Aflatoxin M1 (AFM1) is the hydroxylated metabolite of aflatoxin B1 (AFB1), which is formed in the liver by cytochrome P450 enzymes and can be secreted into the urine, feces, and milk of mammals. AFM1 is a carcinogenic, cytotoxic, teratogenic, mutagenic and genotoxic agent that poses a significant health risk to both humans and animals. This study was conducted to determine the presence of AFM1 in both raw and ultra-high temperature (UHT) cow's milk samples produced in the northern part of Cyprus, and to determine whether it poses a risk to public health. In this survey, a total of 20 UHT cow's milk samples from 2 different milk brands produced in the northern part of Cyprus, and 22 raw cow's milk samples collected from the different dairies were analyzed for the presence of AFM1 by high performance liquid chromatography (HPLC) with fluorescence detector after immunoaffinity cleanup. AFM1 could not be detected in any of the analyzed raw and UHT cow milk samples. The LOD and LOQ values of the HPLC-FLD method were 1.038 μg/kg and 3.145 μg/kg, respectively. The mean recovery and repeatability values of the method were 95.6% and 4.9%, respectively. Although the presence of AFM1 in milk samples produced in the northern part of Cyprus poses no major risk to public health, more milk samples and animal feed must be monitored on a regular basis to decrease potential consumer exposure.