Background: Milk is a perfect complete perishable food that could be adulterated or contaminated by wide varieties of substance throughout the chain of its production. Of these aflatoxins M1 was considered as an important health risk associated with the milk. The objectives of this study are to analyse milk samples, for chemical composition and detection of aflatoxin M1. Method: One hundred milk samples were collected from different sources randomly; 35 samples from farms, 35 samples from sale points and 30 samples from groceries in Gedarif town. Milk-Kana was used for determination of chemical composition (fat, solids not fat, lactose and protein and analysis of freezing point, density and added water) and Unisensor kits were used as screening method for detection of aflatoxin M1 in milk. Results: The chemical composition of milk samples collected from farms, sales points and groceries showed lower content of the fat (3.4±1.3%, 3.4±1.3% and 3.5±1.25%), solids not fat (7.7±1.1%, 8.1±1.7% and 8±1%), lactose (4.2±0.6%, 4.4±0.8% and 4.3±0.5%) and protein (3±0.4%, 3±0.5% and 3±0.4%). The freezing point of milk samples revealed -.503±.079, -.524±.007 and -.522±.099 °C, while the density showed 0.025±0.003, 0.026±0.005 and 0.026±0.003 gm/cm3, respectively. Moreover the added water was found as 10.6±11.6%, 10.6±10.4% and 8.7±8.2% in the samples collected from farms, sale points and groceries, respectively. The occurrence of aflatoxin M1 in raw milk samples were found in 22(27.50%) of the samples. The presence of aflatoxin contamination was high in milk samples collected from sales points (15.0%) followed by farms (11.25%) compared to those obtained from groceries (1.25%). Conclusion: The lower chemical composition of milk ns the high added water suggested adulteration of milk. Moreover the presences of aflatoxin M1 in the milk samples might indicate that the cow milk was contaminated with aflatoxins through feed. Hence this study recommended that good hygienic practices should be considered in Gadarif town. Similarly regulations and lows for selling the milk should be implemented by the official authorities.