We　report　on　a　new　electrochemical　biosensing　strategy　for　the　sensitive　detection　of　hydrogen　peroxide　(H2O2)　in　foodstuff　samples.　It　is　based　on　a　gold　electrode　modified　with　layer　of　graphene　patterned　with　a　multilayer　made　from　an　organic-inorganic　hybrid　nanomaterial.　Initially,　a　layer　of　thionine　(Th)　was　assembled　on　the　surface　of　the　graphene　nanosheets,　and　these　were　then　cast　on　the　surface　of　the　electrode　for　the　alternate　assembly　of　gold　nanoparticles　and　horseradish　peroxidase.　The　large　surface-to-volume　ratio　and　high　conductivity　of　the　nanosheets　provides　a　benign　microenvironment　for　the　construction　of　the　biosensor.　The　use　of　such　a　multilayer　not　only　shortens　the　electron　transfer　pathway　of　the　active　center　of　the　enzyme　due　to　the　presence　of　gold　nanoparticles,　but　also　enhances　the　electrocatalytic　efficiency　of　the　biosensor　toward　the　reduction　of　H2O2.　The　electrochemical　characteristics　of　the　biosensor　were　studied　by　cyclic　voltammetry　and　chronoamperometry.　The　number　of　layers,　the　operating　potential,　and　the　pH　of　the　supporting　electrolyte　were　optimized.　Linear　response　is　obtained　for　the　range　from　0.5　μM　to　1.8　mM　of　H2O2,　the　detection　limit　is　10　nM　(at　S/N　=　3),　and　95%　of　the　steady-state　current　is　reached　within　2　s.　The　method　was　applied　to　sense　H2O2　in　spiked　sterilized　milk　and　correlated　excellently　with　the　permanganate　titration　method.　©　2011　Springer-Verlag.