Analysis of vitamin K 1 and major K 2 variants in rat/human serum and lipoprotein fractions by a rapid, simple, and sensitive UHPLC-MS/MS method.

Autor: Mrštná K; The Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203/8, 50005 Hradec Králové, Czech Republic; The Department of Clinical Biochemistry and Diagnostics, University Hospital Hradec Králové, Sokolská 581, 50005 Hradec Králové, Czech Republic., Matoušová K; The Department of Clinical Biochemistry and Diagnostics, University Hospital Hradec Králové, Sokolská 581, 50005 Hradec Králové, Czech Republic., Krčmová LK; The Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203/8, 50005 Hradec Králové, Czech Republic; The Department of Clinical Biochemistry and Diagnostics, University Hospital Hradec Králové, Sokolská 581, 50005 Hradec Králové, Czech Republic. Electronic address: krcml1aa@faf.cuni.cz., Carazo A; The Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203/8, 50005 Hradec Králové, Czech Republic., Pourová J; The Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203/8, 50005 Hradec Králové, Czech Republic., Mladěnka P; The Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203/8, 50005 Hradec Králové, Czech Republic., Matysová L; The Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203/8, 50005 Hradec Králové, Czech Republic., Švec F; The Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203/8, 50005 Hradec Králové, Czech Republic.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2024 Jan 11; Vol. 1714, pp. 464548. Date of Electronic Publication: 2023 Nov 29.
DOI: 10.1016/j.chroma.2023.464548
Abstrakt: Determination of the various forms of vitamin K, which are involved in coagulation and other physiological processes in humans, is challenging and no standardized method is yet available. Therefore, a reliable and practical method was developed to quantify vitamin K levels in serum and additionally in lipoprotein fractions to clarify its distribution. The LC-MS/MS method for the determination of vitamin K 1 and the three main isoforms of vitamin K 2 (MK-4, MK-7, MK-9) was combined with a gradient ultracentrifugation technique to allow the separation of lipoprotein fractions. The chromatographic separation was carried out on a Kinetex™ C18 column using a mobile phase consisting mainly of methanol. The target analytes were detected by electrospray ionization mass spectrometry. The separation of all four substances was achieved after a simple sample preparation technique based on miniaturized liquid-liquid extraction. Our method of only 8.5 min revealed the levels of the major forms of vitamin K in 59 human and 12 rat sera and confirmed our hypothesis that vitamin K is primarily (about 50 %) found in the high-density lipoprotein fraction. The median concentrations of vitamin K 1 , MK-4, MK-7, and MK-9 were found to be 1.19, 2.98, 0.43, and < 0.71 nmol/L in human serum and 1.74, 6.75, less than 0.2, and less than 0.5 nmol/L in rat serum, respectively.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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