The ubiquitous prevalence of ciprofloxacin (CIP) in aquatic environments has prompted concerns about its durability, ecological effect, and contribution to antimicrobial resistance. To address this difficulty, analytical extraction procedures must be both sensitive and environmentally friendly. This paper describes a green dispersive solid-phase extraction (DSPE) method for CIP determination that uses graphene oxide (GO) derived from sugarcane bagasse as a renewable biomass precursor. GO was synthesized in a single step using a low-temperature thermal technique and then characterized using FTIR, SEM-EDX, and XRD, showing the successful production of oxygen-rich and structurally disordered GO suited for adsorption applications. The DSPE parameters were systematically tuned using Response Surface Methodology (RSM) with a Box-Behnken Design (BBD) to investigate the effects of pH, GO mass, and contact time on CIP adsorption efficiency. The quadratic model well represented adsorption behavior, with a high coefficient of determination (R² = 0.9726) and great prediction dependability. GO mass was found to be the most influential factor, with pH and contact duration serving as secondary parameters that fine-tuned the adsorption process. Under optimal conditions (pH 3.16, GO mass 43.00 mg, contact time 29.94 min), the model projected a 90.46% CIP adsorption efficiency. The experimental validation resulted in an average adsorption of 88.39%, with a small prediction error of 2.34%, demonstrating excellent agreement between anticipated and experimental results. These findings show that sugarcane bagasse-derived GO is a very effective and sustainable adsorbent for DSPE-based CIP analysis, providing a dependable platform for monitoring trace antibiotics in aquatic systems.