Journal
ANALYTICAL BIOCHEMISTRY
Volume 455, Issue -, Pages 35-40Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2014.03.016
Keywords
Trimethylamine-N-oxide; Cardiovascular disease; Mass spectrometry
Funding
- United States National Institutes of Health (NIH)
- Office of Dietary Supplement [R01 HL103866]
- NIH [P20 HL113452]
- American Heart Association Scientist Development Grant [12SDG12050473]
- Center of Innovation Award from AB SCIEX
- Leonard Krieger Fund
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Trimethylamine-N-oxide (TMAO) levels in blood predict future risk for major adverse cardiac events including myocardial infarction, stroke, and death. Thus, the rapid determination of circulating TMAO concentration is of clinical interest. Here we report a method to measure TMAO in biological matrices by stable isotope dilution liquid chromatography tandem mass spectrometry (LC/MS/MS) with lower and upper limits of quantification of 0.05 and >200 mu M, respectively. Spike and recovery studies demonstrate an accuracy at low (0.5 mu M), mid (5 mu M), and high (100 mu M) levels of 98.2, 97.3, and 101.6%, respectively. Additional assay performance metrics include intraday and interday coefficients of variance of <6.4 and <9.9%, respectively, across the range of TMAO levels. Stability studies reveal that TMAO in plasma is stable both during storage at 80 C for 5 years and to multiple freeze thaw cycles. Fasting plasma normal range studies among apparently healthy subjects (n = 349) show a range of 0.73126 mu M, median (interquartile range) levels of 3.45 (2.25-5.79) mu M, and increasing values with age. The LC/MS/MS-based assay reported should be of value for further Studies evaluating TMAO as a risk marker and for examining the effect of dietary, pharmacologic, and environmental factors on TMAO levels. (c) 2014 Elsevier Inc. All rights reserved.
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