An Agilent 1290 Infinity LC System is optimized for the lowest possible extra-column volume, to ensure minimal band broadening and optimal results from a small dimension, Agilent ZORBAX RRHD Eclipse Plus C18, 2.1 × 50 mm, 1.8 µm column. Improvements to isocratic and gradient analyses are evident, with the early eluting peaks from the isocratic analysis showing the greatest improvement of up to a 58% increase in efficiency (k’ = 1.6). Gradient analyses are improved by 24 to 32% with respect to conditional peak capacity (k’ = 1.3 to 5.3). Additionally, there is no strong correlation between flow rate and the effect of extra-column volume, with respect to column performance for isocratic and gradient analyses.
The document describes how to optimize the performance of a small dimension Agilent ZORBAX RRHD Eclipse Plus C18 column (2.1 x 50 mm, 1.8 pm) by minimizing extra-column volume on an Agilent 1290 Infinity LC System. Key optimizations include replacing standard capillaries with Ultra-Low Dispersion Capillaries (0.08 mm id), using a Max-Light Cartridge Flow Cell (V(o} = 0.6 pL), and utilizing the LC System Rack to reduce total extra-column volume by approximately 60%.
The column used is a 2.1 x 50 mm column with a particle size of 1.8 µm.
The optimized configuration has a total extra-column volume of 3.9 pL.
The flow rate for the isocratic analysis is 0.4 mL/min.