Development of a metabolomic approach based on urine samples and direct infusion mass spectrometry
Development of a metabolomic approach based on urine samples and direct infusion mass spectrometry
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DOI：
10.1016/j.ab.2014.07.016
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发表时间：
2014-11-15
影响因子：
2.9
通讯作者：
Luis Gomez-Ariza, Jose
中科院分区：
文献类型：
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作者：
Gonzalez-Dominguez, Raill;Castilla-Quintero, Rocio;Luis Gomez-Ariza, Jose
直接进样质谱法分析尿液时，由于人与人之间尿量的差异，导致尿液含盐量高，样品间变异性大，因而存在离子抑制问题。因此，尿液代谢组学需要仔细选择样品制备程序和标准化策略来处理这些问题。采用直接进样电喷雾质谱仪(DI-ESI-MS)对尿样进行代谢组学分析，包括固相萃取、液-液萃取和样品稀释。此外，还根据电导值和统计处理对结果进行了归一化处理，以最大限度地减少样品的变异性。尿液稀释法和混合模式固相萃取法均能显著降低尿盐含量，提供全面的代谢组指纹图谱。此外，统计数据归一化能够校正样本间的生理变异性，提高了结果的质量。因此，尿样的高通量DI-ESI-MS指纹图谱可以通过简单的前处理程序实现，从而允许在代谢组学中使用这种非侵入性采样。最后，在对阿尔茨海默病转基因小鼠模型(APP/PS1)的尿样进行的初步代谢组学研究中，测试了优化的方法，以说明该方法的潜力。(C)2014 Elsevier Inc.保留所有权利。
The analysis of urine by direct infusion mass spectrometry suffers from ion suppression due to its high salt content and inter-sample variability caused by the differences in urine volume between persons. Thus, urine metabolomics requires a careful selection of the sample preparation procedure and a normalization strategy to deal with these problems. Several approaches were tested for metabolomic analysis of urine samples by direct infusion electrospray mass spectrometry (DI-ESI-MS), including solid phase extraction, liquid liquid extraction, and sample dilution. In addition, normalization of results based on conductivity values and statistical treatment was performed to minimize sample variability. Both urine dilution and solid phase extraction with mixed mode sorbent considerably reduced the salt content in urine, providing comprehensive metabolomic fingerprints. Moreover, statistical data normalization enabled the correction of inter-sample physiological variability, improving the quality of results obtained. Therefore, high-throughput DI-ESI-MS fingerprinting of urine samples can be achieved with simple pretreatment procedures allowing the use of this noninvasive sampling in metabolomics. Finally, the optimized approach was tested in a pilot metabolomic investigation of urine samples from transgenic mice models of Alzheimer's disease (APP/PS1) in order to illustrate the potential of the methodology. (C) 2014 Elsevier Inc. All rights reserved.