Hydrogen isotope separation is a complex task due to the nearly identical physical and thermodynamic properties of isotopes, such as deuterium and protium. Traditional methods, including cryogenic distillation, exhibit limitations such as low selectivity and high energy consumption. Recent advancements utilizing the quantum sieving effect in crystalline porous materials have shown promise under cryogenic conditions, but experimental approaches using larger, more practical sample sizes remain scarce. This study introduces a novel cryogenic breakthrough apparatus designed for hydrogen isotope separation using gram-scale adsorbents. The apparatus supports both refrigerant-based and contact-cooling temperature control methods, with precooling employed to enhance separation efficiency. Results showed that precooling significantly improved deuterium selectivity over hydrogen, and the contact-cooling method demonstrated superior thermal stability and reliability during extended experiments. This system offers a scalable and practical solution for hydrogen isotope separation, addressing current limitations in experimental methodologies and providing valuable insights for both scientific research and industrial applications.