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


Dhananjayan, K and Irrgang, F and Raju, R and Harman, DG and Moran, C and Srikanth, V and Munch, G, Determination of glyoxal and methylglyoxal in serum by UHPLC coupled with fluorescence detection, Analytical Biochemistry, 573 pp. 51-66. ISSN 0003-2697 (2019) [Refereed Article]

Copyright Statement

Copyright 2019 Published by Elsevier Inc.

DOI: doi:10.1016/j.ab.2019.02.014


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 Details

Item Type:Refereed Article
Keywords:blood samples, fluorescence, glyoxal, methylglyoxal, UHPLC, validation
Research Division:Biological Sciences
Research Group:Biochemistry and cell biology
Research Field:Analytical biochemistry
Objective Division:Health
Objective Group:Clinical health
Objective Field:Clinical health not elsewhere classified
UTAS Author:Srikanth, V (Dr Velandai Srikanth)
ID Code:132981
Year Published:2019
Web of Science® Times Cited:12
Deposited By:Menzies Institute for Medical Research
Deposited On:2019-05-29
Last Modified:2020-08-14

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