I have heard many chromatographers describe problems with
HILIC methods they use. Common complaints include long equilibration times and
inconsistent retention when using gradients. Given the nature of the HILIC
retention mechanism, this is not surprising. In HILIC, hydrophilic silanol
groups on the stationary phase surface cause a thick water layer to form, which
allows for the partitioning responsible for analyte retention. The problem with
this retention mode is that the water layer is not a static environment but
rather is constantly changing as the mobile phase changes over the course of a
gradient. Then, during re-equilibration, it takes a significant number of
column volumes to fully regenerate the hydration shell to its original form. It
is this variability in the water layer that gives rise to both long
equilibration times and poor retention precision.
So what solution is there to this kind of problem? It would
be better to rely on a retention mode that does not involve partitioning with a
water layer. With Cogent TYPE-C Silica™ columns, hydrophilic silanols are replaced by
silica hydride groups. Because the latter are moderately hydrophobic, they do
not result in the formation of a thick water layer, as in conventional silica
based phases. Polar compounds can still be retained using these columns in a
mode called Aqueous Normal Phase (ANP). Mechanistic studies involving
measurements of zeta potentials have demonstrated that the retention in ANP is
primarily adsorptive in character rather than due to partitioning. Indeed, the
greater precision and lower equilibration times observed using ANP methods
compared to HILIC are consistent with these findings.
What this means to you is that you can obtain more reliable
data with greater throughput. Solvent savings can also be realized due to the
minimal equilibration. Furthermore, the same column can be used in some
instances for both ANP and reversed phase chromatography, allowing for more
streamlined analyses. With HILIC columns, only polar analytes may be suitable
for retention while more hydrophobic species may elute at the solvent front.
Hence, you can get more versatility out of one column with Cogent TYPE-C Silica™
phases.
Ever-increasing globalization in many industries that rely
on HPLC has created more competition than ever before. In today’s business
environment, laboratories need to maximize their resources in order to stay
competitive. Using Cogent TYPE-C Silica™ columns is one way to help you achieve
these goals.
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