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ELECTROPHESIS
This is a separation technique based on differential migration of charged
solutes in an electric field.
Can be categorized based on the charge type on the analyte. It is known as
cataphoresis if the charge on the analyte is positive and migrates towards
the anode and anaphoresis if the analyte is negatively charged and migrate
towards the anode. The charge of the molecule/analyte may be due to
ionization of specific functional proups on the molecule or they may be
induced by interacting with electrolytes.
For most compounds, the charge on the molecule is dependent on various
factors ans ad pH, nature and concentration of the electrolytes and nature of
the solute. For charges due to ionizable groups, the pKa of individual groups
also play a role e.g weak acidas are unionized in low pH, the reverse is true
for weak acids at a high pH.
What determines high and low pH is the pKa of the acid
When the pH is the same as the pKa of the acid half of the acid present will
be ionized.
pH=pKa + log [A-]/[AH]
At low pH we have unionized acid tan ionized.
When the ratio of the acid increases the pH …..
For weak bases, the reverse is true.
For compounds that are able to carry different charges such as AA, proteins,
the net charge depends on the number of ionizable functional groups, the
pKa and the pH.
The pH at which the amphoteric compund have no net charge is known as
the isoelectric point (PI).
Example of such compounds.
Acid alanine has a carboxyllic group with a pKa of 3,25 and amino group of
pKa 9,7 has a net negative charge at high pH and net positive charge at low
pH. Its isoelectric point is around 6, where the molecule is e...