Simultaneous Determination of Methionine and Homocysteine by on-column derivatization with o-phtaldialdehyde.

Autor: Borowczyk K; University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland. Electronic address: kamilaborowczyk@gmail.com., Chwatko G; University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland., Kubalczyk P; University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland., Jakubowski H; Department of Biochemistry and Biotechnology, University of Life Sciences, 11 Dojazd Str., 60-632 Poznań, Poland; Institute of Bioorganic Chemistry, 12/14 Noskowskiego Str., 61-704 Poznań, Poland; Department of Microbiology, Biochemistry & Molecular Genetics, Rutgers University, New Jersey Medical School, International Center for Public Health, Newark, NJ 07103-3535, USA., Kubalska J; Department of Metabolic Diseases, Endocrinology and Diabetology, Children's Memorial Health Institute, Warszawa, Poland., Głowacki R; University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland.
Jazyk: angličtina
Zdroj: Talanta [Talanta] 2016 Dec 01; Vol. 161, pp. 917-924. Date of Electronic Publication: 2016 Sep 15.
DOI: 10.1016/j.talanta.2016.09.039
Abstrakt: A fast and simple HPLC-based assay has been developed for the simultaneous determination of homocysteine (Hcy) and methionine (Met) in plasma and urine samples, utilizing as small volume of sample as 10μL. The assay uses on-column derivatization with o-phthaldialdehyde. The separation of Hcy and Met was achieved in 14min on a reversed phase C-18 column, followed by fluorescence detection (excitation at 348nm and emission at 438nm for Met; excitation at 370nm and emission at 480nm for Hcy). Linearity of the detector response was observed in the range of 2-60 μmol L -1 for Met and 2-40 μmol L -1 for Hcy. The method was successfully applied for Met and Hcy quantification in human and mouse plasma and urine samples from cystathionine β-synthase deficient and unaffected individuals.
(Copyright © 2016 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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