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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
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Counterflow isotachophoresis in a monolithic column
Bingwen Liu1, Yongzheng Cong, Cornelius F Ivory
1The Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA, USA.
Journal of Separation Science
|June 18, 2014
Summary
Stationary counterflow isotachophoresis (ITP) in monolithic columns significantly reduces analyte dispersion. This method improves processing capacity and offers a robust electrofocusing technique for complex separations.
Area of Science:
- Analytical Chemistry
- Separation Science
- Biophysical Chemistry
Background:
- Isotachophoresis (ITP) is a powerful electrokinetic separation technique.
- Traditional ITP in open capillaries suffers from significant band broadening and dispersion.
- Improving ITP processing capacity and zone stability is crucial for practical applications.
Purpose of the Study:
- To investigate stationary counterflow isotachophoresis (ITP) in a monolithic column.
- To assess the impact of monolith properties on flow profiles and analyte dispersion.
- To demonstrate the enhanced separation performance of monolithic ITP.
Main Methods:
- COMSOL simulations using Brinkman Equation to predict flow profiles in monoliths.
- Fabrication of a poly(acrylamide-co-N,N'-methylenebisacrylamide) monolithic column via UV-initiated polymerization.
- Experimental implementation of stationary counterflow ITP in the monolithic column and open capillary for comparison.
Main Results:
- Monolith permeability dictates flow profile, shifting from parabolic to plug flow as permeability decreases.
- Stationary counterflow ITP in the monolithic column yielded undistorted analyte zones with reduced dispersion.
- Analyte zone width for r-phycoerythrin was reduced by two-thirds compared to open capillary ITP.
Conclusions:
- Stationary counterflow ITP in monolithic columns effectively minimizes dispersion.
- Monolithic ITP offers improved processing capacity and robust analyte focusing.
- This technique presents a practical and efficient electrofocusing method for analytical separations.
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