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OPTIMIZATION OF A FULLY AUTOMATED ELECTROPHORETICALLY MEDIATED MICROANALYSIS SYSTEM FOR CYP1A1 ACTIVITY MONITORING
Farcas, Elena; Servais, Anne-Catherine; Lamalle, Caroline et al.
2016The 16th Edition of National Congress of Pharmacy in Romania
 

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Keywords :
EMMA; Capillary electrophoresis; CYP1A1
Abstract :
[en] Objective of the study Since CYP1A1, a member of cytochrome P450 superfamily, has been described to be over expressed in various types of cancer, our study was focused on the optimization of a fully automated system for the monitoring of this particularly interesting enzyme. Moreover, the potentiality of this approach to screen CYP1A1 inhibitors was investigated. Materials and methods The experiments were carried out on a HP3DCE system using an on-column DAD. The EMMA procedure was performed by injecting a plug containing the co-factor(NADPH) and the substrate(7-ethoxycoumarin) between two plugs of CYP1A1 supersomes. The reaction was triggered by the application of a voltage switch. The voltage was then turned off to allow the metabolic reaction to occur. The separation of the components was then performed. Results Satisfying results were obtained using CYP1A1 at a concentration of 200 pmol/mL, while the incubation time was settled to 15 min. A DoE was performed to find the best mixing conditions. The amount of metabolite obtained was comparable to the one detected after conventional off-line metabolization. The ability of our system to monitor CYP1A1 inhibition was then proven with apigenin, a well-known CYP1A1 inhibitor. Conclusions The present study describes the development of a fully automatized in-capillary method for CYP1A1 activity monitoring and proves the potentialy of our system to be used for the screening of CYP1A1 inhibitors. The advantages of performing inline metabolization assays are mainly the miniaturization and the automatization of the process. Besides, the reagents consumption is drastically reduced due to the injection of few tens of nanoliters.
Research center :
CIRM - Centre Interdisciplinaire de Recherche sur le Médicament - ULiège
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Farcas, Elena ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Servais, Anne-Catherine  ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Lamalle, Caroline
CHIAP, Patrice ;  Centre Hospitalier Universitaire de Liège - CHU > Pharmacologie clinique
Pochet, Lionel
Fillet, Marianne ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
OPTIMIZATION OF A FULLY AUTOMATED ELECTROPHORETICALLY MEDIATED MICROANALYSIS SYSTEM FOR CYP1A1 ACTIVITY MONITORING
Publication date :
01 October 2016
Event name :
The 16th Edition of National Congress of Pharmacy in Romania
Event organizer :
University of Pharmacy Bucharest
Event place :
Bucharest, Romania
Event date :
from 28th of September to 2nd of October
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE]
Available on ORBi :
since 03 October 2016

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