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Drug Permeability Profiling Using the Novel Permeapad® 96-Well Plate



Here, first experiences with a prototype tool for high throughput (passive) permeability profiling, a 96-well plate comprising the Permeapad® membrane, are reported. The permeabilities of a set of drugs were determined and compared to published measures of oral absorption, such as human fraction absorbed (Fa) and in vitro permeability values obtained using other tools.


The tool consists of a 96-well bottom and screen plate with the artificial, phospholipid-based barrier (Permeapad®) mounted between the plates’ lower and upper compartments. The permeability of 14 model compounds including high- and low-absorption drugs, cationic, anionic, zwitterionic and neutral molecules, was determined by quantifying the compounds’ transport over time, deriving the steady-state flux from the linear part of the cumulative curves and calculating the apparent permeability (Papp). The membrane structure was investigated in a high-resolution digital light microscope.


The Permeapad® 96-well plate was found suited to distinguish high and low absorption drugs and yielded a hyperbolic correlation to Fa. The Papp values obtained were congruent with those determined with in-house prepared Permeapad® in the Franz cell set-up. Furthermore, good to excellent correlations were seen with Caco-2 permeability (R2 = 0.70) and PAMPA permeability (R2 = 0.89). Microscopic investigation of the Permeapad® barrier revealed the formation of phospholipid vesicles and myelin figures in aqueous environment.


The Permeapad® 96-well plate permeation set-up is a promising new tool for rapid and reproducible passive permeability profiling.

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Fa :

Fraction absorbed in humans


High-performance liquid chromatography


Parallel artificial membrane permeability assay

Papp :

Apparent permeability


Phosphate buffered saline


Polyvinylidene fluoride


Phospholipid vesicle-based permeation assay


Trifluoroacetic acid


Total polar surface area


Ultra-high-performance liquid chromatography with ultraviolet detection


Unstirred water layer


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Jacobsen, AC., Nielsen, S., Brandl, M. et al. Drug Permeability Profiling Using the Novel Permeapad® 96-Well Plate. Pharm Res 37, 93 (2020).

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Key words

  • 96-well plate
  • artificial barrier
  • high throughput
  • intestinal absorption
  • microplate
  • permeability