Hussain Mohamad Awwad1, Juergen Geisel2, Rima Obeid3. 1. Saarland University Hospital, Department of Clinical Chemistry and Laboratory Medicine Building 57, D-66421, Homburg/Saar, Germany. Electronic address: s9huawwa@stud.uni-saarland.de. 2. Saarland University Hospital, Department of Clinical Chemistry and Laboratory Medicine Building 57, D-66421, Homburg/Saar, Germany. 3. Saarland University Hospital, Department of Clinical Chemistry and Laboratory Medicine Building 57, D-66421, Homburg/Saar, Germany; Aarhus Institute of Advanced Studies, University of Aarhus, Høegh-Guldbergs Gade 6B, Building 1632, Dk-8000, Aarhus C, Denmark. Electronic address: rima.obeid@uks.eu.
Abstract
BACKGROUND: Trimethylamine-N-oxide (TMAO) is produced in the liver from trimethylamine (TMA) and is an important cellular osmolyte and potential atherogenic factor. Taurine is involved in cholesterol metabolism and also serves as a cellular osmolyte. Given their significant biological functions, the development of reliable measurement techniques is crucial to further study their role in health and disease METHODS: A new ultrahigh performance liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of TMA, TMAO, and taurine in plasma and urine. The method consisted of a deproteinization step using methanol/acetonitrile (15:85) that contained 0.2% formic acid and isotope-labeled internal standards. Samples were separated by centrifugation and injected into the UHPLC system. Quantification was conducted using a triple-quadrupole mass spectrometer detector with electrospray ionization interface in positive mode. RESULTS: The limits of detection ranged from 0.08 to 0.12μmol/L. The calibration curves were linear (r≥0.999) over the range examined (0.15-400μmol/L) for all compounds. The inter- and intra-day coefficients of variations were≤14.5% for TMA and ≤8% for TMAO and taurine. TMAO and taurine were found to be stable in EDTA plasma for at least 14 months at -70°C. Mean recoveries ranged from 95% to 109% and the relative matrix effects were≤4.0%. The method was applied to study physiological and pre-analytical factors in plasma and urine samples. CONCLUSIONS: The new UHPLC-MS/MS method has good accuracy, precision, and recovery. The assay combines simple sample processing with a short run time, making it well suited for high-throughput routine clinical or research purposes.
BACKGROUND:Trimethylamine-N-oxide (TMAO) is produced in the liver from trimethylamine (TMA) and is an important cellular osmolyte and potential atherogenic factor. Taurine is involved in cholesterol metabolism and also serves as a cellular osmolyte. Given their significant biological functions, the development of reliable measurement techniques is crucial to further study their role in health and disease METHODS: A new ultrahigh performance liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of TMA, TMAO, and taurine in plasma and urine. The method consisted of a deproteinization step using methanol/acetonitrile (15:85) that contained 0.2% formic acid and isotope-labeled internal standards. Samples were separated by centrifugation and injected into the UHPLC system. Quantification was conducted using a triple-quadrupole mass spectrometer detector with electrospray ionization interface in positive mode. RESULTS: The limits of detection ranged from 0.08 to 0.12μmol/L. The calibration curves were linear (r≥0.999) over the range examined (0.15-400μmol/L) for all compounds. The inter- and intra-day coefficients of variations were≤14.5% for TMA and ≤8% for TMAO and taurine. TMAO and taurine were found to be stable in EDTA plasma for at least 14 months at -70°C. Mean recoveries ranged from 95% to 109% and the relative matrix effects were≤4.0%. The method was applied to study physiological and pre-analytical factors in plasma and urine samples. CONCLUSIONS: The new UHPLC-MS/MS method has good accuracy, precision, and recovery. The assay combines simple sample processing with a short run time, making it well suited for high-throughput routine clinical or research purposes.
Authors: Viktor Kožich; Tamás Ditrói; Jitka Sokolová; Michaela Křížková; Jakub Krijt; Pavel Ješina; Peter Nagy Journal: Br J Pharmacol Date: 2018-11-25 Impact factor: 8.739