A series of novel diglycolamide (DGA) ligands containing cyclohexyl substituent on the amidic N atom were investigated for their solvent extraction and complexation ability toward Sr2+ in HNO3 solution. Compared with the most commonly used N,N,N′,N′-tetraoctyldiglycolamide (TODGA), the introduction of cyclohexyl substituent into the DGA skeleton can significantly enhance the extractability toward Sr2+ in the case of toluene as the diluent. And the more cyclohexyl substituents there are, the more significant the effect is. About 8−80 fold increase of the Sr2+ distribution value for LI−LVI ligands can be achieved than that for TODGA at 2.0 mol/L HNO3. The extractability of these cyclohexyl substituted DGA ligands follows the order of LI ≈ LIII > LII > LIV > LV > LVI > > TODGA. Meanwhile, for the DGA ligands with the same number of cyclohexyl substituents, LIII with moderate alkyl chain length (n-butyl) has higher extractability than LII with short chain (n-ethyl) and LIV with long chain (n-octyl). Slope analysis reveals the formation of 1:3 metal-ligand extracted species during the extraction process, which is also confirmed by the results obtained from ESI-MS, FT-IR and single-crystal X-ray diffraction analyses. Furthermore, the single crystal of DGA ligand with Sr(NO3)2 was obtained for the first time, and the analysis of its structure indicates that each DGA molecule participates in the complexation with the 9-fold coordinated Sr2+ through two amide carbonyl O atoms and one ether O atom, accompanied by two free NO3− in the outer coordination sphere. The extraction of Sr2+ by the novel DGA ligand is a spontaneous and exothermic process of increasing entropy, and follows neutral complexing extraction model. In addition, an extraction test of Sr2+ from simulated high level liquid waste was also carried out and demonstrates a highly efficient and selective separation of Sr2+ over the other co-existing ions.