Abstract: Ultra high performance liquid chromatography (UPLC) is a novel technique in liquid chromatography. It is more efficient, rapid and sensitive than high performance liquid chromatography (HPLC), owing to using smaller particle size in analytical column. Because of these advantages, UPLC has been widely used in many fields. Experiments on rapid separation of caffeine, aspirin and phenacetin by UPLC and quantitative analysis of caffeine in compound aspirin tablets by external standard method were designed. In the experimental teaching, the basic principle, instrument composition and operation, the general methods of optimizing chromatographic conditions and the external standard method were introduced. Through this experiment, students not only better understood the theoretical knowledge, but also learned the operation of UPLC. More importantly, students’ experimental skills have been improved, their scientific thinking ability has been exercised.