A ionexchange chromatographic method with conductivity detection was developed for the determination of acetylcholine（Ach）residue in dairy products and royal jelly. The sample was extracted with ultra-pure water by ultrasonic,and then determined by ion chromatography. The quantification was performed by the external standard,using IonPac CS17(250 mm×4.0 mm i.d.) column as analytical column,and methylsulfonic acid(MSA)as mobile phase at a flow rate of 1.0 mL/min. The column temperature was set at 30 ℃,and the detection cell temperature was set at 35 ℃. The calibration curve was linear between the peak area and the Ach concentration in the range of 0.02-2.0 mg/L with a correlation coefficient of 0.999 9. The limit of detection of Ach was 0.3 mg/kg in milk,yogurt and royal jelly matrices,and 1.0 mg/kg in milk powder matrix.The average recoveries of Ach from spiked dairy products and royal jelly sample ranged from 90% to 98% with RSDs of 3.0%-6.2%.