Monitoring Seizure-Induced Oxidative Stress using Liquid Chromatography and Capillary Electrophoresis with Electrochemical and Fluorescence Detection

Monitoring Seizure-Induced Oxidative Stress using Liquid Chromatography and Capillary Electrophoresis with Electrochemical and Fluorescence Detection

Monitoring Seizure-Induced Oxidative Stress using Liquid Chromatography and Capillary Electrophoresis with Electrochemical and Fluorescence Detection (#49)

Craig Lunte ¹

Department of Chemistry, R.N. Adams Institute for Bioanalytical Chemistry, University of Kansas, Lawrence, KS, United States

Liquid Chromatography (LC) and Capillary electrophoresis (CE) are powerful separation techniques with excellent with complementary characteristics. Excellent detection limits can be achieved for both technique when combined with either electrochemical (EC) or fluorescence detection. In this presentation, our work at implementation of LC and CE in combination with EC and fluorescence detection as applied to studying oxidative stress will be described.

The neurotransmitter catecholamines, gamma aminobutyric acid (GABA) and glutamate are important target analytes. A suite of methods using both LC and CE both in combination with either EC or fluorescence have been developed for specific analytical needs. Each of the methods and its uses will be discussed. Several target analytes that serve as biomarkers of oxidative stress have been identified and analytical methods for these targets developed. Glutathione is a key marker of the oxidative state of a tissue. It is necessary to measure both the oxidized (disulfide) and reduced (thiol) forms of glutathione. Again both LC and CE methods using EC detection have been developed. A method that employs on-capillary stacking to achieve acceptable concentration detection limits and a dual-electrode detector will be described. 8-Oxo-guanine (8-oxoG) and 8-hydroxy-deoxyguanosine (8-OHdG) are biomarkers for DNA oxidation. These biomarkers can also be detected using CEEC with on-capillary concentration. Another important biomarker of oxidative stress is malondialdhyde (MDA), which is a product of lipid peroxidation. MDA must be derivatized to a fluorescent product prior to detection using either LC-fourescence or CE-LIF.

Authors contributing to this presentation.

Lunte, C

Craig E. Lunte is Professor of Bioanalytical Chemistry and Adjunct Professor of Pharmaceutical Chemistry at the University of Kansas. He is Chair of the Chemistry Department. Dr. Lunte is a Fellow of the American Association of Pharmaceutical Scientists and was a Science Foundation Ireland Walton Fellow. He has mentored over 40 graduate students, over 60 undergraduate researchers, and 17 postdoctoral associates. Professor Lunte has been awarded the AAPS Research Achievement Award in Analysis and Pharmaceutical Quality. He has also been awarded the Self Graduate Fellowship Program Mentor Achievement Award and the John C. Wright Graduate Mentor Award. His research centers on development of microdialysis sampling for biomedical studies, the development of capillary electrophoresis techniques, and new methods of electrochemical detection for microcolumn separations. Some current projects include studies of seizure induced oxidative stress, evaluation of anticancer drug neurochemical effects, and the PK/PD of potential new drug compounds.

Monitoring Seizure-Induced Oxidative Stress using Liquid Chromatography and Capillary Electrophoresis with Electrochemical and Fluorescence Detection (#49)

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