Metabolomics is a new branch of systems biology exerting its influence in many aspects. In order to appraise the effects of antibiotics on central carbon metabolism, a CE based method was set up. With this platform, we estimated the organic acid metabolite pools' fluctuation of Escherichia coli and Pseudomonas aeruginosa cultured under 11 different antibiotics' stimuli. Multivariate data analysis showed that different antibiotics had clustered distributions for each strain and could be easily distinguished. Genetic, metabolic and antibiotic mechanism differences could also be deduced by the aid of further correlation analysis. For P. aeruginosa, even synergy action amid antibiotics could be ascertained.