Evaluation and optimization of sample preparation methods for metabolic profiling analysis of Escherichia coli

	Evaluation and optimization of sample preparation methods for metabolic profiling analysis of Escherichia coli
	Wang, Xiyue 1; Yang, Fengxu 2; Zhang, Yuansheng 2; Xu, Guowang 1; Liu, Yang 1; Tian, Jing 2; Gao, Peng 1
刊名	electrophoresis
	2015-09-01
卷号	36 期号:18 页码:2140-2147
关键词	Escherichia coli GC-MS Metabolomics Sample preparation method
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.
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
关键词[WOS]	quantitative metabolomics ; extraction methodologies ; saccharomyces-cerevisiae ; yeast metabolomics ; microorganisms ; chromatography ; metabonomics ; signatures ; genomics ; cultures
收录类别	SCI
语种	英语
WOS记录号	WOS:000361185900002
公开日期	2016-05-09
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/146550]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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
推荐引用方式 GB/T 7714	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.
APA	Wang, Xiyue.,Yang, Fengxu.,Zhang, Yuansheng.,Xu, Guowang.,Liu, Yang.,...&Gao, Peng.(2015).Evaluation and optimization of sample preparation methods for metabolic profiling analysis of Escherichia coli.electrophoresis,36(18),2140-2147.
MLA	Wang, Xiyue,et al."Evaluation and optimization of sample preparation methods for metabolic profiling analysis of Escherichia coli".electrophoresis 36.18(2015):2140-2147.