Abstract: OBJECTIVE To establish a fluorescence analysis method for determining ampelopsin in rabbit plasma. METHODS After intravenous administration with ampelopsin at a single dose, the blood samples were collected and extracted with ethyl acetate. Ampelopsin concentrations were determined with fluorescence detection exciting at 442 nm and emission at 502 nm. RESULTS The calibration curve showed a linear response over the concentration range of 0.5-36.6 μg·mL^-1(r=0.999 6). The limit of detection was 70 ng·mL^-1. The recovery was between 98.78% and 103.57%. The relative standard deviations of intra-day and inter-day were less than 8.25%. CONCLUSION A rapid, simple and accurate method has been successfully applied to a pharmacokinetics study of ampelopsin in rabbits.