Objective: To determine the prevalence of serological markers for hepatitis B in blood donors of Instituto Onco-Hematológico de Anápolis, correlating to age, gender and marital status of the donors with the results of serological screening for hepatitis B. Methods: A retrospective descriptive study was performed by evaluating the data from laboratory tests of markers of hepatitis B serological screening performed in Instituto Onco-Hematológico de Anápolis during the years 2012 and 2013. Data for the months of January to December in both years were collected. The markers were Anti-HBc Total and HBsAg. The estimated population sample (n) was 31,537 peoples. The variables included: age, gender, marital status. The Chi-Square (χ2) or Fisher's exact test were used to analyze categorical variables. Results: There was a low prevalence of markers evaluated, as regards the Anti-HBc reactivity with increasing age. It was observed an association of positivity with serological markers in relation to female and unmarried. Conclusions: Strategies for attracting donors, as well as preventive measures constitute important ways to ensure low virus circulation in the region, ensuring lower costs by performing the serologic screening.