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Separation of Bile Acid 7- and 12-Sulfates by High Performance Liquid Chromatography with Pre-Column Fluorescence Labeling

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Abstract

A method for the separation and characterization of bile acid 7- and 12-sulfates without prior deconjugation by means of high performance liquid chromatography (HPLC) is described. Bile acid 7- and 12-sulfates were derivatized quantitatively into the fluorescent compounds through the hydroxyl group at C-3 by treatment with 1-anthroyl nitrile in the presence of quinuclidine in acetonitrile. Subsequent resolution into each individual sulfate was attaind by HPLC on a Cosmosil 5C18 column using 0.3% potassium phosphate buffer (pH 4.0)-methanol ( 1:3) as a mobile phase. The 3-(l-anthroyl ) derivatives of 7- and 12-sulfates were monitored by fluorescence detection (excitation wavelength 370 nm; emission wavelength 470 nm), the limit of detection being 30 fmol.

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Authors and Affiliations

  1. Pharmaceutical Institute, Tohoku University, Aobayama, Sendai, 980, Japan

    Junichi Goto, Toshiyuki Chikai & Toshio Nambara

Authors
  1. Junichi Goto
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  2. Toshiyuki Chikai
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  3. Toshio Nambara
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Additional information

In this paper following trivial names and abbreviations are used: cholate (CA)=3α,7α, 12α-trihydroxy-5β-cholan-24-oic acid; chenodeoxycholate (CDC)=3α,7α-dihydroxy-5β-cholan-24-oic acid; deoxycholate (DC)=3α, 12α-dihydroxy-5β-cholan-24-oic acid; ursodeoxycholate (UDC)=3α, 7β-di-hydroxy-5β-cholan-24-oic acid; G=glyco; T=tauro; S=sulfate.

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Goto, J., Chikai, T. & Nambara, T. Separation of Bile Acid 7- and 12-Sulfates by High Performance Liquid Chromatography with Pre-Column Fluorescence Labeling. ANAL. SCI. 2, 175–178 (1986). https://doi.org/10.2116/analsci.2.175

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  • DOI: https://doi.org/10.2116/analsci.2.175

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