We present a method that achieves simultaneous preconcentration and separation of analytes using peak-mode isotachophoresis with a single step injection in simple, off-the-shelf microchannels or capillaries. We leverage ions resulting from dissolved atmospheric carbon dioxide to weakly disrupt isotachophoretic preconcentration and induce separation of analyte species. We experimentally study the region between the leading and trailing electrolytes, and individually identify the carbonate and carbamate zones that result from the hydration and carbamation reaction of dissolved atmospheric carbon dioxide, respectively. The width of these zones and the gradient regions between them grow with time and create an electric field gradient that causes analytes to separate. Using this assay, we achieve focusing and separation of a 25 bp DNA ladder in a straight, 34 µm wide microchannel in a single loading step. As a demonstration of the fractionation capabilities of the assay, we show simultaneous preconcentration and separation of a DNA ladder from two proteins, GFP and allophycocyanin.
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