Preconcentration interface coupling liquid chromatography to capillary electrophoresis
View Patent ↗An arrangement for two-dimensional separation of a complex analyte mixture is provided comprising a first separation device for the separation in a first dimension and a second device for the separation in the second dimension. In the area of the interface between the first and the second device a concentration zone is arranged for concentrating the individual sample fractions or components, respectively, leaving the first device before introduction into the second device. The concentration zone is defined in a segment between two or more electrodes seen in flow direction of the sample or sample fractions to be separated. The concentration is achieved by applying a voltage between two electrodes with polarity opposite to the charge of the components of the fraction, sufficiently strong to win the dragging force of the hydrodynamic flow. The introduction into the second device is carried out by turning off the potential difference between the electrodes.
1. An arrangement for two-dimensional separation of a complex analyte mixture comprising a first separation device comprising a liquid chromatography separation column for the separation in a first dimension and a second device comprising a capillary electrophoresis separation device for the separation in the second dimension with an area of interface between the first and the second device, further comprising a concentration zone within the area of interface for concentrating the individual sample fractions or components, respectively, leaving the first device before introduction into the second device, wherein the concentration zone is defined in a segment between two or more electrodes in a flow direction of the sample or sample fractions to be separated, and wherein the arrangement is configured to provide sample flow between the liquid chromatography separation column and the capillary electrophoresis separation device that is constant and without interruption.
2. The arrangement of claim 1 , wherein the two-dimensional separation comprises the combination of nano-reversed phase liquid chromatography with a capillary zone electrophoresis.
3. The arrangement of claim 1 , wherein the analyte mixture comprises peptides.
4. The arrangement of claim 1 , wherein at least one further concentration zone is defined in a segment between a further series of electrodes in the area of the interface between the two devices.
5. The arrangement of claim 4 , wherein the further concentration zone comprises a preconcentration zone or pre-preconcentration zone.
6. The arrangement of claim 1 , characterised in that micro electrodes are arranged before insertion of the individual samples or sample fractions leaving the first device in the second device, the microelectrodes comprising micro channels in contact with a capillary channel connecting the first with the second device.
7. The arrangement of claim 6 , wherein the micro electrodes are used for the preconcentration and/or pre-preconcentration of an analyte.
8. The arrangement of claim 6 , wherein a very narrow aperture contacts the micro channels with a capillary channel connecting the first with the second device.
9. The arrangement of claim 6 , wherein the micro-channels are filled with a polymer or other ion selective material.
10. The arrangement of claim 9 , wherein the polymer comprises polytetrafluoroethylene with sulfonic acid groups interspersed therein.
11. The arrangement of claim 9 , wherein the polymer has an affinity for water and protons.
12. A method for two-dimensional liquid chromatography/electrophoresis/mass spectrometry comprising separating a sample mixture in a reversed phase liquid chromatography followed by a capillary zone electrophoresis, wherein in the area of the interface between the reversed phase liquid chromatography and the capillary zone electrophoresis the fractions leaving the reversed phase liquid chromatography are concentrated before insertion into the following capillary zone electrophoresis, wherein the fractions leaving the reversed phase liquid chromatography are concentrated in at least one segment defined by two electrodes or a series of electrodes by applying a voltage across with polarity opposite to the charge of the components of the fraction sufficiently strong to apply a dragging force of the hydrodynamic flow and in that the insertion into the capillary zone electrophoresis is carried out by turning off the potential difference between the electrodes, wherein the flow of the sample between the liquid chromatography separation column and the capillary electrophoresis separation device is not interrupted.
13. The method of claim 12 , wherein the fractions are concentrated on applying at least one electric field and in that the concentrated fractions are inserted into the following capillary zone electrophoresis in absence or reversion of at least one electric field.
14. The method of claim 12 , wherein the individual components of the fraction separated by the capillary zone electrophoresis are sprayed into a mass spectrometer.