The non-stoichiometric, calcium-deficient hydroxyapatite was prepared through a low-temperature from aqueous solutions method and characterized using Physico-chemical methods. The potential of this hydroxyapatite to adsorb and release insulin from aqueous solutions was evaluated under physiological conditions. The effect of contact time and initial concentration were studied in batch experiments. The adsorption rate reached up to 81±5% in the first half-hour of contact, while the release rate of insulin incubation was about 41 ± 5% after 1 hour. The pseudo-first-order, pseudo-second-order, Elovich equation, Weber and Morris intraparticle diffusion model and Bangham’s pore diffusion model were applied to study the kinetics of the adsorption process. The pseudo-second-order kinetic model provided the best correlation R²(0.998) of the used experimental data compared to the other models. The adsorption of insulin onto hydroxyapatite was correlated well R²(0.998) with the Langmuir model as compared to Freundlich, Temkin and Dubinin–Kaganer–Radushkevich (D-K-R) models, with a maximum adsorption capacity of 24.46 mg/g. The isotherms parameters values of ΔG⁰, b_t and E show that the adsorption process is favorable, spontaneous, exothermic, and controlled by physisorption. The point of zero charge (pHZPC) of hydroxyapatite and the isoelectric point (pI) of insulin indicate that the interaction of insulin molecules with prepared apatite can be well described as an ions exchange reaction.