Reference : OPTIMIZATION OF A FULLY AUTOMATED ELECTROPHORETICALLY MEDIATED MICROANALYSIS SYSTEM F...
Scientific congresses and symposiums : Unpublished conference/Abstract
Human health sciences : Pharmacy, pharmacology & toxicology
http://hdl.handle.net/2268/202263
OPTIMIZATION OF A FULLY AUTOMATED ELECTROPHORETICALLY MEDIATED MICROANALYSIS SYSTEM FOR CYP1A1 ACTIVITY MONITORING
English
Farcas, Elena mailto [Université de Liège > Département de pharmacie > Analyse des médicaments >]
Servais, Anne-Catherine mailto [Université de Liège > Département de pharmacie > Analyse des médicaments >]
Lamalle, Caroline []
CHIAP, Patrice mailto [Centre Hospitalier Universitaire de Liège - CHU > > Pharmacologie clinique >]
Pochet, Lionel []
Fillet, Marianne mailto [Université de Liège > Département de pharmacie > Analyse des médicaments >]
1-Oct-2016
Yes
No
The 16th Edition of National Congress of Pharmacy in Romania
from 28th of September to 2nd of October
University of Pharmacy Bucharest
Bucharest
Romania
[en] EMMA ; Capillary electrophoresis ; CYP1A1
[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.
Centre Interdisciplinaire de Recherche sur le Médicament- CIRM
Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS
Researchers ; Professionals ; Students
http://hdl.handle.net/2268/202263

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