Background: This study aimed to validate the Persian version of Maslach Burnout Inventory for Medical Personnel (MBI-HSS-MP), an instrument developed to capture burnout for health professionals. The specific aims were to psychometrically assess the Persian MBI-HSS-MP in relation to its structure, test-retest reliability, and item properties. Methods: The study setting was all eight hospitals in Qazvin province, Iran (study period from 10 September to 16 November 2020). Health professionals of physicians (n = 106) and nurses (n = 200) participated in the study. The psychometric properties of the 22-item MBI-HSS-MP was then examined for its factor structure via confirmatory factor analysis (CFA) and Rasch models, test-retest reliability, item fit, and differential item functioning (DIF). Results: The MBI-HSS-MP was verified as having a three-factor structure and each item was embedded well in its belonging construct (comparative fit index = 0.941, Tucker-Lewis index = 0.929 derived from CFA results; infit and outfit MnSq = 0.71 to 1.38 derived from Rasch models). Test-retest reliability of each MBI-HSS-MP item was satisfactory and no substantial DIF items were displayed across gender or across health professionals. Conclusion: The MBI-HSS-MP has good psychometric properties to assess burnout accurately among healthcare professionals in the three dimensions of emotional exhaustion, personal accomplishment, and depersonalization.