Sampling BIAS at Channel Junctions in Gated Flow Injection on Chips

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Abstract
The commonly used gated injection scheme was examined and found to suffer from multiple levels of electrokinetic sampling bias, including a new type based on transradial electrokinetic selection (TREKS). TREKS occurs as analytes of differing electrophoretic mobilities migrate around the corner at a channel junction in a microchip. The overall sample bias in gated injection was shown to be time-dependent and resulted in a larger sample bias against components of negative electrophoretic mobility. A new injection procedure for microchip devices based on interstream diffusion at zero potential is proposed. Diffusion of molecules into the separation channel is the main driving force for this type of injection. The new scheme is shown to be useful for injection of complex samples with multiply charged components, such as peptide mixtures. This procedure allows sampling of volumes from 12 to 45 pL, reproducible retention times (RSD < 1.5%), and reproducible peak areas (RSD < 2.3%).