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

journal contribution
posted on 2023-05-20, 04:00 authored by Dhananjayan, K, Irrgang, F, Raju, R, Harman, DG, Moran, C, Srikanth, V, Munch, G
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.

History

Publication title

Analytical Biochemistry

Volume

573

Pagination

51-66

ISSN

0003-2697

Department/School

Menzies Institute for Medical Research

Publisher

Academic Press Inc Elsevier Science

Place of publication

525 B St, Ste 1900, San Diego, USA, Ca, 92101-4495

Rights statement

Copyright 2019 Published by Elsevier Inc.

Repository Status

  • Restricted

Socio-economic Objectives

Clinical health not elsewhere classified

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