The re-crystallization of 2,4-hexadiyn-1,6-dialkylurea, the preparation of co-crystallization and its color development kinetics were studied, which provided theoretical guidance for the commercial application of Vaccine vial monitor. Through the re-crystallization of 2,4-hexadiyn1,6-diethylurea with different solvents and the co-crystallization test of 2,4-hexadiyn-1,6-bis(ethylurea)(KE) and 2,4-hexadiyn-1,6-bis(propylurea)(KPr) with different proportions, the most suitable formula was determined, and its color development kinetics was studied. The optical density was measured by spectrophotometer.The results showed that the solid-state polymerization reaction rates of recrystallizing 2,4-hexadiyne-1,6-bis(ethylurea) with different solubility solvents were different. Through experiments, it is found that the product obtained by recrystallization with the mixed solution of methanol∶mwater=3∶1 and cocrystallization at mKE∶mKPR=2∶1 has better thermal reactivity and distinct color change, and the activation energy of the solid-state polymerization reaction is calculated to be 31.94 kJ/mol. 2,4-hexadiyn-1,6-dialkylurea is a new chemical time and temperature indicator, and its visual response signal is easy to recognize. Matching different kinds of vaccines by re-crystallization and modification of co-crystallization can directly reflect the quality of vaccines in cold chain transportation, preservation and use.