Abstract: A method for determining eight water-soluble sugars in tobacco was developed comprising extraction with 55% acetonitrile under ultrasonic,separation by high performance liquid chromatography with Prevail sugar column under gradient elution with water-acetonitrile,and detection by an evaporative light scattering detector.Thirteen tobacco samples were determined with the developed method.The results showed that:1) the average recoveries of rhamnose,xylose,arabinose,mannose,fructose,glucose,sucrose and maltose were 113.4%,103.7%,109.8%,100.5%,87.1%,115.1%,91.5% and 109.1%,respectively,and their relative standard deviations were less than 4%;2) arabinose,xylose and mannose were not detected in all tobacco samples;3) rhamnose was detected in eleven flue-cured tobacco samples and basically agreed with the reported values,but not detected in the samples of oriental tobacco AG,burley tobacco BH1 and Maryland tobacco C2;4) fructose and glucose were detected in all samples,except for Maryland tobacco,their contents were the highest among the detected sugars and mass ratio between the two was 1 to 1;fructose was the only sugar detectable in Maryland tobacco sample;5) sucrose was detected in eleven tobacco samples not detected in oriental tobacco AG and Maryland tobacco,and maltose was detected in nine samples,not detected in Longyan C2F,oriental tobacco AG,burley tobacco BH1 and Maryland samples,whose contents were higher than those of sucrose in most samples.This method can be used for routine analysis of these eight water-soluble sugars in batches of tobacco samples.