Premium
The effect of temperature on the analysis of metanephrine for catechol‐O‐methyltransferase activity assay by HPLC with electrochemical detection
Author(s) -
Passarinha L. A.,
Bonifácio M. J.,
Queiroz J. A.
Publication year - 2006
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.623
Subject(s) - chemistry , catechol o methyl transferase , catechol , metanephrine , chromatography , amperometry , epinephrine , catecholamine , high performance liquid chromatography , tyramine , substrate (aquarium) , biochemistry , electrochemistry , endocrinology , genotype , urine , electrode , medicine , oceanography , geology , gene
The enzyme catechol‐ O‐ methyltransferase (COMT) plays an important role in the metabolism of catechol estrogens and degradation of the catecholamine neurotransmitters, such as epinephrine. Several analytical methods, mainly high‐performance liquid chromatography with electrochemical amperometric detection, have been reported for the analysis of catecholamines and their metabolites in biological fluids. In this paper we report the relevance of controlling temperature in calibration procedures of metanephrine, an O ‐methylated product of catechol‐ O‐ methyltransferase, using epinephrine as substrate. The results at higher temperatures show shorter retention times of metanephrine, no undue band‐broadening and increased electro signals. This study also showed that, despite different temperatures leading to similarly specific activities of recombinant human COMT as expected, there are additional advantages in flow analytical methods where good sensitivity, efficiency and selectivity is required, mainly in tissues with low levels of COMT activity. Copyright © 2006 John Wiley & Sons, Ltd.