Heavy metals content in tires affects the safety of soil and agricultural products. The digestion method is a pretreatment for determining heavy metals in tire samples, and will affect the efficiency and accuracy of the heavy metal determination. The microwave digestion process and reagents for tire samples are not currently standardized. Therefore, this study attempts to provide an appropriate method of resolution for scholars. All digestion processes were performed in Mars One. We tested 15 different acid mixtures to determine the best reagent type and dose and then investigated the effect of maximum temperature, holding time, and sample grams on the degree of digestion. In summary, the best condition to digest the tire sample was a mixture of 3 ml HNO3 and 7 ml H2SO4, taking 0.1 (± 0.0005) g tire sample, at the maximum digestion temperature of 220 °C for 25 min. The experimental conclusion will provide a reliable experimental method for scientists using MARS One to study heavy metals in tires. At the same time, researchers using the MARS series can also find valuable references in this paper.