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Simultaneous liquid chromatographic determination of vanillylmandelic acid, homovanillic acid, and 5‐hydroxy‐3‐indoleacetic acid in urine, using isocratic elution and electrochemical detection
Author(s) -
Richards D. A.,
Titheradge A. C.
Publication year - 1987
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.1130020307
Subject(s) - vanillylmandelic acid , chemistry , chromatography , homovanillic acid , elution , urine , metabolite , extraction (chemistry) , residue (chemistry) , glassy carbon , electrochemistry , electrode , biochemistry , cyclic voltammetry , receptor , serotonin
A method for the simultaneous measurement of vanillylmandelic acid, homovanillic acid and 5‐hydroxyindoleacetic acid in urine is described. Based on reversed‐phase liquid chromatography with electrochemical detection, the procedure employs isocratic elution, thus making it suitable for use in the less well‐equipped clinical or research laboratory. A simple extraction of the acids from acidified urine into ethyl acetate, is followed by evaporating to dryness a portion of the organic layer, and redissolving the residue in chromatographic mobile phase. Up to 20 samples can be analysed in a single working day. The method is validated and the results obtained are compared with refereènce methods. The cause of contamination of the glassy carbon surface of the working electrode is investigated, and a simple electrochemical pretreatment is described that overcomes this problem. Finally, the extra clinical information that can be derived from multi‐metabolite assays is considered.