The impact of column connection on band broadening in very high pressure liquid chromatography

Stankovich, Joseph J., Gritti, Fabrice, Stevenson, Paul G. and Guiochon, Georges 2013, The impact of column connection on band broadening in very high pressure liquid chromatography, Journal of separation science, vol. 36, no. 17, pp. 2709-2717, doi: 10.1002/jssc.201300175.

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Title The impact of column connection on band broadening in very high pressure liquid chromatography
Author(s) Stankovich, Joseph J.
Gritti, Fabrice
Stevenson, Paul G.ORCID iD for Stevenson, Paul G.
Guiochon, Georges
Journal name Journal of separation science
Volume number 36
Issue number 17
Start page 2709
End page 2717
Total pages 9
Publisher Wiley
Place of publication London, England
Publication date 2013-09
ISSN 1615-9306
Keyword(s) Band broadening
Column connections
Very high pressure liquid chromatography
Void volumes
Summary A series of experiments was conducted to evaluate the degree of band broadening in very high pressure LC due to column connections. Different column manufacturers use slightly different designs for their column fittings. If the same column connections are repeatedly used to attach columns of different origins, different void volumes form between capillary tubes and column inlets. An Agilent Ultra Low Dispersion Kit (tubing id 75 μm) was installed on an Agilent Infinity 1290 ultra HPLC and used to connect successively an Agilent, a Phenomenex, and a Waters column. A series of uracil (unretained) samples were injected and eluted at a wide range of flow rates with a water/acetonitrile mixture as eluent. In order to determine the variance contribution from column connections as accurately as possible a nonretained probe compound was selected because the variance contribution from the column is the smallest for analytes, which have very low k values. Yet, this effect still has an impact on the resolution for moderately retained compounds (k > 2) for narrow-bore columns packed with fine particles, since variance contributions are additive for linear chromatographic systems. Each injection was replicated five times under the same experimental conditions. Then NanoViper column connections (tubing id 75 μm) were used and the same injections were made. This system was designed to minimize connection void volumes for any column. Band variances were calculated as the second central moment of elution peaks and used to assess the degree of band broadening due to the column connections. Band broadening may increase from 3.8 to 53.9% when conventional metal ferrules were used to join columns to connection sites. The results show that the variance contribution from improper connections can generate as much as 60.5% of the total variance observed. This demonstrates that column connections can play a larger role than the column packing with respect to band dispersion.
Language eng
DOI 10.1002/jssc.201300175
Field of Research 030108 Separation Science
Socio Economic Objective 970103 Expanding Knowledge in the Chemical Sciences
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©2013, WIley
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Document type: Journal Article
Collection: School of Life and Environmental Sciences
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