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Measurement of Carnitine, TMAO, -butyrobetaine, d9-carnitine, d9-TMAO, and d9--butyrobetaine Sample preparation and quantification for carnitine and TMAO in healthy volunteers Fifty microliters of plasma and 50 L urine samples were extracted with 450 L and 950 L methanol containing 200 ng/mL isotopically labeled internal standards (d3-carnitine and d9-TMAO) respectively, and the extraction was performed using the Geno/Grinder 2010 (SPEX SamplePrep., Metuchen, NJ, USA) at 1000 rpm for 3 min