Abstract: Aflatoxins contaminate approximately 25% of agricultural products worldwide and have negative public health implications. Little is known on the level of aflatoxins in foodstuffs in Kampala markets and yet open markets in Sub-Saharan Africa have been implicated to have high risk of aflatoxin contamination. A cross-sectional survey was conducted to quantify levels of aflatoxin contamination in major staple foodstuffs in five major markets in Kampala in the month of June, 2014. From each market, 4 incremental samples of approximately 250 g of each food type were picked at random from different stalls to make up a sample of 1000 g. Overall, twenty samples of staple foodstuffs sampled in Kampala markets were analyzed for aflatoxin contamination. Foodstuffs included groundnuts, ground nut paste, cassava flour and maize grains. Total aflatoxins were identified and quantitated using the VICAM AflaTest fluorometric method (VICAM LP, 34 Maple Street, Milford, MA 01757 USA). Aflatoxins were detectable in 16/20 (80%) of the samples and 8/20 (40%) of the samples exceeded FDA/WHO regulatory limit of 20 μg kg–1. All the groundnut paste samples tested 5/5 (100%) had aflatoxin concentration levels more than the 20 μg kg–1. None of the maize grain samples analyzed exceeded the permissible level of 20 μg kg–1. Laboratory analyses indicate that groundnuts and ground nut paste had extremely high alarming aflatoxin levels of up to 940 and 720 μg kg–1, respectively. There is high aflatoxin contamination in groundnuts and groundnut paste. Therefore, interventions should focus on strengthening health inspections, creating awareness and routine monitoring of aflatoxin levels in staple foods in Kampala markets.