无需两性电解质的小分子等电聚焦微量制备方法

摘要

Preparative ampholyte-carrier-free isoelectric focusing electrophoresis was established to separate small zwitterionic compounds.The pH gradient buffer solution was prepared by introducing a series of mixed acid and alkali solution at different pH value into a column containing agarose gel.After loading sample,addi-tional electric field was applied on the column to perform isoelectric focusing electrophoresis.With the continu-ous addition of last buffer solution,the samples were focused at certain pH value zone and eluted from the col-umn one by one,so that they could be fractionatedly collected.The effect of additional electric field on pH gra-dient was investigated.The effects of additional electric field and pH gradient on isoelectric focusing electropho-resis were also studied.Separation results of model samples revealed that small zwitterionic compounds could be separated according to the pI values by this method.%建立了无需两性电解质的等电聚焦微量制备方法，可以分离两性小分子物质。采用混合酸性和碱性溶液在填充琼脂糖凝胶的色谱柱中制备了 pH 值梯度变化的缓冲溶液，上样后，在色谱柱上外加电场进行等电聚焦分离，随着终止缓冲溶液的连续加入，各种样品被聚焦在一定的 pH 值区域并依次洗脱，分段收集。考察了外加电场对 pH 梯度的影响以及外加电场、pH 梯度对等电聚焦分离的影响。模式样品的分离结果表明，所建立的方法可以实现两性小分子物质按照等电点顺序依次分离。