Xylazine (XYL) is veterinary drug administrated for sedative and analgesic purposes. With severe side effects, XYL is not approved for the human use. Recently, XYL has been categorized as a drug of abuse, and has been associated with drug-facilitated sexual assaults. Therefore, the present study described the electrochemical behavior of XYL at novel carbon paste working electrodes integrated with different metallic oxides nanoparticles. Among them, carbon paste matrices fortified with zinc oxide nanoparticles (ZnONPs) showed a noticeable electrocatalytic activity towards the oxidation of XYL with an anodic irreversible oxidation peak at 0.885 V in Britton-Robinson (pH 7.0). Oxidation of XYL molecule at the electrode surface followed diffusion-reaction mechanism through the participation of two-electrons and two protons as confirmed by the molecular orbital calculations and electroanalytical parameters studies. A simple and accurate quantification protocol using square wave voltammetry offered enhanced sensitivity towards XYL within a wide concentration range from 29 to 806 ng mL–1 XYL, with limit of detection 5.44 ng mL–1. The fabricated sensors showed high measurement reproducibility with a prolonged. The ZnONPs/CPEs were introduced for monitoring of XYL residues in veterinary matrices with high accuracy and precision. Moreover, to forensic scenario, a standard addition technique on plasma and urine samples reveled acceptable recovery values, demonstrating the potentiality to forensic and clinical analysis of xylazine intoxication.