Separation and detection of bile acid 3-glucuronides in human urine by liquid chromatography/electrospray ionization-mass spectrometry

Biomedical Chromatography : BMC
S IkegawaJ Goto

Abstract

A method for the separation and detection of bile acid 3-glucuronides by liquid chromatography (LC)/electrospray ionization (ESI)-mass spectrometry (MS) has been developed. On the ESI mode, glucuronides were characterized by an intense pseudo-molecular ion [M-h]- with a doubly charged ion [M-2H]2-, and the ratio of these negative ions were markedly influenced by an acidic component of salt added to a mobile phase, according to a pKa value of an acidic moiety at C-24. Bile acid 3-glucuronides in human urine were extracted with a SepPak C18 cartridge, followed by purification on lipophylic ion exchange gel, piperidinohydroxypropyl Sephadex LH-20. Subsequent resolution into unconjugated, glycine and taurine conjugated bile acid 3-glucuronides was attained by LC on a Develosil ODS-HG-5 column using 20 mM ammonium acetate (pH 7.0)-methanol (10:12, v/v) as a mobile phase, where 3-glucuronides were monitored with a characteristic negative ion [M-H]-. The newly developed method was applied to the quantitatiation of bile acid 3-glucuronides in human urine with satisfactory sensitivity and reliability.

References

Jun 15, 1978·Clinica Chimica Acta; International Journal of Clinical Chemistry·B AlméJ Sjövall
Jul 1, 1978·Clinica Chimica Acta; International Journal of Clinical Chemistry·J GotoT Nambara
Oct 1, 1982·Journal of Biochemistry·H TakikawaT Yamakawa

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Citations

Feb 22, 2002·Steroids·Nariyasu ManoJunichi Goto
May 20, 1998·Steroids·J GotoS Ikegawa
Dec 17, 2009·Journal of Lipid Research·William J Griffiths, Jan Sjövall
Feb 1, 2003·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Nariyasu Mano, Junichi Goto
Sep 20, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yazen AlnoutiCurtis D Klaassen
Jul 28, 2006·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Mélanie VerreaultOlivier Barbier
Oct 24, 2006·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Jocelyn TrottierOlivier Barbier
May 28, 2019·Analytical and Bioanalytical Chemistry·Mainak DuttaAndrew D Patterson
Dec 1, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·A RodaM Baraldini
Oct 26, 2005·Journal of Pharmaceutical and Biomedical Analysis·Masayuki AndoJunichi Goto
Sep 5, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Takaaki GotoJunichi Goto
Oct 13, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Max SchererGerhard Liebisch

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