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Analysis of glutathione and immunoglobuline G inside chromatographic beads using surface-enhanced Raman scattering spectroscopy
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Materials Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Materials Chemistry.
2005 (English)In: Journal of Raman Spectroscopy, ISSN 0377-0486, E-ISSN 1097-4555, Vol. 36, no 5, 394-399 p.Article in journal (Refereed) Published
Abstract [en]

Gold-containing chromatographic beads were used to record spectra of the tripeptide glutathione and the protein immunoglobulin G (IgG) by surface-enhanced Raman scattering (SERS) spectroscopy. SERS spectra were recorded from glutathione in solution at three different pH values, 2, 6 and 10. It was found that the SERS spectra are strongly dependent on pH, which indicates that the adsorption mode of glutathione changes with the pH value of the solution. Assignment of some of the observed bands and a suggestion for adsorption modes at the different pH were made. A SERS spectrum of human IgG in ethanolamine was also recorded. No complete assignment of the bands in this spectrum was performed, but the results indicate adsorption to the gold nanoparticles by aromatic amino acids and cystine residues.

Place, publisher, year, edition, pages
2005. Vol. 36, no 5, 394-399 p.
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:uu:diva-92886DOI: 10.1002/jrs.1304OAI: oai:DiVA.org:uu-92886DiVA: diva2:166198
Available from: 2005-04-19 Created: 2005-04-19 Last updated: 2017-12-14Bibliographically approved
In thesis
1. Application of Raman and Fluorescence Spectroscopy to Single Chromatographic Beads
Open this publication in new window or tab >>Application of Raman and Fluorescence Spectroscopy to Single Chromatographic Beads
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Chromatography is a powerful technique, essential in chemical analyses and preparative separation in industry and research. Many different kinds of chromatographic material are needed, due to the large variety of applications. Detailed methods of characterisation are needed to design new chromatographic materials and understand their properties. In this thesis, confocal Raman spectroscopy and surface enhanced Raman spectroscopy (SERS) have been applied to micrometer-size chromatographic beads, for which these techniques have not been used earlier. New methodology, optimized for use with the chromatographic beads, has been developed and evaluated.

Confocal spectroscopy has been used to determine distributions of functional groups within single chromatographic beads. This distribution is of great importance in determining the chromatographic properties, since the material is porous and the solute molecules can diffuse inside the beads. Most of the confocal experiments have been performed with Raman spectroscopy; fluorescence spectroscopy, using Nd3+ ions or dye-labelled proteins as fluorescence probes, has been used for comparison.

The concentration of adsorbed analytes is very low within the beads. SERS was therefore used to enhance the Raman signal. SERS-active surfaces were prepared by incorporating gold nano-particles into the interior of the bead. TEM measurements showed that the gold nano-particles could be observed throughout, and it was possible to record analyte spectra from different positions within the bead. Enhanced spectra could be obtained both for small test molecules and for larger bio-molecules, although the spectra for the smaller analytes were much more intense.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2005. 59 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 37
Keyword
Chemistry, agarose, chelating group, confocal Raman spectroscopy, confocal scanning laser microscopy, gold nanoparticles, ligand distribution, functional group, neodymium, polymer beads, Raman, SERS, surface-enhanced Raman spectroscopy, Kemi
National Category
Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-5741 (URN)91-554-6210-3 (ISBN)
Public defence
2005-05-13, Häggsalen, The Ångström Laboratory, Lägerhyddsvägen 1, Uppsala, 10:15
Opponent
Supervisors
Available from: 2005-04-19 Created: 2005-04-19Bibliographically approved

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