization mass spectrometry

Abbreviations: BA, bile acid; CA, cholic acid; CDCA, chenodeoxycholic acid; DCA, deoxycholic acid; LC/ESI-MS, liquid chromatography/electrospray ionization mass spectrometry; HDCA, hyodeoxycholic acid; LCA, lithocholic acid; MCA, muricholic acid; NDCA, nordeoxycholic acid; PBA, primary bile acid; SBA, secondary bile acid; SIR, selected ion recording; TBA, total bile acid; UDCA, ursodeoxycholic acid; UPLC, ultra performance liquid chromatography

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