Abstract
Recent developments in chip-based column technology, whereby plate heights corresponding to the domain size can be obtained,1 suggest that it might also be possible to improve the lower minimal plate height limit in packed column HPLC, corresponding to twice the particle size, down to lower values if improved packing procedures can be developed.
In order to allow improved understanding of the order in packed HPLC columns in this work a methodology for immobilizing silica particles is therefore developed based on the polymerization of a monomer and a cross-linker in the interstitial pores of HPLC columns. Subsequent cutting by mechanical cross-sectioning then allows scanning electron microscopy (SEM) based imaging of cross sections of the packed bed over the entire length of the column.
In this way the packing efficiency of in-house packed and commercial HPLC columns comprising the same packing material can be compared. The methodology is developed for native silica used in e.g. hydrophilic interaction liquid chromatography and the information obtained is cross-referenced with external porosity measurements obtained via inversed size exclusion approach.2,3
Results indicate that the home-made columns typically depict a lower total porosity compared to the commercial columns but a higher external porosity, indicating a lower density of the packed bed. This is reflected into losses in efficiency and into more disordered SEM data. The tool can allow fine optimization of packing procedures.
References:
[1] S. Jespers, S. Schlautmann, H. Gardeniers, W. De Malsche, F. Lynen, G. Desmet, Analytical Chemistry 2017, 89, 11605-11613.
[2] H. Guan, G. Guiochon, Journal of Chromatography A 1996, 731, 27-40.
[3] F. Gritti, G. Guiochon, Journal of Chromatography A 2013, 1280, 35-50.
In order to allow improved understanding of the order in packed HPLC columns in this work a methodology for immobilizing silica particles is therefore developed based on the polymerization of a monomer and a cross-linker in the interstitial pores of HPLC columns. Subsequent cutting by mechanical cross-sectioning then allows scanning electron microscopy (SEM) based imaging of cross sections of the packed bed over the entire length of the column.
In this way the packing efficiency of in-house packed and commercial HPLC columns comprising the same packing material can be compared. The methodology is developed for native silica used in e.g. hydrophilic interaction liquid chromatography and the information obtained is cross-referenced with external porosity measurements obtained via inversed size exclusion approach.2,3
Results indicate that the home-made columns typically depict a lower total porosity compared to the commercial columns but a higher external porosity, indicating a lower density of the packed bed. This is reflected into losses in efficiency and into more disordered SEM data. The tool can allow fine optimization of packing procedures.
References:
[1] S. Jespers, S. Schlautmann, H. Gardeniers, W. De Malsche, F. Lynen, G. Desmet, Analytical Chemistry 2017, 89, 11605-11613.
[2] H. Guan, G. Guiochon, Journal of Chromatography A 1996, 731, 27-40.
[3] F. Gritti, G. Guiochon, Journal of Chromatography A 2013, 1280, 35-50.
| Original language | English |
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| Publication status | Published - Jan 2020 |
| Event | 16th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology - Het Pand, Ghent, Belgium Duration: 29 Jan 2020 → 31 Jan 2020 https://kuleuvencongres.be/htc16 |
Conference
| Conference | 16th International Symposium on Hyphenated Techniques in Chromatography and Separation Technology |
|---|---|
| Abbreviated title | HTC-16 |
| Country/Territory | Belgium |
| City | Ghent |
| Period | 29/01/20 → 31/01/20 |
| Internet address |