Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
39, Number 9, September 2001, pp. 361-364
Mobile
Phase Viscosity and Velocity Dependence on Protein Retention Using Nonequilibrium
Chromatographic TechniquesYves
Claude Guillaume, Mireille Thomassin, and Christiane Guinchard Equipe de Chimie Therapeutique, Laboratoire de Chimie Analytique, Faculte
de Medecine Pharmacie, Place Saint Jacques, 25030 Besancon Cedex, France
Nonequilibrium
chromatography (NEC) is an alternative chromatographic procedure for the separation
of macromolecules. The retardation of a protein series is studied using a phosphate
buffer as a mobile phase with various concentrations of glycerol fraction (used
as a viscosity modifier) at different mobile phase velocities and a C1 column
with a very low packing particle diameter as a stationary phase. It is shown
that the two factors (viscosity and velocity) of the mobile phase constituted
important parameters in the retention mechanism of the proteins in NEC. The
retardation velocity domain is divided into two regions. For low velocity regions,
the protein retention decreased with a mobile phase velocity increase. This
retention is enhanced above a critical value of the mobile phase velocity. The
transition between the two well-known NEC methods, slalom chromatography and
hydrodynamic chromatography, is clearly visualized for the first time for the
protein retention of particular values of the mobile phase velocity.
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