Background: Use of additives in food industry is of specific importance. Currently, more than 2,500 additives are added to foods to achieve the desired goals or to extend the product’s shelf life. However, application of these substances has raised concerns about the consumers’ health. Since public information on these substances is inadequate, promoting people’s knowledge can play a key role in this regard. Therefore, knowledge, attitude, and practices of the personnel of Isfahan University of Medical Sciences toward food additives were evaluated. Methods: This cross-sectional descriptive study included 800 participants selected randomly among the personnel of Isfahan University of Medical Sciences in Iran. The participants’ demographic information, knowledge, attitudes, and practices were measured via a self-administered questionnaire. The relationship of participants’ educational level, age, and gender with their knowledge, attitudes, and practices were determined. Results: The participants’ knowledge, attitudes, and practices average scores were calculated as 30.6±1.3, 32±0.9, and 15±1.5 for food additives, respectively. Conclusion: The participants’ knowledge, attitude, and practices were at a poor level. So, people should receive the necessary education and training in terms of food additives as well as food labels in order to prevent misinterpretations regarding food additives.