This study investigates university students’ understanding of the mole concept and its implications for chemistry education, highlighting the critical role of mathematical education. A questionnaire was administered to 303 students from universities in Panama, Mexico, Cuba, Chile, and Spain. The results reveal that only 29.7% of participants recognize the mole as a fundamental unit, while 20.8% confuse the amount of substance with a non-existent “Chemical System.” Only 18.5% correctly identified the substance quantity symbol as “n” and 32.7% were aware that Wilhelm Ostwald introduced the term mole, indicating deficiencies in historical knowledge. The significance of these findings highlights major misconceptions and gaps in both conceptual understanding and historical knowledge, underscoring the urgent need for revised teaching strategies. Addressing these issues is crucial for bridging the gap between theoretical knowledge and practical application, thereby enhancing instructional methods and optimizing chemistry education to improve students’ comprehension of fundamental concepts.