High-performance liquid chromatographic determination of catecholamine metabolites and 5-hydroxyindoleacetic acid in human urine using a mixed-mode column and an eight-channel electrode electrochemical detector

Fumiko Mashige, Akiyuki Ohkubo, Yoshikazu Matsushima, Maiko Takano, Etsuko Tsuchiya, Hideko Kanazawa, Yoshiko Nagata, Nobuharu Takai, Noriko Shinozuka, Ichiro Sakuma

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An HPLC system for the simultaneous determination of acidic catecholamine metabolites, related compounds and 5-hydroxyindoleacetic acid (5-HIAA) in human urine was developed. A mixed-mode (C18/anion-exchange) column with isocratic elution using citrate buffer and an eight-channel electrochemical detector were used. Vanilmandelic acid (VMA), 3,4-dihydroxyphenylacetic acid (DOPAC), 4-hydroxy-3-methoxyphenyllactic acid (vanillactic acid, VLA), homovanillic acid (HVA), vanillic acid (VA) and 5-HIAA in urine were determined simultaneously. Detection limits and inter (n = 5) and intra-assay (n = 5) coefficients of variation were satisfactory. The mean of analytical recoveries (n = 3, ±C.V. (%)) were between 97 ± 3.2 (VMA) and 105 ± 4.8 (VA). Correlations between the analytical results for VMA, HVA and 5-HIAA obtained by an established method and the present method were satisfactory. The mean ±2 S.D. of the excretion rates of VMA, DOPAC, VLA, HVA, 5-HIAA and VA in urine from healthy adult volunteers were 0.61-4.36, 0.13-1.02, 0-0.35, 0.67-6.55, 0.50-5.14 and 0-0.55 mg/g creatinine, respectively.

Original languageEnglish
Pages (from-to)63-68
Number of pages6
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Issue number1
Publication statusPublished - 1994 Aug 5


ASJC Scopus subject areas

  • Chemistry(all)

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