Reversed-phase chromatography of proteins and peptides on compressed continuous beds

Characterization of monolithic columns for HPLC

“…Encapsulated in a tube that prevents the stream of mobile phase from by-passing the porous medium, this block constitutes a monolithic column. Pioneering work by Nakanishi [1 -3], Svec [4 -9], and HjertØn [10,11] led to the breakthrough by Tanaka et al [12 -18] that suddenly brought to the world's attention the unusual and attractive properties of monolithic columns and allowed the preparation of new, superior separation media for chromatography. More than 30 years earlier, the considerable potential advantages of monolithic columns had already been suggested by Knox [19].…”

Stellan Hjertén's contribution to the development of monolithic stationary phases

“…The next step was then dehydration described above followed by reaction with 1,2-epoxyoctane or 1,2-epoxyoctadecane. The resulting hydrophobic particles were then packed in a column and compressed to the final length [10]. The performance demonstrated only with separations of various digests appeared to be decent but without mass spectrometric assignment of the peaks not very useful.…”

“…However, RP chromatography that is very popular mode for the separation of proteins requires stationary phase containing highly hydrophobic ligands to achieve the desired interactions and a mobile phase containing an organic modifier. Since polymerization of hydrophobic monomers in aqueous system is difficult, Hjerten's group prepared the columns in two steps [10]. The first step was the preparation of a gel from a mixture consisting of piperazine diacrylamide, methacrylamide, and 10% allylglycidyl ether using the typical polymerization conditions.…”

“…Monolithic capillary columns with hydrophobic chemistry for the RP separation were prepared using a technique similar to that described above [10]. First, a monolith was prepared by copolymerization of piperazine diacrylamide and methacrylamide.…”

Continuous Beds for Microchromatography: Reversed-Phase Chromatography

“…When a water-soluble monomer with the resolutions for preparing these beds is shown in Table 1 quired ligand group is not easily available, the group (the composition is modified with regard to that decan be introduced following the synthesis of the bed scribed in Ref. (3), as discussed under Results and Dis- (2). In this paper we describe such a procedure for the cussion).…”