HPLC
METHOD DEVELOPMENT |
| Back
to First Page |
| 2 day Hands-on Short Course designed for
chemists with at least 1 year of experience in HPLC |
| Day 1 |
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| 8:45 |
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Registration |
| 9:00 |
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Introduction to HPLC,
Instrumentation (Detectors included) |
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Basic principles, general parameters, calculation of simple retention
factors, system performance analysis. Major instrumentation types, detector suitability
for different applications, system assembly and testing, practical tips. |
| 10:00 |
Coffee |
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| 10:15 |
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Column Selection, Reversed-Phase
HPLC |
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Main column parameters, system suitability, surface chemistry, eluent
type and composition effects. |
| 11:00 |
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pH effect on HPLC separations (Part
I) |
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pH effect on HPLC retention, column properties, method development
strategy |
| 12:00 |
Lunch |
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| 1:00 |
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Hands-on Lab Experiments |
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System setup, column selection, cleaning, testing and equilibration.
PDA wavelength selection, peak purity, etc. General pH effect on the retention of acidic
and basic components. Effect of the eluent composition on the retention of ionizable
components (organic type and content, type and concentration of pH modifier, pKa
shift). Development of the separation of unknown mixtures. |
| 4:00 |
Coffee |
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| 4:15 |
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Short discussion (submit
questions for discussion on day 2) |
| Day 2 |
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| 9:00 |
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LC/MS |
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Types of LC/MS interfaces, principles, suitability of different
interfaces for certain analyses, buffer selection. |
| 10:00 |
Coffee |
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| 10:15 |
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HPLC Method Development
and method validation |
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Strategy of initial approach, starting experiments, fine tuning,
checking method stability and ruggedness. Method validation considerations. |
| 11:00 |
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pH effect on HPLC separations (Part
II) |
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Fine tuning of the HPLC separation with pH adjustment. Buffer
concentration effect. Influence of counteranions on the retention of basic pharmaceutical
components. |
| 12:00 |
Lunch |
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| 1:00 |
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Hands-on Lab Experiments |
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Optimization of HPLC separation using pH adjustment, variation of the
salt concentration. Chaotropic effect. Effect of different salts. |
| 5:30 |
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Open discussion.
Discussion of the questions submitted on the first day. Troubleshooting tips. |
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