Abstract: One of the essential steps in the manufacture of biodiesel is its purification from the glycerol by-product. The produced biodiesel should have a low glycerol content which is regarded as one of the important needed pointers for passing the international biodiesel standards. Low cost Deep Eutectic Solvents (DES) have been tested for their solvation properties. In this work, DESs were used to remove all free glycerol and reduce total glycerol from palm oil-based biodiesel. Liquid-liquid extraction experiments were conducted to explore the effect of DES structure on the glycerol separation. All synthesized DESs were able to remove all free glycerol, successfully. The optimum molar ratio of DES to biodiesel was found to be 1:1 for all DESs.