20.500.12556/RUL-138399 Simultaneous quantification of intracellular concentrations of clinically important metabolites of folate-homocysteine cycle by LC-MS/MS Inadequate folate status is detrimental to human development. Deficiency has been implicated in congenital birth defects and cancer, whereas excess has been linked to various negative neurocognitive development outcomes. We developed a method for translational studies involving lymphoblastoid cell models for studying role of folates in vital cell processes. We describe a simple, sensitive, and fast liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of intracellular concentrations of clinically important metabolites of folate-homocysteine cycle; namely, folic acid (FA), 5-methyltetrahydrofolate (5-Me-THF), and homocysteine (Hcy). The method was validated for specificity, linearity, limits of quantification, repeatability, reproducibility, matrix effects, and stability. Method had a wide linear range between 0.341 and 71.053 ng Hcy/mg protein for Hcy, 0.004–0.526 ng FA/mg protein for FA and 0.003–0.526 ng 5-Me-THF/mg protein for 5-Me-THF. The method overcomes challenges associated with the quantification of endogenous molecules, poor stability, and extremely small amounts of the analytes. The method was successfully applied to evaluate the effects of FA and 5-Me-THF treatment of cells in vitro mimicking supplement therapy with various metabolically active species, and showed that 5-Me-THF is more effective than FA in increasing intracellular levels of the biologically active form of folate. LC-MS/MS folic acid 5-methyltetrahydrofolate homocysteine cell lysate folate markers homocistein folna kislina folatni markerji vitamini true false true Angleški jezik Slovenski jezik Članek v reviji 2022-07-19 15:56:13 2022-07-19 15:56:18 2022-09-15 04:16:43 0000-00-00 00:00:00 2020 0 0 12 str. Vol. 605, art. 113830 2020 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.164.1:616.074 0003-2697 10.1016/j.ab.2020.113830 25786371 1-s2.0-S0003269720303626-main.pdf 1-s2.0-S0003269720303626-main.pdf 1 B13D8045D5130A8336282D5CDF0AC7C6 f44bd0729a22abe62e3ca78e20a56c09371a63d064aaeaa74e18d6a75b896941 9aeac935-076a-11ed-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=158705 https://www.sciencedirect.com/science/article/pii/S0003269720303626 1 51682125-076a-11ed-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=158703 Fakulteta za farmacijo Medicinska fakulteta 0 0 0