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

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

Research output: Chapter in Book/Report/Conference proceedingChapter

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
Title of host publicationJournal of Chromatography - Biomedical Applications
Pages117-125
Number of pages9
VolumeVol. 382
Publication statusPublished - 1986

Fingerprint

Mixed Function Oxygenases
Dopamine
Rats
Tissue
Plasmas
Liquids
Norepinephrine
High performance liquid chromatography
Assays
High Pressure Liquid Chromatography
Plasma (human)
Scintillation Counting
Scintillation counters
Reverse-Phase Chromatography
Enzymes
Human Activities
Catecholamines
Detectors
Substrates

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Medicine
  • Analytical Chemistry

Cite this

Rácz, K., Kuchel, O., Debinski, W., & Buu, N. T. (1986). Improved liquid chromatographic determination of dopamine-β-hydroxylase activity in tissues and plasma. In Journal of Chromatography - Biomedical Applications (Vol. Vol. 382, pp. 117-125)

Improved liquid chromatographic determination of dopamine-β-hydroxylase activity in tissues and plasma. / Rácz, K.; Kuchel, O.; Debinski, W.; Buu, N. T.

Journal of Chromatography - Biomedical Applications. Vol. Vol. 382 1986. p. 117-125.

Research output: Chapter in Book/Report/Conference proceedingChapter

Rácz, K, Kuchel, O, Debinski, W & Buu, NT 1986, Improved liquid chromatographic determination of dopamine-β-hydroxylase activity in tissues and plasma. in Journal of Chromatography - Biomedical Applications. vol. Vol. 382, pp. 117-125.
Rácz K, Kuchel O, Debinski W, Buu NT. Improved liquid chromatographic determination of dopamine-β-hydroxylase activity in tissues and plasma. In Journal of Chromatography - Biomedical Applications. Vol. Vol. 382. 1986. p. 117-125
Rácz, K. ; Kuchel, O. ; Debinski, W. ; Buu, N. T. / Improved liquid chromatographic determination of dopamine-β-hydroxylase activity in tissues and plasma. Journal of Chromatography - Biomedical Applications. Vol. Vol. 382 1986. pp. 117-125
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