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.

中文翻译：

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从Schizochytrium sp。纯化高纯度二十二碳六烯酸。SH103使用制备型HPLC

高纯度的多不饱和脂肪酸（> 95％）对于合成用于临床应用的特殊的亲分解脂质介体（SPM）（如保护素，RESOLVIN和MARES）至关重要。迄今为止，已经通过鱼油的各种制造步骤生产了高纯度（> 95％）的二十碳五烯酸（EPA； C20：5n3）和二十二碳六烯酸（DHA； C22：6n3）。在这项研究中，我们优化了制备型高效液相色谱（HPLC）工艺，以从油性微藻Shizochytrium sp。中纯化出高纯度DHA乙酯（DHAee;> 98％）。SH103含有至少34％的DHA。通过改变流动相速度，负载量和流动相组成，通过反相色谱法确定DHA的纯度和收率。在半制备规模上，以0.5 mL / min的流动相速度，10 mg / mL的上样量以及甲醇/水的流动相组成（96：4，v / v）获得了等度洗脱中的最佳DHA分离DHA为98.5％。将该分离物按比例放大至制备柱，得到99.0％的DHA馏分，产率为79.8％。该结果表明，当将制备柱按比例放大到工业柱时，可以从微藻中产生大量高纯度DHA。