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    新型磁性分子印迹聚合物快速萃取-高效液相色谱法测定牛血清白蛋白

    Determination of Bovine Serum Albumin by High Performance Liquid Chromatography After Rapid Extraction Using a New Magnetic Molecularly Imprinted Polymer

    • 摘要: 以牛血清白蛋白(BSA)为模板分子,多巴胺为功能单体,磁性四氧化三铁-石墨烯-多壁碳纳米管(Fe3O4-G/MWNT)为载体,制备了一种新型磁性分子印迹聚合物材料(Fe3O4-G/MWNT@MIP),用于萃取牛血清白蛋白。用10 mmol·L-1的磷酸盐缓冲溶液(pH 6.0,含1.0 mmol·L-1 NaCl)将血清样品稀释10倍,加入20.0 mg的Fe3O4-G/MWNT@MIP,振摇平衡20 min,弃去液体,用5.0 mL的10 mmol·L-1 NaCl溶液(pH 6.0)洗涤固体,再用2.0 mL的0.5 mmol·L-1 NaCl溶液(pH 5.0)解吸20 min,将洗脱液移至超滤管中,以12 000转·min-1离心20 min后,取超滤液进行高效液相色谱分析。结果表明:Fe3O4-G/MWNT@MIP对BSA具有良好的选择性,最大吸附容量达56.79 mg·g-1,重复使用20次后,最大吸附容量仅降低了7%。方法成功于应用于小牛血清中牛血清白蛋白的测定,加标回收率在90.0%~120%之间,测定值的相对标准偏差(n=7)小于3.0%。

       

      Abstract: A novel magnetic molecularly imprinted polymer (Fe3O4-G/MWNT@MIP) was prepared with bovine serum albumin (BSA) as the template molecule, dopamine as the functional monomer, and magnetic ferroferric oxide-graphene-multiwalled carbon nanotubes (Fe3O4-G/MWNT) as carriers, and used for extraction of BSA. After the serum sample was diluted 10-fold with 10 mmol·L-1 phosphate buffer solution (pH 6.0, containing 1.0 mmol·L-1 NaCl), 20.0 mg of Fe3O4-G/MWNT@MIP was added, and the mixture was shaken for 20 min. The liquid was discarded, and the solid was washed with 5.0 mL of 10 mmol·L-1 NaCl solution (pH 6.0), then desorbed with 2.0 mL of 0.5 mmol·L-1 NaCl solution (pH 5.0) for 20 min. The eluate was transferred into a ultrafitration tube and centrifuged at 12 000 r·min-1 for 20 min, and the ultrafitrate was taken for high performance liquid chromatography analysis. The results showed that, Fe3O4-G/MWNT@MIP had good selectivity to BSA, and the maximum adsorption capacity was 56.79 mg·g-1. After repeated use for 20 times, the maximum adsorption capacity only reduced by 7%. The method was successfully applied to determination of BSA in calf serum. Values of recovery obtained by standard addition method were between 90.0% and 120%, and the relative standard deviations (n=7) of the measured values were less than 3.0%.

       

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