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Development of the Tracer-Pulse method for adsorption studies of analyte mixures in liquid chromatography utilizing mass spectrometric detection
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Surface Biotechnology.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Surface Biotechnology. Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Surface Biotechnology. Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry.
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2008 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 80, no 6, 2105-2112 p.Article in journal (Refereed) Published
Abstract [en]

The tracer-pulse method provides the real adsorption data points directly from simple, straightforward calculations and is therefore a superior method for multicomponent adsorption isotherm determination in HPLC. Only one important problem has restricted its use so far: the tracer peaks are invisible using any conventional detection principle. We present a solution to this problem with an approach with a firm base in analytical chemistry, utilizing stable isotopes and mass spectrometric detection. The new approach was used for the determination of binary adsorption isotherms, and a systematic investigation was made of its main sources of error. With this modification, the tracer method can be a prime choice for future characterizations of multicomponent separation systems and of competitive drug binding studies.

Place, publisher, year, edition, pages
2008. Vol. 80, no 6, 2105-2112 p.
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:uu:diva-96956DOI: 10.1021/ac702399aISI: 000254017000035PubMedID: 18290673OAI: oai:DiVA.org:uu-96956DiVA: diva2:171704
Available from: 2008-04-04 Created: 2008-04-04 Last updated: 2017-12-14Bibliographically approved
In thesis
1. Development of Methods for Phase System Characterization in Liquid Chromatography
Open this publication in new window or tab >>Development of Methods for Phase System Characterization in Liquid Chromatography
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The aim of this thesis is first and foremost to improve the fundamental knowledge of nonlinear and preparative separation theory by focusing on some of the remaining “white spots” on the theoretical chromatographic map. Secondly, the acquired knowledge is used to develop, validate and execute new methods for phase characterization in liquid chromatography. The methodology used in this thesis is a combination of experiments, fundamental nonlinear theory and systematic computer simulations.

A fundamental knowledge of the molecular interactions between the compounds to be separated and the separation media requires the determination of adsorption isotherms over a broad concentration range to give a complete picture of all interactions in the separation system - weak as well as strong. In addition, such adsorption data is essential for optimization in preparative chromatography.

For the first time, it has been experimentally shown that the injected molecules are not present in the detected peak when a small excess of molecules are injected into a chromatographic system equilibrated with a constant stream of identical molecules. Several experimental procedures for this method were developed such as (i) the optimal injection strategy and (ii) different labeling methods for visualizing the injected molecules. Remarkable phenomena in the single-component case, such as invisible peak deformation and deformed (invisible) frontal chromatograms, are reported, investigated, and explained. This phenomenon has asides from its future practical implementation, also a large didactic value.

The accuracy of the ECP method is experimentally improved, and used to characterize the separation of protolytic compounds at different pH on modern commercially available silica and hybrid silica column packing materials. That investigation enables us to answer why basic compounds give a much more compact preparative peak profile at pH 11 than they yields at lower pH.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2008. 69 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 416
Keyword
Analytical chemistry, Liquid chromatography, Adsorption isotherms, Adsorption energy distribution, Tracer pulse method, Elution by characteristic points, Peak deformation, Validation, Hybrid column material, Alkaline separation, Injection-profiles, Analytisk kemi
Identifiers
urn:nbn:se:uu:diva-8597 (URN)978-91-554-7145-3 (ISBN)
Public defence
2008-04-25, B41, BMC, Husargatan 3, Uppsala, 13:15 (English)
Opponent
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Available from: 2008-04-04 Created: 2008-04-04 Last updated: 2009-03-20Bibliographically approved

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Sjöberg, Per J. R.

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