One-step Concentration of Analytes Based on Dynamic Change in PH in Capillary Zone Electrophoresis

A novel strategy for one-step concentration of analytes during capillary electrophoresis (CE) is presented. A short platinum wire was inserted into the 75-microm-i.d. separation capillary. When a high voltage was applied for CE separation, a sharp pH gradient along the capillary was created dynamically by the electrolysis of water in the running buffer. Concentration of a large volume of injected analytes was accomplished by the change in analyte charge due to the dynamic pH gradient. Depending on the polarity of the applied potential and the direction of electroosmotic flow, either anions or cations can be concentrated. Several hundredfold concentration factors were achieved. Fluorescence imaging by a CCD camera was used to monitor 10 cm of the capillary near the platinum wire during the concentration process. The observations are consistent with a sweeping mechanism.