Abstract: To optimize the chlorophyll extraction method in plant physiology experiments, a freeze grinder was used instead of a mortar in the traditional method with green pineapple as the experimental material and using chlorophyll content as the indicator. Based on a single factor experiment, a Box-Behnken experimental design was carried out, and response surface optimization experiments were conducted for liquid solid ratio, extraction time, and extraction frequency. The results indicated that the primary and secondary relationships of the three factors were as follows: extraction time>extraction frequency>liquid-solid ratio. Based on the response surface optimization experiment results and actual operation, the optimal extraction process parameters were determined as follows: a liquid solid ratio of 15 m/L, extraction time of 45 s, extraction frequency of 65 Hz, and chlorophyll content of (1.7270±0.0197) mg/g. The optimization method was accurate and reliable. The consumption of extraction reagents and operation time accounted for 1/5 and 1/3 of that of the traditional method, respectively, and the chlorophyll content has increased by 32.00% compared with that of the traditional method. This method can be applied to teaching in plant physiology experiments, enriching the experimental content and cultivating and improving students’ scientific thinking and practical innovation abilities. It can also be used for extraction in batches to improve efficiency and achieve the goal of saving reagents and time.

Key words: chlorophyll, extraction method, experimental improvement, plant physiology experiment, response surface methodology