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A simplified routine assay for urinary 3-methoxy-4-hydroxyphenylglycol

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Summary

Urinary 3-methoxy-4-hydroxyphenylglycol (MHPG) level may provide us with a biochemical index for central noradrenergic activity. Previous methods for assaying MHPG in urine often lacked specificity, sensitivity, cost-effectiveness or convenience. We now describe a simplified routine assay for urinary MHPG by high-pressure liquid chromatography using electrochemical detection. For convenience and cost-effectiveness within a typical batch assay of total MHPG, 0.5 mL of urine sample, 50 μL of glusulase and 4 μg iso-vanillyl alcohol (internal standard) were used to hydrolyze conjugated MHPG at 37 °C overnight. Alternatively, for a same-day operation, it is efficient to hydrolyze the sample at 50 °C for 3 hours. Each sample was separated isocratically on a reversed-phase column (Ultracarb 5 ODS) at 25 °C with the flow rate at 1 mL/min. Intra- and inter-assay coefficients of variation were found to be 4.0% (n=10) and 5.0% (n=27), respectively, for MHPG at a mean concentration of 1.9 mg/L. Sulfatase or glucuronidase can be substituted for glusulase to obtain either sulfate- or glucuronide-conjugated MHPG. This procedure requires smaller sample amounts and less preparation time without compromising sensitivity and reproducibility.

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Author notes

  1. J. K. Yao

    Present address: Western Psychiatric Institute and Clinic, VA Pittsburgh Healthcare System (Highland Drive), and University of Pittsburgh Medical Center, Pittsburgh, PA, USA

Authors and Affiliations

  1. Western Psychiatric Institute and Clinic, VA Pittsburgh Healthcare System (Highland Drive), and University of Pittsburgh Medical Center, Pittsburgh, PA, USA

    P. Zhu, D. J. Wilds & D. P. van Kammen

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  1. J. K. Yao
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  2. P. Zhu
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  3. D. J. Wilds
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  4. D. P. van Kammen
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Yao, J.K., Zhu, P., Wilds, D.J. et al. A simplified routine assay for urinary 3-methoxy-4-hydroxyphenylglycol. J. Neural Transmission 104, 967–975 (1997). https://doi.org/10.1007/BF01285564

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  • DOI: https://doi.org/10.1007/BF01285564

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