Determination of plasma homocysteine with a UHPLC-MS/MS method: Application to analyze the correlation between plasma homocysteine and whole blood 5-methyltetrahydrofolate in healthy volunteers

Abstract

An ultra-high performance liquid chromatography tandem mass spectrometry method was developed for determination of homocysteine (HCY) in human plasma. The HCY was derivatized with 2-chloro-1-methylquinolinium tetrafluoroborate and isolated using solid-phase extraction. Derivatization, isolation and detection procedures were optimized. Satisfactory linearity was obtained with determination coefficients (r² ) >0.999. The intra- and inter-day precisions were in the interval of 1.2-5.1% and accuracy was within ±7%. Mean recoveries were close to 100%. The limit of detection and the limit of quantification were 0.46 and 1.38 μmol/L, respectively. The method was then applied to investigate the relationship between plasma HCY and whole blood 5-methyltetrahydrofolate levels in healthy volunteers. The results revealed that the plasma level of HCY was significantly negatively correlated to whole blood 5-methyltetrahydrofolate in volunteers.

Keywords: 5-methyltetrahydrofolate; UHPLC-MS/MS; homocysteine.