Purpose: Usability is one of the key concerns in the development of mobile learning applications. The aim of this paper is to design and validate a usability evaluation questionnaire (UEQ) for mobile learning applications. Design/methodology/approach: The UEQ was developed in four stages: selecting primary studies and extracting usability problems, thematic analysis, creating UEQ items and validation and reliability using confirmatory factor analysis (CFA). Findings: CFA to derive the model fit was computed using AMOS to test the construct validity. The model-fit values were within their respective expected acceptance levels. To assess the reliability of the instrument item loadings, the internal consistency coefficients such as Cronbach's alpha, McDonald's Omega and composite reliability were considered. Indicator loadings ranged between 0.735 and 0.933, fulfilling the threshold of above 0.7. Originality/value: The study provides a novel UEQ for mobile learning applications, which can be used by developers and in academic research to assess mobile learning applications.