Analycal locity 455 (2014) 35-40 Contents lists available at ScienceDitest Analytical Biochemistry journal homepage: www.elsevier.com/locate/yabio Measurement of trimethylamine-N-oxide by stable isotope dilution liquid chromatography tandem mass spectrometry orn Zeneng Wang’, Bruce S. Levison*, Jennie E. Hazen, Lillian Donahue, Xin-Min Li*, Stanley L. Hazen *” {Deparment of Celular & Moller Medic, tere Research tae Cleelnd Cine. Cleveland OH 4195 US Deparment of Cardiovascular Medi, Here and Vasc nse, clvlan Cn Ceveand, GH 44S, USA ARTICLE INFO ABSTRACT Received 22 November 2013 Avaable online 1 Ap 2014 ‘Timethylamine-A-onide (IMAO) levels in Blood predict future dak for major adverse cardac events Including myocardial infarction, stroke, and death. Thus, the rapid determination of circulating IMAO Concentration is of einicl interest. Here we report 2 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 pM. respectively Spike and recovery studies dem crate an accuracy at lov (05 uM), mid (5 pM), and high (100M) levee of $8.2. 97.3, and 101.5%, respectively. Adéitinal assay performance metrics include intraday and interday coeliiensof variance of 6.4 and <8, respectively, across the range of TMAO levels. Stability studies reveal that TMAO in Sinerpecmeruy, plasma is stable both during storage at —80°C for 5 yeats a to mltiple freeze thaw [12] AK stom. JA Oe Laie, Tiimetylamine oxi: a erminal electron ‘epee reason of Bae) Gen Woah 1198) 12] E Stenberg, OW. Siyrold AR. Strom, Tmetylmine oie respiration in etineeaspoe J Bate 149 (1962 22-26