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Determination of glyoxal and methylglyoxal in serum by UHPLC coupled with fluorescence detection

Dhananjayan, K, Irrgang, F, Raju, R, Harman, DG, Moran, C, Srikanth, V and Munch, G 2019 , 'Determination of glyoxal and methylglyoxal in serum by UHPLC coupled with fluorescence detection' , Analytical Biochemistry, vol. 573 , pp. 51-66 , doi: 10.1016/j.ab.2019.02.014.

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Abstract

Glyoxal (GO) and methylglyoxal (MGO) are two important biomarkers in diabetes. Analytical methods for determination of GO and MGO in serum samples are either HPLC with UV-Vis (low sensitivity) or MS/MS (expensive) detection. These disadvantages have hampered the introduction of these biomarkers as a routine analyte for diabetes diagnostics into the clinical laboratory. In this study, we introduce a UHPLC method with fluorescence detection for the measurement of GO and MGO in serum samples by pre-column derivatization at neutral pH with 5, 6-diamino-2,4-dihydroxypyrimidine sulfate (DDP) to form lumazines. The method was validated as per FDA guidelines. Using this method, we have determined GO and MGO in a variety of animal serum samples, and for example, determined the GO and MGO concentration in adult bovine serum to be 852 ± 27 and 192 ± 10 nmol/L, respectively. In human serum, GO and MGO levels in non-diabetic subjects (n = 14) were determined to be 154 ± 88 and 98 ± 27 nmol/L, and in serum samples from subjects with diabetes (n = 14) 244 ± 137 and 190 ± 68 nmol/L, respectively. In addition, interference studies showed that physiological serum components did not lead to an artificial increase in the levels of GO and MGO.

Item Type: Article
Authors/Creators:Dhananjayan, K and Irrgang, F and Raju, R and Harman, DG and Moran, C and Srikanth, V and Munch, G
Keywords: blood samples, fluorescence, glyoxal, methylglyoxal, UHPLC, validation
Journal or Publication Title: Analytical Biochemistry
Publisher: Academic Press Inc Elsevier Science
ISSN: 0003-2697
DOI / ID Number: 10.1016/j.ab.2019.02.014
Copyright Information:

Copyright 2019 Published by Elsevier Inc.

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