Abstract: To rapidly determine seven synthetic sweeteners (Acesulfame potassium, Sodium cyclamate, Saccharin sodium, Aspartame, Neotame, Sucralose and Stevioside) and six natural sweeteners (Erythritol, Xylitol, Mannitol, Sorbitol, Maltitol and Isomaltitol) in gum-based chewing tobacco, an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed. Samples of gum-based chewing tobacco were extracted by shaking with n-hexane and water. The seven synthetic sweeteners were separated by reversed phase column using gradient methanol/water as mobile phase. The six natural sweeteners were separated by special sugar column using gradient acetonitrile/water solution as mobile phase. All the sweeteners were analyzed by UPLC-MS/MS under the negative ion monitoring mode. The results showed that:1) The linear correlation of the thirteen sweeteners were good (R²>0.994), the limits of detection and quantitation were 6.8×10^-6-0.30 and 2.3×10^-5-1.0 μg/kg, respectively; the linear ranges of all the sweeteners ranged from 0.001 to 1.0 μg/mL. 2) The recoveries of the sweeteners were from 80.2% to 97.6%. 3) The precisions of intra- and inter-day were all less than 10% (n=6). This method is suitable for the rapid qualitative and quantitative analysis of sweeteners in gum-based chewing tobacco.