Microchip electrophoresis at elevated temperatures and high separation field strengths

We report free-solution microchip electrophoresis performed at elevated temperatures and high separation field strengths. We used microfluidic devices with 11 cm long separation channels to conduct separations at temperatures between 22 (ambient) and 45°C and field strengths from 100 to 1000 V/cm. To evaluate separation performance, N-glycans were used as a model system and labeled with 8-aminopyrene-1,3,6-trisulfonic acid to impart charge for electrophoresis and render them fluorescent. Typically, increased diffusivity at higher temperatures leads to increased axial dispersion and poor separation performance; however, we demonstrate that sufficiently high separation field strengths offset the impact of increased diffusivity in order to maintain separation efficiency. Efficiencies for these free-solution separations are the same at temperatures of 25, 35, and 45°C with separation field strengths ≥ 500 V/cm.