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Magnetic Fe3O4-graphene was synthesized.Fe3O4-graphene nanocomposite was used for NSAIDs extraction in human plasma and urine.Low LOQ and high extraction recovery were obtained.Development of UHPLC-PDA method for NSAIDs analysis.A novel, rapid, simple graphene/Fe3O4 based dispersive magnetic solid phase extraction was developed for the simultaneous separation/preconcentration and determination of non steroidal anti-inflammatory drugs with ultra high performance liquid chromatography coupled with photodiode array detection. Several parameters influencing the extraction efficiency of the investigated analytes such as the extraction time, the amount of graphene/Fe3O4, the sample pH, the ionic strength and the elution solvent were evaluated and optimized. Under optimal conditions, the linearity was in the range of 0.002–25 μg/mL for furprofen, diclofenac and ketoprofen, 0.003–25 for flurbiprofen, naproxen and fenbufen, 0.004–25 for indoprofen with a good coefficient of determination (R2> 0.9991) for each analyte. The inter-and- intra day accuracy (BIAS%) for human plasma and urine ranged between −7.15% to 6.20% and −5.17% to 4.87%, respectively.The precision (RSD%) in human plasma and urine was less than 8.67% and 8.92%, respectively. The proposed method was applied to the determination of NSAIDs in human plasma and urine.