Department of Chemical Engineering, Vrije Universiteit Brussel (VUB), Brussels, Belgium.
Future Industries Institute, University of South Australia, Adelaide, Australia.
J Sep Sci. 2023 Sep;46(18):e2300439. doi: 10.1002/jssc.202300439. Epub 2023 Jul 28.
Polymer monolithic stationary phases are designed as a continuous interconnected globular material perfused by macropores. Like packed column, where separation efficiency is related to particle diameter, the efficiency of monoliths can be enhanced by tuning the size of both the microglobules and macropores. This protocol described the synthesis of poly(styrene-co-divinylbenzene) monolithic stationary phases in capillary column formats. Moreover, guidelines are provided to tune the macropore structure targeting high-throughput and high-resolution monolith chromatography. The versatility of these columns is exemplified by their ability to separate tryptic digests, intact proteins, and oligonucleotides under a variety of chromatographic conditions. The repeatability of the presented column fabrication process is demonstrated by the successful creation of 12 columns in three different column batches, as evidenced by the consistency of retention times (coefficients of variance [c.v.] = 0.9%), peak widths (c.v. = 4.7%), and column pressures (c.v. = 3.1%) across the batches.
聚合物整体固定相设计为一种连续的相互连接的球形材料，由大孔灌注而成。与分离效率与粒径有关的填充柱一样，通过调整小微球和大孔的尺寸，可以提高整体固定相的效率。本方案描述了在毛细管柱形式下合成聚苯乙烯-二乙烯基苯整体固定相。此外，还提供了针对高通量和高分辨率整体固定相色谱的大孔结构调整的指南。这些柱子的多功能性体现在它们能够在各种色谱条件下分离胰蛋白酶消化物、完整蛋白质和寡核苷酸。通过在三个不同的柱批次中成功制备 12 根柱，证明了所提出的柱制造过程的可重复性，这表现在保留时间（变异系数 [c.v.]=0.9%）、峰宽（c.v.=4.7%）和柱压（c.v.=3.1%）在批次之间的一致性。
