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New approaches in analytical and preparative free fluid electrophoresis

English title New approaches in analytical and preparative free fluid electrophoresis
Applicant Thormann Wolfgang
Number 146161
Funding scheme Project funding
Research institution Institut für Klinische Pharmakologie Universität Bern
Institution of higher education University of Berne - BE
Main discipline Biophysics
Start/End 01.09.2013 - 31.03.2017
Approved amount 342'657.00
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Keywords (10)

stereoselective drug metabolism; enantiomeric resolution; isoelectric focusing; capillary electrophoresis; electrophoresis in free solution; isotachophoresis; electrokinetic capillary chromatography; permeability marker; computer simulation of electrophoresis; alcohol marker

Lay Summary (German)

Lead
Unter Elektrophorese versteht man den Transport von geladenen Teilchen in einem flüssigen Medium unter Einfluss eines elektrischen Feldes. Wegen unterschiedlichen Wanderungsgeschwindigkeiten lassen sich unterschiedliche Ionen im elektrischen Feld trennen. Angewandt wird die Elektrophorese vor allem als Analyseverfahren in der Biologie und Medizin (DNA- und Protein-Analysen). Das Projekt befasst sich mit Elektrophorese in freier Lösung, d.h. ohne Gel das die Lösung stabilisiert.
Lay summary

Die Forschungsvorhaben umfassen drei Teile: A) Computersimulationen von elektrophoretischem Transport in Kapillaren, B) Entwicklung von kapillarelektrophoretischen Analyseverfahren zur Bestimmung von Alkoholmarker und Zuckereinheiten in biologischen Proben, und C) Entwicklung elektrophoretischer Verfahren zur stereoselektiven Trennung von Stoffen und deren Anwendung in der Untersuchung der Stereoselektivität des Medikamentenabbaus. Damit werden einerseits Grundlagen des elektrophoretischen Transports erarbeitet und andererseits Verfahren entwickelt, die zur Bestimmung von Markersubstanzen im Blut und der Erforschung des Medikamentenabbaus benötigt werden.

Die erarbeiteten Daten werden zum besseren Verständnis der elektrophoretischen Stofftrennung und Analysen beitragen und neue Methoden für die klinische, pharmakologische und forensische Laboranalytik hervorbringen. Die Arbeiten werden mit schweizerischer und internationaler Beteiligung (USA, Tschechische Republik, Australien und Ungarn) ausgeführt und zum regen Austausch innerhalb der wissenschaftlichen Kreisen führen.
Direct link to Lay Summary Last update: 15.04.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Modeling of formation and prevention of a pure water zone in capillary isoelectric focusing with narrow pH range carrier ampholytes.
Takácsi-Nagy Anna, Kilár Ferenc, Thormann Wolfgang (2017), Modeling of formation and prevention of a pure water zone in capillary isoelectric focusing with narrow pH range carrier ampholytes., in Electrophoresis, 38(5), 677-688.
Monitoring of cefepime in human serum and plasma by micellar electrokinetic capillary chromatography: Improvement of sample preparation and validation by liquid chromatography coupled to mass spectrometry.
Šestáková Nela, Theurillat Regula, Sendi Parham, Thormann Wolfgang (2017), Monitoring of cefepime in human serum and plasma by micellar electrokinetic capillary chromatography: Improvement of sample preparation and validation by liquid chromatography coupled to mass spectrometry., in Journal of separation science, 40(8), 1805-1814.
Cardiopulmonary effects and anaesthesia recovery quality in horses anaesthetized with isoflurane and low‐dose S‐ketamine or medetomidine infusions
Menzies M Paula Larenza, Ringer Simone K, Conrot Aude, Theurillat Regula, Kluge Katharina, Kutter Annette PN, Jackson Michelle, Thormann Wolfgang, Bettschart‐Wolfensberger Regula (2016), Cardiopulmonary effects and anaesthesia recovery quality in horses anaesthetized with isoflurane and low‐dose S‐ketamine or medetomidine infusions, in Veterinary Anaesthesia and Analgesia, 43(6), 623-634.
Microassay for ketamine and metabolites in plasma and serum based on enantioselective capillary electrophoresis with highly sulfated γ-cyclodextrin and electrokinetic analyte injectionCE and CEC
Theurillat Regula, Sandbaumhüter Friederike A., Bettschart-Wolfensberger Regula, Thormann Wolfgang (2016), Microassay for ketamine and metabolites in plasma and serum based on enantioselective capillary electrophoresis with highly sulfated γ-cyclodextrin and electrokinetic analyte injectionCE and CEC, in ELECTROPHORESIS, 37(9), 1129-1138.
Microfluidic Isoelectric Focusing of Amyloid Beta Peptides Followed by Micropillar-Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry.
Mikkonen Saara, Jacksén Johan, Roeraade Johan, Thormann Wolfgang, Emmer Åsa (2016), Microfluidic Isoelectric Focusing of Amyloid Beta Peptides Followed by Micropillar-Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry., in Analytical chemistry, 88(20), 10044-10051.
Pharmacokinetics of ketamine and three metabolites in Beagle dogs under sevoflurane vs. medetomidine comedication assessed by enantioselective capillary electrophoresis
Sandbaumhüter Friederike A., Theurillat Regula, Bektas Rima N., Kutter Annette P.N., Bettschart-Wolfensberger Regula, Thormann Wolfgang (2016), Pharmacokinetics of ketamine and three metabolites in Beagle dogs under sevoflurane vs. medetomidine comedication assessed by enantioselective capillary electrophoresis, in Journal of Chromatography A, 1467, 436-444.
Polysaccharide capsule composition of pneumococcal serotype 19A subtypes: Unaltered among subtypes and independent of the nutritional environment.
Brugger Silvio D, Troxler Lukas J, Rüfenacht Susanne, Frey Pascal M, Morand Brigitte, Geyer Rudolf, Mühlemann Kathrin, Höck Stefan, Thormann Wolfgang, Furrer Julien, Christen Stephan, Hilty Markus (2016), Polysaccharide capsule composition of pneumococcal serotype 19A subtypes: Unaltered among subtypes and independent of the nutritional environment., in Infection and immunity, 84, 3152-3160.
Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods
Pelcová Marta, Řemínek Roman, Sandbaumhüter Friederike A., Mosher Richard A., Glatz Zdeněk, Thormann Wolfgang (2016), Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods, in Journal of Chromatography A, 1471, 192-200.
Therapeutic drug monitoring of cefepime with micellar electrokinetic capillary chromatography: Assay improvement, quality assurance, and impact on patient drug levelsElectrodriven Separations
Theurillat Regula, Joneli Jeannine, Wanzenried Ursula, Schiess Jeannette, Hurni Monika, Weber Thomas, Sendi Parham, Thormann Wolfgang (2016), Therapeutic drug monitoring of cefepime with micellar electrokinetic capillary chromatography: Assay improvement, quality assurance, and impact on patient drug levelsElectrodriven Separations, in Journal of Separation Science, 39(13), 2626-2632.
Validation of CE modeling with a contactless conductivity array detector.
Caslavska Jitka, Koenka Israel Joel, Hauser Peter C, Thormann Wolfgang (2016), Validation of CE modeling with a contactless conductivity array detector., in Electrophoresis, 37(5-6), 699-710.
Clinical and pharmacokinetic evaluation of S-ketamine for intravenous general anaesthesia in horses undergoing field castration.
Casoni Daniela, Spadavecchia Claudia, Wampfler Beat, Thormann Wolfgang, Levionnois Olivier L (2015), Clinical and pharmacokinetic evaluation of S-ketamine for intravenous general anaesthesia in horses undergoing field castration., in Acta veterinaria Scandinavica, 57, 21-21.
Computer simulation of electrophoretic aspects of enantiomer migration and separation in capillary electrochromatography with a neutral selector
Thormann Wolfgang (2015), Computer simulation of electrophoretic aspects of enantiomer migration and separation in capillary electrochromatography with a neutral selector, in Electrophoresis, 36, 773-783.
Computer simulations of sample preconcentration in carrier-free systems and isoelectric focusing in microchannels using simple ampholytes.
Mikkonen Saara, Thormann Wolfgang, Emmer Åsa (2015), Computer simulations of sample preconcentration in carrier-free systems and isoelectric focusing in microchannels using simple ampholytes., in Electrophoresis, 36(19), 2386-95.
Effects of medetomidine and its active enantiomer dexmedetomidine on N-demethylation of ketamine in canines determined in vitro using enantioselective capillary electrophoresis.
Sandbaumhüter Friederike A, Theurillat Regula, Thormann Wolfgang (2015), Effects of medetomidine and its active enantiomer dexmedetomidine on N-demethylation of ketamine in canines determined in vitro using enantioselective capillary electrophoresis., in Electrophoresis, 36, 2703-2712.
Impact of Taylor-Aris diffusivity on analyte and system zone dispersion in CZE assessed by computer simulation and experimental validation.
Caslavska Jitka, Mosher Richard A, Thormann Wolfgang (2015), Impact of Taylor-Aris diffusivity on analyte and system zone dispersion in CZE assessed by computer simulation and experimental validation., in Electrophoresis, 36(14), 1529-38.
Optimized on-line enantioselective capillary electrophoretic method for kinetic and inhibition studies of drug metabolism mediated by cytochrome P450 enzymes.
Řemínek Roman, Glatz Zdeněk, Thormann Wolfgang (2015), Optimized on-line enantioselective capillary electrophoretic method for kinetic and inhibition studies of drug metabolism mediated by cytochrome P450 enzymes., in Electrophoresis, 36(11-12), 1349-57.
A contactless Conductivity Detector Array for Capillary Electrophoresis
Stojkovic J., Koenka I.J., Thormann Wolfgang, Hauser Peter C. (2014), A contactless Conductivity Detector Array for Capillary Electrophoresis, in Electrophoresis, 482.
Determination of Genetic Transferrin Variants in Human Serum by Capillary Zone Electrophoresis
Caslavska Jitka, Joneli Jeannine, Wanzenried Ursula, Schiess Jeannette, Lanz Christian, Thormann Wolfgang (2014), Determination of Genetic Transferrin Variants in Human Serum by Capillary Zone Electrophoresis, in Journal of Separation Science, 1663.
Dynamic High-Resolution Computer Simulation of Isotachophoretic Enantiomer Separation and Zone Stability
Caslavska Jitka, Breadmore Michael C., Thormann Wolfgang (2014), Dynamic High-Resolution Computer Simulation of Isotachophoretic Enantiomer Separation and Zone Stability, in Electrophoresis, 625.
The mood-stabilizer lithium prevents hippocampal apoptosis and improves spatial memory in experimental meningitis.
Liechti Fabian D, Stüdle Nicolas, Theurillat Regula, Grandgirard Denis, Thormann Wolfgang, Leib Stephen L (2014), The mood-stabilizer lithium prevents hippocampal apoptosis and improves spatial memory in experimental meningitis., in PloS one, 9(11), 113607-113607.
Analysis of genetic variants of transferrin in human serum after desialylation by capillary zone electrophoresis and capillary isoelectric focusing.
Caslavska Jitka, Lanz Christian, Burda Patricie, Tobler Micha, Thormann Wolfgang, Analysis of genetic variants of transferrin in human serum after desialylation by capillary zone electrophoresis and capillary isoelectric focusing., in Journal of separation science.
Development of a Method for Analysis of Ketamine and Norketamine Enantiomers in Equine Brain and Cerebrospinal Fluid by Capillary Electrophoresis
Theurillat Regula, Larenza M. Paula, Feige Karsten, Bettschart-Wolfensberger Regula, Thormann Wolgang, Development of a Method for Analysis of Ketamine and Norketamine Enantiomers in Equine Brain and Cerebrospinal Fluid by Capillary Electrophoresis, in Electrophoresis.
Dynamic Computer Simulation of Electrophoretic Enantiomer Migration Order and Separation in presence of a neutral cyclodextrin
Thormann Wolfgang, Chankvetadze L., Gumustas M., Chankvetadze B., Dynamic Computer Simulation of Electrophoretic Enantiomer Migration Order and Separation in presence of a neutral cyclodextrin, in Electrophoresis.
Effect of the α2 -receptor agonists medetomidine, detomidine, xylazine, and romifidine on the ketamine metabolism in equines assessed with enantioselective capillary electrophoresis.
Sandbaumhüter Friederike A, Theurillat Regula, Bettschart-Wolfensberger Regula, Thormann Wolfgang, Effect of the α2 -receptor agonists medetomidine, detomidine, xylazine, and romifidine on the ketamine metabolism in equines assessed with enantioselective capillary electrophoresis., in Electrophoresis.
New insights into head-column field-amplified sample stacking: Part I. Detailed study of electrokinetic injection of a weak base across a short water plug.
Šesták Jozef, Thormann Wolfgang, New insights into head-column field-amplified sample stacking: Part I. Detailed study of electrokinetic injection of a weak base across a short water plug., in Journal of chromatography. A.
Separation of hydroxynorketamine stereoisomers using capillary electrophoresis with sulfated β-cyclodextrin and highly sulfated γ-cyclodextrin.
Sandbaumhüter Friederike A, Theurillat Regula, Thormann Wolfgang, Separation of hydroxynorketamine stereoisomers using capillary electrophoresis with sulfated β-cyclodextrin and highly sulfated γ-cyclodextrin., in Electrophoresis.

Collaboration

Group / person Country
Types of collaboration
Prof. F. Kilar, University of Pecs Hungary (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Professor Peter C. Hauser, University of Basel Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Dr. M.C. Breadmore, University of Tasmania Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
Dr. R.A. Mosher, Tucson, AZ United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Professor Regula Bettschart-Wolfensberger, Vetsuisse Faculty, University of Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
PD Dr. P. Burda, University Children's Hospital, Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
Dr. P. Gebauer, Institute of Analytical Chemistry, Brno Czech Republic (Europe)
- in-depth/constructive exchanges on approaches, methods or results
PD Dr. Markus Hilty, University of Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
International PhD Sudents/Postdoc Meeting 2016 of the German Pharmaceutical Society (DPhG) Talk given at a conference Pharmacokinetics of Ketamine and three Metabolites in Beagle Dogs under Sevoflurane vs. Medetomidine Comedication 16.03.2016 Grünenthal, Aachen, Germany Sandbaumhüter Friederike Andrea;
International PhD Sudents/Postdoc Meeting 2016 of the German Pharmaceutical Society (DPhG) Poster Pharmacokinetics of Ketamine and three Metabolites in Beagle Dogs under Sevoflurane vs. Medetomidine Comedication 16.03.2016 Grünenthal, Aachen, Germany Thormann Wolfgang; Sandbaumhüter Friederike Andrea;
CE Forum 2015 Poster Effect of Medetomidine on Ketamine N-Demethylation in vitro analyzed by Enantioselective Capillary Electrophoresis 05.10.2015 Tübingen, Germany Thormann Wolfgang; Sandbaumhüter Friederike Andrea;
12th International Interdisciplinary Meeting on Bioanalysis (CECE 2015) Poster Evaluation of Limits for direct NADPH Application in on-line Capillary Electrophoretic Methods for Drug Metabolism Studies 21.09.2015 Brno, Czech Republic Thormann Wolfgang;
10th Balaton Symposium on High-Performance Separation Methods Poster Modeling of the Development of a Pure Water Zone in Capillary Isoelectric Focusing. 01.09.2015 Siófok, Hungary Thormann Wolfgang;
22th International Symposium on Electro- and Liquid Phase Separation Techniques (ITP2015) and 8th Nordic Separation Science Symposium (NoSSS2015) Poster Practice in NADPH Cofactor Application in on-line Drug Biotransformation by Capillary Electrophoresis 30.08.2015 Helsinki, Finland Thormann Wolfgang;
22th International Symposium on Electro- and Liquid Phase Separation Techniques (ITP2015) and 8th Nordic Separation Science Symposium (NoSSS2015) Poster Impact of Taylor-Aris Diffusivity on Analyte and System Zone Dispersion in Capillary Zone Electrophoresis Assessed by Computer Simulation and Experimental Validation. 30.08.2015 Helsinki, Finland Caslavsky Furrer Jitka; Thormann Wolfgang;
22th International Symposium on Electro- and Liquid Phase Separation Techniques (ITP2015) and 8th Nordic Separation Science Symposium (NoSSS2015) Poster Effect of Medetomidine on Ketamine N-Demethylation in vitro analyzed by Enantioselective Capillary Electrophoresis 30.08.2015 Helsinki, Finland Sandbaumhüter Friederike Andrea; Thormann Wolfgang;
22th International Symposium on Electro- and Liquid Phase Separation Techniques (ITP2015) and 8th Nordic Separation Science Symposium (NoSSS2015) Talk given at a conference Therapeutic Drug Monitoring by CE for Patient Care 30.08.2015 Helsinki, Finland Thormann Wolfgang;
22th International Symposium on Electro- and Liquid Phase Separation Techniques (ITP2015) and 8th Nordic Separation Science Symposium (NoSSS2015) Poster Microassay for Ketamine and ist Metabolites in Plasma based on Enantioselective Capillary Electrophoresis with Electrokinetic Sample Injection. 30.08.2015 Helsinki, Finland Sandbaumhüter Friederike Andrea; Thormann Wolfgang;
22th International Symposium on Electro- and Liquid Phase Separation Techniques (ITP2015) and 8th Nordic Separation Science Symposium (NoSSS2015) Talk given at a conference Combining Microchannel Sample Preparation, Capillary Electrophoresis and Mass Spectrometry for Analysis of Biosamples 30.08.2015 Helsinki, Finland Thormann Wolfgang;
63rd ASMS Conference on Mass Spectrometry and Allied Topics Poster Microfluid Isoelectric Focusing combined with MALDI- and nano-ESI-MS. 31.05.2015 St. Louis, Missouri, United States of America Thormann Wolfgang;
Swiss Society for Infectious Diseases Annual Meeting Poster In Pneumococcal Meningitis Lithium is Neuroprotective. 28.08.2014 Aarau, Switzerland Thormann Wolfgang;
Swiss Society for Infectious Diseases Annual Meeting Poster In Pneumococcal Meningitis Lithium is Neuroprotective 28.08.2014 Aarau, Switzerland Thormann Wolfgang;
Swiss Pharma Science Day Poster Effect of Medetomidine on Ketamine N-Demethylation in Canine Liver Microsomes analysed by Enatioselective Capillary Electrophoresis 20.08.2014 Bern, Switzerland Thormann Wolfgang; Sandbaumhüter Friederike Andrea;
24th European Congress of Clinical Microbiology and Infectious Diseases Poster Lithium protects Neurons from Apoptosis in experimental Pneumococcal Meningitis 10.05.2014 Barcelona, Spain Thormann Wolfgang;
30th International Symposium on MIcroscale Bioseparations (MSB 2014) Poster Optimized on-line Capillary Electrophoretic Method for CYP3A4 mediated Enantioselective N-Demethylation of Ketamine 27.04.2014 Pecs, Hungary Thormann Wolfgang;
30th International Symposium on MIcroscale Bioseparations (MSB 2014) Poster Modeling of Sampling Strategies for Capillary Isoelectric Focusing 27.04.2014 Pecs, Hungary Thormann Wolfgang;


Associated projects

Number Title Start Funding scheme
129706 New approaches in analytical and preparative free fluid electrophoresis 01.04.2010 Project funding

Abstract

Electrophoresis is a key technology in genomics, proteomics and metabolomics, in clinical, forensic and pharmaceutical analysis, in biomedical research and many other fields of application. Our research laboratory specializes in the exploration, development and application of novel approaches in analytical and preparative free fluid electrophoresis. Electrophoresis in free solution can be applied to small molecular mass compounds, ions, peptides, proteins and carbohydrates. A continuation of the work conducted during the previous grant periods is proposed as three parts. Part A comprises fundamental work dealing with electrokinetic transport and separations in capillaries and microchannels using predictions obtained by computer simulation and, whenever possible, experimental validation with setups featuring multiple detectors along the separation column. It is anticipated that our dynamic PC model for electrophoresis, GENTRANS, is employed to study (i) aspects of isoelectric focusing, namely focusing and isotachophoretic mobilization of amphoteric analytes with pI values outside and within the pH gradient established by carrier ampholytes, and focusing in mixtures of good and poor carrier ampholytes, and ii) aspects of zone electrophoretic and isotachophoretic enantiomer separations in presence of a chiral selector. The underlying transport processes can thereby be studied and visualized under realistic time scales and complexity that were hitherto inaccessible. In part B it is proposed to develop (i) capillary electrophoretic conditions for ultrahigh-resolution of transferrin isoforms which will permit the determination of carbohydrate-deficient transferrin in sera of patients showing genetic variants and sera with large and broad trisialo-transferrin peaks, analysis of congenital disorders of glycosylation and cerebrospinal fluid leakages, (ii) capillary electrophoretic assays for determining plasma apolipoprotein J isomers and its sialic acids in the context of alcohol markers, (iii) capillary electrophoretic conditions for determination of urinary marker sugars which are administered to investigate intestinal permeability, and (iv) possibilities for using capillary electrophoretic techniques to characterize capsular carbohydrates of bacteria. Part C deals with stereoselective aspects of electrokinetic separations. Proposed investigations include (i) computer simulation study and application of electrophoretically mediated microanalysis for enzymatic formation of drug metabolites followed by their stereoselective analysis using sulfated cyclodextrins, ii) the characterization of zone electrophoretic and isotachophoretic enantiomer separations in presence of multiply charged cyclodextrins such as sulfated cyclodextrins by computer simulation using SIMUL5complex and experimental validation in different instrumental setups, and (iii) the development and application of stereoselective capillary electrophoretic assays for ritalin and its major metabolites in body fluids, including saliva and plasma. The data obtained will contribute to the better understanding of electrokinetic separations and will pave the way for improved and widespread use of free fluid electrophoretic technology in proteomics, pharmaceutical and pharmacological sciences, clinical analysis and forensic case work. The proposed work will include collaborations with scientists in the Czech Republic, USA, Australia, Hungary and Switzerland.
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