L1 /
13
CH3041
HPLC III
• The sample (» 0.1 mL) is filtered &
injected into a closed loop. The
loop is swept out
under pressure (200 atm) onto the column & the column is eluted with solvent (10 mL/min).
under pressure (200 atm) onto the column & the column is eluted with solvent (10 mL/min).
•The
MP is usually MeOH/H2O or CH3CN/H2O
the content can range from polar at 100% H2O to essentially non-polar at 100% organic – hence the name reverse phase.
the content can range from polar at 100% H2O to essentially non-polar at 100% organic – hence the name reverse phase.
• Degassed & filtered mobile phase is pumped
through with either isocratic (constant
composition) or gradient elution.
Increasing eluent strength ensures good, fast
separations.
• Eluent strength
is increased by increasing
the proportion of the organic solvent.
the proportion of the organic solvent.
