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.
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.
Mashige
F
F
Department of Laboratory Medicine, Faculty of Medicine, University of Tokyo, Japan.
Ohkubo
A
A
Matsushima
Y
Y
Takano
M
M
Tsuchiya
E
E
Kanazawa
H
H
Nagata
Y
Y
Takai
N
N
Shinozuka
N
N
Sakuma
I
I
D015102:3,4-Dihydroxyphenylacetic Acid / Q000652:urine; D000328:Adult; D002395:Catecholamines / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D004563:Electrochemistry; D006719:Homovanillic Acid / Q000031:analogs & derivatives / Q000652:urine; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000652:urine*; D012016:Reference Values; D012680:Sensitivity and Specificity; D014641:Vanillic Acid / Q000652:urine; D014642:Vanilmandelic Acid / Q000652:urine
D002395:Catecholamines; D015102:3,4-Dihydroxyphenylacetic Acid; C003753:vanillactic acid; D006897:Hydroxyindoleacetic Acid; D014642:Vanilmandelic Acid; D014641:Vanillic Acid; D006719:Homovanillic Acid