Introduction and objective Acrylamide is a “probably human carcinogen” monomer that can form in heated starchy food as a result of a reaction between asparagine and reducing sugars via Maillard reaction. The main source of acrylamide in human diet are potato products, cereal products and coffee. Tobacco smoke may be another significant source of exposure to acrylamide. The aim of our study was to determine acrylamide content in cigarettes available on the Polish market and to estimate the exposure to acrylamide originating from tobacco smoke in smokers in Poland. Material and Methods The material was cigarettes of the top five brands bought in Poland and tobacco from non-smoked cigarettes. Acrylamide content in cigarettes mainstream smoke was determined by LC-MS/MS. Exposure assessment was carried out using analytical data of acrylamide content in cigarettes and the mean quantity of cigarettes smoked daily by smokers in Poland, assuming body weight at 70 kg. Results The mean content of acrylamide was 679.3 ng/cigarette (range: 455.0 – 822.5 ng/cigarette). The content of acrylamide was evidenced to correlate positively with total particulate matter (TPM) content in cigarettes. The estimated average exposure to acrylamide from tobacco smoke in adult smokers in Poland is 0.17 μg/kg b.w./day. Conclusions Our results demonstrate that tobacco smoke is a significant source of acrylamide and total exposure to acrylamide in the population of smokers, on average, is higher by more than 50% in comparison with non-smokers. Our estimation of exposure to acrylamide from tobacco smoke is the first estimation taking into account the actual determined acrylamide content in the cigarettes available on the market.