Reversed-phase high-performance liquid chromatographic separation and quantification of individual human bile acids

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Abstract

The reversed-phase high-performance liquid chromatographic separation and quantification of individual bile acids is described. Taurine- and glycine-conjugated bile acids were separated and detected directly by an ultraviolet absorbance detector operating at 200 nm. Simultaneous quantitation of at least 100 ng of each conjugated bile acid is possible. Carboxylic (free and glycine-conjugated) bile acids were esterified with p-bromophenacyl-bromide. The reaction, using N,N-diisopropylethylamine as catalyst, yields quantitatively the strongly absorbing p-bromophenacyl esters whch can be determined by absorbance measurement at 254 nm. Simultaneous quantitation of less than 20 ng of each bile acid is possible. The present method is applied to the quantitation of individual bile acids in ten human gallbladder bile samples.
Original languageEnglish
Pages (from-to)277-286
Number of pages10
JournalJOURNAL OF CHROMATOGRAPHY A
Volume183
Publication statusPublished - 1980

Keywords

  • Acetophenones
  • Bile Acids and Salts
  • Chromatography, High Pressure Liquid
  • Esterification
  • Ethylamines
  • Gallbladder
  • Humans
  • Spectrophotometry, Ultraviolet

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