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A quick, convenient and economical method for the reliable determination of methylglyoxal in millimolar concentrations: the N-acetyl-l-cysteine assay

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Abstract

The determination of methylglyoxal (MG) concentrations in vivo is gaining increasing importance as high levels of MG are linked to various health impairments including complications of diabetes. In order to standardize the measurements of MG in body fluids, it is necessary to precisely determine the concentration of MG stock solutions used as analytical standards. The “gold standard” method for the determination of MG concentration in the millimolar range is an enzyme-catalyzed endpoint assay based on the glyoxalase I catalyzed formation of S-lactoylglutathione. However, as this assay used purified glyoxalase I enzyme, it is quite expensive. Another method uses a derivation reaction with 2,4-dinitrophenylhydrazine, but this substance is explosive and needs special handling and storage. In addition, precipitation of the product methylglyoxal-bis-2,4-dinitrophenylhydrozone during the reaction limits the reliability of this method. In this study, we have evaluated a new method of MG determination based on the previously published fast reaction between MG and N-acetyl-l-cysteine at room temperature which yields an easily detectable condensation product, N-α-acetyl-S-(1-hydroxy-2-oxo-prop-1-yl)cysteine. When comparing these three different assays for the measurement of MG concentrations, we find that the N-acetyl-l-cysteine assay is the most favorable, providing an economical and robust assay without the need for the use of hazardous or expensive reagents.

Comparison of the three different spectrophotometrical methods of MG determination

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Abbreviations

2,4-DNPH:

2,4-Dinitrophenylhydrazine

MG:

Methylglyoxal

ε :

Molar absorption coefficient

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Acknowledgments

We gratefully acknowledge the financial support of the NHMRC (grant IDs: 436797, 606543, 1046227). We also thank Dr. Michael Muller for expert advice.

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Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, University of Western Sydney, Locked Bag 1797, Penrith South DC, 1797, Australia

    Rebekka Wild, Lezanne Ooi & Gerald Münch

  2. Department of Medicine, Monash University, Wellington Road, Clayon, VIC, 3168, Australia

    Velandai Srikanth

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  1. Rebekka Wild
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  2. Lezanne Ooi
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  3. Velandai Srikanth
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  4. Gerald Münch
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Correspondence to Gerald Münch.

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Wild, R., Ooi, L., Srikanth, V. et al. A quick, convenient and economical method for the reliable determination of methylglyoxal in millimolar concentrations: the N-acetyl-l-cysteine assay. Anal Bioanal Chem 403, 2577–2581 (2012). https://doi.org/10.1007/s00216-012-6086-4

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  • DOI: https://doi.org/10.1007/s00216-012-6086-4

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