Stacking in a continuous sample flow interface in capillary electrophoresis

Using a tee connector in a commercial capillary electrophoresis instrument, the effect of field amplified sample injection from both flowing and static sample volumes was investigated. It is shown that under identical conditions (40min electrokinetic injection at 5kV from a sample volume of 295μL) the limit of detection using the continuous sample flow interface is 4 times lower than from a static vial. The relationship between different flow rates and injection voltages on the injected sample amount was also investigated using a 2D axisymmetric simulation (COMSOL 4.3b) and verified experimentally, confirming conditions under which there is near-quantitative injection of the sample target ions. Using electrokinetic injection at 30kV and a flow rate of 558nL/s the same enhancement from an even smaller volume of 184μL could be achieved in 5.5min than could be achieved from 295μL and a 40min injection. This sensitivity enhancement factor corresponded to four orders of magnitude improvement compared to a hydrodynamic injection. This is the first report showing that a continuous sample flow interface combined with stacking methods under conditions approaching quantitative injection from the entire sample volume has the potential to be more sensitive than a static system.