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Porous graphitic carbon chromatography-tandem mass spectrometry for the detection of isoprostanes in human cerebrospinal fluid
Stockholm University, Faculty of Science, Department of Analytical Chemistry.
Stockholm University, Faculty of Science, Department of Analytical Chemistry.
2005 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 827, no 1, 39-43 p.Article in journal (Refereed) Published
Abstract [en]

F2-isoprostanes are produced by the non-enzymatic peroxidation of arachidonic acid in membrane phospholipids. This paper describes a new method for the determination of all four classes of F2-isoprostanes in human cerebrospinal fluid (CSF) involving separation on a 1 mm × 150 mm porous graphitic carbon (PGC) column and detection by triple quadrupole mass spectrometry in negative-ion electrospray mode. The sample pre-treatment consisted of an ultrafiltration step, following which 300 μl of CSF sample could be injected directly onto a 1 mm × 10 mm PGC guard column functioning as a trap for the analytes. The loading solvent was Milli-Q water at 125 μl/min. After 3 min, the sample was switched into the separation column. The F2-isoprostanes were separated in 20 min using a linear solvent gradient comprising water, methanol, acetonitrile and ammonium hydroxide at a pH of 9.5 and a flow of 50 μl/min The limit of detection (calculated as 3S/N) was approximately 40 pM (14 pg/ml). The assay was linear within the examined range (18–450 pg/ml), using CSF spiked with iPF2α-III standard (r2 > 0.995). Repeatability data were calculated for CSF spiked to 90 pg/ml and the relative standard deviation (RSD) obtained was 3% (n = 6).

Place, publisher, year, edition, pages
2005. Vol. 827, no 1, 39-43 p.
Keyword [en]
Isoprostanes; Porous graphitic carbon; Cerebrospinal fluid; Oxidative stress
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:su:diva-23522DOI: 10.1016/j.jchromb.2005.02.023OAI: oai:DiVA.org:su-23522DiVA: diva2:192468
Note
Part of urn:nbn:se:su:diva-322Available from: 2004-12-21 Created: 2004-12-21 Last updated: 2017-12-13Bibliographically approved
In thesis
1. Determination of biomarkers for lipid peroxidation and oxidative stress: Development of analytical techniques and methods
Open this publication in new window or tab >>Determination of biomarkers for lipid peroxidation and oxidative stress: Development of analytical techniques and methods
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Oxidative stress can be defined as a state of disturbance in the pro-oxidant/antioxidant balance in favour of the former, leading to potential damage. Processes associated with oxidative stress involve reactive oxygen species and radicals and can result in elevated levels of oxidatively modified or toxic molecules that can cause cellular malfunction, and even cell death. Destruction of membrane lipids, lipid peroxidation, caused by reactive oxygen species and radicals has been coupled to many diseases and also normal ageing.

The measurement of low molecular weight biomarkers of oxidative stress present in complex matrices such as brain tissue, plasma, urine or cerebrospinal fluid is a delicate and difficult task and there is a need for improved analytical tools in this field of research.

The major foci of this thesis and the work underlying it are the development of analytical techniques and methods for determining biomarkers for oxidative stress and lipid peroxidation. Aspects of particular concern include the effects of sample treatments prior to analysis, evaluation of the developed methods with respect to possible artefacts, and the scope for results to be misinterpreted. The specific research goals and issues addressed are detailed in five papers, which this thesis is based upon.

Paper I focuses on malondialdehyde, describing and evaluating two new simplified sample pre-treatment regimes for the determination of malondialdehyde in rat brain tissue by capillary electrophoresis with UV detection. The effects of sample storing and handling are also considered.

Paper II describes the synthesis, characterization and implementation of a new internal standard for the determination of malondialdehyde in biological samples using electrophoretic or chromatographic separation techniques. The usefulness of the internal standard is demonstrated in analyses of rat brain tissue samples.

Paper III presents a method for the determination of 4-hydroxynon-2-enal in brain tissue from rats employing micellar electrokinetic chromatography separation and laser-induced fluorescence detection.

Paper IV is focused on the development of a new methodology for determining the stereoisomeric F2-isoprostanes in human urine samples employing chromatographic separation on porous graphitic carbon and detection by electrospray ionization-tandem mass spectrometry. The results from this study conflict with the hypothesis that peripheral isoprostanes are elevated in patients with Alzheimer’s disease.

Paper V describes porous graphitic carbon chromatography-tandem mass spectrometry for the determination of isoprostanes in human cerebrospinal fluid. A new simplified sample pre-treatment regime, involving a column switching technique, is presented that allows direct injection of a relatively large volume of CSF into the chromatographic system.

Place, publisher, year, edition, pages
Stockholm: Institutionen för analytisk kemi, 2005. 91 p.
Keyword
capillary electrophoresis, mass spectrometry, electrospray, porous graphitic carbon, oxidative stress, lipid peroxidation, biomarker, malondialdehyde, hydroxynonenal, isoprostane
National Category
Analytical Chemistry
Identifiers
urn:nbn:se:su:diva-322 (URN)91-7265-988-2 (ISBN)
Public defence
2005-01-27, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 12 A, Stockholm, 14:00
Opponent
Supervisors
Available from: 2004-12-21 Created: 2004-12-21Bibliographically approved

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