Steviol glycoside is increasingly being used as a non-caloric natural sweetener. The aim of this study was to assess whether steviol glycoside affects ethanol absorption in rats and to compare its effect with the effect of sucrose and artificial non-caloric sweetener cyclamate. The study included 144 male Witsar rats which were divided into 4 groups each being intragastrically administered 1500 mg/kg of ethanol in the form of the differently sweetened v/v=16% alcoholic solutions: non-sweetened (C), sucrose sweetened (S), cyclamate sweetened (CY), steviol glycoside sweetened (ST). The rats were blood sampled from the tail vein at t=10,20,40,60,90,120 min after ethanol administration. Pharmacokinetic parameters determined included: blood ethanol concentration at 10 min (C10), maximum blood ethanol concentration (CMAX) and area under concentration vs. time curve (AUC). All pharmacokinetic values significantly differed between groups. Post hoc analysis revealed C10 and CMAX were both significantly higher in C and ST group than in S and CY group. On the other hand, AUC was significantly higher in C than in all other groups. Steviol glycoside affects ethanol absorption uniquely compared to other sweeteners studied. This research represents first description of pharmacokinetic interaction of ethanol and steviol glycoside in an in vivo model.