Abstract: To overcome the limitations of current sweetness measurement methods for cigarette smoke and achieve objective quantification of sensory sensation, a HEK293 cell line overexpressing human sweet taste receptors T1R2/T1R3 was established, and by employing calcium imaging technology to monitor real-time changes in intracellular calcium signals induced by sweet substances, quantitative characterization of sweetness response was realized. On this basis, the sweetness values of ten representative sweet substances previously reported in smoke, including furfural and furanone, were systematically determined. Furthermore, the influence of the total particulate matter (TPM) matrix on the sweetness performance of these compounds was investigated. The results showed that: 1) Most sweet substances exhibited varying degrees of reduced sweetness values in TPM matrix, which may be attributed to the competitive or non-competitive inhibition on sweet receptors by other complex smoke components. 2) Partial substances (fenugreek lactone and 3,5-dimethyl-1,2-cyclopentanedione) maintained stable or slightly enhanced sweetness in the TPM matrix, suggesting that the smoke matrix exerted differential effects on sweetness of different sweet substances and might even involve synergistic effects. These findings suggest that cigarette flavor regulation should focus on components that exhibit stable or enhanced sweetness in the smoke matrix.