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A new method for the determination of peak distribution across a two-dimensional separation space for the identification of optimal column combinations

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Abstract

For the identification of the optimal column combinations, a comparative orthogonality study of single columns and columns coupled in series for the first dimension of a microscale two-dimensional liquid chromatographic approach was performed. In total, eight columns or column combinations were chosen. For the assessment of the optimal column combination, the orthogonality value as well as the peak distributions across the first and second dimension was used. In total, three different methods of orthogonality calculation, namely the Convex Hull, Bin Counting, and Asterisk methods, were compared. Unfortunately, the first two methods do not provide any information of peak distribution. The third method provides this important information, but is not optimal when only a limited number of components are used for method development. Therefore, a new concept for peak distribution assessment across the separation space of two-dimensional chromatographic systems and clustering detection was developed. It could be shown that the Bin Counting method in combination with additionally calculated histograms for the respective dimensions is well suited for the evaluation of orthogonality and peak clustering. The newly developed method could be used generally in the assessment of 2D separations.

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Acknowledgments

The authors would like to thank Harald Möller-Santner and Eike Logé from Sciex for the loan of the Eksigent NanoLC 425 system. Furthermore, we would like to thank Juergen Maier-Rosenkranz for organizing the packing of PGC columns. In addition the authors would like to thank Altmann Analytik GmbH & Co. KG and Merck KGaA for providing the HILIC column.

Notes

For the automated data evaluation, based on the in this work presented method, a Python 3.x based Script (2D-Distribution.py) was developed. The script as well as an introduction is a part of the supplementary material and is available at https://uni-duisburg-essen.sciebo.de/index.php/s/tWXvtNIluTCTZv8.

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Authors and Affiliations

  1. Institut für Energie- und Umwelttechnik e. V., IUTA (Institute of Energy and Environmental Technology), Bliersheimer Str. 58-60, 47229, Duisburg, Germany

    Juri Leonhardt, Thorsten Teutenberg & Oliver Gassner

  2. Instrumental Analytical Chemistry, University of Duisburg-Essen, Universitätsstr. 5, 45141, Essen, Germany

    Juri Leonhardt, Greta Buschmann & Torsten C. Schmidt

Authors
  1. Juri Leonhardt
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  2. Thorsten Teutenberg
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  3. Greta Buschmann
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  4. Oliver Gassner
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  5. Torsten C. Schmidt
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Correspondence to Thorsten Teutenberg.

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The research in this manuscript did not involve human participants and/or animals. All authors of this manuscript were informed and agreed for submission.

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The authors declare that they have no conflict of interest.

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Leonhardt, J., Teutenberg, T., Buschmann, G. et al. A new method for the determination of peak distribution across a two-dimensional separation space for the identification of optimal column combinations. Anal Bioanal Chem 408, 8079–8088 (2016). https://doi.org/10.1007/s00216-016-9911-3

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  • DOI: https://doi.org/10.1007/s00216-016-9911-3

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