基于大气压光离子化质谱联用流动注射分析的新型代谢组学方法在阿尔茨海默病研究中的应用。

Application of a novel metabolomic approach based on atmospheric pressure photoionization mass spectrometry using flow injection analysis for the study of Alzheimer's disease.

机构信息

Department of Chemistry and CC.MM, Faculty of Experimental Science, University of Huelva, Campus de El Carmen, 21007 Huelva, Spain; Campus of Excellence International ceiA3, University of Huelva, Spain; Research Center of Health and Environment (CYSMA), University of Huelva, Campus de El Carmen, 21007 Huelva, Spain.

出版信息

Talanta. 2015 Jan;131:480-9. doi: 10.1016/j.talanta.2014.07.075. Epub 2014 Aug 21.

DOI:10.1016/j.talanta.2014.07.075

PMID:25281130

Abstract

The use of atmospheric pressure photoionization is not widespread in metabolomics, despite its considerable potential for the simultaneous analysis of compounds with diverse polarities. This work considers the development of a novel analytical approach based on flow injection analysis and atmospheric pressure photoionization mass spectrometry for rapid metabolic screening of serum samples. Several experimental parameters were optimized, such as type of dopant, flow injection solvent, and their flows, given that a careful selection of these variables is mandatory for a comprehensive analysis of metabolites. Toluene and methanol were the most suitable dopant and flow injection solvent, respectively. Moreover, analysis in negative mode required higher solvent and dopant flows (100 µl min(-1) and 40 µl min(-1), respectively) compared to positive mode (50 µl min(-1) and 20 µl min(-1)). Then, the optimized approach was used to elucidate metabolic alterations associated with Alzheimer's disease. Thereby, results confirm the increase of diacylglycerols, ceramides, ceramide-1-phosphate and free fatty acids, indicating membrane destabilization processes, and reduction of fatty acid amides and several neurotransmitters related to impairments in neuronal transmission, among others. Therefore, it could be concluded that this metabolomic tool presents a great potential for analysis of biological samples, considering its high-throughput screening capability, fast analysis and comprehensive metabolite coverage.

摘要

大气压光电离在代谢组学中的应用并不广泛，尽管它具有同时分析不同极性化合物的巨大潜力。本工作考虑开发一种基于流动注射分析和大气压光电离质谱的新分析方法，用于血清样品的快速代谢筛选。优化了几个实验参数，如掺杂剂的类型、流动注射溶剂及其流速，因为仔细选择这些变量对于全面分析代谢物是强制性的。甲苯和甲醇分别是最合适的掺杂剂和流动注射溶剂。此外，与正模式相比（分别为 50 μl min(-1) 和 20 μl min(-1)），负模式需要更高的溶剂和掺杂剂流速（分别为 100 μl min(-1) 和 40 μl min(-1)）。然后，优化后的方法用于阐明与阿尔茨海默病相关的代谢变化。结果证实，二酰基甘油、神经酰胺、神经酰胺-1-磷酸和游离脂肪酸增加，表明膜去稳定化过程，以及脂肪酸酰胺和几种与神经元传递受损相关的神经递质减少等。因此，可以得出结论，考虑到高通量筛选能力、快速分析和全面的代谢物覆盖范围，这种代谢组学工具具有很大的生物样品分析潜力。