Aflatoxin B1 contamination adversely affects human health by impairing long-term physical and cognitive development. Several crops have been associated with aflatoxin B1 contamination and corn is one of them. In the Anuradhapura district of the North Central Province of Sri Lanka, corn is one of the main agricultural produce. Due to poor farming practices in this area, it is possible that aflatoxin B1 is somehow transported from soil to the corn ears. This study was carried out to detect and quantify aflatoxin B1 in corn and corn-grown soils in Anuradhapura. Corn (n = 60) and corn-grown soil (n = 60) samples were randomly collected from 20 minor-scale corn-grown fields with three random replicates. Each sample was prepared for the measurement of aflatoxin B1 levels using the Enzyme-Linked Immunosorbent Assay (ELISA). Though 20 ppb is considered as the poisonous or deleterious level for corn consumption, there were toxin contaminations of up to 60–70 ppb in the corn kernel, while majority of soil had 350–400 ppb of aflatoxin B1 levels. Fifteen corn samples had exceeded the acceptable level while 22 samples were free of aflatoxin B1 and 23 samples were within the acceptable level. The results showed that the presence of aflatoxin B1 in corn is not habitually distributed throughout Anuradhapura district and it increased with the soil aflatoxin B1 concentration. It appears that there is a relationship between corn kernel and corn-grown soil aflatoxin B1 levels.