Abstract: The present research were tempted to investigate whether Aflatoxin is an additive factor in development of HCC through detecting its metabolite Aflatoxin M1 in serum and urine of HCC and cirrhotics in Egypt. Present study comprised (46) Hepatocellular Carcinoma (HCC) patients with mean age (56.28+8.08), 30 males and 16 females, (12) cirrhotic patients with mean age (47.83±18.20), 7 males and 5 females and (12) sex and age matched healthy controls. All were exposed to, liver function tests, abdominal ultrasonography and detection of Aflatoxin metabolite M1 in serum and urine by means of the reverse phase HPLC device. Aflatoxin M1 was detected in sera of HCC group, cirrhotics and controls (57.8%) (5.61±17.21 ng mL-1), (91.7%) (19.23±20.42 ng mL-1) and (50%) (0.66±0.84 ng mL-1), respectively and in urine (41.3%) (3.82±8.03 ng mL-1) (91.7%) (43.22±45.02 ng mL-1) and (50%) (0.98±1.4 ng mL-1), respectively representing significant increase in the serum of the cirrhotic group (p<0.02) and a high significant increase in urine of the cirrhotic group (p<0.0001). Among HCC group patients, there is high significant value of M1 concentration in urine of upper Egypt residents compared to those of lower Egypt (p<0.002). The mean value of Aflatoxin M1 concentration among females of the HCC group was significantly higher than that among males (p = 0.006). There is higher statistical significance of aflatoxin prevalence and concentration in serum and urine of cirrhotics than HCC patients and controls and in concentration in urine of HCC patients from upper than lower Egypt.