A 2 m long open tubular column with a copolymer layer fabricated on the inner surface of silica capillary of 50 μm id has been prepared. A ligand with a terminal halogen was bound to the capillary inner surface via silanol-isocyanate reaction and an initiator moiety was introduced by reacting with the terminal halogen. Then a thick layer of linear copolymer chains was generated on the inner surface of capillary by reversible addition-fragmentation transfer polymerization of styrene and N-phenyl acrylamide. The resultant open tubular column exhibited a remarkably high separation efficiency for the separation of a synthetic mixture of five peptides in liquid capillary chromatography (in the pressure mode). The effect of monomer mixing ratio (styrene versus N-phenyl acrylamide) on the chromatographic separation efficiency has been studied. The column prepared with the optimum mixing ratio produced the number of theoretical plates (N) of 391,200 per column under the optimized elution pressure. The column to column repeatability estimated as relative standard deviation in plate count, retention factor, asymmetry factor, and resolution was found better than 3.0%.

中文翻译：

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用于液相色谱分离多肽的 2 m 长、内径为 50 μm 的开放式毛细管柱的制备和评价

已经制备了一个 2 m 长的开放管柱，在 50 μm id 的硅胶毛细管的内表面上制造了共聚物层。具有末端卤素的配体通过硅烷醇-异氰酸酯反应结合到毛细管内表面，并且通过与末端卤素反应引入引发剂部分。然后通过苯乙烯和N-苯基丙烯酰胺的可逆加成-断裂转移聚合在毛细管内表面生成一层厚厚的线性共聚物链。所得开管柱在液体毛细管色谱（压力模式）中分离五种肽的合成混合物时表现出非常高的分离效率。已经研究了单体混合比（苯乙烯与 N-苯基丙烯酰胺）对色谱分离效率的影响。以最佳混合比制备的色谱柱在优化的洗脱压力下产生的理论塔板数 (N) 为每柱 391,200。发现以塔板数、保留因子、不对称因子和分辨率的相对标准偏差估算的柱间重复性优于 3.0%。