An efficient LC–MS/MS method for simultaneous determination of four substances in the Hcy metabolism pathway: supplementary materials

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.