Improving peak capacity in fast online comprehensive two-dimensional liquid chromatography with post-first-dimension flow splitting.

Abstract

The use of flow splitters between the two dimensions in online comprehensive two-dimensional (2D) liquid chromatography (LC × LC) has not received very much attention, in comparison with their use in 2D gas chromatography (GC × GC), where they are quite common. In principle, splitting the flow after the first dimension column and performing online LC × LC on this constant fraction of the first dimension effluent should allow the two dimensions to be optimized almost independently. When there is no flow splitting, any change in the first-dimension flow rate has an immediate impact on the second dimension. With a flow splitter, one could, for example, double the flow rate into the first dimension column and perform a 1:1 flow split without changing the sample loop size or the sampler's collection time. Of course, the sensitivity would be diminished, but this can be partially compensated through the use of a larger injection; this will likely only amount to a small price to pay for this increased resolving power and system flexibility. Among other benefits, we found a 2-fold increase in the corrected 2D peak capacity and the number of observed peaks for a 15-min analysis time, using a post-first-dimension flow splitter. At a fixed analysis time, this improvement results primarily from an increase in the gradient time, resulting from the reduced system re-equilibration time, and, to a smaller extent, it is due to the increased peak capacity achieved by full optimization of the first dimension.