制备混合模式固定相用于高效液相色谱法中肽和蛋白质的分离。
Preparation of mixed-mode stationary phase for separation of peptides and proteins in high performance liquid chromatography.
机构信息
Department of Chemistry, College of Science, Taif University, P.O. Box 11099, Taif, 21944, Saudi Arabia.
Department of Chemistry, The University of Haripur, Haripur, 22620, KPK, Pakistan.
出版信息
Sci Rep. 2022 Mar 8;12(1):4061. doi: 10.1038/s41598-022-08074-7.
Porous silica particles were prepared by sol-gel method with some modification to get wide-pore particles. These particles were derivatized with N-phenylmaleimide-methylvinylisocyanate (PMI) and styrene by reversible addition fragmentation chain transfer (RAFT) polymerization to prepare N-phenylmaleimide embedded polystyrene (PMP) stationary phases. Narrow bore stainless steel column (100 × 1.8 mm i.d) was packed by slurry packing method. The chromatographic performance of PMP column was evaluated for the separation of synthetic peptides mixture composed of five peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine enkephalin) and tryptic digest of human serum albumin (HAS) respectively. Number of theoretical plates as high as 280,000 plates/m were obtained for peptides mixture at optimum elution condition. Separation performance of the developed column was compared with commercial Ascentis Express RP-Amide column and it was observed that separation performance of PMP column was better than commercial column in terms of separation efficiency and resolution.
采用溶胶-凝胶法并进行一些改进来制备多孔硅粒子，得到大孔粒子。这些粒子通过可逆加成-断裂链转移（RAFT）聚合与 N-苯基马来酰亚胺-甲基乙烯基异氰酸酯（PMI）和苯乙烯进行衍生化，制备 N-苯基马来酰亚胺嵌入聚苯乙烯（PMP）固定相。采用淤浆填充法填充窄孔径不锈钢柱（100×1.8mm id）。分别以合成的五肽混合物（Gly-Tyr、Gly-Leu-Tyr、Gly-Gly-Tyr-Arg、Tyr-Ile-Gly-Ser-Arg、Leucine enkephalin）和人血清白蛋白（HAS）的胰蛋白酶消化物为对象，评价了 PMP 柱的色谱性能。在最佳洗脱条件下，对于肽混合物，可获得高达 280000 块/米的理论塔板数。与商业 Ascentis Express RP-Amide 柱相比，开发柱的分离性能，在分离效率和分辨率方面，均优于商业柱。
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