Abstract: To quantify water-soluble sugars in natural tobacco flavors, the analytical method described in YC/T 447—2012 was optimized, and an HPLC-ELSD method with water extraction was developed to determine 10 water-soluble sugars in natural tobacco flavors, namely rhamnose, stevioside, fructose, sorbitol, mannose, galactose, glucose, inositol, sucrose, maltose. The results showed that: 1) The sample solution with water extraction presented better stability compared with the control method with 0.02 mol/L NaOH extraction. 2)The retention time of the analytes was more stable after adding aqueous ammonia into the mobile phase. 3)The stabilities of ELSD peak areas of the ananlytes were improved at a gas flow rate of 2.20 L/min and a drift tube temperature of 80 ℃. 4)The LODs of the 10 sugars were in the range of 4.19-12.30 μg/mL and the spiked recoveries ranged from 80.20% to 106.00% with good intra-day (RSD≤4.05%) and inter-day (RSD≤6.25%) precisions. 5) The contents of these sugars in 239 natural tobacco flavors were determined with the developed method, and the top five sugars were glucose, fructose, sucrose, maltose and sorbitol.