Chromatographia

, Volume 81, Issue 4, pp 575–584 | Cite as

Visualizing Events Under Solvent-Gradient and Insights Thereof

Original

Abstract

This paper describes graphical methods to visualize variations in mobile phase composition inside a chromatographic column under solvent-gradient conditions, and how these variations affect movement of analyte bands. Based on the visualization techniques, the report explains how variations in composition “experienced” by an analyte, which is moving, are different from variations experienced by a static point inside the column. For example, if a linear gradient is created by the system, every point inside the column will experience the same linear variation of composition, although at different times, assuming that the solvent-gradient created at the mixer propagates through the system intact and undisturbed. For an analyte, on the other hand, the variation could be strongly non-linear because normally it travels with varying speed along the column during a gradient run. And because different analytes travel with different speeds, solvent-gradient experienced by the analytes will not be the same, even under the same method condition. Although these events are captured in the fundamental equation of solvent-gradient chromatography and generally understood by researchers working with gradient theories, the visualization methods discussed here may provide a clearer imagery of events for wider appreciation.

Keywords

Solvent-gradient Visualization Travel-time 

Notes

Acknowledgements

Helpful discussions with Dr Fabrice Gritti, Dr Martin Gilar and Dr Allen Caswell of Waters Corporation are acknowledged. Thanks to Prof Gert Desmet of Vrije Universiteit Brussel for valuable comments.

Compliance with Ethical Standards

Conflict of interest

The author declares that he has no conflict of interest.

Ethical statement

The research described in this publication does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    Synder LR, Dolan JW (2007) High-performance gradient elution. Wiley, HobokenGoogle Scholar
  2. 2.
    Freiling EC (1955) Ion exchange as a separations method. ix. gradient elution theory. J Am Chem Soc 77:2067CrossRefGoogle Scholar
  3. 3.
    Freiling EC (1957) Gradient elution theory. J Phys Chem 61:543CrossRefGoogle Scholar
  4. 4.
    Snyder LR, Dolan JW, Gant JR (1979) Gradient elution in high-performance liquid chromatography: I. theoretical basis for reversed-phase systems. J Chromatogr 165:3–30Google Scholar
  5. 5.
    Schoenmakers PJ, Billiet HAH, Tijssen R, Galan LD (1978) Gradient selection in reversed-phase liquid chromatography. J Chromatogr A 149:519–537CrossRefGoogle Scholar
  6. 6.
    Jandera P, Churacek J (1974) Gradient elution in liquid chromatography: I. the influence of the composition of the mobile phase on the capacity ratio (retention volume, band width, and resolution) in isocratic elution -theoretical considerations. J Chromatogr 91:207–221Google Scholar
  7. 7.
    Poppe H, Paanakker J, Bronckhorst M (1981) Peak width in solvent-programmed chromatography: I. General description of peak broadening in solvent programmed elution. J Chromatogr 204:77–84Google Scholar
  8. 8.
    Gritti F, Guiochon G (2010) Performance of columns packed with the new shell kinetex-c18 particles in gradient elution chromatography. J Chromatogr A 1217:1604–1615CrossRefGoogle Scholar
  9. 9.
    Nikitas P, Pappa-Louisi A (2005) Expressions of the fundamental equation of gradient elution and a numerical solution of these equations under any gradient profile. Anal Chem 77:5670–5677CrossRefGoogle Scholar
  10. 10.
    Tyteca E, Priat A, Rudaz S, Desmet G, Guillarme D (2014) Retention modeling and method development in hydrophilicinteraction chromatography. J Chromatogr A 1337:116–127CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Waters CorporationMilfordUSA

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