Influenced by the new crown epidemic, online teaching has entered the public’s view, and exploring the hybrid teaching between online and offline has become the hot content of university Chemistry course practice nowadays. This paper systematically studies the willingness of participants, influencing factors, support systems, and the evaluation of the effects of the knowledge interaction in the university Chemistry SPOC hybrid teaching mode. The teaching effect evaluation index system of the new mode is constructed using the AHP method of covariance correction. Taking part of the textbook of the university Chemistry course as an example of course design, a two-month teaching practice is carried out in the Chemistry major of X university through the 12xue platform to explore the advantages and disadvantages of the SPOC online and off-line blended teaching mode in the teaching of university Chemistry. The experimental results show that the adoption of the SPOC blended teaching mode in university Chemistry teaching practice has a certain effect on improving students’ Chemistry performance, with a performance improvement of 43.95 percentage points. And it helps students improve their independent learning abilities. The exploration of this teaching method provides a new way for college Chemistry teachers to enhance their teaching methods.