Objectives: To assess the effects of exposure to 6 chemical agents on the permeability of latex gloves by dye permeability test and to qualitatively evaluate the microscopic changes in the ultrastructure of the gloves. Method and Materials: The middle fingers of 35 medium-sized, nonsterile latex gloves were used. The chemical agents tested were eugenol, 5% NaOCl, 17% EDTA, 0.2% chlorhexidine gluconate, Cresophene (Septodent), and Chlorispray (Anios). Following treatment for 15 minutes with each chemical agent, glove fingers were filled with 10 mL of 0.02% erythrosine dye solution. Then the outer glove surface was washed with 10 mL of distilled water at intervals of 15, 30, 45, and 60 minutes. A spectrophotometer was used at 530-nm wavelength to determine the percentage of absorption from each collected washing solution. The results were compared with the values obtained from untreated gloves. Additionally, small pieces of the glove samples were examined by SEM to determine whether any ultrastructural changes occurred upon exposure to the chemicals. Results: The permeability of gloves was increased by exposure to Chlorispray and Cresophene, but 5% NaOCl, 17% EDTA, and 0.2% chlorhexidine gluconate had no effect. Major surface changes were noticed in NaOCl, EDTA, Cresophene, and Chlorispray groups, while eugenol and chlorhexidine gluconate had minimal or no effect. Conclusion: Damaging effects of chemical agents on latex gloves for penetration and infection control should be considered by the dental practitioner. Keywords: chemical agents, latex gloves, permeability