Abstract: The objective of this study was to extract cembratriene-diols in tobacco flowers environment-friendly and to establish the optimum process conditions of supercritical CO₂ extraction. The single factor test and response surface analysis were used to explore the influence of extraction time, extraction temperature, extraction pressure and the entrainer flow on the extraction amount of cembratriene-diols in tobacco flowers. The central composite design was carried on the basis of single factor experiment. The optimal extraction conditions were found to be extraction at 42 ℃ and 13.9 MPa for 120 min using ethanol as the entrainer at a flow velocity of 0.67 g/min. Under this condition, the predicted extraction amount of cembratriene-diols was 6.065 mg/g based on the response surface model. In this study, the actual extraction amount and extraction rate of cembratriene-diols were 5.978 mg/g and 97.1%, respectively. This suggested that efficient extraction of cembratriene-diols in tobacco can be achieved using the central composite design and response surface analysis.