title: Determination of fexofenadine in Hank's balanced salt solution by high-performance liquid chromatography with ultraviolet detection: application to Caco-2 cell permeability studies.
abstract: The drug-transporting proteins can affect the pharmacokinetics and pharmacodymanics of many drugs, resulting in an erratic and unpredictable pharmacological response. The Caco-2 monolayer is routinely applied to investigate the carrier-mediated transport of drugs. Therefore, the selection of a marker compound able to characterize the activity of such transporters is crucial. Fexofenadine (FEX), a P-gp/OATP substrate, can be considered a suitable probe. However, in order to use be used as a marker compound, it is mandatory to develop an analytical method able to quantify this drug during the in vitro permeability assay. An HPLC method with ultraviolet detection was developed; the mobile phase consisted of phosphate buffer (pH 3.2) containing 10 m m of sodium octanosulphonate and acetonitrile (60:40) and the flow rate was set at 1.2 mL/min. Fexofenadine was eluted at 40°C, the retention time was about 4.6 min. The LOD and LOQ values were 1.9 and 6.2 ng/mL, respectively. Verapamil and ketoconazole, the most common P-gp inhibitors, were eluted as distinct peaks of that corresponding to fexofenadine The method was successfully applied to quantify the amount of FEX transported across the Caco-2 monolayer and could be an additional tool for those investigating the role of membrane transporters on drug absorption.
authors.affiliation: Universidade Federal de São João del-Rei, Campus Centro-Oeste, Brazil.
authors.forename: Marco Antônio dos Reis
authors.initials: MA
authors.lastname: Júnior
authors.forename: Ana Cláudia Miranda
authors.initials: AC
authors.lastname: de Faria
authors.forename: Eudes da Silva
authors.initials: Eda S
authors.lastname: Velozo
authors.forename: Teresa
authors.initials: T
authors.lastname: Dalla Costa
authors.forename: Frank Pereira
authors.initials: FP
authors.lastname: de Andrade
authors.forename: Whocely Victor
authors.initials: WV
authors.lastname: de Castro
mesh_terms: D018938:Caco-2 Cells; D002477:Cells / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D026901:Membrane Transport Proteins / Q000378:metabolism; D010539:Permeability; D016593:Terfenadine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism
chemical_list: D003470:Culture Media; D026901:Membrane Transport Proteins; D016593:Terfenadine; C093230:fexofenadine
keywords: Caco-2; HPLC-UV; fexofenadine; permeability