DICP OpenIR
Evaluation and optimization of sample preparation methods for metabolic profiling analysis of Escherichia coli
Wang, Xiyue1; Yang, Fengxu2; Zhang, Yuansheng2; Xu, Guowang1; Liu, Yang1; Tian, Jing2; Gao, Peng1
KeywordEscherichia Coli Gc-ms Metabolomics Sample Preparation Method
Source PublicationELECTROPHORESIS
2015-09-01
DOI10.1002/elps.201400567
Volume36Issue:18Pages:2140-2147
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordQUANTITATIVE METABOLOMICS ; EXTRACTION METHODOLOGIES ; SACCHAROMYCES-CEREVISIAE ; YEAST METABOLOMICS ; MICROORGANISMS ; CHROMATOGRAPHY ; METABONOMICS ; SIGNATURES ; GENOMICS ; CULTURES
AbstractIn 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.
Language英语
WOS IDWOS:000361185900002
Citation statistics
Cited Times:5[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/146550
Collection中国科学院大连化学物理研究所
Affiliation1.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
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.
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