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目的探索海带渣中二十碳五烯酸(EPA)提取纯化及含量测定的方法,为深入研究海带渣中EPA提供技术方法。方法以海带渣为原料,在45℃条件下用3倍量无水乙醇提取2次得到粗脂,采用氢氧化钾乙醇皂化和盐酸酸化制备游离混合脂肪酸,并以EPA含量为指标,用正交试验探讨尿素包合法富集海带渣中EPA的工艺条件。高效液相色谱法(HPLC)测定游离EPA的含量。结果皂化后游离EPA的相对含量为5.13%。尿素包合反应最佳工艺为:95%乙醇为溶剂,尿脂比3∶1,包合温度为-15℃的条件下包合2次,每次包合15h。尿包后EPA的相对含量达24.46%。结论该工艺简单易操作,可用于海带渣中EPA的提取纯化及含量测定。
Objective To explore the extraction, purification and determination of eicosapentaenoic acid (EPA) in kelp residue, and to provide technical methods for the further study of EPA in kelp residue. Methods The crude kelp residue was obtained by extracting kerogen residue twice with 3 times the amount of absolute ethanol at 45 ℃. The free fatty acids were prepared by saponification with potassium hydroxide ethanol and acidification with hydrochloric acid. The content of EPA was determined by orthogonal The experiment discussed the urea enrichment process of enrichment of EPA in kelp residue. Determination of free EPA by high performance liquid chromatography (HPLC). Results The relative content of free EPA after saponification was 5.13%. The best urea inclusion reaction process: 95% ethanol as solvent, urine ratio of 3: 1, inclusion temperature of -15 ℃ under the conditions of inclusion 2 times, each inclusion 15h. The relative content of EPA after urination reached 24.46%. Conclusion The process is simple and easy to operate and can be used for the extraction, purification and determination of EPA in kelp residue.