Abstract: The purpose of this paper is to find the optimal conditions for glucosinolate extraction by response surface method and the effect of this extraction process on the nutritional content of rapeseed meal, which can provide information for promoting the valorization of rapeseed meal. The single-factor experiment was designed with extraction time, temperature and pH as treatment factors. The extraction of glucosinolate was carried out under the condition of material-liquid ratio of 1:10. The significant factors were optimized by responding to the surface design to get optimal extraction conditions of glucosinolate, and change of content of some nutrients was determined. Crude protein, fat and total sugar were determined according to GB/T 6432-2018, GB 5009.6-2016 and GB 5009.8-2016, respectively, and the glucoside content was determined by palladium chloride. The result of univariate experiments showed that the extraction time and temperature had significant influences on the extraction amount of glucosinolate. Since the extraction pH had little effect, the response surface design of two factors and three levels was selected. The response surface of the above factors was enhanced by Design Expert of central composite experimental design, and the software showed that the best conditions were extraction time of 28.87 min and extraction temperature of 90℃. The maximum extraction amount of glucosinolate was expected to be 35.01 mg/g. The experimental results showed that when the extraction time was 28 min and the extraction temperature was 90℃, the extraction amount of glucosinolate in rapeseed meal reached 34.32 mg/g, and the relative error with the theoretical value was only 2%. Meanwhile, the protein content of rapeseed meal after extraction was 42.76%. Ethanol can be used for extraction of glucosinolate from rapeseed meal and high level of protein was retained in the rapeseed meal after extraction. This study can provide valuable information for the valorization of rapeseed meal.