Abstract: The method of determination of glucose, fructose and sucrose in tobacco and tobacco products was developed by high performance liquid chromatography with a refractive index detector. Tobacco samples were extracted for 60 min by shaking with 0.01 mol/L sodium hydroxide solution. The chromatographic separation was conducted on a cation-exchange column packed with highly cross-linked sulfonated styrenedivinylbenzene resin in the Pb2+ ionic form. The eluent was pure water in isocratic gradient. The external standard method was applied in quantitation. The linearity of the method was good with correlation coefficients above 0.999 in all cases. The recoveries were between 98.9 %-101.8 % and the precision was between 0.44 %-0.90 %. The results determined by the developed method were consistent with those determined by the ion chromatography method. The method is applicable to the determination of fructose, glucose and sucrose in various tobacco and tobacco product samples.