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Development of a molecularly imprinted polymer based solid-phase extraction of local anaesthetics from human plasma
Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Chemistry, Department of Physical and Analytical Chemistry.
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2004 In: Analytica Chimica Acta, ISSN 0003-2670, Vol. 526, no 2, 147-154 p.Article in journal (Refereed) Published
Place, publisher, year, edition, pages
2004. Vol. 526, no 2, 147-154 p.
Identifiers
URN: urn:nbn:se:uu:diva-97811OAI: oai:DiVA.org:uu-97811DiVA: diva2:172890
Available from: 2008-11-20 Created: 2008-11-20Bibliographically approved
In thesis
1. Development of Liquid-based Separation Techniques using Tailored Surfaces for Analysis of Biological Samples
Open this publication in new window or tab >>Development of Liquid-based Separation Techniques using Tailored Surfaces for Analysis of Biological Samples
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Development and improvement of analytical techniques are vital in analytical chemistry research. This thesis describes the development and use of tailored surfaces for bioanalytical applications.

In sample preparation, solid phase extraction is often used and the development of a protocol for extraction on a molecular imprinted polymer (MISPE) directly from plasma sample is presented. Molecular imprinted polymers (MIP) offer selective sorbents for the imprinted analyte. MISPE has mainly been used in organic phase but in this thesis the development of a protocol for direct extraction of the analyte form an aqueous phase is described.

For analysis of complex samples a separation step is often needed. The growing interest in analysis of biological samples and analysis of the human proteome and potential biomarkers has increased the interest in developing new separation techniques. Capillary electrophoresis (CE) has evolved into an important technique for use in analysis of body fluids. In this thesis a novel polyamine coating named PolyE 323 tailored for minimizing the adsorption of basic proteins to the surface is introduced. A straightforward coating protocol, with four simple rinsing steps, was developed. The coating was highly reproducible and useable over a wide pH range. Successful protein separations on PolyE-323-coated capillaries coupled to electrospray ionization mass spectrometry (ESI-MS) were demonstrated.

The coated capillaries were also used in studies of protein content of aqueous humor samples from cataract patients as a complement to capillary liquid chromatography. In the studies presented the protein content of aqueous humor samples from two clinical groups was compared. By using capillary liquid separation techniques coupled to matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and MS/MS in combination with isobaric tags for relative and absolute quantitation (iTRAQ) the identity and relative concentrations of proteins in the samples were evaluated. Earlier studies of the proteins in these kinds of samples have mainly been done with techniques using immunological detection where the proteins of interest were chosen in advance. In this thesis it was shown that liquid-based separation techniques are a good complement and by using the mass spectrometry approach presented the protein content of the samples could be evaluated without bias.

Place, publisher, year, edition, pages
Uppsala: Universitetsbiblioteket, 2008. 48 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 577
Keyword
aqueous humor, biological samples, capillary electrophoresis, coatings, drugs, liquid chromatography, molecular imprinted polymers, peptides, proteins, pseudoexfoliation
National Category
Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-9399 (URN)978-91-554-7350-1 (ISBN)
Public defence
2008-12-11, B42, BMC, Husargatan 3, Uppsala, 10:15
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Supervisors
Available from: 2008-11-20 Created: 2008-11-20 Last updated: 2012-02-08Bibliographically approved

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