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Since cefixime is considered a widely used antibiotic in many countries, it was important to develop a method for its removal from water as a part of water purification using an adsorption technique with different nanoparticles. In order to optimize its removal conditions, a fractional factorial design was applied to screen experimental factors including pH, contact time, and amounts of nanoparticles. A three-factor, two-level I-optimal design was designed using Derringer\u2019s desirability algorithm, in which the optimal removal conditions of cefixime were found to be Fe<jats:sub>3<\/jats:sub>O<jats:sub>2<\/jats:sub> dose (13\u00a0mg\/L), pH (5.9), and contact time (180\u00a0min) at room temperature. An HPLC method was developed for monitoring the adsorption process using a Kinetex C<jats:sub>18<\/jats:sub> stationary phase (100\u00a0mm L \u00d7 4.6\u00a0mm i.d., particle size 5\u00a0\u03bcm, USA), and a mobile phase consisting of phosphate buffer (adjusted to pH 6.8) and methanol in the ratio of 75: 25 which was pumped at a flow rate of 1 mL\/min at room temperature with UV detection at 288\u00a0nm. The proposed HPLC method was validated according to ICH guidelines and was assessed using the greenness tools Analytical Eco-scale (AES), GAPI, and AGREE. The removal and analysis procedures were successfully applied to simulated wastewater samples containing cefixime with a bias of 1.9%. 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