The Tohoku Journal of Experimental Medicine
Online ISSN : 1349-3329
Print ISSN : 0040-8727
ISSN-L : 0040-8727
Simultaneous Determination of Urinary Vanillylmandelic Acid and 5-Hydroxyindoleacetic Acid
佐藤 辰男和田 保男前橋 賢
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1963 年 80 巻 1 号 p. 1-8

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A method for simultaneous quantitative determination of vanillylmandelic acid (VMA) and 5-hydroxyindoleacetic acid (5-HIAA) in urine by means of highvoltage paper electrophoresis was described. The extraction with ethyl acetate, coloration with diazotized p-nitroaniline, elution and spectrophotometric determination was carried out in the same way as used for VMA. VMA was separated at first from other urinary compounds including 5-HIAA by electrophoresis using pH 3.6 buffer, then isolation of 5-HIAA was carried out chromatographically on the same filter paper to the right-angle direction. For this purpose, filter paper which was cut into a crosswise form was employed. This method is highly specific and less time consuming, so that it may be useful in studies on catecholamine and serotonin metabolism. The recovery rate was about 86 per cent for 5-HIAA and 94 per cent for VMA, and the error of measurement was about 5 per cent respectively by this method. Estimation revealed that the excretion of 5-HIAA in human urine per day ranged from 1.9 to 5.1 mg. with an average of 3.04±1.07 mg. in 20 normal subjects, and that the urinary excretion ratio of 5-HIAA/VMA was about 1:1.22.
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