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.
|Publication status||Published - 10 Apr 2000|
- High-pressure liquid chromatography
- Histidine decarboxylase
Ortiz, J., Gómez, J., Torrent, A., Aldavert, M., & Blanco, I. (2000). Quantitative radioisotopic determination of histidine decarboxylase using high-performance liquid chromatography. Analytical Biochemistry, 280, 111-117. https://doi.org/10.1006/abio.2000.4494