Evaluation of extraction processes for intracellular metabolite profiling of mammalian cells: matching extraction approaches to cell type and metabolite targets
Evaluation of extraction processes for intracellular metabolite profiling of mammalian cells: matching extraction approaches to cell type and metabolite targets
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DOI：
10.1007/s11306-010-0216-9
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发表时间：
2010-09-01
期刊：
影响因子：
3.6
通讯作者：
Dickson, Alan J.
中科院分区：
文献类型：
--
作者：
Sellick, Christopher A.;Knight, David;Dickson, Alan J.
在这项研究中，我们报告了中国仓鼠卵巢（CHO）细胞内代谢物的全球代谢组学谱生成技术的优化。我们评估了一系列不同的提取方法应用于CHO细胞的有效性，CHO细胞已使用先前优化的方法淬灭。测试的提取方法包括冷甲醇、热乙醇、酸、碱和甲醇/氯仿及其组合。代谢物的提取使用两次100%甲醇萃取，然后最后用水萃取回收最大范围的代谢物。对于大多数代谢物，以这种方式产生的提取物具有最大的回收率和高重现性。因此，这是从单一样品中获得全球代谢概况的最佳提取方法。然而，可能还需要另一种平行提取方法（例如碱）来最大限度地回收代谢物（例如非极性代谢物）的范围。
In this study we report on the optimisation of the technologies for generation of a global metabolomics profile for intracellular metabolites in Chinese hamster ovary (CHO) cells. We evaluated the effectiveness of a range of different extraction methods applied to CHO cells which had been quenched using a previously optimised approach. The extraction methods tested included cold methanol, hot ethanol, acid, alkali and methanol/chloroform plus combinations of these. The extraction of metabolites using two 100% methanol extractions followed by a final water extraction recovered the largest range of metabolites. For the majority of metabolites, extracts generated in this manner exhibited the greatest recovery with high reproducibility. Therefore, this was the best extraction method for attaining a global metabolic profile from a single sample. However, another parallel extraction method (e.g. alkali) may also be required to maximise the range of metabolites recovered (e.g. non-polar metabolites).