Wide injection zone compression in gradient reversed-phase liquid chromatography

Journal of Chromatography. a
Martin GilarJames W Jorgenson

Abstract

Chromatographic zone broadening is a common issue in microfluidic chromatography, where the sample volume introduced on column often exceeds the column void volume. To better understand the propagation of wide chromatographic zones on a separation device, a series of MS Excel spreadsheets were developed to simulate the process. To computationally simplify these simulations, we investigated the effects of injection related zone broadening and its gradient related zone compression by tracking only the movements of zone boundaries on column. The effects of sample volume, sample solvent, gradient slope, and column length on zone broadening were evaluated and compared to experiments performed on 0.32mm I.D. microfluidic columns. The repetitive injection method (RIM) was implemented to generate experimental chromatograms where large sample volume scenarios can be emulated by injecting two discrete small injection plugs spaced in time. A good match between predicted and experimental RIM chromatograms was observed. We discuss the performance of selected retention models on the accuracy of predictions and use the developed spreadsheets for illustration of gradient zone focusing for both small molecules and peptides.

References

Jan 11, 2005·Journal of Separation Science·Gerard RozingHenk H Lauer
Feb 14, 2006·Journal of Chromatography. a·Uwe D NeueLloyd R Snyder
Jul 12, 2007·Journal of Separation Science·Hongfeng Yin, Kevin Killeen
Dec 8, 2007·Journal of Chromatography. a·Fabrice Gritti, Georges Guiochon
Oct 8, 2008·Journal of Chromatography. a·P Nikitas, A Pappa-Louisi
Jul 17, 2009·Journal of Chromatography. a·Fabrice Gritti, Georges Guiochon
May 7, 2010·Journal of Chromatography. a·Uwe Dieter Neue, Hans-Joachim Kuss
May 18, 2013·Rapid Communications in Mass Spectrometry : RCM·Haihong ZhouThomas P Roddy
Mar 19, 2014·Journal of Chromatography. a·Paul G StevensonNeil W Barnett

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Citations

Jan 26, 2016·Journal of Chromatography. a·Fabrice Gritti
Jul 16, 2016·Trends in Analytical Chemistry : TRAC·James P Grinias, Robert T Kennedy
Jul 17, 2016·Journal of Pharmaceutical and Biomedical Analysis·Zoltán MártaPál Tamás Szabó

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