earticle

영어

Metabolomics require the gain of reproducible, robust, reliable, and homogeneous biological data sets. Intracellular metabolite is a reflection of all the metabolic functions of an organism under any particular growth condition. For the development of robust experimental procedure, extraction of the metabolome to quantitative analysis of the metabolites is required. E. coli was grown in chemostat culture so that cellular metabolism could be held in reproducible, steady-state conditions under a range of precisely defined growth conditions, thus enabling sufficient replication of samples. This investigation indicates that 60% cold (-40°C) methanol solution is the most appropriate method to quench metabolism. In this study, metabolites were extracted from E. coli using six different commonly used procedures including acid or alkaline treatments, high-temperature extraction in the presence of ethanol or methanol, lysis with chloroform-methanol, and extraction by hot water. Intracellular metabolites were extracted by the six methods from cells grown under identical conditions. The intracellular metabolite profiles were generated using HPLC system coupled with an ion trap mass spectrometer equipped with a turbo-ion spray source.