Institute of Food Science Research (CIAL), CSIC, Nicolas Cabrera, Campus de Cantoblanco, Madrid, Spain.
Electrophoresis. 2011 Jun;32(13):1765-77. doi: 10.1002/elps.201100019. Epub 2011 May 30.
In this work, four different metabolite purification approaches are investigated prior to metabolomics of human HT29 colon cancer cells. Namely, methanol deproteinization, ultrafiltration and two SPE methods using C18 and polymer-based cartridges were studied. The extracts were characterized via a metabolomic approach based on the application of CE TOF MS (CE-MS). CE-MS analysis time was less than 20 min per sample and allowed the simultaneous and reproducible analysis of more than 80 metabolites in a single run with a minimum consumption of sample and reagents. Metabolome analysis revealed in some cases important differences among the studied metabolite purification procedures. No significant differences were observed in the metabolite profile using C18 and polymer-based cartridges, or between ultrafiltration and methanol deproteinization. However, important differences were observed in the metabolomic profiles obtained from SPE and methanol deproteinization samples. These results demonstrate the crucial role of the metabolite purification strategy in metabolomics since it can bias (and in some cases mislead) the conclusions achieved by the metabolomic study.
在这项工作中，我们研究了四种不同的代谢物纯化方法，然后对人 HT29 结肠癌细胞进行代谢组学分析。分别考察了甲醇沉淀法、超滤法和两种基于 C18 和聚合物的固相萃取（SPE）方法。通过基于 CE-TOF MS（CE-MS）的代谢组学方法对提取物进行了表征。CE-MS 分析时间为每个样品不到 20 分钟，允许在单次运行中同时且可重复地分析 80 多种代谢物，同时消耗最少的样品和试剂。代谢组学分析显示，在所研究的代谢物纯化程序中，某些情况下存在重要差异。使用 C18 和聚合物基的 SPE 和超滤与甲醇沉淀法之间没有观察到代谢物图谱的显著差异。然而，从 SPE 和甲醇沉淀法样品中获得的代谢组学图谱存在重要差异。这些结果表明，代谢物纯化策略在代谢组学中起着至关重要的作用，因为它会影响（并且在某些情况下会误导）通过代谢组学研究得出的结论。
