Resulted　from　the　severe　side　effects,　the　development　of　inexpensive,　simple　and　sensitive　method　for　amethopterin　(ATP,　an　antineoplastic　drug)　is　very　important　but　it　still　remains　a　challenge.　In　this　work,　low　cost　nanohybrid　composed　of　carbon　nanobowl　(CNB)　and　β-cyclodextrins　(β-CD)　(CNB-CD)　was　prepared　with　a　simple　autopolymerization　way　and　applied　as　electrode　material　to　develop　a　novel　electrochemical　sensor　of　ATP.　Scanning-/transmission-electron　microscopy,　Fourier　transform　infrared　spectrum,　photographic　image　and　electrochemical　technologies　were　utilized　to　characterize　morphologies　and　structure　of　the　as-prepared　CNB　and　CNB-CD　materials.　On　the　basic　of　the　coordination　advantages　from　CNB　(prominent　electrical　property　and　surface　area)　and　β-CD　(superior　molecule-recognition　and　solubility　capabilities),　the　CNB-CD　nanohybrid　modified　electrode　exhibits　superior　sensing　performances　toward　ATP,　and　a　low　detection　limit　of　0.002　μM　coupled　with　larger　linearity　of　0.005–12.0　μM　are　obtained.　In　addition,　the　as-prepared　sensor　offers　desirable　repeatability,　stability,　selectivity　and　practical　application　property,　confirming　that　this　proposal　may　have　important　applications　in　the　determination　of　ATP.　©　2024　The　Authors