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Quantitative Radioisotopic Determination of Histidine Decarboxylase Using High-Performance Liquid Chromatography

✍ Scribed by Jordi Ortiz; Jordi Gómez; Anna Torrent; Marta Aldavert; Isaac Blanco

Publisher
Elsevier Science
Year
2000
Tongue
English
Weight
78 KB
Volume
280
Category
Article
ISSN
0003-2697

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✦ Synopsis

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 nonradiolabeled histamine as internal standard, which is quantified by ultraviolet detection. This simple procedure allows highly sensitive and accurate determinations of histamine synthesis.

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