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Mobile Phase Optimization Strategies For Reversed Phase HPLC
Mobile Phase Optimization Strategies For Reversed Phase HPLC
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Speakers
Kevin Schug
Tony Taylor
Technical Director
CHROMacademy
Moderator
Dave Walsh
Editor In Chief
LCGC Magazine
MeOH
MeCN
MobilePhase_01.flv
We will discuss the pH of the mobile phase and choice of buffer shortly
Simulation to Optimise %B
1. Use modelling for predicting separation
2. This DryLab model was achieved using 2 injections at 30% B
and 50% B
Mixture of 6 Neutral Compounds
MobilePhase_02.flv
Altering Selectivity
1. What if separation
isnt achieved with
the OPTIMUM
isocratic
composition from
the scouting
gradient?
2. Adjust %B either
side of optimum by
5-10% B
3. Use an alternative
solvent to adjust
selectivity
MobilePhase_03.flv
MobilePhase_04.flv
MobilePhase_05.flv
MobilePhase_06.flv
Simulation to Optimise pH
1. Use modelling for predicting separation
2. This DryLab model was achieved using 5 runs injections at:
pH 2.9 / 3.0 / 3.5 / 5.0 / 6.5
Mixture of Acidic and Neutral Analytes
MobilePhase_07.flv
MobilePhase_08.flv
Amphetamine peak 7
Amphetamine peak 7
pH 2.5
pH 7.0
MobilePhase_09.flv