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[Display omitted] • Developed an ecologically safe chitosan-carbon nanofiber-based membrane. • Functionalized membrane used as bioelectrode for detection of oxytocin. • The developed bioelectrode showed an LOD of 24.98 ± 11.37 pg/mL of oxytocin. • The material has potential as a disposable alternative to conventional electrodes. Bioelectrodes with low carbon footprint can provide an innovative solution to the surmounting levels of e-waste. Biodegradable polymers offer green and sustainable alternatives to synthetic materials. Here, a chitosan-carbon nanofiber (CNF) based membrane has been developed and functionalized for electrochemical sensing application. The surface characterization of the membrane revealed crystalline structure with uniform particle distribution, and surface area of 25.52 m2/g and pore volume of 0.0233 cm3/g. The membrane was functionalized to develop a bioelectrode for the detection of exogenous oxytocin in milk. Electrochemical impedance spectroscopy was employed to determine oxytocin in a linear concentration range of 10 to 105 ng/mL. The developed bioelectrode showed an LOD of 24.98 ± 11.37 pg/mL and sensitivity of 2.77 × 10-10 Ω / log ng mL−1/mm2 for oxytocin in milk samples with 90.85–113.34 percent recovery. The chitosan-CNF membrane is ecologically safe and opens new avenues for environment-friendly disposable materials for sensing applications. [ABSTRACT FROM AUTHOR]