Within-day reproducibility of an HPLC-MS-Based method for metabonomic analysis: Application to human urine
Within-day reproducibility of an HPLC-MS-Based method for metabonomic analysis: Application to human urine
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DOI：
10.1021/pr070183p
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发表时间：
2007-08-01
影响因子：
4.4
通讯作者：
Wilson, Ian D.
中科院分区：
文献类型：
--
作者：
Gika, Helen G.;Theodoridis, Georgios A.;Wilson, Ian D.
不言而喻，在支持“系统生物学”的领域，如基因组学，蛋白质组学和代谢组学的研究是关键依赖于可靠的分析数据的产生。使用基于LC-MS的方法进行代谢表型分析目前处于相对早期的开发阶段，确保数据质量的方法仍在开发中。作为LC-MS在代谢组学中应用研究的一部分，已使用标准“质量控制”（QC）样品研究了LC-MS的日内重现性，包括正和负电喷雾电离（ESI）。结果表明，系统上的前几次进样不具有代表性，应丢弃，重现性主要取决于信号强度。根据这些结果，开发了人尿液代谢组学分析的分析方案，并根据逐步评估的数据制定了拟定验收标准。还评估了人尿液的短期样本稳定性。样品在自动进样器中在4 ℃下稳定至少20 h，同时排队进行分析。在-20 ℃或-80 ℃下储存长达1个月的样品在随后的LC-MS分析中无法区分。总体而言，通过仔细监测QC数据，可以证明LC-MS的“日内”重现性足以确保全球代谢分析应用中的数据质量。
Self-evidently, research in areas supporting "systems biology" such as genomics, proteomics, and metabonomics are critically dependent on the generation of sound analytical data. Metabolic phenotyping using LC-MS-based methods is currently at a relatively early stage of development, and approaches to ensure data quality are still developing. As part of studies on the application of LC-MS in metabonomics, the within-day reproducibility of LC-MS, with both positive and negative electrospray ionization (ESI), has been investigated using a standard "quality control" (QC) sample. The results showed that the first few injections on the system were not representative, and should be discarded, and that reproducibility was critically dependent on signal intensity. On the basis of these findings, an analytical protocol for the metabonomic analysis of human urine has been developed with proposed acceptance criteria based on a step-by-step assessment of the data. Short-term sample stability for human urine was also assessed. Samples were stable for at least 20 h at 4 degrees C in the autosampler while queuing for analysis. Samples stored at either -20 or -80 degrees C for up to 1 month were indistinguishable on subsequent LC-MS analysis. Overall, by careful monitoring of the QC data, it is possible to demonstrate that the "within-day" reproducibility of LC-MS is sufficient to ensure data quality in global metabolic profiling applications.