Abstract: A simple and rapid method has been developed for determination of chlorogenic acid in tobacco by RP-HPLC and the extracting process was optimized. The quantitative results were obtained by the internal standard method. The results indicated that the optimum extraction process is as follows: the content of ethanol is 60%,the temperature of ultrasound is 50℃,the pH of ethanol solution is 4, the time of extraction is 45 minutes. The method has good linearity with correlation coefficient of 0.9999.The limit of detection based on signal-to-noise ratio of 3 was 1.4×10^-2 mg/10 mL. The recovery of chlorogenic acid in tobacco was 101.5%,with RSD of 1.58%.