Purification of high purity docosahexaenoic acid from Schizochytrium sp. SH103 using preparative-scale HPLC

• Published in: Applied biological chemistry Vol. 63; no. 1; p. 56
• Main Authors: Oh, Chi-Eun, Kim, Gap-Jin, Park, Seung-Jin, Choi, Seunghoon, Park, Min-Joo, Lee, O-Mi, Seo, Jeong-Woo, Son, Hong-Joo
• Format: Journal Article
• Language: English
• Published: Singapore Springer Singapore 01.12.2020; 한국응용생명화학회
• Subjects: Applied Microbiology; Biological Techniques; Bioorganic Chemistry; Chemistry; Chemistry and Materials Science; docosahexaenoic acid; eicosapentaenoic acid; fish oils; high performance liquid chromatography; methanol; microalgae; reversed-phase liquid chromatography; Schizochytrium; 농학; High purity DHA; Purification; Preparative HPLC; sp
• ISSN: 2468-0834; 2468-0842
• DOI: 10.1186/s13765-020-00542-w

High purity polyunsaturated fatty acids (> 95%) are essential for the synthesis of specialized pro-resolving lipid mediators (SPMs), such as protectins, resolvins, and maresins, which are used for clinical application. To date, high purity (> 95%) eicosapentaenoic acid (EPA; C20:5n3) and docosahexaenoic acid (DHA; C22:6n3) have been produced through various manufacturing steps using fish oil. In this study, we optimized preparative high performance liquid chromatography (HPLC) process to purify high-purity DHA ethyl ester (DHAee; > 98%) from oleaginous microalgae Shizochytrium sp. SH103 containing at least 34% DHA content. The purity and yield of DHA were determined by reverse phase chromatography with changing the mobile phase velocity, loading amount, and mobile phase composition. On a semi-preparative scale, optimal DHA separation in isocratic elution was obtained with a mobile phase velocity of 0.5 mL/min, a loading amount of 10 mg/mL, and mobile phase composition of methanol/water (96:4, v/v), wherein the purity of DHA was 98.5%. This separation was scaled up to a preparative column, resulting in 99.0% DHA fraction with a yield of 79.8%. This result suggests that a large amount of high purity DHA can be produced from microalgae when scaling up a preparative column to an industrial column.