INTRODUCTION: Food handlers' hands serve as a vehicle for potential foodborne pathogenic contamination which constitutes a public health risk. In Ghana, there are always constant reports of outbreak of foodborne diseases in schools. However, determinants of hand hygiene practice among educational institutions food handlers are little known in Ghana. The study, therefore, aimed to assess the determinants of hand hygiene practice at critical times among educational institutions food handlers in the Sagnarigu Municipality of Ghana.METHODS: This was a cross-sectional survey among educational institutions food handlers in the Sagnarigu Municipality. Two hundred and six food handlers were selected through convenience sampling in the educational institutions. At the same time, structured questions that were developed from previous studies were used to assess food handlers' hand hygiene practice at critical times. Data were analyzed using descriptive statistics, bivariate and multivariate logistic regression models. All variables with their respective confidence interval (95%) and adjusted odds ratios were declared significant at p-value less than 0.05.RESULTS: The hand hygiene practice at critical times among the food handlers was relatively good 66.0% (95% CI: 59.1, 72.5%) and is independently predicted by training on food safety [AOR = 24.39, (95% CI: 2.24 – 245.45) p=0.007], sufficient knowledge of hand hygiene [AOR = 16.34, (95% CI: 3.92 – 68.05) p<0.001] and positive attitude towards hand hygiene [AOR = 82.08, (95% CI: 14.21-474.26) p<0.001]. Only 17.0% of the food handlers demonstrated proper handwashing techniques whereas a good number (85.4%) of the food handlers did not wash their hands after touching money.CONCLUSION: Good hand hygiene practice at critical times was influenced by training on food safety, sufficient knowledge of hand hygiene, and positive attitudes towards hand hygiene. These findings demonstrate the significant role of food handlers' hand hygiene knowledge, attitudes, and training on food safety in improving hand hygiene practices. Concerned stakeholders and organizations should focus on WASH interventions that seek to improve educational institutions food handlers’ knowledge and attitudes towards hand hygiene, coupled with training on food safety.