Department of Chemistry, University of Malakand, Khyber Pakhtunkhwa, Pakistan.
Department of Chemistry, Inha University, Incheon, South Korea.
J Sep Sci. 2021 Apr;44(7):1430-1439. doi: 10.1002/jssc.202100001. Epub 2021 Feb 10.
A particle-based stationary phase has been prepared for the separation of five synthetic peptides and a mixture containing tryptic digest of cytochrome C in liquid chromatography. Particles originating from silica monolith were differentially sedimented to obtain 1-2 μm particles. A stationary phase was achieved by the coating of poly(styrene-methacrylic acid-N-phenylacrylamide) copolymer onto the particles via reversible addition-fragmentation chain transfer polymerization reaction. Stainless steel column (30 cm long and 1 mm internal diameter) was packed with stationary phase. Very high separation efficiency (ca. 351 000 plates/m) was achieved for five commercial peptides with a percent relative standard deviation of less than 1%. Protocol for the synthesis and modification of silica monolith particles has been well optimized with a good reproducibility both in particle and pore size. The column resolved about 21 peptide components from a mixture containing tryptic digest of cytochrome C, under the elution conditions of acetonitrile/15 mM ammonium format (65/35 v/v%) with a flow rate of 28 μL/min.
一种基于颗粒的固定相已被制备用于在液相色谱中分离五种合成肽和包含细胞色素 C 胰蛋白酶消化物的混合物。源自硅胶整体的颗粒通过差速沉降来获得 1-2μm 的颗粒。通过可逆加成-断裂链转移聚合反应将聚苯乙烯-甲基丙烯酸-N-苯丙烯酰胺共聚物涂覆到颗粒上，从而实现固定相。不锈钢柱（30cm 长，1mm 内径）用固定相填充。对于五种商业肽，实现了非常高的分离效率（约 351000 板/m），相对标准偏差小于 1%。已经对硅胶整体颗粒的合成和修饰方案进行了很好的优化，颗粒和孔径的重现性都很好。在流速为 28μL/min 的洗脱条件下，使用乙腈/15mM 甲酸铵（65/35v/v%），该柱从包含细胞色素 C 胰蛋白酶消化物的混合物中分离出约 21 个肽组分。
