Theophylline (1,3-dimethyl-2,3,6,7-tetrahydro-1-hydrogen-purine-6,2-dione) is a bronchodilator drug used in the treatment of asthma and chronic obstructive pulmonary disease (COPD). Due to the narrow therapeutic strip of this drug, it is important to measure its blood level.To achieve a simple, safe and sensitive method based on dispersive solid phase extraction (DSPE) using magnetite nanoparticles covered with nanocellulose and beta-cyclodextrin for preconcentration and measurement of theophylline in aqueous samples and its validation, this method was used. In this research, variables affecting the extraction such as pH, ionic strength, and amount of adsorbent, temperature, time, limit volume and type of solvent were optimized. After extraction, the adsorbent was easily separated from the aqueous sample using an external magnetic field without filtration or centrifugation. The results showed that optimum conditions were obtained with pH=5, 1 mg of sodium chloride, 0.2 g of adsorbent with a capacity of 0.53 mg/g, temperature of 40 degrees Celsius, 20 minutes of contact between the adsorbent and the sample and 5 mL of methanol as the desorption solvent. In optimal conditions, the suitable concentration factor is 12, low detection limit (0.17 mg/L), wide linear range and in accordance with the therapeutic strip of the drug (5-50 mg/L), good correlation coefficient (R2=0.9991) and good accuracy (RSD=1.03%) was obtained.Urine sample was used to validate the above method. Also, it was found that the proposed method is suitable, efficient and usable for measuring of theophylline. Manuscript profile