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Unified Quantification of Homocysteine and Folate in Human Serum or PlasmaUsing Liquid Chromatography/Tandem Mass Spectrometry

Published

Author(s)

B C. Nelson, Mary B. Satterfield, Lorna T. Sniegoski, Michael J. Welch

Abstract

Hyperhomocysteinemia is an established, independent risk factor for cardiovascular disease. Folate deficiency is emerging as a potential risk factor for cardiovascular disease. The intake of folate may or may not reduce the risk of cardiovascular events through the capacity of folate to lower circulating homocysteine levels in the body. A unified extraction and quantification procedure based on stable, isotope-dilution liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed for the simultaneous determination of total homocysteine (tHcy) and folate [5-methyltetrahydrofolic acid (5MT) and folic acid (FA)] levels in human serum or plasma. Analytes are quantitatively isolated from samples (500 L) prior to LC/MS/MS analysis using a novel two-step stabilization process combined with C18 solid phase extraction. The method exhibits excellent linearity over four orders of magnitude for each analyte with limits of detection of 0.02 ng/mL, 0.003 mol/L and 0.01 ng/mL for FA, tHcy and 5MT, respectively. The limits of quantification are 0.06 ng/mL, 0.01 mol/L and 0.03 ng/mL for FA, tHcy and 5MT, respectively. The accuracy and practical applicability of the method were proven by applying the method to the quantitative determination of each analyte in a new NIST serum Standard Reference Material (NIST SRM 1955 - Homocysteine and Folate in Frozen Human Serum) and in a small subset of normal donor plasma samples.
Citation
Analytical Chemistry

Keywords

cardiovascular disease, endothelial cell disfunction, folate, homocysteine, liquid chromatography, serum, standard reference material, tandem mass spectrometry

Citation

Nelson, B. , Satterfield, M. , Sniegoski, L. and Welch, M. (2021), Unified Quantification of Homocysteine and Folate in Human Serum or PlasmaUsing Liquid Chromatography/Tandem Mass Spectrometry, Analytical Chemistry (Accessed March 5, 2024)
Created October 12, 2021