Microbial metabolomics with gas chromatography/mass spectrometry

被引:260
作者
Koek, MM
Muilwijk, B
van der Werf, MJ
Hankemeier, T
机构
[1] TNO Qual Life, Dept Analyt Sci, NL-3704 HE Zeist, Netherlands
[2] Leiden Univ, LACDR Analyt Biosci, NL-2333 CC Leiden, Netherlands
关键词
D O I
10.1021/ac051683+
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
An analytical method was set up suitable for the analysis of microbial metabolomes, consisting of an oximation and silylation derivatization reaction and subsequent analysis by gas chromatography coupled to mass spectrometry. Microbial matrixes contain many compounds that potentially interfere with either the derivatization procedure or analysis, such as high concentrations of salts, complex media or buffer components, or extremely high substrate and product concentrations. The developed method was extensively validated using different microorganisms, i.e., Bacillus subtilis, Propionibacterium freudenreichii, and Escherichia coli. Many metabolite classes could be analyzed with the method: alcohols, aldehydes, amino acids, amines, fatty acids, (phospho-) organic acids, sugars, sugar acids, (acyl-) sugar amines, sugar phosphate, purines, pyrimidines, and aromatic compounds. The derivatization reaction proved. to be efficient (> 50% transferred to derivatized form) and repeatable (relative standard deviations < 10%). Linearity for most metabolites was satisfactory with regression coefficients better than 0.996. Quantification limits were 40-500 pg on-column or 0.1-0.7 mmol/g of microbial cells (dry weight). Generally, intrabatch precision (repeatability) and interbatch precision (reproducibility) for the analysis of metabolites in cell extracts was better than 10 and 15%, respectively. Notwithstanding the nontargeted character of the method and complex microbial matrix, analytical performance for most metabolites fit the requirements for target analysis in bioanalysis. The suitability of the method was demonstrated by analysis of E. coli samples harvested at different growth phases.
引用
收藏
页码:1272 / 1281
页数:10
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