Abstract: In order to improve the research and utilization of chemical components in air-dried tobacco, two fluorescent small molecules, 2-methylquinoline (QND) and 3-indole (YD), were isolated from Yunnan local air-dried tobacco. Then, their spectral analysis, pharmacological toxicity and imaging analysis were studied. The results showed that two small fluorescent molecules possessed high fluorescence quantum yields, stable response to environmental pH changes and complementary trend. The cell experiments had also showed that the two molecules were low cytotoxicity and good cell imaging, which showed the feasibility of application in the biological field. In summary, these results might show a good application prospect for the comprehensive application of air-dried tobacco resources and might explore the new usages of tobacco.