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

  • Oh, Chi-Eun1
  • Kim, Gap-Jin2
  • Park, Seung-Jin1
  • Choi, Seunghoon1
  • Park, Min-Joo1
  • Lee, O-Mi3
  • Seo, Jeong-Woo4
  • Son, Hong-Joo1, 1
  • 1 Pusan National University, Miryang, 50463, Republic of Korea , Miryang (South Korea)
  • 2 PharmaMega Inc., Busan, 48055, Republic of Korea , Busan (South Korea)
  • 3 Plant Quarantine Technology Center, Animal and Plant Quarantine Agency, Gimcheon, 39660, Republic of Korea , Gimcheon (South Korea)
  • 4 Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup, 56212, Republic of Korea , Jeongeup (South Korea)
Published Article
Applied Biological Chemistry
Springer Singapore
Publication Date
Sep 16, 2020
DOI: 10.1186/s13765-020-00542-w
Springer Nature


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.

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