Isopropylammonium formate as a mobile phase modifier for liquid chromatography
Matthew P Collins1, Ling Zhou, Suzanne E Camp
1Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA.
Journal of Chromatographic Science
|June 22, 2012
Summary
Isopropylammonium formate (IPAF) is a novel ionic liquid suitable as a mobile phase in reversed-phase liquid chromatography. It effectively replaces organic solvents for separating various compounds and proteins.
Area of Science:
- Analytical Chemistry
- Green Chemistry
Background:
- Alkylammonium formates (AAFs) are emerging as environmentally friendly alternatives to traditional organic solvents.
- Room temperature ionic liquids (RTILs) offer unique solvent properties for chromatographic applications.
Purpose of the Study:
- To synthesize and characterize isopropylammonium formate (IPAF) as a potential mobile phase for reversed-phase liquid chromatography (LC).
- To evaluate IPAF's solvent properties and its efficacy in separating diverse analytes, including proteins.
Main Methods:
- Synthesis and characterization of IPAF.
- Determination of LC polarity (P') and solvent strength (S) parameters.
- Separation of a test mixture (niacinamide, acetophenone, p-nitroaniline) using IPAF-water mobile phases.
- Van Deemter plot analysis with various LC columns.
- Temperature-dependent separation of cytochrome c and tryptophan.
Main Results:
- IPAF exhibits LC polarity (P'=6.0) and solvent strength (S=2.4), comparable to methanol and acetonitrile.
- Successful separation of a model mixture using IPAF as a mobile phase component.
- Van Deemter profiles indicate optimal mixing with water depends on column dimensions and particle size.
- IPAF acts as a non-denaturing solvent for protein separation, outperforming methanol at varying temperatures.
Conclusions:
- IPAF is a viable, greener alternative to organic solvents in reversed-phase LC.
- Further research into AAF mobile phases holds promise for semi-preparative separation of native proteins.
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