Objective The Migraine-Specific Quality of Life Questionnaire version 2.1 (MSQ) has been shown to have good psychometric performance in measuring headache impact in migraine patients, but its properties specifically in chronic migraine (CM) patients are unknown. The objective of this study was to evaluate the psychometric properties of the MSQ in a group of CM patients undergoing prophylactic treatment. Methods Measurement properties of the MSQ were examined using two international, multicenter, randomized clinical trials evaluating onabotulinumtoxinA as headache prophylaxis in CM patients (N = 1,376). Confirmatory factor analysis (CFA) was used to test the latent structure of the MSQ in CM patients. The reliability, convergent and discriminant validity, and responsiveness of the MSQ were assessed. Results CFA confirmed the currently proposed three-factor MSQ latent structure across the two studies. Good reliability was observed for all three MSQ scales, across studies and time points. MSQ scale scores strongly correlated with the scores of the Headache Impact Test-6 (HIT-6). Analysis of knowngroups validity indicated that MSQ scale scores discriminated between groups of patients differing in their 28-day headache frequency were as follows\10, 10-14, and C15 days, and the sample-derived quartiles of the total cumulative hours of headache were as follows \140, 140 to \280, 280 to \420, and C420 h (p \ 0.0001), across both studies and time points. MSQ change scores were higher in magnitude in groups experiencing greater decline in headache frequency (p \ 0.001). Conclusion The MSQ is a psychometrically valid tool that can be used to reliably measure the impact of migraine among CM patients.