An efficient LC-MS/MS method for simultaneous determination of four substances in the Hcy metabolism pathway.

Aim: This study aimed to develop and validate an efficient LC-MS/MS method for the simultaneous determination of Hcy, Cys, Met and 5-methyltetrahydrofolate in human serum and to apply this method to patients with coronary artery disease. Methodology and results: Serum samples were prepared by reduction with dithiothreitol followed by protein precipitation. The analytical run time was 2.2 min. The linearity was good in the range of 2-100 μmol/l-1 for Hcy and Met, 10-500 μmol/l-1 for Cys, and 1-50 ng/ml-1 for 5-methyltetrahydrofolate. Conclusion: An accurate and precise method that was rapid, robust and with high-throughput for the routine clinical monitoring of patients with coronary artery disease was developed.