离子液体双水相萃取及检测牛血中血红蛋白  

Extraction and detection of hemoglobin in bovine blood by ionic liquid-based aqueous Bbiphasic system

作　　者：姜健 刘波 李刚 刘宇 王香竹 JIANG Jian;LIU Bo;LI Gang;LIU Yu;WANG Xiang-zhu(College of Chemical Engineering and Machinery,Eastern Liaoning University,Dandong 118003,China;Research Centre for Analytical Sciences,Northeastern University,Shenyang 110819,China)

机构地区：[1]辽东学院化工与机械学院,辽宁丹东118000 [2]东北大学分析科学研究中心,辽宁沈阳110819

出　　处：《化学研究与应用》2022年第9期2142-2148,共7页Chemical Research and Application

摘　　要：为了探究溴化1-丁基-3-甲基咪唑([C_(4)mim]Br)离子液体形成双水相及萃取生物活性物质的能力,采用[C_(4)mim]Br离子液体浓溶液与Na_(2)CO_(3)溶液形成双水相,并进行了牛血红蛋白的萃取试验。结果表明:[C_(4)mim]Br离子液体可与Na_(2)CO_(3)形成双水相。[C_(4)mim]Br溶液(91.87%)与Na_(2)CO_(3)溶液(0.208 g·mL^(-1))以体积比1∶5形成的双水相,分相后离子液体相(上相)体积变为原体积的2.30倍,相界面清晰,分相速度快。采用浊度滴定法测定了该双水相体系双节线,其拟合方程为:ω_(1)=-0.2036 ln(ω_(2)+0.0064)-0.2742,σ^(2)=0.0038。在静态条件下利用该双水相体系萃取牛血红蛋白,采用光纤光谱仪连续测定离子液体相中血红蛋白的吸光度,吸光度值在8 min内达到稳定。分析了该体系快速萃取牛血红蛋白的机理,紫外-可见吸收光谱及圆二色光谱(CD)分析结果表明萃取前后血红蛋白的二级结构未发生改变。浓度在0.010~0.200 mg·mL^(-1)之间时,该双水相体系对牛血红蛋白萃取率在93.40%~99.62%之间。建立了用该双水相体系萃取并检测牛血中血红蛋白的方法并测定了新鲜抗凝牛血中血红蛋白的含量,双水相上相中血红蛋白吸光度的线性方程为:A=1.814 c+0.024,n=7,R^(2)=0.9959,线性范围为0.010~0.450 mg·mL^(-1)。三份新鲜抗凝牛血中血红蛋白含量在102.0~118.6 mg·mL^(-1)之间,回收率在99.4%~105.0%,该离子液体双水相体系适用于新鲜抗凝牛血中血红蛋白含量的测定。In order to explore the ability of 1-butyl-3-methylimidazole bromide([C_(4)mim]Br)ionic liquid to form aqueous two-phase systems and extract bioactive substances,[C_(4)mim]Br solution and Na_(2)CO_(3)solution are used to form aqueous two-phase system which used to extract hemoglobin in bovine blood.The results show that[C_(4)mim]Br and Na_(2)CO_(3)can form ionic liquid-based aqueous biphasic systems.The aqueous two-phase system is formed by[C_(4)mim]Br solution(91.87%)and Na_(2)CO_(3)solution(0.208 g·mL^(-1))with a volume ratio of 1∶5.After phase separation,the volume of the ionic liquid phase(upper phase)became 2.30 times of the original volume.The phase interface was clear and the phase separation speed was fast.The binodal curve of the aqueous two-phase system is determined by turbidimetric titration.The empirical mathematical description of the binodal curve is:ω_(1)=-0.2036 ln(ω_(2)+0.0064)-0.2742,σ^(2)=0.0038.Under static condition,the absorbance value of bovine hemoglobin in ionic liquid phase continuously determined by optical fiber spectrometer reached stable within 8 min when the aqueous two-phase system was used to extract bovine hemoglobin.The mechanism of rapid extraction of bovine hemoglobin by this system is analyzed.The analysis results of UV-Vis absorption spectrum and CD spectrum showed that the secondary structure of hemoglobin was not changed before and after extraction.When the concentration of bovine hemoglobin is 0.010~0.200 mg·mL^(-1),the extraction efficiency is 93.40%to 99.62%.A method was established for the extraction of hemoglobin in bovine blood and determination the content of hemoglobin in fresh anticoagulant bovine blood.The linear equation of the absorbance of hemoglobin in the upper phase of the aqueous two-phase system is as follows:A=1.814 c+0.024,n=7,R^(2)=0.9959,and the linear range was 0.010~0.450 mg·mL^(-1).The hemoglobin content in three fresh anticoagulant bovine blood range from 102.0 to 118.6 mg·mL^(-1),and the recovery is 99.4%to 105.0%.The ionic liquid-based

关 键 词：溴化1-丁基-3-甲基咪唑 Na_(2)CO_(3) 离子液体双水相 牛血红蛋白 萃取 检测 

分 类 号：O658.2[理学—分析化学]