Extraction parameters for metabolomics from cultured cells.
Extraction parameters for metabolomics from cultured cells.
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DOI：
10.1016/j.ab.2015.01.003
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发表时间：
2015-04-15
影响因子：
2.9
通讯作者：
Locasale, Jason W.
中科院分区：
文献类型：
--
作者：
Ser, Zheng;Liu, Xiaojing;Ngoc Nu Tang;Locasale, Jason W.
代谢物的成功提取是代谢物分析的关键步骤。通过优化代谢物提取，可以提高代谢组学研究的范围和定量能力。我们考虑了八种单独的提取方案，用于从培养的哺乳动物细胞中制备代谢物提取物。考虑的参数包括提取前的温度、pH 值和细胞洗涤。使用高分辨率液相色谱质谱 (LC-HRMS) 平台研究了对代谢物回收的影响，该平台可测量不同化学类别的代谢物，包括氨基酸、脂质和糖衍生物。提取过程中考虑的温度或常用添加剂甲酸的存在对代谢物的测量离子强度影响最小。然而，在代谢物提取之前清洗样品，无论是用水还是 PBS（这两种通常被认为的做法），都会对细胞内和细胞外代谢物的测量强度产生巨大影响。这些发现共同对代谢物分析的提取条件进行了系统评估。
The successful extraction of metabolites is a critical step in metabolite profiling. By optimizing metabolite extraction, the range and quantitative capacity of metabolomics studies can be improved. We considered eight separate extraction protocols for the preparation of a metabolite extract from cultured mammalian cells. Parameters considered included temperature, pH, and cell washing before extraction. The effects on metabolite recovery were studied using a high resolution liquid chromatography mass spectrometry (LC-HRMS) platform that measures metabolites of diverse chemical classes including among others amino acids, lipids, and sugar derivatives. The temperature considered during the extraction or the presence of formic acid, a commonly used additive, was shown to have minimal effects on the measured ion intensities of metabolites. However, washing of samples before metabolite extraction whether with water or PBS (both commonly considered practices) exhibited dramatic effects on measured intensities of both intra- and extra-cellular metabolites. Together these findings present a systematic assessment of extraction conditions for metabolite profiling.
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通讯作者：
Ressom HW
影响因子：
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作者：
Lorenz, Matthew A.;Burant, Charles F.;Kennedy, Robert T.
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作者：
Wang, Xiaodong;Wang, Yongcheng
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