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Numerous studies have been undertaken to develop and provide validity evidence for using scales to measure students’ motivation in science learning. This study aims to adapt the Science Motivation Questionnaire II (SMQ II) in Chinese cultural context. In two large samples (total N = 1124), we evaluated its psychometric properties: internal structure validity, and criterion-related validity. In addition, the measurement invariance results revealed the structure of the SMQ II-C is equivalent within subgroups (e.g. gender, grade level). Overall, this study suggests the SMQ II-C is a robust instrument to evaluate Chinese students’ science motivation. Significant gender differences were found among all five subscales of science motivation.