Quantitative radioisotopic determination of histidine decarboxylase using high-performance liquid chromatography

Jordi Ortiz, Jordi Gómez, Anna Torrent, Marta Aldavert, Isaac Blanco

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7 Citations (Scopus)

Abstract

We have developed a procedure for the accurate measurement of histidine decarboxylase in tissues expressing low levels of enzymatic activity. Briefly, histamine is enzymatically synthesized from [ 3 H]-labeled histidine, followed by purification using high-performance liquid chromatography (HPLC) and quantitation by liquid scintillation counting. This method presents three advantages over previous techniques. First, prior to HPLC purification, excess precursor [ 3 H]histidine is removed on an anion-exchange resin. Second, purification by HPLC is considerably more selective than that of classical cation-exchange gravity columns or organic solvent extractions. Finally, the accuracy of this method is improved by including non- radiolabeled histamine as internal standard, which is quantified by ultraviolet detection. This simple procedure allows highly sensitive and accurate determinations of histamine synthesis. (C) 2000 Academic Press.
Original languageEnglish
Pages (from-to)111-117
JournalAnalytical Biochemistry
Volume280
DOIs
Publication statusPublished - 10 Apr 2000

Keywords

  • High-pressure liquid chromatography
  • Histamine
  • Histidine
  • Histidine decarboxylase

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