The efficient degradation and accurate quantification of tetracycline in environment and food samples is pivotal for ensuring public health and safety by monitoring potential contamination and maintaining regulatory standards. Hence, in this study, photocatalytic degradation of tetracycline and its electrochemical detection in environment and food samples based on dual-functional silver-doped zinc ferrite nanoparticles embedded chitosan-functionalized carbon nanofibers fabricated on a screen-printed carbon electrode (AgZFO/CHIT-CNF/SPCE) is presented. A hydrothermal method was used in the synthesis of Ag-doped ZFO, and chitosan was functionalized on the CNF surface using a swift and cost-effective chemical modification process of carboxyl groups. Various techniques, such as XRD, HRTEM, elemental mapping, EIS, XPS, FTIR, VSM, BET, UV–Vis DRS, and Raman spectroscopy were used to analyze the characteristics of the prepared nanocomposite. Cyclic voltammetry and differential pulse voltammetry were used to evaluate the surface-controlled electrocatalytic properties of AgZFO/CHIT-CNF towards tetracycline. Electrochemical tests revealed that the proposed electrode exhibited excellent sensitivity for detecting tetracycline. The fabricated electrode had a low detection limit of 1 nM and a wide linear range (0.2–53.2 μM). The sensor also demonstrated exceptional selectivity, stability, and reusability. The practical feasibility evaluated with real samples, including chicken feed, shrimp, milk, soil, and wastewater, resulted in high recovery values.