Separation of Bile Acid TV-Acetylglucosaminides by High-Performance Liquid Chromatography with Precolumn Fluorescence Labeling

Toshifumi Niwa, Kazuhiro Fujita, Junichi Goto, Toshio Nambara

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

A sensitive and reliable method for the separation and characterization of bile acid N-acetylglucosaminides has been developed by precolumn labeling followed by high-performance liquid chromatography (HPLC) with fluorescence detection. The 3-iV-acetylglucosaminides of unconjugated, glycine- and taurine-conjugated bile acids were derivatized into fluorescent esters through a primary hydroxyl group on the sugar moiety by treatment with 9-anthroyl cyanide in the presence of quinuclidine in acetonitrile. Subsequent separation into each 3-N-acetylglucosaminide was achieved by reversed-phase chromatography on a Cosmosil 5Cm column using a 0.3% potassium phosphate buffer (pH 7.0)/methanol (1:4, v/v) as a mobile phase. The derivatized 3-Ar-acetylglucosaminides were monitored by fluorescence detection (excitation wavelength 362 nm; emission wavelength 470 nm), the limit of detection being lOOfmol. The chromatographic separation of 3- and 7-N-acetylglucosaminides of ursodeoxycholates is also described.

Original languageEnglish
Pages (from-to)659-662
Number of pages4
Journalanalytical sciences
Volume8
Issue number5
DOIs
Publication statusPublished - 1992 Jan 1

Keywords

  • 9-anthroyl cyanide
  • Bile acid
  • N-acetylglucosaminide
  • fluorescence detection
  • high-performance liquid chromatography
  • precolumn labeling

ASJC Scopus subject areas

  • Analytical Chemistry

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