Abstract: Objective: To develop the method of pretreatment and analysis for content determination of jujuboside A and B in Zizyphi Spinosi Semen ( ZSS) , to study the contents of jujuboside A and B in ZSS from different regions.Methods:Accelerated solvent extraction ( ASE) was used as method of extracting jujuboside A and B from ZSS and the conditions of ASE were optimized after parameters of ELSD were optimized and HPLC analysis method was tested, contents of jujuboside A and B in ZSS from seven different origins were determined.Results: The optimal ASE conditions: two-step extraction, optimum conditions of the first step were 60% ethanol, 1 times / min ( extraction frequency) , 100 bar ( extraction pressure) , 100℃ ( extraction temperature) , optimum conditions of the second step were 60% ethanol, 5 times / min, 120 bar, 100℃, the order of jujuboside contents in ZSS from different origins as follows: He Bei Baiping > He Bei Zanhuang >Shan Dong > He Bei Fuping > Liao Ning > Laos > Myanmar.Conclusion: HPLC- ELSD combined with ASE method is very suitable for determination of jujubosides A and B in ZSS with high recovery rate and good accuracy; ZSS produced in He Bei and Shan Dong in contents of jujubosides A and B is far higher than other ZSS, and these two ZSS can be used as high quality raw materials for researching and developing health care products.