Introduction: Safety culture is an important factor for improving safety in university. This study aimed to examines the construct validity and reliability of the elements of safety culture in research universities in Malaysia. Methods: A measuring instrument was developed and analyzed for reliability using an exploratory factor analysis approach. The reliability analysis was determined using Cronbach’s Alpha. About 298 postgraduates’ students from five research universities were selected randomly. An exploratory factor analysis was performed using the principal component method with varimax rotation, Kaiser Meyer Olkin, Bartlett’s test of Sphericity and Cronbach’s alpha were obtained. Statistical analysis was carried out using the Statistical Package for Social Science (SPSS) and Analysis Moment of Structures (AMOS) version 24.0 (IBM SPSS-AMOS V24.0) software. Results: Findings indicated that the Keiser-Meyer-Olkin for all elements of safety culture (training, leadership, management commitment and communication) was in the range 0.799 and 0.916, in which the value was greater than 0.70, while Barlett’s test with a p value of 0.000. Factor loadings were greater than 0.60 in all elements of safety culture. Cronbach’s alpha coefficient was at the range of reliability between 0.8 < α < 0.9. Conclusion: It can be concluded the instrument was found to be a potential mechanism for measuring safety culture in research universities in Malaysia.