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Clinical Chemistry 22: 1955-1961, 1976;
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Clinical Chemistry, Vol 22, 1955-1961, Copyright © 1976 by American Association for Clinical Chemistry

Diazotization of catecholamines and their analogs and metabolites for urinary screening tests: chemical aspects

G Vauquelin, R Verheyden, G Ebinger and AD Strosberg

Coupling of diazotized p-nitroaniline to catecholamines and their metabolites in urine has been proposed for use in screening for secreting neuroblastoma in childhood. We have coupled diazotized p- nitroaniline to catecholamines, derivatives, and metabolites and examined the reaction products by thin-layer chromatography and physico- chemical methods (ultraviolet spectra, mass spectroscopy, nuclear magnetic resonance). We conclude that during diazotization, products containing a p-hydroxybenzyl alcohol or a p-hydroxybenzoic acid structure (e.g., vanillic acid, vanilmandelic acid, 3-methoxy-4- hydroxyphenyl-ethyleneglycol, metanephrine, normetanephrine, synephrine, and isoproterenol) react with a diazonium cation, with release of their alcohol or acid moiety. Therefore the mentioned qualitative screening methods are very nonspecific. In contrast, thin- layer chromatographic screening methods provide complete separation and unambiguous identification of those metabolites and are to be preferred for use in detecting secreting neuroblastoma in childhood.




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Copyright © 1976 by the American Association for Clinical Chemistry.