Development and validation of a simple UHPLC-MS/MS method for the simultaneous determination of trimethylamine N-oxide, choline, and betaine in human plasma and urine

Journal of Pharmaceutical and Biomedical Analysis
Andrew J OcqueThomas D Nolin

Abstract

A simple, sensitive, and precise ultra-high performance liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of trimethylamine N-oxide, choline, and betaine in human plasma and urine. Sample preparation involved protein precipitation with methanol containing internal standards. Chromatographic separation was achieved using an Acquity BEH Amide (2.1mm×50mm, 1.7μm) analytical column with gradient elution of solvent A (10mM ammonium formate, pH 3.5) and solvent B (acetonitrile). The flow rate was 0.4mL/min and the total run time was 5min. Detection of analytes was performed using heated electrospray ionization (positive mode) and selected reaction monitoring. Excellent linearity was observed over the standard curve concentration ranges of 0.010-5.00μg/mL (plasma) and 1.00-150μg/mL (urine) for all analytes. The intra- and inter-day accuracy and precision for all quality controls were within ±10%. Excellent recovery was observed. The method is rapid, accurate and reproducible, and was successfully applied to a pilot study of markers of atherosclerosis in patients with kidney disease who underwent successful kidney transplantation.

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Citations

Sep 27, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jia LiuYuxin Yin
Jun 10, 2016·The American Journal of Clinical Nutrition·Richard D SembaMark J Manary
Jul 30, 2017·Molecular Nutrition & Food Research·Ralf KrügerBernhard Watzl
Aug 1, 2015·Journal of the American Society of Nephrology : JASN·Jason R StubbsAlan S Yu
Jan 23, 2019·Clinical Journal of the American Society of Nephrology : CJASN·Jason R StubbsThomas D Nolin
Jun 20, 2018·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Cassandra JohnsonJason R Stubbs
Oct 23, 2019·Clinical Chemistry and Laboratory Medicine : CCLM·Liam M Heaney
Oct 14, 2017·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Yongxin LiCong Yu
Oct 25, 2017·Scientific Reports·Eke G GruppenRobin P F Dullaart
Oct 16, 2019·Journal of Pharmaceutical and Biomedical Analysis·Jessica FioriRoberto Gotti
Jan 16, 2021·Biomedical Chromatography : BMC·Elham Sadat Hosseini, Kourosh Tabar Heydar
May 28, 2021·Journal of Separation Science·Elham Sadat Hosseini, Kourosh Tabar Heydar

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