E19-37: How Liquid Chromatography Works: Separation Principles Explained in Chromatograms

One-Day Course 
Wednesday, Nov. 20; 8:30am – 5:00pm

Dr. Martin Gilar, Waters Corp.,  Milford, MA

The course attendees will learn about principles of chromatographic modes such as reversed-phase, hydrophilic interaction, ion-exchange and size exclusion chromatography. Special emphasis will be given to reversed phase mode. MS Excel will be employed to illustrate retention and dispersion processes on columns. Excel generated chromatograms will be utilized to explain the principles of band broadening on column and due to extra-column effects. Couse attendees will be able to use the spreadsheets, vary separation parameters and instantly observe the effects in chromatograms. Basic pitfalls of method development and method transfer will be covered.

Chromatography practitioners, lab personal, students, researchers, who are interested in deeper understanding of chromatographic principles, small molecule specialists who are new in the world of biopolymer separations and vice versa.  The attendees are recommended to bring their own laptop computers with installed MS Excel. The attendees will be provided with excel sheets based chromatographic simulators. They will be able to perform their own in-silico chromatographic experiments.

1.  Principles of chromatographic separation (RP, HILIC, IEX, SEC, SPE modes)
2. Retention, band broadening on column, particle size effect on efficiency
3. Simulated chromatograms –column efficiency and extra column dispersion explained
4.  Simulated chromatograms – isocratic retention versus gradient chromatography
5. Effects of extra column dispersion on the observed separation
6. Chromatographic effects of large volume injection and incompatible solvents
7.  Comparison of Liquid Chromatography and Solid Phase Extraction – aren’t they the same thing?
8.  Chromatographic differences of small molecules versus biopolymers
9.. Simulated chromatograms-effect of gradient delay on method transfer
10. Simple method development performed with MS excel spreadsheet
11. Conclusion of the course. Questions and answers. Self-guided chromatographic simulations

Dr. Martin Gilar received his Ph.D. in analytical chemistry from Institute of Chemical Technology in Prague (1996). He spent postdoc years in Hybridon Inc. (1996-1998) and Northeastern University in Boston (1998) developing separation methods for antisense oligonucleotides and fraction collector for DNA molecules. Since 1998 he has worked at Waters Corp. in Milford, Massachusetts, participating in column, sample preparation and instrument research. Dr. Martin Gilar is a Scientific Fellow in the Core Research group at Waters Corporation. He has more than 25 years of experience in the separation sciences, including chromatography, electrophoresis, and mass spectrometry. His research interest is the analysis of biopolymers, 2D LC and chromatographic simulations. He has published over 55 peer reviewed papers.