title: Simultaneous Determination of Methionine and Homocysteine by on-column derivatization with o-phtaldialdehyde.
abstract: 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-1for Met and 2-40 μmol L-1for 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.
authors: University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland. Electronic address: kamilaborowczyk@gmail.com.
authors: Kamila
authors: K
authors: Borowczyk
authors: University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland.
authors: Grażyna
authors: G
authors: Chwatko
authors: University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland.
authors: Paweł
authors: P
authors: Kubalczyk
authors: 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.
authors: Hieronim
authors: H
authors: Jakubowski
authors: Department of Metabolic Diseases, Endocrinology and Diabetology, Children's Memorial Health Institute, Warszawa, Poland.
authors: Jolanta
authors: J
authors: Kubalska
authors: University of Łódź, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland.
authors: Rafał
authors: R
authors: Głowacki
mesh_terms: D000328:Adult; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006710:Homocysteine / Q000097:blood / Q000737:chemistry / Q000652:urine; D006712:Homocystinuria / Q000097:blood* / Q000652:urine*; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008715:Methionine / Q000097:blood / Q000737:chemistry / Q000652:urine; D051379:Mice; D008822:Mice, Transgenic; D009764:o-Phthalaldehyde / Q000737:chemistry*
chemical_list: D006710:Homocysteine; D009764:o-Phthalaldehyde; D008715:Methionine
keywords: Biological fluids; High performance liquid chromatography; Hyperhomocysteinemia; On-column derivatization