In this work, UiO-66 with defects was successfully prepared by a synthesis strategy of using benzoic acid as a modulator and postsynthetic acid treatment. The defective frameworks can be confirmed by N2 adsorption–desorption analysis and 1H NMR. It is observed that this strategy can effectively enlarge the surface area and pore volume of stable UiO-66 through the removal of coordinated benzoate ligands. The resulting defective UiO-66 shows high BET surface area and total pore volume (1890m2/g and 0.88cm3/g), which are both larger than those of defect-free sample (1200m2/g and 0.49cm3/g) as well as all the reported UiO-66s with defects so far to the best of our knowledge. This MOF exhibits significantly improved capture ability (366mg/g) toward Safranine T (ST) compared with the defect-free UiO-66 (39mg/g) and most of reported adsorbents. Meanwhile, it also shows high selectivity for ST over Crystal Violet (CV) due to the size-exclusion effect constructed from the defects in the framework. The results show that this work provides a promising strategy to rationally design novel MOFs for separating large molecules.