In this study, a two-step modification of sepiolite for adsorption enhancement was investigated. The cetyltrimethylammonium bromide (CTAB) was utilized for the organic modification process after a heat modification. To develop the optimal modification condition, adsorption of Acid Orange II onto modified sepiolite was investigated with respect to heat temperature and adsorbent dosage. The temperature of 200 °C and 100% cation exchange capacity (CEC) was deemed as the optimal condition. The impacts of operation conditions on adsorption procedure, including pH, adsorbent dosage and adsorption duration, were comprehensively discussed. The adsorption of Acid Orange II by sepiolite is in accordance with the quasi-secondary kinetic model. Moreover, the results of intraparticle diffusion indicate that the intraparticle diffusion was the dominant adsorption force in the initial adsorption period. The adsorption process was obeyed with the Langmiur adsorption model. The results from regeneration procedure suggest that the superior regeneration obtained with 0.8 mol/L NaOH concentration.