CHAPTER 13

Hypercrosslinked Polystyrene as
Column Packing Material in HPLC

1. MACROPOROUS POLYSTYRENE VERSUS SILICA-BASED

HPLC PACKINGS
It was recognized by the end of the 1970s that the resolving power of
column liquid chromatography can be substantially enhanced by using very
small particles as the column packing materials, preferably with diameters in
the 3–10 mm range. Additional optimization of the porous structure of
these adsorbing particles allows reduction of the height equivalent to
theoretical plate to two to three particle diameters, thus attaining high
column efficiency of about 40,000–100,000 theoretical plates per meter
column length. This was the beginning of the era of high-performance
liquid chromatography (HPLC), which has become the leading analytical
technique today.
Over 500 HPLC packings have been described in the literature. Never­
theless, as the result of years of development, only a limited number of types
of stationary phases remain on the market. Most of the conventional HPLC
separations today are performed using monodisperse silica gel 3 or 5 mm
microbeads, especially those grafted with C4, C8, or C18 alkyl chains, as
well as with cyano-propyl or amino-propyl groups. The last two bonded
silicas and bare silica are used in normal phase (NP) HPLC, where the
mobile phase (usually hexane with small amounts of isopropyl alcohol) is
less polar than the stationary phase. Even more popular is the reversed phase
(RP) mode, which uses polar eluents (mostly water or methanol with such
additives as acetonitrile, methanol, or tetrahydrofuran (THF)) in combina­
tion with nonpolar alkyl-bonded stationary phases.
The drawbacks of silica-based materials are well known. Most serious
among them are reduced hydrolytic stability in aqueous and aqueous-
organic media, which practically eliminates the possibility of regenerating
a contaminated column by an acidic or alkaline wash and dramatically
reduces the lifetime of a column used outside of the 2–8 pH range.

Comprehensive Analytical Chemistry, Volume 56  2011 Elsevier B.V.

ISSN 0166-526X, DOI 10.1016/S0166-526X(10)56013-7 All rights reserved. 503