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Electrochemical sensors have long suffered from issues such asnonspecific adsorption, poor anti-interference ability, and internal and externaldisturbances. To address these challenges, we developed a facile electrochemicalmethod, which integrated a ratiometric strategy with self-cleaning electrodes. In thenovel sensing system, the self-cleaning electrode was realized via forming ahydrophobic layer on carbonized ZIF-67@ZIF-8 (cZIF) by polydimethylsiloxane(PDMS) precursor vaporization. As for ratiometry, it is worth to mention that themeasurements were conducted by adding an interior reference (methylene blue)directly into electrolyte solution, which is more facile andflexible to operatecompared with conventional ones. Sensing performance of the self-cleaning electrodeas well as the newly established ratiometric strategy was explored fully, and it turnedout that PDMS@cZIF nanocomposites provided decent electrocatalytic ability,superhydrophobic property, and stability. Furthermore, the ratiometric strategysignificantly elevated the robustness and reproducibility of electrochemical sensing.Simultaneous detection of Adr, 5-HT, and Trp was performed under the optimum experimental conditions with wide linearranges and low detection limits. Finally, the original ratiometric electrochemical sensor was successfully applied for monitoring the three target molecules in biological samples.