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

摘要

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%.