Chromatographic Methods   Capillary Electrophoresis

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Chromatographic Methods Capillary Electrophoresis

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1. Chapter 23 Chromatographic Methods & Capillary Electrophoresis

2. Capillary Electrophoresis in Medicine

3. 23.1 Ion-Exchange Chromatography 1) Structures

4. Ion-exchange selectivity R-K+ + Li + ? R-Li+ + K+ selectivity coefficient: 23.1 Ion-Exchange Chromatography

5. b) Hydrated radius : hydrated radius? or Ionic charge? ? bind to ion-exchange resin? order of selectivity: 23.1 Ion-Exchange Chromatography

6. 23.1 Ion-Exchange Chromatography c) Gradient elution with increasing ionic strength (ionic concentration) to separate one ion from another by ion-exchange chromatography.

7. 3) What is deionized water ? Preconcentration trace analysis ? [s] is extremely low important for environmental problem 23.1 Ion-Exchange Chromatography -5

8. 23.1 Ion-Exchange Chromatograph

9. 23.2 Ion Chromatography a high-performance version of ion-exchange chromatography, with a key modification that removes eluent ions before detecting analyte ions. in semiconductor industry: to monitor anions & cations level at 0.1-ppb levels in deionized water. in environmental analysis:

10. 23.2 Ion Chromatography (4) Anions are separated by ion exchange & detected by their electrical conductivity. Problem : It is difficult to detect the conductivity change when analyte ions are eluted. Suppressed-ion anion chromatography : Remove the unwanted electrolyte prior to conductivity measurement.

11. 23.2 Ion Chromatography

12. 23.3 Molecular Exclusion Chrom. Molecules are separated according to their “SIZE”.

13. 23.4 Affinity Chromatography Isolate a single compound from a complex mixture. a. specific binding to s.p. b. unbind : change pH or change ionic strength

14. 23.5 What is Capillary Electrophoresis? a) Electrophoresis is the migration of ion in an electric field.

15. The greater the charge on the ion, the faster it migrates. The greater the size of the molecule, the slower it migrates. Different ions migrate at different speeds, so they separate. 23.5 What is Capillary Electrophoresis?

16. Electropherogram Sensitivity

17. b) Capillary electrophoresis : extremely high resolution in a narrow capillary tube (only B term in the van Deemter eqn.) No s.p. ? C = 0 Open tubular column ? A = 0 23.5 What is Capillary Electrophoresis?

21. Hydrodynamic flow induced by a pressure difference, and creates broad band

24. 23.7 Types of capillary electrophoresis

25. 23.7 Types of capillary electrophoresis (1) Capillary zone electrophoresis : cations > neutrals (unseparated) > anions (2) Micellar electrokinetic capillary electrophoresis separate neutral molecules as well as ions

27. Neutral molecules reach the detector at a time between tmc (the time for micelles to reach detector) and t0 (absence of micelles). soluble in the micelle ? ? time inside the micelle ? ? migration time ?, i.e. tr: nonpolar solutes > polar 23.7 Types of capillary electrophoresis

28. (3) Capillary gel electrophoresis Macromolecules are separated by sieving Small molecules travel faster than large molecules through the gel. (opposite in molecular exclusion chromatography) Is used to sequence DNA (opening at p510) 23.7 Types of capillary electrophoresis

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