Capillary Electrophoresis Flashcards
what is electrophoresis?
separation technique based on different rates of migration of charged species in E field
cations move toward cathode, anions move towards anode
In what direction do cations and anions move in electrophoresis?
cations move toward cathode, anions move towards anode
The cathode has a ____ charge. The anode has a ____ charge.
cathode = negative anode = positive
Migration of molecules in the E field will depend upon factors relating to (2):
physical characteristics of electrophoresis system
solute molecule characteristics (size, charge)
migration describes ____ and ____ of the molecule’s movement
direction
rate
describe the general design/features of a capillary electrophoresis system
2 separate solvent reservoirs, connected to high-voltage power supply (1 is anode, 1 is cathode)
capillary runs between them, with detector in middle (collect data on molecule movement between reservoirs)
What type of capillary is used for CE? What other detectors can this be used with?
narrow bore capillaries
made of fused silica (or modified silica)
absorbance, fluorescence, MS, conductivity…
When a particle is placed in an E field, what happens?
accelerates towards electrode with opposite charge
What is ‘Vep?’
electrophoretic velocity (cm/s) (rate of migration of particle in E field)
What does ‘Uep’ stand for
electrophoretic mobility (cm/sV)
The electrophoretic mobility is proportional to:
the charge density (q/rs)
What factors affect mobility of a charged analyte in CE? (6)
strength of E field temperature pH of separation buffer buffer ion type/ionic strength size/shape of analyte molecules charge of analyte molecules
What happens to the buffer liquid in CE system when a high voltage is applied across the fused silica capillary? What is this called?
buffer liquid starts to migrate towards cathode
Veo; electroosmotic flow velocity
T/F: electroosmotic flow velocity and electrophoretic velocity have the same unit
true
What causes the electroosmotic flow velocity?
charge distribution at silica/buffer interface + creation of electric ‘double layer’ (fixed layer + mobility/diffuse layer)
apply voltage -> cations in mobility layer migrate to cathode, carries water with them