Stability and repeatability of capillary columns based on porous monoliths of poly(butyl methacrylate-co-ethylene dimethacrylate)

Abstract

Monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns have been prepared via either thermally or photochemically initiated polymerization of the corresponding monomers and the repeatability of their preparation has been explored. Three separate batches of five columns each were prepared using thermal and photochemical initiation for a total of thirty columns. All thirty capillary columns were tested in liquid chromatography-electrospray ionizationmass spectrometry mode for the separation of a model mixture of three proteins -ribonuclease A, cytochrome c and myoglobin. Excellent repeatability of retention times was observed for the proteins as evidenced by relative standard deviation (RSD) values of less than 1.5%. Somewhat broader variations with RSD values of up to 10% were observed for the pressure drop in the columns. The stability of retention times was also monitored using a single monolithic column and no significant shifts in either retention times or back pressure was observed in a series of almost 2200 consecutive protein separations.

Basic Chromatographic Properties of Polyethylene Glycol-type, Polymer-based Monolithic Columns

“…such as 50 -100 μm because of shrinkage of the polymer. 9,[16][17][18][19][20][21][22][23][24][25] In this experiment, in spite of the wider 0.53 mm i.d. capillary, PEG-type monolithic polymers were formed homogeneously.…”

Optimization of the porous structure and polarity of polymethacrylate‐based monolithic capillary columns for the LC‐MS separation of enzymatic digests

“…In contrast, methacrylate-based monolithic columns only gave satisfactory results in the separation of proteins [30], but their use in the separation of peptides has met with limited success due to the weak retention and limited resolution of the columns [28,31].…”

Monolithic porous polymer stationary phases in polyimide chips for the fast high-performance liquid chromatography separation of proteins and peptides

“…Different liquid precursors have been used for the preparation of polymer-based monolithic stationary phases [16][17][18][21][22][23]. Monomers such as acrylamide [23], styrene, divinylbenzene [24][25], as well as various acrylates [26], methacrylates [27][28][29][30], and norbornene [31] have all been used. The morphology of the monolithic polymer is strongly affected by both the composition of the polymerization mixture and the conditions under which the polymerization process is carried out [21,28,29].…”