论文部分内容阅读
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NaB r浓度30 mmol.L-1、CTAB浓度85 mmol.L-1,反萃取液NaB r浓度32 mmol.L-1,萃取及反萃取均为30℃,萃取及反萃取均搅拌5 m in;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KC l浓度26.5mmol.L-1、AOT浓度30 mmol.L-1,反萃取液KC l浓度27.5 mmol.L-1,萃取及反萃取均为30℃,萃取搅拌5 m in,反萃取搅拌9 m in。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验两者的ATU回收率分别为80.70%和82.15%、比活性分别为2 389.54及2 459.98 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 CTAB-n-butanol-n-octane reverse micellar system and the AOT-isooctane reverse micellar system Hirudin extraction effect. Methods The extraction rate of antithrombin activity unit (ATU) was used as an index, and the activity of extract was determined by the method of thrombin titration. The effects of salt concentration, pH, stirring time, surfactant concentration and other factors were investigated by orthogonal test. The reverse micelle extraction process of hirudin reference solution of CTAB-n-butanol-n-octane system and AOT-isooctane system was optimized, and the purification and purification of hirudin crude extract was optimized. Results The optimum extraction conditions of hirudin CTAB reverse micellar system were pH10.0, pH2.0, 30 mmol.L-1 NaBr, 85 mmol.L-1 CTAB and NaBr Concentration of 32 mmol.L-1, extraction and back-extraction were 30 ℃, extraction and back-extraction were stirred 5 min; AOT reverse micelles system extraction process: extraction pH 2.0, stripping pH 9.0, extract KC Concentration of 26.5mmol.L-1, AOT concentration of 30mmol.L-1, KC l concentration of the stripping solution 27.5mmol.L-1, extraction and back-extraction were 30 ℃, extraction and stirring 5min, m in. The recovery of ATU was 82.95% for the former and 85.53% for the latter. The recovery rates of the crude extracts of hirudin were 80.70% and 82.15% respectively, and their specific activities were 2 389.54 and 2 459.98 ATU.mg-1 . Conclusions The two kinds of reverse micelle systems have better hirudin extraction efficiency, while the latter is slightly better than the former.