Development of a Robust and Repeatable UPLC-MS Method for the Long-Term Metabolomic Study of Human Serum
Development of a Robust and Repeatable UPLC-MS Method for the Long-Term Metabolomic Study of Human Serum
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DOI：
10.1021/ac8019366
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发表时间：
2009-02-15
影响因子：
7.4
通讯作者：
Kellt, Douglas B.
中科院分区：
文献类型：
--
作者：
Zelena, Eva;Dunn, Warwick B.;Kellt, Douglas B.
基于蛋白质沉淀和超高效液相色谱-飞行时间质谱（UPLC-TOF-MS），开发了一种用于进行人血清非靶向代谢组学分析的方法。该方法专门设计用于满足长期代谢组学研究的要求，持续时间超过3年，随后对其耐用性和重复性进行了全面评价。在这里，我们描述了观察到的仪器性能随着时间的推移和它的改进与分析块的长度的调整漂移。针对来自30名健康个体的一组血清样品进一步验证了用于我们目的的最佳设置。我们还评估了来自同一生产商但来自不同生产批次的具有相同固定相化学性质的色谱柱的重现性。这些结果使作者能够准备SON用于“适合目的”的长期UPLC-MS代谢组学研究，例如正在HUSERMET项目中使用的研究。这种方法允许采集数据，并随后对多个月或多年收集的数据进行比较。
A method for performing untargeted metabolomic analysis of human serum has been developed based on protein precipitation followed by Ultra Performance liquid Chromatography and Time-of-Flight mass spectrometry (UPLC-TOF-MS). This method was specifically designed to fulfill the requirements of a long-term metabolomic study, spanning more than 3 years, and it was subsequently thoroughly evaluated for robustness and repeatability. We describe here the observed drift in instrumental performance over time and its improvement with adjustment of the length of analytical block. The optimal setup for our purpose was further validated against a set of serum samples from 30 healthy individuals. We also assessed the reproducibility of chromatographic columns with the same chemistry of stationary phase from the same manufacturer but from different production batches. The results have allowed the authors to prepare SON for "fit for purpose" long-term UPLC-MS metabolomic studies, such as are being employed in the HUSERMET project. This method allows the acquisition of data and subsequent comparison of data collected across many months or years.