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A rapid assay method for catechol‐ O ‐methyltransferase activity by flow injection analysis
Author(s) -
Aoyama Nozomi,
Tsunoda Makoto,
Nakagomi Kazuya,
Imai Kazuhiro
Publication year - 2002
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.156
Subject(s) - chemistry , normetanephrine , catechol o methyl transferase , chromatography , catechol , metabolite , flow injection analysis , substrate (aquarium) , norepinephrine , fluorescence , ethylenediamine , quantitative analysis (chemistry) , detection limit , biochemistry , organic chemistry , endocrinology , dopamine , medicine , urine , allele , oceanography , physics , quantum mechanics , gene , geology
A rapid assay employing flow injection analysis (FIA) to determine the activity of purified catechol‐ O ‐methyltransferase (COMT) from porcine liver is described. The method was based on the determination of normetanephrine, the 3‐ O ‐methyl metabolite of the substrate norepinephrine. Excess norepinephrine was removed from the incubation mixture by alumina extraction twice to allow normetanephrine to be subjected to flow injection analysis, coulometrical oxidation, fluorogenic reaction with ethylenediamine and fluorescence detection. K m and V max values for COMT obtained with the system were 503 µ M and 4.51 nmol/min/mg protein, respectively. The method is suitable for screening of COMT inhibitors or activators, as a large number of samples, up to 200, can be processed in one working day. Copyright © 2002 John Wiley & Sons, Ltd.