Assay for dopamine beta-hydroxylase in human plasma and rat serum by high-performance liquid chromatography with fluorimetric detection.
A highly sensitive assay for dopamine beta-hydroxylase activity in human plasma and rat serum by high-performance liquid chromatography with fluorimetric detection is described. Norepinephrine, formed enzymatically from the substrate dopamine, and epinephrine (internal standard), after clean-up with a cation-exchange cartridge (Toyopak IC-SP M), are converted into the corresponding fluorescent compounds by reaction with 1,2-diphenylethylenediamine. The derivatives are separated by reversed-phase chromatography on TSK gel ODS-80TM. The detection limit for norepinephrine formed enzymatically is 5 pmol per assay tube.
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
M K
MK
Lee
H
H
Nohta
Y
Y
Ohkura
D000328:Adult; D000818:Animals; D002374:Catalase; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004299:Dopamine beta-Hydroxylase / Q000037:antagonists & inhibitors / Q000097:blood*; D004837:Epinephrine / Q000097:blood; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D051381:Rats; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D014439:Tyramine / Q000494:pharmacology
D007202:Indicators and Reagents; D002374:Catalase; D004299:Dopamine beta-Hydroxylase; D014439:Tyramine; D004837:Epinephrine