The aim of this study, therefore, provides information about Aflatoxin levels in raw cow’s milk in Injibara Town of Awi Administrative zone by using HPLC-FLD. A good linearity of standard calibration was found for AFM1 at a range of 0.5–7 µg/L. Regression coefficient (R2) values were 0.9999, whereas slope and intercept were 2.5278 and 0.1012, respectively. The average recoveries for the spiked samples were range from76.62– 90.98 % and the RSD values ranged between 2.55–7.36 %. The results showed that 15 % of samples (3 out of 20) were contaminated with AFM1 in the range of 0.046–0.22 µg/L. The average contamination level was 0.121 µg/L. The determined mean values of AFM1 in the collected milk samples were above the standard limit of the European Commission and lower than the level established by United States regulations. Further monitoring of Aflatoxin in milk samples from different regions of the country is justified to conclusively determine the actual safe/risks and possibly low Aflatoxins-risk milk production areas.