Determination of catechol-O-methyltransferase activity by high-performance liquid chromatography with electrochemical detection.
A rapid and simple assay for catechol-O-methyltransferase (COMT) activity by high-performance liquid chromatography with electrochemical detection is described. The method is based on the measurement of 3-O- and 4-O-methylated products (3-methoxy-4-hydroxy- and 4-methoxy-3-hydroxybenzylamine, respectively) of the substrate 3,4-dihydroxybenzylamine. These compounds are determined from the incubation mixture after removal of protein injecting aliquot into the liquid chromatograph. The detection limit is 1 pmol of product. This method is very suitable for screening of COMT activity as well as for determining the meta/para product ratios.
D000818:Animals; D001921:Brain / Q000201:enzymology; D002394:Catechol O-Methyltransferase / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000031:analogs & derivatives / Q000378:metabolism; D004563:Electrochemistry; D005260:Female; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D051381:Rats
C022133:3,4-dihydroxybenzylamine; D002394:Catechol O-Methyltransferase; D004298:Dopamine