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

J 2015

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

LANGMAJEROVÁ, Monika, Roman ŘEMÍNEK, Marta PELCOVÁ, František FORET, Zdeněk GLATZ et. al.

Basic information

Original name

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

Name in Czech

Kombinace on-line kapilární elektroforézy s hmotnostně spektroemtrickou detekcí pro studium metabolismu léčiv

Authors

LANGMAJEROVÁ, Monika (203 Czech Republic, belonging to the institution), Roman ŘEMÍNEK (203 Czech Republic, belonging to the institution), Marta PELCOVÁ (203 Czech Republic, belonging to the institution), František FORET (203 Czech Republic, belonging to the institution) and Zdeněk GLATZ (203 Czech Republic, guarantor, belonging to the institution)

Edition

Electrophoresis, Verlag Chemie, 2015, 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: 2.482

RIV identification code

RIV/00216224:14310/15:00080656

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1002/elps.201400394

UT WoS

000356004200017

Keywords (in Czech)

kapilární elektroforéza; hmotnostní spektrometrie; enzymatická reakce

Keywords in English

capillary electrophoresis; mass spectrometry; enzymatic reaction

Abstract

ORIG CZ

V originále

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 isoform 2C9. As a result, the method is conceptually applicable for the screening of any other cytochromes P450 isoform and its substrates and inhibitors after adapting the incubation and separation conditions.

In Czech

Byla vyvinuta nová metoda pro studium metabolismu léčiv pomocí CE-MS metody zahrnující enzymatickou reakci v prostoru kapiláry.

Links

GBP206/12/G014, research and development project

Name: Centrum pokročilých bioanalytických technologií

Citovat

LANGMAJEROVÁ, Monika, Roman ŘEMÍNEK, Marta PELCOVÁ, František FORET and Zdeněk GLATZ. Combination of on-line capillary electrophoretic assay with mass spectrometry detection for the study of drug metabolism by cytochromes P450. Electrophoresis. Verlag Chemie, 2015, vol. 36, 11-12, p. 1365-1373. ISSN 0173-0835. Available from: https://dx.doi.org/10.1002/elps.201400394.

@article{1219254,
   author = {Langmajerová, Monika and Řemínek, Roman and Pelcová, Marta and Foret, František and Glatz, Zdeněk},
   article_number = {11-12},
   doi = {http://dx.doi.org/10.1002/elps.201400394},
   keywords = {capillary electrophoresis; mass spectrometry; enzymatic reaction},
   language = {eng},
   issn = {0173-0835},
   journal = {Electrophoresis},
   title = {Combination of on-line capillary electrophoretic assay with mass spectrometry detection for the study of drug metabolism by cytochromes P450},
   volume = {36},
   year = {2015}
}

TY  - JOUR
ID  - 1219254
AU  - Langmajerová, Monika - Řemínek, Roman - Pelcová, Marta - Foret, František - Glatz, Zdeněk
PY  - 2015
TI  - Combination of on-line capillary electrophoretic assay with mass spectrometry detection for the study of drug metabolism by cytochromes P450
JF  - Electrophoresis
VL  - 36
IS  - 11-12
SP  - 1365-1373
EP  - 1365-1373
PB  - Verlag Chemie
SN  - 01730835
KW  - capillary electrophoresis
KW  - mass spectrometry
KW  - enzymatic reaction
N2  - 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 isoform 2C9. As a result, the method is conceptually applicable for the screening of any other cytochromes P450 isoform and its substrates and inhibitors after adapting the incubation and separation conditions.
ER  -

LANGMAJEROVÁ, Monika, Roman ŘEMÍNEK, Marta PELCOVÁ, František FORET and Zdeněk GLATZ. Combination of on-line capillary electrophoretic assay with mass spectrometry detection for the study of drug metabolism by cytochromes P450. \textit{Electrophoresis}. Verlag Chemie, 2015, vol.~36, 11-12, p.~1365-1373. ISSN~0173-0835. Available from: https://dx.doi.org/10.1002/elps.201400394.

Detailed Information on Publication Record