2010
In-capillary nanoreactor based on transverse diffusion of laminar flow profiles for cytochrome P450 2C9 isoform reaction with diclofenac
ŘEMÍNEK, Roman; Aleš MÁDR a Zdeněk GLATZZákladní údaje
Originální název
In-capillary nanoreactor based on transverse diffusion of laminar flow profiles for cytochrome P450 2C9 isoform reaction with diclofenac
Autoři
ŘEMÍNEK, Roman; Aleš MÁDR a Zdeněk GLATZ
Vydání
Baltimore, MD, USA, Book of Abstracts, 17th International Symposium on Electro- and Liquid Phase-Separation Techniques ITP 2010, od s. 62-62, 1 s. 2010
Nakladatel
Prof. Ziad El Rassi, Oklahoma State University, Stillwater, OK, SA
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Spojené státy
Utajení
není předmětem státního či obchodního tajemství
Označené pro přenos do RIV
Ne
Organizační jednotka
Přírodovědecká fakulta
Klíčová slova česky
CYP 2C9, diklofenak, on-line reakce, kapilární elekroforéza
Klíčová slova anglicky
CYP 2C9, diclofenac, on-line reaction, capillary electrophoresis
Štítky
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 13. 12. 2010 08:20, prof. RNDr. Zdeněk Glatz, CSc.
V originále
A fused silica capillary as an important part of capillary electrophoresis system represents a favourable instrument not only for highly effective separations but also for creating nanoscale reaction chambers. Today commonly used methodology of electrophoretically mediated microanalysis does not allow simple changing of an incubation mixture composition, nevertheless, foreclosing in-capillary reactions from screening studies. To solve this limitation, Krylov et al. introduced the generic methodology for mixing the reactants inside the capillary based on transverse diffusion of laminar flow profiles (TDLFP) [1]. Conceptually, solutions of incubation mixture components are injected by pressure as a series of consecutive plugs which have parabolic profiles due to the laminar nature of flow inside the capillary. Resulting longitudinal interfaces between the plugs then enable the plugs mixing by transverse diffusion within minutes. In this study a method for cytochrome P450 2C9 (CYP2C9) reaction with diclofenac as a probe substrate was developed. 30 min incubation time and sandwich layout, where a plug of CYP2C9 is surrounded by two plugs of diclofenac and NADPH mixture, were selected as appropriate incubation conditions for final method within the optimizing. Although bare silica capillary was used for performing the analyses of samples consisting of enzyme preparations, strong rinsing procedure with 1 M H3PO4 and 1 M NaOH applied between following runs provides good method’s repeatability with RSD values of 1.52 % and 4.83 % (n = 6) for migration times and peak areas of 4’-hydroxydiclofenac, respectively. Since methodology of TDLFP allows simple implementation of changes in incubation mixture composition, introduced method represents a promising tool for in-capillary inhibition studies of CYP2C9.
Česky
Byla zavedena metoda pro on-line reakci cytochromu P450 2C9 se specifickým substrátem diklofenakem.
Návaznosti
| GAP206/10/0057, projekt VaV |
| ||
| LC06023, projekt VaV |
| ||
| MSM0021622413, záměr |
|