Improved liquid chromatographic determination of dopamine-β-hydroxylase activity in tissues and plasma

K. Racz, O. Kuchel, W. Debinski, N. T. Buu

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

We describe a new assay for dopamine-β-hydroxylase (DβH) activity in human and rat plasma and rat tissues using reversed-phase high-performance liquid chromatography with electrochemical detection. Human and rat plasma DβH activity was measured directly, without extraction of the enzyme. The DβH from rat tissues was extracted on Concanavalin A-Sepharose before the assay to avoid interference from the presence of tissue catecholamines. Dopamine, the natural substrate of DβH, was utilized at optimal (enzyme-saturating) concentration. The reation product, norepinephrine, was isolated on Dowex AG 50W-X4 (H+ from) column. An internal standard, [3H]norepinephrine, was included to correct for the loss of norepinephrine during the procedure. This method allows for the first time the determination of DβH activity in small volumes of rat and human plasma (5-20 μl) and tissues. The procedure can be easily set up in any laboratory equipped with a high-performance liquid chromatography, an electrochemical detector, and a liquid scintillation counter.

Original languageEnglish
Pages (from-to)117-125
Number of pages9
JournalJournal of Chromatography - Biomedical Applications
VolumeVol. 382
Publication statusPublished - Jan 1 1986

ASJC Scopus subject areas

  • Chemistry(all)

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