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@article{898961, author = {Řemínek, Roman and Glatz, Zdeněk}, article_location = {Německo}, article_number = {20}, keywords = {Sweeping; Cytochrome P450; Diclofenac; Drug metabolism.}, language = {eng}, issn = {1615-9306}, journal = {Journal of Separation Science}, title = {Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination}, volume = {33}, year = {2010} }
TY - JOUR ID - 898961 AU - Řemínek, Roman - Glatz, Zdeněk PY - 2010 TI - Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination JF - Journal of Separation Science VL - 33 IS - 20 SP - 3201-3206 EP - 3201-3206 PB - WILEY-VCH Verlag GmbH & Co. SN - 16159306 KW - Sweeping KW - Cytochrome P450 KW - Diclofenac KW - Drug metabolism. N2 - 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. ER -
ŘEMÍNEK, Roman a Zdeněk GLATZ. Study of Atypical Kinetic Behaviour of Cytochrome P450 2C9 Isoform with Diclofenac at Low Substrate Concentrations by Sweeping-MEKC Combination. \textit{Journal of Separation Science}. Německo: WILEY-VCH Verlag GmbH \&{} Co., 2010, roč.~33, č.~20, s.~3201-3206. ISSN~1615-9306.
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