Abstract: Chromatography is a crucial component of investigative experiments in the Animal Nutrition experimental course for students majoring in Animal Science. Under the guidance of their instructors, a student team independently designed an experimental protocol by integrating a scientific research project into practical production needs. They successfully established a high-performance liquid chromatography method for determining the content of GMP and IMP in feed flavor enhancer. After sample pretreatment, isocratic elution was performed on an Athena C₁₈ column, using a mobile phase consisting of methanol and 0.05 mol/L potassium dihydrogen phosphate solution (2∶98, v/v) at a flow rate of 0.6 mL/min. The column temperature was maintained at 25 °C, and the detection wavelength was set to 250 nm with an injection volume of 10 μL. The results indicated a good linear relationship between the concentrations of GMP and IMP and their peak areas over the range of 0.05~2.00 mg/mL (R²=0.9999). The limits of detection were 0.4 μg/mL for IMP and 0.8 μg/mL for GMP, respectively. Furthermore, the method exhibited high recovery rates (up to 98%), with relative standard deviations (RSDs) of 0.81% and 0.75% for IMP and GMP, respectively. This experimental project is not only simple to perform with reasonable analysis time, but also demonstrates high precision and excellent recovery rates. It effectively stimulates students’ interest in learning and spirit of inquiry, while fostering their innovative capabilities and scientific literacy.