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Simultaneous determination of 3,4-dihydroxymandelic acid, 4-hydroxy-3-methoxymandelic acid, 3,4-dihydroxyphenylglycol, 4-hydroxy-3-methoxyphenylglycol, and their precursors, in human urine by HPLC with electrochemical detection

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Summary

A method has been developed for the quantification of urinary 3,4-dihydroxymandelic acid (DOMA), 4-hydroxy-3-methoxymandelic acid (VMA), 3,4-dihydroxyphenylglycol (DHPG), and 4-hydroxy-3-methoxyphenyglycol (MHPG). Separation and determination of these compounds in biological samples was previously thought to be very difficult. In this work the separation has been achieved by reversed-phase high-performance liquid chromatography with step-wise gradient elution with three mobile phases. The conditions for coulometric detection have been optimized for effective determination of these compounds. In analysis of a sample of human urine, after a simple deproteinization proceudre, DOMA, VMA, DHPG, and MHPG were separated from interferences and quantified successfully; the average levels of these compounds in six different samples were 33.87±1.03, 1202±41.3, 31.3±1.92, and 80.6±2.15 μg (24 h)−1, respectively. Their precursors E, MN, DOPA, DA, NE, DOPAC, HVA, 3MT, and NMN, and the indolamine 5HT and its metabolite 5HIAA (a list of abbreviations is given at the end of the paper) can also be determined simultaneously in the same chromatographic run. The overlapping peak of DHPG was resolved by deconvolution.

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Abbreviations

DA:

3-Hydroxytyramine

DHCA:

3,4-dihydroxyhydrocinnamic acid

DHPG:

3,4-Dihydroxyphenylglycol

DOMA:

3,4-Dihydroxymandelic acid

DOPA:

3,4-Dihydroxyphenylalanine

DOPAC:

3,4-Dihydroxyphenylacetic acid

E:

Epinephrine

5HIAA:

5-Hydroxyindole-3-acetic acid

5HT:

5-Hydroxytryptamine

HVA:

4-Hydroxy-3-methoxyphenylacetic acid

2-Me-DOPA:

2-methyl-3-(3,4-dihydroxyphenyl)-alanine

MHPG:

4-Hydroxy-3-methoxyphenylglycol

3MT:

3-Methoxytyramine

MN:

Metanephrine

NE:

Norepinephrine

N-Me-DA:

N-Methyldopamine

NMN:

Normetanephrine

6-OH-DOPA:

6-Hydroxydopa

VA:

Vanillic acid

VMA:

4-Hydroxy-3-methoxymandelic acid

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Authors and Affiliations

  1. Pharmaceutical Institute, School of Medicine, Keio University, Shinano-Machi 35, Shinjuku-ku, 160, Tokyo, Japan

    G.-M. Cao & T. Hoshino

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  1. G.-M. Cao
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  2. T. Hoshino
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Cao, GM., Hoshino, T. Simultaneous determination of 3,4-dihydroxymandelic acid, 4-hydroxy-3-methoxymandelic acid, 3,4-dihydroxyphenylglycol, 4-hydroxy-3-methoxyphenylglycol, and their precursors, in human urine by HPLC with electrochemical detection. Chromatographia 47, 396–400 (1998). https://doi.org/10.1007/BF02466470

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