This research article was published Journal of Food Safety and Hygiene in 2022Consumption of aflatoxins contaminated foods has led to detrimental health effects worldwide, with even more severe cases in African countries including Tanzania. A cross sectional study was conducted to assess awareness and aflatoxins contamination of sesame seeds in Lindi and Mtwara regions. Subsequently, a total of 70 sesame seed samples were randomly purchased from local markets for aflatoxin determination using High Performance Liquid Chromatography (HPLC). Qualitative data were analyzed using SPSS 20 for descriptive and correlation analysis. Results show that 82.4% of the respondents were not aware of aflatoxin contamination of agricultural produce. Awareness was negatively correlated to the levels of education (p=-0.309) and positively correlated with gender whereby men were more aware than women (p=0.03). On the other hand, 37 out of 70 sesame seeds samples were contaminated with total aflatoxins at a range of 0.009 ng/g to 5.557 ng/g. Although none of these samples exceeded the Tanzania maximum limits of 10 ng/g for total aflatoxins, 2 samples exceeded the maximum limit of 4 ng/g set by the European Union. Furthermore, Aflatoxin B1 was detected in 13 samples moreover the concentration was below the Tanzania and EU maximum limit of 5 ng/g and 2 ng/g respectively. Though the contamination was below the national maximum limits and limited to one agro-ecological zone and season, these findings provide useful insights on aflatoxins contamination of sesames seeds from the two main growing regions in Tanzania.