A validated simple LC-MS/MS method for quantifying trimethylamine N-oxide (TMAO) using a surrogate matrix and its...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Chemistry

Background:

• Trimethylamine N-oxide (TMAO) is a metabolite from dietary choline and carnitine via gut microbes.
• Elevated plasma TMAO levels are linked to increased risks of cardiovascular and chronic kidney diseases.
• Accurate quantification of TMAO is crucial for clinical applications and disease monitoring.

Purpose of the Study:

• To develop and validate a simple, accurate, and sensitive quantitative method for TMAO.
• To establish a reliable LC-MS/MS assay for TMAO measurement in biological samples.
• To support clinical pharmacology studies by providing a robust TMAO quantification tool.

Main Methods:

• Development of a quantitative method using liquid chromatography tandem mass spectrometry (LC-MS/MS).
• Construction of an eight-point calibration curve in an artificial surrogate matrix (1-5,000 ng/mL).
• Validation of the method for linearity, precision, accuracy, and cross-validation with human plasma.

Main Results:

• Excellent linearity achieved with R² > 0.996.
• Intra-day and inter-day assays demonstrated satisfactory precision (1.65-7.15%) and accuracy (96.36-111.43%).
• The method was successfully cross-validated and applied to a clinical pharmacology study.

Conclusions:

• The developed LC-MS/MS method provides accurate and sensitive quantification of TMAO.
• This validated method is suitable for clinical research, including disease progression monitoring and drug effect evaluation.
• The assay's robustness supports its application in clinical pharmacology studies.