AI Article Synopsis
- Researchers developed a reliable LC-MS/MS method for quantifying various forms of vitamin K, including MK-4, MK-7, and MK-9, in human serum and lipoprotein fractions, addressing challenges in standardization.
- The method utilized gradient ultracentrifugation to separate lipoprotein fractions and involved a fast sample preparation technique, achieving results in just 8.5 minutes.
- Findings showed that approximately 50% of vitamin K is present in the high-density lipoprotein fraction, with specific median concentrations of vitamin K forms identified in both human and rat sera.
Article Abstract
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 and the three main isoforms of vitamin K (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, 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.
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| http://dx.doi.org/10.1016/j.chroma.2023.464548 | DOI Listing |
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