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-Ar-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-acetyiglucosaminide was achieved by reversed-phase chromatography on a Cosmosil 5C18 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 100 fmol. The chromatographic separation of 3- and 7-N-acetylglucosaminides of ursodeoxycholates is also described.
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In this paper following trivial names and abbreviations are used: cholic acid=3α,7α, 12α-trihydroxy-5β-choIan-24-oic acid; chenodeoxycholic acid=3α,7α-dihydroxy-5β-cholan-24-oic acid; deoxycholic acid=3α,12α-dihydroxy-5β-cholan-24-oic acid; ursodeoxycholic acid=3α,7β-dihydroxy-5β-cholan-24-oic acid; lithocholic acid=3α-hydroxy-5β-cholan-24-oic acid.
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Niwa, T., Fujita, K., Goto, J. et al. Separation of Bile Acid N-Acetylglucosaminides by High-Performance Liquid Chromatography with Precolumn Fluorescence Labeling. ANAL. SCI. 8, 659–662 (1992). https://doi.org/10.2116/analsci.8.659
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DOI: https://doi.org/10.2116/analsci.8.659