Capillary electrophoresis Flashcards
Is CE chromatography?
No, even though it is about separation, chromatography is about separation based on distribution between two phases. In CE no stationary phase and no partitioning
What is electrophoresis?
Separation of charged analytes in an applied electric field
Why do ions move at different speed?
It depends on their charge and hydrodynamic radius (size)
What is a challenge in CE?
Joule heating
Heat released from the current flowing through the tube, proportional to diameter of the tube and applied electric field. It causes a loss of resolution
How can we minimise Joule heating?
Narrower tubes (but detectability goes down)
Less conducting buffer solutions
Cold temperatures (higher density of water in cold temp means less convection)
What are the components of CE?
Empty fused silica capillaries
High voltage power supply
(cheap equipment)
What are advantages of CE over HPLC?
High efficiency
Wide pH range
Low solvent consumption
Ability to study equilibria in solution
What are disadvantages of CE over HPLC?
Stability issues
High detection limits
Limited selectivity
What is the most commonly used capillary material?
Fused silica
What is BGE?
Background electrolyte
What is EOF?
Electroosmotic flow
What is the wall/surface of the fused silica capillary like?
It has OH groups, when pH is higher than pKa the H+ leaves so negatively charged O- on wall and H+ in BGE
Is the surface/wall negatively charged when pH is larger than pKa?
Yes
Where is the EOF moving?
Towards the cathode
What does the cross-sectional flow profile look like?
Flat unlike HPLC
How can you control the EOF?
Temperature, electric field, buffer pH, ionic strength of buffer
What are the ions movements based on?
Charge and size