注射用高纯度蛋黄卵磷脂制备新工艺的研究

A New Method for the Preparation of High Purity Egg Yolk Lecithin Used for Injection

本文探索建立了一条以新鲜蛋黄为原料,以乙醇为唯一提取溶剂,制备高纯度注射用蛋黄卵磷脂的新工艺。首先,用4倍重量的96%(v/v)乙醇作为沉淀剂将新鲜蛋黄中的蛋白质沉淀为滤饼,得到初级提取液;然后,再用4倍重量的96%(v/v)乙醇对滤饼抽提3次,提取残余卵磷脂,得到次级提取液。合并提取液,置9℃下静置分层,除去大部分蛋黄油。所得上清液上硅胶柱,用86%(v/v)的乙醇作为洗脱剂,纯化蛋黄卵磷脂,经高效液相色谱检测纯度达98%。并对提取过程进行了优化,得到最佳工艺条件:35℃的96%乙醇溶液,添加量为1∶4(w/w)提取蛋黄液,180 rpm转速下搅拌10 min。抽滤后,用60℃的96%乙醇提取滤饼3次,料液比为1∶4,静置15 min。最佳工艺条件下,蛋黄卵磷脂的提取率可达93.38%。

A new process was explored and established for the preparation of high purity egg yolk lecithin used for injec-tion using fresh egg yolk as raw material,ethanol as the only extraction solvent. First of all,4 parts 96% （v/v） ethanol was used to precipitate the protein from the fresh egg yolk. The egg yolk protein as filter cake and the primary extract as filtrate were collected separately;after that, the remaining lecithin in filter cake was extracted by 4 parts 96% （v/v） eth-anol for 3 times as the secondary extract. Then the primary and secondary extract were combined and stored at 9 ℃ over night to remove most of the egg yolk oil. Finally,the resulting supernatant was separated via silica gel column with 86% （v/v） ethanol as eluent, and the purity of egg yolk lecithin fraction, detected by HPLC, reached to 98 %. Meanwhile, the extraction process was optimized as follows ：for precipitation,35 ℃ ,the ratio of 96% （v/v） ethanol to fresh egg yolk was 4：1 （w/w） ,with 180 rpm stirring speed for 10 min;for extraction,60 ℃ ,the ratio of 96% （v/v） ethanol to filter cake was 4：1 ,standing 15 min for 3 times. Under optimized conditions,the extraction yield of egg yolk lecithin reached to 93.38%.