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

J 2016

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

PELCOVÁ, Marta, Roman ŘEMÍNEK, Friederike A SANDBAUMHÜTER, Richard A MOSHER, Zdeněk GLATZ et. al.

Basic information

Original name

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

Authors

PELCOVÁ, Marta (203 Czech Republic, guarantor, belonging to the institution), Roman ŘEMÍNEK (203 Czech Republic, belonging to the institution), Friederike A SANDBAUMHÜTER (276 Germany), Richard A MOSHER (840 United States of America), Zdeněk GLATZ (203 Czech Republic, belonging to the institution) and Wolfgang THORMANN (756 Switzerland)

Edition

Journal of Chromatography A, Holandsko, Elsevier, 2016, 0021-9673

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 3.981

RIV identification code

RIV/00216224:14310/16:00088196

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1016/j.chroma.2016.10.002

UT WoS

000387520400022

Keywords (in Czech)

kapilární elektroforéza; enzymov\á reakce; elektroforeticky zprostředkovaná analýza; difuze; simulace; Taylor-Aris disperze;

Keywords in English

capillary electrophoresis; enzyme reaction; electrophoretically mediated microanalysis; diffusion; simulation; Taylor-Aris dispersion

Abstract

ORIG CZ

V originále

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.

In Czech

Byly provedeny simulace difuzního a elektromigračního míchání enzymové reakce cytochromu P450 3A4 s ketamínem.

Links

GA16-06106S, research and development project

Name: Vysoce efektivní systém založený na kapilární elektroforéze pro screening inhibitorů beta-sekretázy jako terapeutického cíle pro Alzheimerovu chorobu (Acronym: Alzheimer)

Investor: Czech Science Foundation

MUNI/A/1265/2015, interní kód MU

Name: Podpora biochemického výzkumu v roce 2016

Investor: Masaryk University, Category A

Citovat

PELCOVÁ, Marta, Roman ŘEMÍNEK, Friederike A SANDBAUMHÜTER, Richard A MOSHER, Zdeněk GLATZ and Wolfgang THORMANN. Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods. Journal of Chromatography A. Holandsko: Elsevier, 2016, vol. 1471, November, p. 192-200. ISSN 0021-9673. Available from: https://dx.doi.org/10.1016/j.chroma.2016.10.002.

@article{1355796,
   author = {Pelcová, Marta and Řemínek, Roman and Sandbaumhüter, Friederike A and Mosher, Richard A and Glatz, Zdeněk and Thormann, Wolfgang},
   article_location = {Holandsko},
   article_number = {November},
   doi = {http://dx.doi.org/10.1016/j.chroma.2016.10.002},
   keywords = {capillary electrophoresis; enzyme reaction; electrophoretically mediated microanalysis; diffusion; simulation; Taylor-Aris dispersion},
   language = {eng},
   issn = {0021-9673},
   journal = {Journal of Chromatography A},
   title = {Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods},
   volume = {1471},
   year = {2016}
}

TY  - JOUR
ID  - 1355796
AU  - Pelcová, Marta - Řemínek, Roman - Sandbaumhüter, Friederike A - Mosher, Richard A - Glatz, Zdeněk - Thormann, Wolfgang
PY  - 2016
TI  - Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods
JF  - Journal of Chromatography A
VL  - 1471
IS  - November
SP  - 192-200
EP  - 192-200
PB  - Elsevier
SN  - 00219673
KW  - capillary electrophoresis
KW  - enzyme reaction
KW  - electrophoretically mediated microanalysis
KW  - diffusion
KW  - simulation
KW  - Taylor-Aris dispersion
N2  - 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.
ER  -

PELCOVÁ, Marta, Roman ŘEMÍNEK, Friederike A SANDBAUMHÜTER, Richard A MOSHER, Zdeněk GLATZ and Wolfgang THORMANN. Simulation and experimental study of enzyme and reactant mixing in capillary electrophoresis based on-line methods. \textit{Journal of Chromatography A}. Holandsko: Elsevier, 2016, vol.~1471, November, p.~192-200. ISSN~0021-9673. Available from: https://dx.doi.org/10.1016/j.chroma.2016.10.002.

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