Quantitative profiling of 1-Carbon Metabolites, Amino Acids and Precursors, and Plasmalogens in human plasma using Ultra-High-Pressure Liquid Chromatography coupled with Tandem Mass Spectrometry and robotic compound extraction

Amino acids (AAs) and one-carbon (1-C) compounds are involved in a range of key metabolic pathways, and mediate numerous health and disease processes in the human body. Previous assays have quantified a limited selection of these compounds. Here, we describe an analytical method for the simultaneous quantification of 37 1-C metabolites, amino acids, precursors, and plasmalogens using reversed-phase ultra-high pressure liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Compound extraction from human plasma was tested manually before being robotically automated. The final analytical panel was validated on human plasma samples. Our automated and multiplexed method holds promise for application to large cohort studies.