【摘要】 目的:研究以菲牛蛭为原料,二-(2-乙基已基)磷酸(D2EHPA)作流动载体,脱水山梨醇脂肪酸酯(Span 80)为乳化剂,辛烷与D2EHPA混合构成膜溶液,稀HCl溶液作内水相的水蛭素乳状液膜体系萃取。方法:用正交试验优化膜相、内水相体积比(MIPVR),内、外水相的pH,流动载体浓度等水蛭素对照品萃取条件,然后用水蛭素粗提液进行纯化实验。结果:萃取的最优条件为:MIPVR 10∶3,内水相pH 2.6,外水相pH 3.4,流动载体质量分数2%;当水蛭素初始浓度为1抗凝血酶活力单位(ATU).mL-1时,对照品ATU回收率为83.06%,粗提液达到82.99%,样品比活力为3 289.48 ATU.mg-1,不连续聚丙烯酰胺凝胶电泳及光谱扫描结果表明纯度与对照品相当。结论:乳状液膜体系萃取水蛭素工艺较简单,实验样品纯度及ATU回收率均较高。更多还原

【Abstract】 Objective: To study the extraction system of hirudin emulsion liquid membrane with the Poecilobdella manillensis as raw material,di-(2-ethylhexyl) phosphate(D2EHPA) as carrier,Span 80 as emulsifier,octane and D2EHPA mixed to constitute membrane solution,diluted HCl solutions as internal aqueous phase.Method: Using the orthogonal experiment to optimize the extraction conditions of hirudin reference substance such as membrane phase,internal aqueous phase volume ratio(MIPVR),external aqueous phase pH,internal aqueous phase pH,mobile carrier concentration and so on,and then using hirudin crude extracts to do purifying experiment,and gaining experimental samples.Result: The optimal conditions of hirudin extraction were as follows: MIPVR 10∶3,internal aqueous phase pH 2.6,external aqueous phase pH 3.4,the mass fraction of carrier D2EHPA 2%.In the optimal extraction conditions,when the initial concentration of hirudin was one anti-thrombin activity units(ATU)·mL-1,ATU recovery rate of the reference substance was 83.06%.In the purifying experiment of crude extracts,ATU recovery rate was 82.99%,and the specific activity of sample was 3 289.48 the ATU·mg-1.Discontinuous polyacrylamide gel electrophoresis and spectral scanning,the results showed that the purity and reference substance were considerable.Conclusion: The method of preparation hirudin was relatively simple,the purity of the experimental samples and ATU recovery were both high.更多还原