The Instrumental Analytical Chemistry course, taught in the fifth semester across several programs at the Faculty of Exact Sciences of the National University of La Plata, places a significant emphasis on high-performance liquid chromatography (HPLC). This versatile analytical technique is essential for the separation, identification, and quantification of compounds in various matrices with widespread applications spanning analytical chemistry, biochemistry, pharmaceuticals, forensics, environmental science, and the food industry. Despite its importance, practical HPLC training faces challenges related to the expense of instrumentation and consumables, the significant time investment required for experiments, and the technical expertise needed for operation. To address these limitations, simulation software offers an effective approach to teaching the fundamental concepts of HPLC and demonstrating the influence of different variables on chromatographic separations. This study explores the use of the freely available Practical HPLC simulator v1.0 as a supplementary tool for hands-on HPLC activities in the regular curriculum. The pedagogical framework for the designed activities is Kolb's experiential learning theory, which guides a sequence involving an introductory session on the simulator's features and capabilities, followed by teacher-led group simulations and an evaluation of learning through individual and group problem-solving tasks. Initial findings from the implementation of the HPLC simulator indicate a notable enhancement in students' comprehension of the foundational principles of the technique. Furthermore, the simulator enriches practical HPLC exercises by providing clearer explanations, improving knowledge retention via practical engagement and increasing student motivation and interest, thereby fostering greater participation. In conclusion, this innovation offers a valuable and flexible tool for the comprehensive training of students by integrating theoretical knowledge, practical application, and problem-solving skills. Consequently, this initiative presents a significant opportunity to optimize student education in liquid chromatography, more effectively equipping students for future academic and professional endeavors.