Corynebacterium　glutamicum　(C.　glutamicum) is　a　well-known　workhorse for　the　industrial　production　of　amino　acids.　Different　carbon,　nitrogen,　and　sulfur　source　may　force　the　bacterium　to　produce　specific　metabolites.　In　this　work,　a　method　of　high-speed　MEKC　with　LIF　detection　was　developed　to　rapidly　analyze　the　amino　acid　metabolites　released　by　C.　glutamicum,　which　is　fed　with　different　culture　mediums.　Corynebacterium　glutamicum　was　cultured　in　microbial　fuel　cells　to　monitor　its　metabolism　process　and　collect　its　metabolites.　In　the　CE　system,　a　microliter-scale　sample　reservoir　was　designed　and　applied　to　perform　tiny　volume　sample　injection.　With　the　assistance　of　microwave,　the　derivatization　time　for　amino　acids　with　FITC　was　greatly　shortened　to　6 min.　Under　the　optimized　condition,　the　eight　candidate　amino　acids　of　metabolites　could　be　rapidly　separated　within　2 min.　The　whole　analysis　process　for　real　samples,　from　sampling　to　determination,　could　be　shortened　to　less　than　10 min.　The　results　showed　that　C.　glutamicum　could　produce　additional　l-lysine　and　l-valine　as　the　metabolites　when　fed　with　glucose　and　l-methionine,　respectively.　The　method　proved　that　culture　mediums　used　to　feed　C.　glutamicum　had　great　effect　on　the　bacterium＇s　metabolites.　©　2019　WILEY-VCH　Verlag　GmbH　&　Co.　KGaA,　Weinheim