Detailed Information on Publication Record
2007
Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography
KONEČNÝ, Jiří, Jan JUŘICA, Josef TOMANDL and Zdeněk GLATZBasic information
Original name
Study of recombinant cytochrome P450 2C9 activity with diclofenac by micellar electrokinetic capillary chromatography
Name in Czech
Studium reakce rekombinantního cytochromu P450 2C9 s dikofenakem pomocí MEKC
Authors
KONEČNÝ, Jiří (203 Czech Republic, belonging to the institution), Jan JUŘICA (203 Czech Republic, belonging to the institution), Josef TOMANDL (203 Czech Republic, belonging to the institution) and Zdeněk GLATZ (203 Czech Republic, guarantor, belonging to the institution)
Edition
Electrophoresis, Weinheim, VHC, 2007, 0173-0835
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10600 1.6 Biological sciences
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
Impact factor
Impact factor: 3.609
RIV identification code
RIV/00216224:14310/07:00020039
Organization unit
Faculty of Science
UT WoS
000246116100009
Keywords in English
drug metabolism; cytochrome P450 2C9; diklofenac; MEKC
Tags
International impact, Reviewed
Změněno: 6/6/2012 08:44, doc. RNDr. Josef Tomandl, Ph.D.
V originále
Cytochrome P450 2C9 (CYP2C9) is one of the most important isoform in human liver involved in the metabolism of a large number of therapeutic agents. The aim of this communication is to demonstrate the applicability of capillary electrophoresis for the determination of the enzymatic activity of CYP2C9 with diclofenac as a probe substrate. Micellar electrokinetic capillary chromatography (MEKC) with SDS as a pseudostationary phase was used for this purpose. Compared to other assays, the MEKC based method is rapid, can be automated, and requires only small quantity of enzymes and substrate. Moreover, the enzymatic reaction can be monitored with high sensitivity and reproducibility even when the reaction mixture is used for the analysis without any pre-treatment. The kinetic study on the given enzymatic reaction was also performed since basic characterization of drug biotranformation generally begins with the enzyme kinetic analysis of metabolite formation. As a result Michaelis constant and maximum reaction velocity were evaluated, the values Km 3.44 uM, respectively 19.78 nmol.min-1.nmol-1 were in agreement with the literature data. On the other hand the slight deviation from typical Michaelis-Menten kinetics with a weak positive cooperativity was found at diclofenac concentration below 2 uM. The same atypical kinetic behaviour of CYP2C9 was also observed by other authors.
In Czech
Byla vyprcována metoda pro stanovení aktivity CYP P450 2C9 pomocí MEKC. Metoda byla rovněž použita pro stanovení kinetických parametrů daného enzymu.
Links
| GA203/06/0047, research and development project |
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| LC06023, research and development project |
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| MSM0021622413, plan (intention) |
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