Chromatographic Properties of Silica-Based Monolithic HPLC Columns
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The Chromolith SpeedRODâ ¢ (EM Science, Gibbstown NJ) is a commercially available silica-based monolithic column. This work investigated the chromatographic properties of the 50x4.60 mm (ODS) SpeedRODâ ¢. Data fit to the van Deemter equation (mean square error=0.834) indicated that the van Deemter model was valid for monolithic columns. An effective particle size of 4 Î¼m for the SpeedRODâ ¢ column was assigned by comparing the minimum of van Deemter curves with a series of particulate columns having various particle diameters. Separation Impedance (E), an empirically derived measure of column performance, was calculated as an alternate method of evaluating column efficiency. Data collected using this model confirmed monolithic columns behaves as a (more efficient) 3 Î¼m column.
A series of experiments were designed to compare the effects of mobile phase strength and mobile phase viscosity between the SpeedRODâ ¢ column and a particulate column. The results indicated both solvent strength and viscosity have effects on the monolithic column at the optimum linear velocity.
A fast (90 s) HPLC method was developed using the SpeedRODâ ¢ column and a seven-component test mixture with a large range of hydrophobicities. The precision for both retention time and peak area was measured at high linear velocities (8 mL/min) and the percent relative standard deviation (RSD) calculated. Column to column reproducibility (n=6) was measured. The overall percent RSD ranged from 0.25% to 4.56% for retention time and from 1.08% to 6.77% for peak area. Run to run reproducibility (n=15) was measured for all six columns. Averages ranged for retention time from 0.89% to 5.09% RSD and for peak area from 4.65% to 6.18% RSD.
Applications for the SpeedRODâ ¢ column with various sample types were developed and discussed. These methods demonstrated the effectiveness of the SpeedRODâ ¢ at fast flow rates.
- Doctoral Dissertations