Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F10%3A00044832" target="_blank" >RIV/00216224:14310/10:00044832 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination

Popis výsledku v původním jazyce

In view of the fact that several studies have shown that diclofenac hydroxylation by cytochrome P450 2C9 deviated from Michaelis-Menten kinetics at low substrate concentrations, sweeping combined with MEKC was applied for the kinetic study of this pharmacologically important reaction. A 50 um fused silica capillary (56 cm effective length) was used to carry out all separations. 70 mM SDS in 20 mM phosphate 20 mM tetraborate buffer pH 8.6 was used as the background electrolyte. Injection was accomplishedby the application of 50 mbar (5 kPa) pressure to the sample vial for 52 s. Separation was performed at 22 kV (positive polarity), with a capillary temperature of 25 C and detection at 200 nm. The higher sensitivity of the sweeping-MEKC combination compared to the simple MEKC method enabled this reaction to be fitted to a Hill kinetic model and confirmed the findings of other authors. A Michaelis constant of 2.91 uM, maximum reaction velocity of 9.16 nmol/min/nmol and Hill coefficient o

Název v anglickém jazyce

Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination

Popis výsledku anglicky

In view of the fact that several studies have shown that diclofenac hydroxylation by cytochrome P450 2C9 deviated from Michaelis-Menten kinetics at low substrate concentrations, sweeping combined with MEKC was applied for the kinetic study of this pharmacologically important reaction. A 50 um fused silica capillary (56 cm effective length) was used to carry out all separations. 70 mM SDS in 20 mM phosphate 20 mM tetraborate buffer pH 8.6 was used as the background electrolyte. Injection was accomplishedby the application of 50 mbar (5 kPa) pressure to the sample vial for 52 s. Separation was performed at 22 kV (positive polarity), with a capillary temperature of 25 C and detection at 200 nm. The higher sensitivity of the sweeping-MEKC combination compared to the simple MEKC method enabled this reaction to be fitted to a Hill kinetic model and confirmed the findings of other authors. A Michaelis constant of 2.91 uM, maximum reaction velocity of 9.16 nmol/min/nmol and Hill coefficient o

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)<br>V - Vyzkumna aktivita podporovana z jinych verejnych zdroju

Rok uplatnění

2010

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Separation Science

ISSN

1615-9306

e-ISSN

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Svazek periodika

33

Číslo periodika v rámci svazku

20

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

6

Strana od-do

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Kód UT WoS článku

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EID výsledku v databázi Scopus

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