A highly sensitive liquid chromatography electrospray ionization mass spectrometry method for quantification of TMA, TMAO and creatinine in mouse urine.
A highly sensitive liquid chromatography electrospray ionization mass spectrometry method for quantification of TMA, TMAO and creatinine in mouse urine.
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DOI：
10.1016/j.mex.2017.09.004
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发表时间：
2017
期刊：
影响因子：
1.9
通讯作者：
Shephard EA
中科院分区：
文献类型：
--
作者：
Veeravalli S;Karu K;Phillips IR;Shephard EA
我们的方法描述了小鼠尿中三甲胺(TMA)、三甲胺N-氧化物(TMAO)和肌酐的定量。该方法结合了TMA与溴乙酸乙酯的衍生化和在ACEC18柱上的LC色层分离。将流出的污水连续电喷入线性离子陷阱质谱仪(LTQ)，该质谱仪在为目标分析物及其内标(IS)设置的选择离子监测(SIM)模式下运行。所有验证参数均在分析方法验证指南的可接受范围内。日内和日间精密度和准确度变异系数为3.1%，TMA和TMAO的回收率为97-104%。开发的方法使用两个步骤的程序。首先用5分钟的梯度CAP-LC-SIMS分析了TMA和TMAO，然后在相同的实验条件下分析了肌酸。该方法灵敏、灵敏、重现性好，可高通量地检测TMA、TMAO和肌酐，柱上浓度分别为28、115和1 ng/mL。该方法适用于雌、雄小鼠尿液中TMA、TMAO和肌酐的分析。该方法的主要优点是：所需的尿样体积小，克服了在规定的时间收集足够数量的尿液的困难。无需样品前处理。用同样的CAP-LC-MS方法对TMA、TMAO和肌酐进行定量。
Our method describes the quantification in mouse urine of trimethylamine (TMA), trimethylamine N-oxide (TMAO) and creatinine. The method combines derivatization of TMA, with ethyl bromoacetate, and LC chromatographic separation on an ACE C18 column. The effluent was continuously electrosprayed into the linear ion trap mass spectrometer (LTQ), which operated in selective ion monitoring (SIM) modes set for targeted analytes and their internal standards (IS). All validation parameters were within acceptable ranges of analytical method validation guidelines. Intra- and inter-day assay precision and accuracy coefficients of variation were <3.1%, and recoveries for TMA and TMAO were 97–104%. The method developed uses a two-step procedure. Firstly, TMA and TMAO are analyzed without a purification step using a 5-min gradient cap-LC- SIMs analysis, then creatinine is analyzed using the same experimental conditions. The method is robust, highly sensitive, reproducible and has the high-throughput capability of detecting TMA, TMAO and creatinine at on-column concentrations as low as 28 pg/mL, 115 pg/mL and 1 ng/mL, respectively. The method is suitable for analysis of TMA, TMAO and creatinine in both male and female mouse urine. The key benefits of the method are: The small sample volume of urine required, which overcomes the difficulties of collecting sufficient volumes of urine at defined times. No sample pre-treatment is necessary. The quantification of TMA, TMAO and creatinine using the same cap-LC-MS method.