In this work, metal-organic framework compound CPL-1 of a formula {[Cu₂(pzdc)₂(pyz)]⋅2H₂O}_n (pzdc = pyrazine-2,3-dicarboxylate, pyz = pyrazine) possessing kinetic quantum sieving effect on H₂/D₂ separation was selected to prepare two new chromatography stationary phase materials [email protected]γ-Al₂O₃ and MnCl₂@[email protected]γ-Al₂O₃ by loading CPL-1 on γ-Al₂O₃ particles with size 80–100 mesh through repeating in situ crystallization process, and afterwards by loading MnCl₂ on [email protected]γ-Al₂O₃ via impregnation approach. The two optimized materials were thoroughly characterized by IR, XRD, SEM, EDS and TG/DTA method. The experimental results showed that, under liquid nitrogen temperature by using just 1-m chromatographic column and cheap He gas as carrier gas, the MnCl₂@[email protected]γ-Al₂O₃ can solve the problem of long separation time and trailing phenomenon existing in the [email protected]γ-Al₂O₃ column, can realize highly effective hydrogen isotope H₂/D₂ separation with good separation resolution (R = 1.62) and approximate Gauss distribution of chromatographic peak, can be used for both qualitative and quantitative analysis of hydrogen isotope H₂/D₂ with short separation time of 6.5 min.