Preparation of Highly Purified Stearidonic Acid from Echium Oil via an Enzymatic Method Combined with Preparative High Performance Liquid Chromatography

Stearidonic acid (SDA), an n-3 polyunsaturated fatty acid (PUFA), can be obtained from plant origin oils and it can be a good source of PUFA for vegetarians. SDA can be easily converted to longer PUFA such as docosahexaenoic acid and eicosapentaenoic acid. Highly purified stearidonic acid (SDA) was prepared successfully from echium oil via an enzymatic method combined with preparative high performance liquid chromatography. In the 1st step, SDA enrichment was accomplished using Candida rugosa lipase and 39.5% of SDA was obtained in the fatty acid fraction. Subsequently, the 1st reaction mixture was used for the 2nd enzymatic esterification without any separation process. The 2nd esterification was conducted for further SDA enrichment in a packed-bed reactor using Lipozyme RM IM from Rhizomucor miehei and the SDA content increased in a very short residence time. Ethanol was selected as an appropriate alcohol to react as an acyl receptor, and the other conditions for SDA enrichment were optimized at 20 degrees C of temperature, and 1:4 of molar ratio (i.e., fatty acid to ethanol). Under these conditions, 51.6% of SDA was obtained in the fatty acid fraction after a residence time of 15 min. Finally, highly purified SDA (purity, >99%) was obtained by prep-HPLC using the SDA-rich fraction obtained from the two-step lipase-catalyzed esterification.