Muhamadali Howbeer, Simoens Kenneth, Xu Yun, Nicolai Bart, Bernaerts Kristel, Goodacre Royston
Department of Biochemistry and Systems Biology, Institute of Systems Molecular and Integrative Biology, University of Liverpool, Biosciences Building, Liverpool L69 7ZB, UK.
School of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK.
Metabolites. 2020 Sep 22;10(9):379. doi: 10.3390/metabo10090379.
In the past two decades, metabolomics has proved to be a valuable tool with many potential applications in different areas of science. However, there are still some challenges that need to be addressed, particularly for multicenter studies. These challenges are mainly attributed to various sources of fluctuation and unwanted variations that can be introduced at pre-analytical, analytical, and/or post-analytical steps of any metabolomics experiment. Thus, this study aimed at using TK24 as the model organism in a cross-laboratory experiment in Manchester and Leuven to evaluate the reproducibility of a standard sample preparation method, and determine the optimal sample format (cell extract or quenched biomass) required to preserve the metabolic profile of the cells during cross-lab sample transportation and storage. Principal component analysis (PCA) scores plot of the gas chromatography-mass spectrometry (GC-MS) data from both laboratories displayed clear growth-dependent clustering patterns which was in agreement with the Procrustes analysis findings. In addition, the data generated in Manchester displayed tight clustering of cell pellets (quenched biomass) and metabolite extracts, confirming the stability of both sample formats during the transportation and storage period.
在过去二十年中,代谢组学已被证明是一种有价值的工具,在不同科学领域有许多潜在应用。然而,仍有一些挑战需要解决,特别是对于多中心研究。这些挑战主要归因于在任何代谢组学实验的分析前、分析中和/或分析后步骤中可能引入的各种波动源和不必要的变化。因此,本研究旨在以TK24作为模式生物,在曼彻斯特和鲁汶进行跨实验室实验,以评估标准样品制备方法的重现性,并确定在跨实验室样品运输和储存过程中保持细胞代谢谱所需的最佳样品形式(细胞提取物或淬灭生物量)。来自两个实验室的气相色谱 - 质谱(GC-MS)数据的主成分分析(PCA)得分图显示出明显的生长依赖性聚类模式,这与普洛斯特分析结果一致。此外,在曼彻斯特生成的数据显示细胞沉淀(淬灭生物量)和代谢物提取物紧密聚类,证实了两种样品形式在运输和储存期间的稳定性。