Determination of glyoxal and methylglyoxal in serum by UHPLC coupled with fluorescence detection.
Anal Biochem
; 573: 51-66, 2019 05 15.
Article
em En
| MEDLINE
| ID: mdl-30796906
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⯱â¯27 and 192⯱â¯10â¯nmol/L, respectively. In human serum, GO and MGO levels in non-diabetic subjects (nâ¯=â¯14) were determined to be 154⯱â¯88 and 98⯱â¯27â¯nmol/L, and in serum samples from subjects with diabetes (nâ¯=â¯14) 244⯱â¯137 and 190⯱â¯68â¯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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Aldeído Pirúvico
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Cromatografia Líquida de Alta Pressão
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Corantes Fluorescentes
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Glioxal
Tipo de estudo:
Diagnostic_studies
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Guideline
Limite:
Aged
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Aged80
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Animals
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Female
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Humans
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Male
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Middle aged
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article