Homogeneous reversed-phase agarose thermogels for electrochromatography

A method for the derivatization of agarose by covalent attachment of hydrophobic ligands for reversed-phase (RP) chromatographic separation and ionic groups for generation of electroosmosis under electrochromatographic conditions in the capillaries or microfluidic channels filled with the thermogel of this agarose derivative is described. The product renders a capability of reversible thermogelation. The thermogels formed provide sufficient hydrophobicity and electroosmosis for the separations of the analytes under RP mobile-phase conditions and electric field applied. The gels may be used repeatedly without loss of resolution. They are thermally replaceable and UV transparent (providing possibility in column/in-gel detection), require no covalent attachment to the capillary inner wall (or microchip channel), and are suitable for isocratic or gradient operation in the aqueous–organic mobile phases.