Combination of on-line capillary electrophoretic assay with mass spectrometry detection for the study of drug metabolism by cytochromes P450

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F15%3A00080656" target="_blank" >RIV/00216224:14310/15:00080656 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1002/elps.201400394" target="_blank" >http://dx.doi.org/10.1002/elps.201400394</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/elps.201400394" target="_blank" >10.1002/elps.201400394</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Combination of on-line capillary electrophoretic assay with mass spectrometry detection for the study of drug metabolism by cytochromes P450

Popis výsledku v původním jazyce

A new CE-MS method with enzymatic reaction inside the capillary was developed for the study of drug metabolism by cytochromes P450. This automated method, based on the transverse diffusion of laminar flow profiles methodology, is comprised of the injection of substrates and enzyme, their mixing, incubation and separation of the reaction products, all performed by CE, and their detection, identification and quantification by MS. The developed and validated method was finally used to conduct a kinetic study of cytochrome P450 isoform 2C9 or human liver microsomes with diclofenac in order to demonstrate its practical functionality. All the estimated kinetic values ? apparent Michaelis constants and apparent maximum reaction velocities were in agreement with literature data obtained using other techniques. In addition, the consumption of reactants was in the tens of nL per analysis. The method?s usability was further demonstrated on tolbutamide, the other probe substrate of cytochrome P450

Název v anglickém jazyce

Combination of on-line capillary electrophoretic assay with mass spectrometry detection for the study of drug metabolism by cytochromes P450

Popis výsledku anglicky

A new CE-MS method with enzymatic reaction inside the capillary was developed for the study of drug metabolism by cytochromes P450. This automated method, based on the transverse diffusion of laminar flow profiles methodology, is comprised of the injection of substrates and enzyme, their mixing, incubation and separation of the reaction products, all performed by CE, and their detection, identification and quantification by MS. The developed and validated method was finally used to conduct a kinetic study of cytochrome P450 isoform 2C9 or human liver microsomes with diclofenac in order to demonstrate its practical functionality. All the estimated kinetic values ? apparent Michaelis constants and apparent maximum reaction velocities were in agreement with literature data obtained using other techniques. In addition, the consumption of reactants was in the tens of nL per analysis. The method?s usability was further demonstrated on tolbutamide, the other probe substrate of cytochrome P450

Druh

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

CEP obor

CE - Biochemie

OECD FORD obor

—

Projekt

<a href="/cs/project/GBP206%2F12%2FG014" target="_blank" >GBP206/12/G014: Centrum pokročilých bioanalytických technologií</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2015

Kód důvěrnosti údajů

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

Název periodika

Electrophoresis

ISSN

0173-0835

e-ISSN

—

Svazek periodika

36

Číslo periodika v rámci svazku

11-12

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

9

Strana od-do

1365-1373

Kód UT WoS článku

000356004200017

EID výsledku v databázi Scopus

—