PVC固载亲水性离子液体选择性分离血红蛋白研究

Preparation of bonded hydrophilic ionic liquid on PVC for the selective isolation of hemoglobin

将N-甲基咪唑嫁接到聚氯乙烯(PVC)链上制备了亲水性离子液体1-乙烯基-3-甲基咪唑氯代盐-PVC复合物,并用红外光谱、核磁共振氢谱和表面电荷分析对其进行了表征,咪唑基团对氯乙烯基中氯的取代率为2.8%.蛋白质吸附研究表明,在特定条件下该固载型离子液体对血红蛋白具有吸附选择性,据此建立了固相萃取分离血红蛋白的方法.考察了吸附时间、离子强度、洗脱剂及其浓度等对分离纯化效率的影响.在最优实验条件下,该固载型离子液体-PVC复合物对30μg·mL-1血红蛋白的吸附效率为91%,洗脱效率为75%,吸附容量为22.7μg·mg-1.用1.0%(m/V)SDS洗脱后血红蛋白活性约为原始溶液中的71%.SDS-PAGE凝胶电泳证明其有效地从人全血样品中分离出纯度较高的血红蛋白.

A polyvinyl chloride （PVC） supported hydrophilic ionic liquid （PVC-mimC1） was prepared by immobilizing N-methylimidazole （N-mim） onto the PVC chains. It was characterized by FT-IR, 1H NMR and surface charge analysis and the results indicted that 2.8% of Cl on the PVC chain were substituted by N-mim. A distinct feature of the bonded ionic liquid is its selectivity for the adsorption of hemoglobin at certain conditions, and a novel solid phase extraction procedure for the isolation of hemoglobin was proposed. A thorough scrutiny has been made to investigate the effects of a series of experimental parameters on the separation efficiency, including adsorption time, ionic strength, stripping reagent and its concentration. An adsorption efficiency of 91% was achieved for a hemoglobin solution of 30.0μg·mL-1 by 15.0 mg of PVC-mimCl, and 75% of the retained hemoglobin could be readily recovered by elution with a 1.0% SDS solution. The sorption capacity for hemoglobin was derived to be 22.7μg·mg-1. 71% of the activity of hemoglobin was maintained after pretreated by using the PVC-mimCl hybrid and eluted with 1.0% （m/V） SDS solution. Hemoglobin in human whole blood was isolated by this bonded hydrophilic ionic liquid and satisfactory results were obtained by SDS-PAGE assay.