Protocols for finding the most orthogonal dimensions for two-dimensional high performance liquid chromatography

Bassanese, Danielle N., Holland, Brendan J., Conlan, Xavier A., Francis, Paul S., Barnett, Neil W. and Stevenson, Paul G. 2015, Protocols for finding the most orthogonal dimensions for two-dimensional high performance liquid chromatography, Talanta, vol. 134, pp. 402-408, doi: 10.1016/j.talanta.2014.11.037.

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Title Protocols for finding the most orthogonal dimensions for two-dimensional high performance liquid chromatography
Author(s) Bassanese, Danielle N.ORCID iD for Bassanese, Danielle N.
Holland, Brendan J.
Conlan, Xavier A.ORCID iD for Conlan, Xavier A.
Francis, Paul S.ORCID iD for Francis, Paul S.
Barnett, Neil W.
Stevenson, Paul G.ORCID iD for Stevenson, Paul G.
Journal name Talanta
Volume number 134
Start page 402
End page 408
Total pages 7
Publisher Elsevier
Place of publication Amsterdam, The Netherlands
Publication date 2015
ISSN 1873-3573
Keyword(s) Bins method
HPLC simulation
Potassium permanganate chemiluminescence
Stationary phase optimisation
Two-dimensional high performance liquid chromatography
Potassium permanganate
Two-dimensional high performance liquid
Science & Technology
Physical Sciences
Chemistry, Analytical
Summary The selection of two high performance liquid chromatography (HPLC) columns with vastly different retention mechanisms is vital for performing effective two-dimensional (2D-) HPLC. This paper reports on a systematic method to select a pair of HPLC columns that provide the most different separations for a given sample. This was completed with the aid of a HPLC simulator that predicted retention profiles on the basis of real experimental data, which is difficult when the contents of sample matrices are largely-or completely-unknown. Peaks from the same compounds must first be matched between chromatograms to compare the retention profiles and optimised 2D-HPLC column selection. In this work, two methods of matching peaks between chromatograms were explored and an optimal pair of chromatography columns was selected for 2D-HPLC. First, a series of 17 antioxidants were selected as an analogue for a coffee extract. The predicted orthogonality of the standards was 39%, according to the fractional surface coverage 'bins' method, which was close to the actual space utilisation of the standard mixture, 44%. Moreover, the orthogonality for the 2D-HPLC of coffee matched the predicted value of 38%. The second method employed a complex sample matrix of urine to optimise the column selections. Seven peaks were confidently matched between chromatograms by comparing relative peak areas of two detection strategies: UV absorbance and potassium permanganate chemiluminescence. It was found that the optimal combinations had an orthogonality of 35% while the actual value was closer to 30%.
Language eng
DOI 10.1016/j.talanta.2014.11.037
Field of Research 030108 Separation Science
Socio Economic Objective 970103 Expanding Knowledge in the Chemical Sciences
HERDC Research category C1 Refereed article in a scholarly journal
Grant ID DP140100439
Copyright notice ©2015, Elsevier
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