Objectives: The main purpose of the study was to investigate the association between sex and the level of information stress, as mediated by affective temperaments. Material and Methods: The sample consisted of 231 healthy Caucasian adults (150 women and 81 men) recruited from a general population. The participants’ age ranged 18–56 years (M±SD = 25.07±6.36). Affective temperaments were assessed using the Temperament Evaluation of Memphis, Pisa, Paris and San Diego Auto-questionnaire (TEMPS-A). To assess the level of information stress, the Information Stress Questionnaire (ISQ) was used. Results: Information stress displayed low to medium positive correlations with depressive, cyclothymic, irritable and anxious temperaments, and a negative correlation with the hyperthymic temperament. The female group was characterized with significantly higher age, information stress, and anxious temperament values, and with a significantly lower irritable temperament value, when compared to males. Cyclothymic temperament, anxious temperament and hyperthymic temperament were found to be significant predictors of information stress. The mediation analysis showed a significant direct effect of sex on information stress. The anxious temperament was a significant mediator of the relationship between sex and information stress. Conclusions: The results showed the relationship between sex and information stress, including the role of anxious temperament as a mediator. Int J Occup Med Environ Health. 2019;32(5):635–44