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

ŘEMÍNEK, Roman and Zdeněk GLATZ. Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination. Journal of Separation Science. Německo: WILEY-VCH Verlag GmbH & Co., vol. 33, No 20, p. 3201-3206. ISSN 1615-9306. 2010.

Basic information

• Original name: Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination
• Name in Czech: Studium atypického kinetického chování cytochromu P450 2C9 s diclofenac při nízkých koncentracích substrátu pomocí Sweeping-MEKC
• Authors: ŘEMÍNEK, Roman (203 Czech Republic) and Zdeněk GLATZ (203 Czech Republic, guarantor).
• Edition: Journal of Separation Science, Německo, WILEY-VCH Verlag GmbH & Co. 2010, 1615-9306.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Germany
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 2.631
• RIV identification code: RIV/00216224:14310/10:00044832
• Organization unit: Faculty of Science
• UT WoS: 000284181900013
• Keywords (in Czech): Sweeping; Cytochrom P450; Diclofenac; Metabolismus léčiv.
• Keywords in English: Sweeping; Cytochrome P450; Diclofenac; Drug metabolism.
• Tags: International impact, Reviewed

Abstract

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 accomplished by 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 of 1.66 were found.

Abstract (in Czech)

Byla provedena studie atypického kinetického chování cytochromu P450 2C9 s diclofenac při nízkých koncentracích substratu pomocí Sweeping-MEKC

Links

• GAP206/10/0057, research and development project: Name: Miniaturizovaný systém pro "on-line" studie metabolismu léčiv založený na kapilární elektroforéze

Investor: Czech Science Foundation, Miniaturized on-line drug metabolism system based on capillary electrophoresis

• LC06023, research and development project: Name: Integrované bioanalytické technologie pro mikroanalýzy a diagnostiku s využitím LIF a hmotnostní spektrometrie

Investor: Ministry of Education, Youth and Sports of the CR

• MSM0021622413, plan (intention): Name: Proteiny v metabolismu a při interakci organismů s prostředím

Investor: Ministry of Education, Youth and Sports of the CR, Proteins in metabolism and interaction of organisms with the environment