Work

Development and Application of a UPLC–MRM–MS Method for Quantifying Trimethylamine, Trimethylamine-N-Oxide, and Related Metabolites in Individuals with and Without Metabolic Syndrome

Year: 2025

Type: article

Abstract: Trimethylamine-N-oxide (TMAO) is associated with various chronic diseases. TMAO is a downstream oxidative metabolite of trimethylamine (TMA) that is generated by the gut microbiota from dietary choline, carnitine, and betaine. Current analytical methods predominantly target TMAO only, due to the challenge of efficiently extracting and quantifying TMA. The present study demonstrates a simple and rapid UPLC-MRM-MSMS method for concurrent quantification of TMAO, TMA, and related precursors (choline, betaine and various carnitines) following a methanol extraction from plasma. Pure methanol resulted in a higher extractability of TMA (up to two-fold) compared to both pure acetonitrile and various methanol/acetonitrile mixtures. The quantification method showed high linearity within the tested range of 0.0625-100 μmol/L (correlation co-efficient >0.999), intra- (n=3) and inter-day (n=9) precision of 2-8% along with an average recovery of above 94% for all metabolites (TMAO, TMA, choline, betaine, L-carnitine, acetyl-L-carnitine and propionyl-L-carnitine). The method applicability was confirmed through a comparison of TMAO and its precursor concentrations in plasma samples of overnight-fasted subjects with (n=12) and without (n=21) metabolic syndrome.

Source: Separations

DOI: 10.3390/separations12020053

Authors: Mohammed Hefni, Cornelia M. Witthöft

Institutions: Mansoura University, Linnaeus University

Language: English

Topic: Metabolomics and Mass Spectrometry Studies

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $3,137