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GenPharmTox BioTech AG
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Absorption

Caco-2 Permeability Assay

Caco-2 cells are a well-established in vitro model to predict intestinal absorption of drugs and chemicals in humans. Plated Caco-2 cells on a membrane support allow the study of drug transport from the apical side to the basolateral side (A to B transport) as well as from the basolateral side to the apical side (B to A transport). The apical and basolateral sides of Caco-2 cells represent the luminal and blood sides, respectively, of the gastrointestinal tract in vivo. In addition, the transport kinetics can be investigated.
The apparent permability (Papp) of the test item is calculated and compared with that of reference compounds of known permeability.


Quantity of test item required: 15 mg
Turnaround time of draft report: 20 working days after study initiation


Transport

  • triplicates
  • Caco-2 cells (monolayer in Transwell plates)
  • one concentration of test item (50µM)
  • one time point (120 min)

Kinetics

  • triplicates
  • Caco-2 cells (monolayer in Transwell plates)
  • one to four concentrations of test item (12.5, 25, 50, 100µM)
  • one to six time points (0, 15, 30, 60, 90, 120 min)
  • positive controls

Controls

  • propranolol (high perrmeability, transcellular transport)
  • hydrocortisone (moderate permeability, transcellular transport)
  • ranitidine (low permeability, paracellular transport)
  • vinblastine inhibited by verapamil (P-glycoprotein mediated transport)
  • measurement of transepithelial electrical resistance (TEER) to check for membrane integrity

Analytics

  • HPLC-DAD/FLD or LC-MS/MS or Radiodetection:
    detection of parent compound

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GenPharmTox BioTech AG • phone: +49 (0) 89 89 55 59-0 • fax: +49 (0) 89 89 55 59-18 • eMail: info@genpharmtox.de

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