An efficient method for the purification of arachidonic acid from fungal single‐cell oil (ARASCO)

Abstract PUFA, such as arachidonic acid (AA), have several pharmaceutical applications. An efficient method was developed to obtain high‐purity arachidonic acid (AA) from ARASCO, a single‐cell oil from Martek (Columbia, MD). The method comprises three steps. In the first step, AA was enriched from saponified ARASCO oil by low‐temperature solvent crystallization using a polar, aprotic solvent, which gave a FA fraction containing 75.7% AA with 97.3% yield. The second step involved enriching AA content via lipase‐catalyzed selective esterification of FA with lauryl alcohol. When a mixture of 1 g FA/lauryl alcohol (2∶1 mol/mol), 50 mg Candida rugosa lipase, and 0.33 g water was incubated at 50°C for 24 h with stirring at 400 rpm, the AA content in the unesterified FA fraction was as much as 89.3%, with ca. 90% yield. Finally, a solvent extraction procedure, in which acetonitrile was the extracting solvent, was used to enrich AA from FA fraction dissolved in n ‐hexane. The best results were obtained when 2 g FA was dissolved in 80 mL hexane and extracted twice, each time with 20 mL acetonitrile at −20°C, by allowing 2 h storage. This step gave a FA fraction containing 95.3% AA with 81.2% yield. By using this three‐step process the AA content in the saponified single‐cell oil (ARASCO) was increased from 38.8 to 95.3% with a total yield of ca. 71%.