Improvement of chromatographic performances of in-situ synthesized hybrid C₈ silica monoliths by reduction of structural radial heterogeneities

Several modifications of a previously described protocol are proposed to improve the performances of in-situ synthesized C₈ hybrid silica monoliths. Our attention was focused on reducing the sources of radial heterogeneity that may be responsible for the poor efficiencies observed in the hydrodynamic elution mode. It was demonstrated that a decrease in the temperature of the capillary during the filling step equally to that of the polymerization mixture (0 °C), associated with a decrease of the gelation temperature to 20 °C along with a new pre-treatment of the capillary's internal walls [with a mixture of tetraethoxysilane (TEOS)/EtOH (1/3, v/v)] allows (i) increasing the radial homogeneity of the monolith, thus further enhancing the performances in the nano-liquid chromatography (nano-LC) mode, (ii) improving the capillary to capillary reproducibility in terms of permeability and efficiencies. In fact, the average minimum plate height H_min was lowered from 24 to 14 μm and the capillary-to-capillary reproducibility of the synthesis was widely improved by factors two and three of reduction on the calculated standard deviation, respectively for both the efficiency in the nano-LC mode and the permeability. At last, the improved radial homogeneity and anchoring of the synthesized monoliths allowed increasing the inner diameter of the capillary (up to 150 μm) without any significant loss in efficiency. Finally, long term stability of the as-obtained monolithic stationary phases in terms of retention and efficiency was studied. In addition, the evaluation of their chromatographic behaviour was also achieved with the Tanaka test and the results were compared to those already published for commercial monoliths (Chromolith) as well as for particulate stationary phases.