Development and validation of a simple UHPLC-MS/MS method for the simultaneous determination of trimethylamine N-oxide, choline, and betaine in human plasma and urine
Development and validation of a simple UHPLC-MS/MS method for the simultaneous determination of trimethylamine N-oxide, choline, and betaine in human plasma and urine
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DOI：
10.1016/j.jpba.2015.02.040
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发表时间：
2015-05-10
影响因子：
3.4
通讯作者：
Nolin, Thomas D.
中科院分区：
文献类型：
--
作者：
Ocque, Andrew J.;Stubbs, Jason R.;Nolin, Thomas D.
建立了一种简便、灵敏、准确的超高效液相色谱-串联质谱法同时测定人血浆和尿液中氧化三甲胺、胆碱和甜菜碱的方法。样品制备涉及用含内标物的甲醇进行蛋白质沉淀。使用Acquity BEH Amide（2.1 mm × 50 mm，1.7 μ m）分析柱，用溶剂A（10 mM甲酸铵，pH 3.5）和溶剂B（乙腈）进行梯度洗脱，实现色谱分离。流速为0.4 mL/min，总运行时间为5 min。使用加热电喷雾电离（正离子模式）和选择反应监测进行分析物检测。对于所有分析物，在0.010-5.00 μ g/mL（血浆）和1.00-150 μ g/mL（尿液）的标准曲线浓度范围内观察到良好的线性。所有质控品的日内和日间准确度和精密度均在+/-10%范围内。观察到良好的恢复。该方法快速、准确、重复性好，已成功应用于肾移植患者动脉粥样硬化标志物的初步研究。(C)2015 Elsevier B. V.版权所有。
A simple, sensitive, and precise ultra-high performance liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of trimethylamine N-oxide, choline, and betaine in human plasma and urine. Sample preparation involved protein precipitation with methanol containing internal standards. Chromatographic separation was achieved using an Acquity BEH Amide (2.1 mm x 50 mm, 1.7 mu m) analytical column with gradient elution of solvent A (10 mM ammonium formate, pH 3.5) and solvent B (acetonitrile). The flow rate was 0.4 mL/min and the total run time was 5 min. Detection of analytes was performed using heated electrospray ionization (positive mode) and selected reaction monitoring. Excellent linearity was observed over the standard curve concentration ranges of 0.010-5.00 mu g/mL (plasma) and 1.00-150 mu g/mL (urine) for all analytes. The intra- and inter-day accuracy and precision for all quality controls were within +/- 10%. Excellent recovery was observed. The method is rapid, accurate and reproducible, and was successfully applied to a pilot study of markers of atherosclerosis in patients with kidney disease who underwent successful kidney transplantation. (C) 2015 Elsevier B.V. All rights reserved.