title: Flow-through enzyme immobilized amperometric detector for the rapid screening of acetylcholinesterase inhibitors by flow injection analysis.
abstract: A commercially available thin-layer flow-through amperometric detector, with the sensing block customized in an original design, was applied to the screening of drug compounds known as acetylcholinesterase (AChE) inhibitors. AChE from electric eel was covalently immobilized onto a cysteamine modified gold disk adjacent to a silver disk working electrode. On-line studies were performed by flow injection analysis (FIA) in PBS buffer pH 7.4. Seven commercially available AChE inhibitors used in the medical field, namely neostigmine, eserine, tacrine, donepezil, rivastigmine, pyridostigmine and galantamine as well as two natural compounds, quercetin and berberine, were investigated. The same trend of inhibitory potency as described in the literature was observed. Of particular interest and in addition to the determination of the IC50 values, this flow-through system allowed the study of both, the stability of the enzyme-inhibitor complex and the kinetic of the enzyme activity recovery.
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium. Electronic address: mvdeput@ulb.ac.be.
authors.forename: Marie
authors.initials: M
authors.lastname: Vandeput
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium.
authors.forename: Cobra
authors.initials: C
authors.lastname: Parsajoo
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Vanheuverzwijn
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Patris
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Yüzüncü Yıl University, Campus Zeve, 65080 Van, Turkey.
authors.forename: Yavuz
authors.initials: Y
authors.lastname: Yardim
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium.
authors.forename: Alexandre
authors.initials: A
authors.lastname: le Jeune
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Sarakbi
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium.
authors.forename: Dominique
authors.initials: D
authors.lastname: Mertens
authors.affiliation: Laboratory of Instrumental Analysis and Bioelectrochemistry, Faculty of Pharmacy, Université Libre de Bruxelles, Boulevard du Triomphe, Campus Plaine, CP 205/06, 1050 Brussels, Belgium. Electronic address: jmkauf@ulb.ac.be.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Kauffmann
mesh_terms: D002800:Cholinesterase Inhibitors / Q000032:analysis*; D055664:Electrochemical Techniques / Q000295:instrumentation*; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry*; D017022:Flow Injection Analysis / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized
keywords: Acetylcholinesterase; Drug screening; Enzyme inhibition; FIA; Silver electrode