Background: Achievement goal instructions are instructions that assign the learners' achievement goals beforehand, such as mastery-approach goal instructions that emphasize "to learn as much as possible" and performance-approach goal instructions that emphasize "to be the best player". Achievement goal instructions can induce specific goals in learning, but it is unclear which achievement goal instruction is best for motivation, cognition, and emotion in game-based learning. Aims: The purpose of this paper is to investigate 1) how achievement goal instructions affect motivation (i.e., achievement goals), cognition (i.e., mental effort and performance), and emotion (i.e., achievement emotions) in chemistry game-based learning and 2) whether prior achievement goals moderate the effects of achievement goal instructions. Sample: Participants were secondary school students (N = 450). Methods: In a 2 x 2 factorial design with the factors mastery-approach goal instructions (yes, no) and performance-approach goals (yes, no), participants were randomly assigned to one of the four conditions: mastery-approach goal instructions condition, performance-approach goal instructions condition, combined mastery-approach and performance-approach goal instructions condition, and control condition. Results: Robust regression analysis revealed that mastery-approach goal instructions and performance-approach goal instructions did not interact. Mastery-approach goal instructions had no effects on mastery-approach goals. Performance-approach goal instructions promoted higher performance-approach goals and higher mental effort but lower posttest performance. Prior mastery-approach goals moderated the effects of achievement goal instructions on mental effort. Conclusions: We conclude that achievement goal instructions in game-based learning affect cognitive and motivational outcomes differently. Educators would do well to consider achievement goal instructions and learners' prior mastery-approach goals.