Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
47, Number 6, July 2009, pp. 418-431
Recent Applications of Organic Monoliths in Capillary
Liquid Chromatographic Separation of Biomolecules
Rania Bakry, Christian W. Huck, and Günther
K. Bonn
Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens
University, Innrain 52a, 6020-Innsbruck, Austria
Monolithic columns are an attractive alternative
to traditional particulate solid phases for capillary liquid
chromatography. A monolith is a continuous interconnected skeleton
with large through-pores. This structure reduces the diffusion
path and provides high permeability, resulting in excellent separation
efficiency. The integral structure enhances the mechanical strength,
while the large through-pores (a few µm) have very low
flow impedance. This combination allows smaller diameter monolithic
columns to be operated at higher flow-rates, simultaneously increasing
both sensitivity and throughput. Polymeric monoliths were first
described back in the 1960s, but the first successful ones designed
for protein separations appeared much later, in the late 1980s.
Organic monoliths are based upon copolymerization of a monofunctional
and a bifunctional (uncommonly trifunctional) organic precursor
in the presence of a suitable initiator and porogenic solvents.
During the last 15 years, a vast number of different monomers
and crosslinkers have been introduced and copolymerized using
different polymerization techniques and initiators. Various mechanisms,
including thermally- and UV-initiated free radical polymerization,
as well as ring opening metathesis copolymerizations, have been
demonstrated for the preparation of monolithic columns. In this
review, we summarize the recent application of different organic
monoliths, including styrene-, acrylate-, methacrylate-, and
acrylamide for the liquid separation of biomolecules (e.g., proteins,
peptides, and oligonucleotides).
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