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Analytical and Bioanalytical Chemistry

, Volume 408, Issue 3, pp 971–981 | Cite as

Fully automatic flow-based device for monitoring of drug permeation across a cell monolayer

  • Lucie Zelená
  • Sara S. Marques
  • Marcela A. Segundo
  • Manuel Miró
  • Petr Pávek
  • Hana SklenářováEmail author
  • Petr Solich
Research Paper

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.

Keywords

Fully automated system Permeation study P-glycoprotein transporter Rhodamine 123 Sequential injection analysis 

Notes

Acknowledgments

The authors gratefully acknowledge the financial support of the Ministry of Education of the Czech Republic, project no. SVV 260 184, Grant Agency of the Charles University, project no. 159415, and the Grant Agency of the Czech Republic, project no. 15-10781S/P206. This work received financial support from the European Union (FEDER funds through COMPETE) and Portuguese National Funds (FCT, Fundação para a Ciência e Tecnologia) through project UID/Multi/04378/2013. M.M. is grateful to the Spanish Ministry of Science and Competitiveness for financial support through projects CTM2014-56628-C3-3-R and CTM2014-61553-EXP. Special thanks to Mgr. Lucie Hyršová for cell cultivation and kinetic profiles evaluation.

Compliance with ethical standard

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

216_2015_9194_MOESM1_ESM.pdf (69 kb)
ESM 1 (PDF 68 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Lucie Zelená
    • 1
  • Sara S. Marques
    • 1
    • 2
  • Marcela A. Segundo
    • 2
  • Manuel Miró
    • 1
    • 3
  • Petr Pávek
    • 4
  • Hana Sklenářová
    • 1
    Email author
  • Petr Solich
    • 1
  1. 1.Department of Analytical Chemistry, Faculty of Pharmacy in Hradec KrálovéCharles University in PragueHradec KrálovéCzech Republic
  2. 2.UCIBIO, REQUIMTE, Department of Chemistry, Faculty of PharmacyUniversity of PortoPortoPortugal
  3. 3.FI-TRACE group, Department of ChemistryUniversity of Balearic IslandsPalma de MallorcaSpain
  4. 4.Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec KrálovéCharles University in PragueHradec KrálovéCzech Republic

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