Determination of trimethylamine N-oxide in human plasma by ultra performance liquid chromatography-tandem mass spectrometry
Objective To establish an ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determining the content of trimethylamine N-oxidein in human blood.Methods Add an appropriate amount of internal standard to human plasma samples as required,extract and rapidly precipitate proteins using methanol,centrifuge at high speed,take the supernatant and centrifuge at high speed again before measuring on the machine.The hydrophilic BEH HILIC(100 mm× 2.1 mm,1.7 μm)column was selected for gradient elution with 10 mmol/L ammonium acetate acetonitrile mobile phase,UPLC-MS/MS was used for determination,electrospray ionization and multiple reaction monitoring mode were selected,the quantitative ion pair(m/z)was 76.10/58.10,and the internal standard method was used for quantification.Result The linear relationship between the content of trimethylamine N-oxide in methanol solution within the range of 4-500 μg/L(converted to plasma concentration of 0.266-33.28 μmol/L)was good,the linear regression equation was y=0.077 44x+0.005 89,with a correlation coefficient of 0.999 8.At 2.663,6.656,13.314 μmol/L of the three additive level,the recovery rate of the sample was 89.5％-93.6％.The relative standard deviation(RSD)was 3.10％-5.25％,and the detection limits was 0.026 6 μmol/L.Conclusion The established method has simple and fast preprocessing,high accuracy and precision,wide linear range,and is suitable for the detection of trimethylamine N-oxidein in human plasma.
Ultra performance liquid chromatography-tandem mass spectrometryPlasmaTrimethylamine N-oxide
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