Abstract: In the experiment for determination of reaction rate constant of sucrose hydrolysis and activation energy, the circular polarimeter is usually used as the instrument of measuring optical rotation. How to use the instrument has always been a sticking point in experimental teaching. It is often difficult for students to correctly grasp the essentials of the instrument’s operation only through the teacher’s on-site demonstration. In order to solve this problem, a kind of virtual simulation software for determination of sucrose hydrolysis reaction rate constant and activation energy was designed and developed. At the same time, it was used in experimental teaching by combining virtual and real teaching methods. Through classroom feedback and the analysis of students’ experimental results, it is found that using the software can promote students’ in-depth understanding of experimental principles as well as operation points and improve their experimental performance.