A　simple,　reliable　and　reproducible　method　for　the　separation　and　determination　of　five　β-casomorphins　(β-CMs,　namely　TPGN,　PGPI,　TPGI,　TPGP　and　TPPG)　based　on　glass　microfluidic　chip　electrophoresis　and　laser-induced　fluorescence　detection　is　first　described　in　here.　The　microfluidic　chip　electrophoresis　and　laser-induced　fluorescence　detection　system　consisted　of　a　home-made　glass　＂double-T＂　microchip　and　a　simple　LIF　detector　with　excitation　and　emission　wavelengths　of　473　and　525　nm,　respectively.　Fluorescein　isothiocyanate　(FITC)　was　used　as　the　precolumn　derivatization　reagent　to　label　fluorophore　on　five　β-CMs,　and　the　optimum　conditions　of　FITC-derivatization　reaction　and　MCE　separation　were　investigated　in　detail.　Under　optimum　conditions,　five　β-CMs　were　completely　separated　and　detected　within　30　min　with　a　detection　limit　of　18.7-75.1　nmol/L　and　an　RSD　(n=5)　of　3.0-5.9%,　respectively.　The　proposed　method　has　been　successfully　used　to　detect　β-CMs　in　real　cheese　sample　with　a　recovery　of　89-109%,　suggesting　that　our　method　is　sensitive　and　reliable.　These　features,　as　well　as　its　low　cost,　operation　convenience,　stability　and　reusability,　make　it　a　promising　alternative　to　β-CMs　detection　methods.　Copyright　©　2011　WILEY-VCH　Verlag　GmbH　&　Co.　KGaA,　Weinheim.