Design and Practice of Undergraduate Biochemistry Experiments Based on the DOK Model: A Case Study of HPLC Quantification of TMZ in Mouse Tissues

While biochemistry laboratory instruction continuously evolves to enhance student learning, effectively cultivating higher‐order thinking skills in complex experimental settings remains an ongoing challenge. By applying the Depth of Knowledge (DOK) model, we designed and implemented a comprehensive experiment titled “Determination of Temozolomide Content in Mouse Tissues Using High‐Performance Liquid Chromatography (HPLC)” for undergraduate bioscience majors. This experiment helps students understand drug tissue distribution mechanisms while filling a curricular gap in teaching pharmacokinetic analysis in complex biological samples. Based on the DOK framework, we designed a four‐level task chain—Recall, Skill, Strategy, and Innovation—shifting the pedagogical focus from basic technical training to the systematic cultivation of higher‐order thinking skills. Educational evaluation revealed high student satisfaction and demonstrated that the DOK‐based approach significantly improved students' higher‐order cognitive abilities, particularly in strategic analysis and innovation, compared to traditional instruction. Consequently, this study provides an effective practical case and a transferable paradigm for systematically fostering higher‐order thinking in science laboratory education.