Specific ion-exchange chromatography and fluorimetric assay for urinary 3-O-methyldopamine

Journal of Chromatography
Y Dalmaz, L Peyrin

Abstract

A technique for the selective extraction of 3-O-methyldopamine, normetanephrine and metanephrine from a single urine sample has been investigated. After hydrolysis of the conjugates, the diluted mixture is passed through a Dowex 50W-X2 column and the methoxylated amines are eluted by means of concentrated ammonia. The eluate, containing metanephrine, normetanephrine and 3-O-methyldopamine is evaporated, and a solution of the residue in borate buffer is fractionated under strictly controlled conditions on an Amberlite CG-50 column. The three amines so separated are estimated by specific fluorimetric methods. The extraction recovery is 80 +/- 3% for pure solutions and 78 +/- 4% for 3-O-methyldopamine added to urine. The fluorimetric procedure, carried out under well-defined conditions, allows the estimation of 10 ng of 3-O-methethyldopamine. The spectral characteristics of the fluorescent derivative are similar to those obtained with dopamine, so that it can be assumed that iodine oxidation of 3-O-methyldopamine demethylates this compound and oxidises the resulting dopamine to the dopamine fluorophore (5,6-dihydroxy-indole). Of the compounds that might interfere in the fluorimetric procedure, dopamine, DOPA and alpha-methyl-DOPA ...Continue Reading

References

Citations

Jan 1, 1980·Journal of Neural Transmission·J M Cottet-EmardJ Bonnod
Jan 1, 1979·Journal of Neural Transmission·Y DalmazJ Dutruge
Nov 1, 1981·Archives internationales de physiologie et de biochimie·L PeyrinM Goffart
May 1, 1978·Archives internationales de physiologie et de biochimie·Y Dalmaz, L Peyrin
Dec 1, 1978·Archives internationales de physiologie et de biochimie·J M PequignotR Flandrois

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