In this study, supercritical carbon dioxide is chosen for the extraction and purification of omega-3 fatty acids from Hevea brasiliensis (rubber) oil because this method is environmental friendly and the extracted oil has improved nutritional and functional values. In addition, extraction using this method also can prevent organic solvent residual inside the extracted oil. During the extraction using supercritical carbon dioxide, the oil yield is increased with the increase of temperature and pressure. The best operating condition is pressure of 30 MPa, and temperature of 80 C giving 21.4714% oil yield. The analysis of fatty acids composition is using gas-chromatography. The extracted rubber seeds oil is converted to fatty acids methyl esters (FAME) make it more accessible for chromatogram analysis. From the analysis, the omega-3 fatty acids inside rubber seeds oil especially alpha-linolenic acid (ALA) is the highest compare to soybean oil and olive oil. Beside this, rubber seeds oil also contains oleic acid which is omega-9 and linoleic acid is omega-6. This is the reason rubber seeds oil can be regard as oleic-linolenic seed oil. Alpha-linolenic acid is the precursor to convert polyunsaturated fatty acids (PUFAs) to docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Oleic acid (Omega-9) has the potential for heart protection and cancer prevention but still in the state of prove. On the other hands, the linoleic acid, the element of omega-6 has the benefits to treat to treat arthritis and autoimmune disease. Therefore, rubber seeds oil has very high potential to be developed into difference products in difference industries such as pharmaceutical, cosmetics, oleochemical and foods.