Pre-Column (HPLC) Fluorescence Labelling of Glucuronides

  • H. Lingeman
  • G. W. M. Meussen
  • C. van der Zouwen
  • W. J. M. Underberg
  • A. Hulshoff
Part of the Methodological Surveys in Biochemistry and Analysis book series (MSBA, volume 16 A)


A selective and sensitive HPLC method has been developed for the determination of glucuronides in plasma and urine. Prior to chromatography the carboxylic acid function of the glucuronic acid moiety is converted into a fluorescent amide by a method selective for the carboxyl group. The acidic function is first activated with 2-bromo-1-methylpyridinium iodide and then reacted with the fluorescence label [N-(1-dim ethyl aminonaphthalenesulphonyl)cadaverine or N-(1-naph-thyl)ethylenediamine] to yield the amide. The derivatization products are separated by a two-column system in combination with a column-switching technique. For α-naphthyl-β-D-glucuronide, β-oestradiol-17-β-D-glucuronide and some other glucuronides, consideration is given to chromatographic properties, intrinsic fluorescence sensitivities (both of reagents and of derivatives) and clean-up procedures.


Glucuronic Acid Dansyl Derivative Carboxylic Acid Function Sensitive HPLC Method Tetramethylammonium Bromide 
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Copyright information

© Springer Science+Business Media New York 1986

Authors and Affiliations

  • H. Lingeman
    • 1
  • G. W. M. Meussen
    • 1
  • C. van der Zouwen
    • 1
  • W. J. M. Underberg
    • 1
  • A. Hulshoff
    • 1
  1. 1.Pharmaceutical Laboratory, Department of Analytical PharmacyState University of UtrechtUtrechtThe Netherlands

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