The work aims to prepare microbial time-temperature indicators (TTIs) for monitoring the quality of food products during storage and transportation. Lactobacillus acidophilus was selected as the microorganism of the time-temperature indicator, and nine mixed pH indicator solutions were prepared for screening. The TTI was prepared with suitable mixed pH indicator solutions, and the indicators were evaluated by adding different concentrations of Lactobacillus acidophilus and stored under different isothermal conditions (5, 15 and 25 ℃), and the kinetics and temperature dependence were investigated with suitable indicator parameters as response values. Five of the nine mixed pH indicator solutions had significant color changes, one of which was used to prepare the TTI, and the activation energy (Ea) was 84.9 kJ/mol with the total color difference value (∆E) as the response parameter. Five mixed pH indicator solutions with significant color changes can be used as indicators for the TTI, and the prepared microbial-type TTI has potential to be applied in shelf life prediction of food with an activation energy of quality index from 59.9 kJ/mol to 109.9 kJ/mol.