Workers of synthetic leather industries may be exposed to the mixture solvents, such as N,N-dimethylformamide (DMF) and methyl ethylketone (MEK). DMF has been considered as a readily skin-absorbed chemical in literature. The object of this study was to investigate the permeation of DMF/MEK through the re-exposed neoprene gloves using decontamination methods. Deionized water, methanol, ethanol, 2-propanol and acetone were the collection medium and flowed through the closed-loop system of ASTM F739 cells. The concentrations of DMF/MEK in the collection medium were analyzed using GC/FID. The permeation rates and breakthrough time for DMF/MEK were statistically significant (p<0.05) for test collection medium. As conducting the permeation test, it should be noted to determine the collection medium. The results of DMF/MEK mixtures can be interpreted using the solubility parameter theory. The glove materials were tested as received and after exposures to DMF/MEK for five times. The steady state permeation rates were not significantly different, but the breakthrough time was shorter than the first test. The result of decontamination experiments showed that the used protective gloves heated at a higher temperature can be an appropriate decontaminated method.