Monolithic capillary columns prepared by ring-opening metathesis polymerisation

Miniaturisation has been one of the main trends in HPLC over the past 10 years. To cope with the increasing requirements in the field of proteomics and drug discovery, separation media with smaller inner diameters, higher separation efficiency and better sensitivity are needed. Monolithic stationary phases represent an attractive approach for the fabrication of capillary columns, due to their ease of preparation without the requirement of sophisticated packing procedures or the manufacturing of end frits. Monolithic capillary columns were prepared from silanised fused-silica capillaries of 50 - 200 µm inner diameter by Ring-Opening Metathesis Polymerisation (ROMP) [2]. Monolithic columns based on norborn-2-ene (NBE) and 1,4,4a,5,8,8a-hexahydro-1,4,5,8,-exo,endo-dimethanonaphthalene (DMN-H6) are copolymerised with Grubbs-type initiator Cl2(PCy3)2Ru=CHPh using suitable porogenic systems [3]. We will show that the synthesised monoliths allow rapid and highly efficient separation of proteins and peptides by reversed phase high performance liquid chromatography. Furthermore, variations in polymerisation parameters allow the preparation of tailor-made separation media. The effect of a modified manufacturing process on batch-to-batch reproducibility of capillary monoliths will be shown by different protein and peptide mixtures. [1] Sinner, F. M., Buchmeiser, M. R., Angew. Chem. 112, 1491 (2000) [2] Mayr, B., Hölzl, G., Eder, K., Buchmeiser, M. R., Huber, C. G., Anal. Chem. 74, 6080 (2002) [3] Sinner, F. M., Buchmeiser, M. R., Macromolecules 33, 5777 (2000)