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题名: Evaluation and optimization of sample preparation methods for metabolic profiling analysis of Escherichia coli
作者: Wang, Xiyue1;  Yang, Fengxu2;  Zhang, Yuansheng2;  Xu, Guowang1;  Liu, Yang1;  Tian, Jing2;  Gao, Peng1
关键词: Escherichia coli ;  GC-MS ;  Metabolomics ;  Sample preparation method
刊名: ELECTROPHORESIS
发表日期: 2015-09-01
DOI: 10.1002/elps.201400567
卷: 36, 期:18, 页:2140-2147
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: In this study, exemplified by Escherichia coli, a sample preparation method was developed and evaluated with more concerns on cell pellets washing, disruption, and metabolite extraction for global metabolite profiling analysis using GC-MS. We found that loss of some intracellular metabolites occurred inevitably, no matter which washing strategy was used. But PBS washing resulted in the least leakage of intercellular metabolites among the six tested methods. In addition, bead-beating showed better repeatability and high throughput than sonication for cell pellet disruption. Total six extraction solvents, including chloroform/methanol/water (2:0.21:0.79, v/v/v), methyl tert-butyl ether/methanol/water (2:0.21:0.79), acetonitrile/methanol/water (2:2:1), water/isopropanol/methanol (3:2:5), pure methanol, and methanol/water (1:1), were evaluated for metabolite extraction efficiency. We demonstrated that the numbers of extracted metabolites were almost the same, but methyl tert-butyl ether/methanol/water and chloroform/methanol/water could achieve the best recoveries in view of MS intensities. This study implies the necessity to evaluate the potential effects of serial sample preparation procedures prior to metabolomic analysis.
关键词[WOS]: QUANTITATIVE METABOLOMICS ;  EXTRACTION METHODOLOGIES ;  SACCHAROMYCES-CEREVISIAE ;  YEAST METABOLOMICS ;  MICROORGANISMS ;  CHROMATOGRAPHY ;  METABONOMICS ;  SIGNATURES ;  GENOMICS ;  CULTURES
语种: 英语
WOS记录号: WOS:000361185900002
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/146550
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian, Peoples R China
2.Dalian Polytech Univ, Sch Bioengn, Dalian, Peoples R China

Recommended Citation:
Wang, Xiyue,Yang, Fengxu,Zhang, Yuansheng,et al. Evaluation and optimization of sample preparation methods for metabolic profiling analysis of Escherichia coli[J]. ELECTROPHORESIS,2015,36(18):2140-2147.
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