Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods

Pelcová, Marta; Řemínek, Roman; Sandbaumhüter, Friederike Andrea; Mosher, Richard A; Glatz, Zdeněk; Thormann, Wolfgang (2016). Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods. Journal of chromatography A, 1471, pp. 192-200. Elsevier Science 10.1016/j.chroma.2016.10.002

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The establishment of an efficient reaction mixture represents a crucial part of capillary electrophoresis based on-line enzymatic assays. For ketamine N-demethylation to norketamine mediated by the cytochrome P450 3A4 enzyme, mixing of enzyme and reactants in the incubation buffer at physiological pH was studied by computer simulation. A dynamic electrophoretic simulator that encompasses Taylor-Aris diffusivity which accounts for dispersion due to the parabolic flow profile associated with pressure driven flow was utilized. The simulator in the diffusion mode was used to predict transverse diffusional reactant mixing occurring during hydrodynamic plug injection of configurations featuring four and seven plugs. The same simulator in the electrophoretic mode was applied to study electrophoretic reactant mixing caused by voltage application in absence of buffer flow. Resulting conclusions were experimentally verified with enantioselective analysis of norketamine in a background electrolyte at low pH. Furthermore, simulations visualize buffer changes that occur upon power application between incubation buffer and background electrolyte and have an influence on the reaction mixture.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Laboratory for Clinical Pharmacology
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases

UniBE Contributor:

Pelcová, Marta; Sandbaumhüter, Friederike Andrea and Thormann, Wolfgang

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

0021-9673

Publisher:

Elsevier Science

Language:

English

Submitter:

Wolfgang Thormann

Date Deposited:

14 Mar 2017 15:48

Last Modified:

14 Mar 2017 15:48

Publisher DOI:

10.1016/j.chroma.2016.10.002

PubMed ID:

27720173

Uncontrolled Keywords:

Capillary electrophoresis; Diffusion; Electrophoretically mediated microanalysis; Enzyme reaction; Simulation; Taylor-Aris dispersion

BORIS DOI:

10.7892/boris.93155

URI:

https://boris.unibe.ch/id/eprint/93155

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