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

• Published in: Journal of chromatography. Biomedical applications 1994-08, Vol.658 (1), p.63-68
• Main Authors: Mashige, Fumiko, Ohkubo, Akiyuki, Matsushima, Yoshikazu, Takano, Maiko, Tsuchiya, Etsuko, Kanazawa, Hideko, Nagata, Yoshiko, Takai, Nobuharu, Shinozuka, Noriko, Sakuma, Ichiro
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Non peptidic neurotransmitters, polyamines; Other biological molecules
• ISSN: 0378-4347
• DOI: 10.1016/0378-4347(94)00227-4

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 (C 18/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.