title: Fully automatic flow-based device for monitoring of drug permeation across a cell monolayer.
abstract: A novel flow-programming setup based on the sequential injection principle is herein proposed for on-line monitoring of temporal events in cell permeation studies. The permeation unit consists of a Franz cell with its basolateral compartment mixed under mechanical agitation and thermostated at 37 °C. The apical compartment is replaced by commercially available Transwell inserts with a precultivated cell monolayer. The transport of drug substances across epithelial cells genetically modified with the P-glycoprotein membrane transporter (MDCKII-MDR1) is monitored on-line using rhodamine 123 as a fluorescent marker. The permeation kinetics of the marker is obtained in a fully automated mode by sampling minute volumes of solution from the basolateral compartment in short intervals (10 min) up to 4 h. The effect of a P-glycoprotein transporter inhibitor, verapamil as a model drug, on the efficiency of the marker transport across the cell monolayer is thoroughly investigated. The analytical features of the proposed flow method for cell permeation studies in real time are critically compared against conventional batch-wise procedures and microfluidic devices.
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05, Hradec Králové, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Zelená
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05, Hradec Králové, Czech Republic.|UCIBIO, REQUIMTE, Department of Chemistry, Faculty of Pharmacy, University of Porto, 4051-401, Porto, Portugal.
authors.forename: Sara S
authors.initials: SS
authors.lastname: Marques
authors.affiliation: UCIBIO, REQUIMTE, Department of Chemistry, Faculty of Pharmacy, University of Porto, 4051-401, Porto, Portugal.
authors.forename: Marcela A
authors.initials: MA
authors.lastname: Segundo
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05, Hradec Králové, Czech Republic.|FI-TRACE group, Department of Chemistry, University of Balearic Islands, Carretera de Valldemossa km 7.5, 07122, Palma de Mallorca, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Miró
authors.affiliation: Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05, Hradec Králové, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Pávek
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05, Hradec Králové, Czech Republic. Hana.Sklenarova@faf.cuni.cz.
authors.forename: Hana
authors.initials: H
authors.lastname: Sklenářová
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05, Hradec Králové, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Solich
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D001331:Automation / Q000379:methods*; D001692:Biological Transport; D004847:Epithelial Cells / Q000737:chemistry / Q000378:metabolism*; D017022:Flow Injection Analysis / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007700:Kinetics; D020112:Rhodamine 123 / Q000737:chemistry / Q000378:metabolism; D014700:Verapamil / Q000737:chemistry / Q000378:metabolism*
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D020112:Rhodamine 123; D014700:Verapamil
keywords: Fully automated system; P-glycoprotein transporter; Permeation study; Rhodamine 123; Sequential injection analysis