摘要：
目的 比较溴化十六烷基三甲铵(CTAB)-正丁醇-正辛烷反胶束体系及丁二酸-2-乙基己基酯磺酸钠(AOT)-异辛烷反胶束体系的水蛭素萃取效果.方法 以抗凝血酶活性单位(ATU)的萃取率为指标,用凝血酶滴定改进法测定萃取物的活性,正交实验对盐浓度、pH,搅拌时间、表面活性剂浓度等因素进行考察,优化CTAB-正丁醇-正辛烷体系及AOT-异辛烷体系的水蛭素对照品溶液反胶束萃取工艺,并进行水蛭素粗提液的分离纯化实验.结果 水蛭素CTAB反胶束体系最佳萃取工艺为:萃取pH10.0,反萃取pH 2.0,萃取液NaBr浓度30 mmol·L-1,CTAB浓度85 mmol·L-1,反萃取液NaBr浓度32 mmol·L,萃取及反革取均为30°C,萃取及反萃取均搅拌5 min;AOT反胶束体系最佳萃取工艺为:萃取pH 2.0,反萃取pH 9.0,萃取液KCI浓度26.5mmol·L-1,AOT浓度30 mmol·L-1,反革取液KCl浓度27.5 mmol·L-1,萃取及反革取均为30°C,萃取搅拌5min,反革取搅拌9 min.ATU回收率前者为82.95%,后者为85.53%;水蛭素粗提液分离纯化实脸两者的ATU回收率分别为80.70%和82.15%、比活性分别为2389.54及2459.98 ATU·mg-1.结论 两种反胶束体系的水蛭素萃取效果都较好,而后者又稍优于前者.%OBJECTIVE To compare the extraction effects of cetyl-trimethyl-ammnonium bromide ( CTAB)-normal butyl alcohol-octane reverse micelle and succinate-(2-ethylhexyl) sodium sulphosuccinate ( AOT)-isooctane reverse micelle for hirudin. METHODS Using the extraction rate of antithrombin activity units (ATU) as the index, the modified titration method of thrombin was used to detect the activity of the extracts. The optimal condition of the reverse micelle systems of CTAB-normal butyl alcohol-octane and AOT-isooctane was determined by the orthogonal test with factors such as salt concentration, Ph, mixing time, surfactant concentration and so on. And then the separation and purification test of hirudin crude solution was performed. RESULTS The optimal conditions of the reverse micelle system of CTAB with n-octane as extraction solvent and distilled water as anti-extraction solvent were: extraction Ph of 10.0, anti-extraction Ph of 2.0, concentration of extractant NaBr of 30 mmol ? L-1, CTAB concentration of 85 mmol ? L-1, concentration of anti-extractant NaBr of 32 mmol ? L -1, temperature and stirring time for extraction and anti-extraction of 30 °C and 5 min, respectively. The optimal conditions of the reverse micelles system of AOT with isooctane as extraction solvent and distilled water as anti-extraction solvent were; extraction Ph of 2.0, anti-extraction Ph of 9.0, concentration of extractant KC1 of 26.5 mmol ? L-1, AOT concentration of 30 mmol ? L-1, concentration of anti-extractant KC1 of 27.5 mmol ? L -1, temperature for extraction and anti-extraction of 30 °C, stirring time for the twp processes of 5 min and 9 min, respectively. The ATU back extraction rates of the two systems were 82.95% and 85.53% , respectively. The ATU back extraction rates of the two systems in the separation and purification of the crude extracts of hirudin were 80.70% and 82. 15%. The specific activities were 2 389. 54 and 2 459. 98 ATU ? Mg~', respectively. CONCLUSION The two reverse micelle extraction systems of hirudin show good results, in which the latter one is slightly better than the former.
