Microchip free flow planar reversed phase electrochromatography with monolithic stationary phase

Area of Science:

• Analytical Chemistry
• Separation Science
• Microfluidics

Background:

• Traditional separation techniques often face limitations in resolution and throughput.
• Microfluidic devices offer miniaturization and enhanced control for separation processes.

Purpose of the Study:

• To develop and validate a novel microchip free flow planar RP electrochromatography (microFF-PRPEC) system.
• To evaluate the separation efficiency of microFF-PRPEC for both small molecules (dyes) and large biomolecules (proteins).

Main Methods:

• In situ polymerization of poly(butyle methyacrylate-co-ethylene dimethacrylate) monolithic material within a microchamber.
• UV-initiated polymerization using specific porogens and 2-acrylamide-2-methyl-propanesulfonic acid (AMPS) to induce electroosmotic flow (EOF).
• Application of a transverse voltage for separation in a continuous flow system.

Main Results:

• Successful separation of rhodamine B and methyl green with a resolution of 4.6, surpassing micro free flow electrophoresis (microFFE).
• Achieved high-resolution separation (9.4) of lysozyme and ribonuclease B, demonstrating versatility for protein analysis.
• The developed microFF-PRPEC system exhibited superior resolution compared to microFFE under optimal conditions.

Conclusions:

• Microchip free flow planar RP electrochromatography (microFF-PRPEC) is a promising technique for microscale continuous sample preparation.
• The developed method offers significantly improved separation resolution for diverse analytes.
• This technology holds potential for advanced analytical and preparative applications in various scientific fields.