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
8:45 Registration
9:00 Introduction to HPLC, Instrumentation (Detectors included)
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
10:15 Column Selection, Reversed-Phase HPLC
Main column parameters, system suitability, surface chemistry, eluent type and composition effects.
11:00 pH effect on HPLC separations (Part I)
pH effect on HPLC retention, column properties, method development strategy
12:00 Lunch
1:00 Hands-on Lab Experiments
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
4:15 Short discussion (submit questions for discussion on day 2)
Day 2
9:00 LC/MS
Types of LC/MS interfaces, principles, suitability of different interfaces for certain analyses, buffer selection.
10:00 Coffee
10:15 HPLC Method Development and method validation
Strategy of initial approach, starting experiments, fine tuning, checking method stability and ruggedness. Method validation considerations.
11:00 pH effect on HPLC separations (Part II)
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
1:00 Hands-on Lab Experiments
Optimization of HPLC separation using pH adjustment, variation of the salt concentration. Chaotropic effect. Effect of different salts.
5:30 Open discussion. Discussion of the questions submitted on the first day. Troubleshooting tips.
To REGISTER: Call: (973) 761-9042 Email: kazakeyu@shu.edu