Free‐flow electrophoresis of proteins in solutions of borax and mannitol was investigated. The borax solution represents an equimolar boric acid‐NaOH buffer with a pH of 9.2. On addition of mannitol the pH of the buffer decreases, with increasing polyol concentration, accompanied by a slight decrease in conductivity. Using a 2.5 mM borax solution, buffers with pH from 9.2 to 4.5 can be obtained. Under the influence of the electric field the pH and conductivities of these buffers change in the near‐membrane regions of the separation chamber. Despite the changes the buffers are suitable for electrophoresis of proteins in a free‐flow apparatus. The separation of bovine hemoglobin as well as a mixture of bovine hemoglobin, human serum albumin and cytochrome C in borax‐mannitol solutions is demonstrated at 80–100 mA constant current and a residence time of only 2.0–2.6 min.
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