With the increase in biodiesel production, which generates about 10 wt% glycerol as the main byproduct, numerous added-value applications for glycerol have been investigated. One such application is its use as a plasticizer in polymers. In the current investigation, glycerol was used to produce flexible polyvinyl chloride (PVC) which was compared with standard flexible PVC obtained with plasticizer dioctyl phthalate (DOP). Flexible PVC sheets containing 50 phr of each plasticizer, in addition to stearic acid and heat stabilizer, were manufactured by means of a conventional rolling mill. Test specimens were subjected to density, hardness, tear and tensile testing. Exudation tests and migration stability tests in water, ethanol, and gasoline were also carried out. Density, hardness, tensile modulus and tear strength values of the flexible PVC containing glycerol were similar to the standard flexible PVC containing DOP. However, tensile strength and strain at break of the former were about 50% higher than the latter.