Abstract: The present invention relates a high-purity purification method for omega-3 highly unsaturated fatty acids. More specifically, it relates to a high-purity purification method for omega-3 highly unsaturated fatty acids which is both environmentally friendly and easy to implement and comprises the steps of: a) preparing a fatty acid ethyl ester (FAEE) by ethanolysis of a natural oil or fat, using ethanol, in the presence of an enzyme catalyst extracted from at least one microorganism selected from the group consisting of Candida genus, Rhizopus genus, Mucor genus, Aspergillus genus and Pseudomonas genus; b) subjecting the said prepared fatty acid ethyl ester to preliminary distillation using short-path distillation (SPD) device between 100 and 200° C. and between 0.001 and 10 mmHg; c) forming a concentrated fatty acid by subjecting the ethyl ester, which has been subjected to the preliminary distillation, to reduced-pressure fractional distillation at between 100 and 200° C. and between 0.