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.

Základní údaje

Originální název

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

Autoři

PELCOVÁ, Marta (203 Česká republika, garant, domácí), Roman ŘEMÍNEK (203 Česká republika, domácí), Friederike A SANDBAUMHÜTER (276 Německo), Richard A MOSHER (840 Spojené státy), Zdeněk GLATZ (203 Česká republika, domácí) a Wolfgang THORMANN (756 Švýcarsko)

Vydání

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

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 3.981

Kód RIV

RIV/00216224:14310/16:00088196

Organizační jednotka

Přírodovědecká fakulta

DOI

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

UT WoS

000387520400022

Klíčová slova česky

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

Klíčová slova anglicky

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

Štítky

AKR, rivok

Změněno: 13. 4. 2017 22:23, Ing. Andrea Mikešková

Anotace

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.

Česky

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

Návaznosti

GA16-06106S, projekt VaV

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

Investor: Grantová agentura ČR, Vysoce efektivní systém založený na kapilární elektroforéze pro screening inhibitorů beta-sekretázy jako terapeutického cíle pro Alzheimerovu chorobu

MUNI/A/1265/2015, interní kód MU

Název: Podpora biochemického výzkumu v roce 2016

Investor: Masarykova univerzita, Podpora biochemického výzkumu v roce 2016, DO R. 2020_Kategorie A - Specifický výzkum - Studentské výzkumné projekty

Citovat

PELCOVÁ, Marta, Roman ŘEMÍNEK, Friederike A SANDBAUMHÜTER, Richard A MOSHER, Zdeněk GLATZ a 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, roč. 1471, November, s. 192-200. ISSN 0021-9673. Dostupné z: 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 a 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, roč.~1471, November, s.~192-200. ISSN~0021-9673. Dostupné z: https://dx.doi.org/10.1016/j.chroma.2016.10.002.

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