The authors present findings on the third round of the Czech validation of the Motivated Strategies for learning questionnaire (MSLQ), originally developed by Pintrich et al. (1991). The validation only covered an area designed to access motivation in self-regulated learning. Data was collected from a sample of university students in regular classroom settings. Principal component analysis (PCA) was conducted with eigenvalues exceeding 1. An inspection of the scree plot, discontinuity in variance, Monte Carlo parallel analysis and Cronbach's alphas were performed to assess the psychometric properties. The results were further supported by the confirmatory factor analysis with no post hoc model modifications needed. The analysis confirmed the first and second round validation structure bringing a 3-factor model and indicated that the revised MSLQ is an acceptable measure of motivation in self-regulated learning.