|Title||Acidic acetonitrile for cellular metabolome extraction from Escherichia coli.|
|Publication Type||Journal Article|
|Year of Publication||2007|
|Authors||Rabinowitz, JD, Kimball, E|
|Date Published||2007 Aug 15|
|Keywords||Acetonitriles, Amino Acids, Carbon, Chemical Fractionation, Escherichia coli, Gas Chromatography-Mass Spectrometry, Metabolism, Nucleotides, Solvents|
Cellular metabolome analysis by chromatography-mass spectrometry (MS) requires prior metabolite extraction. We examined a diversity of solvent systems for extraction of water-soluble metabolites from Escherichia coli. Quantitative yields of approximately 100 different metabolites were measured by liquid chromatography-tandem MS and displayed in clustered heat map format. Many metabolites, including most amino acids and components of central carbon metabolism, were adequately extracted by a broad spectrum of solvent mixtures. For nucleotide triphosphates, however, mixtures of acidic (0.1 M formic acid-containing) acetonitrile/water (80:20) or acetonitrile/methanol/water (40:40:20) gave superior triphosphate yields. Experiments involving isotopic tracers revealed that the improved triphosphate yields in the acidic acetonitrile were in part due to reduced triphosphate decomposition, which is a major problem when extracting with other solvent systems such as methanol/water. We recommend acidic solvent mixtures containing acetonitrile for extraction of the E. coli metabolome.
|Alternate Journal||Anal. Chem.|