In silico model-based exploration of the applicability of parallel artificial membrane permeability assay (PAMPA) to screen chemicals of environmental concern

Abstract

Parallel Artificial Membrane Permeability Assay (PAMPA) is an in vitro laboratory method for screening the transmembrane permeability of chemicals. Stemming from medicinal chemistry, PAMPA has the potential for use in the cost-effective high-throughput evaluation of chemicals of environmental concern. However, many chemicals of environmental concern differ substantially from pharmaceuticals in hydrophobicity and volatility. Here, we develop an in silico mass balance model to explore the impacts of chemical properties on chemical mass transfer in PAMPA and PAMPA’s applicability to hydrophobic or volatile chemicals of environmental concern. The model’s performance is evaluated by agreement between predicted and measured permeabilities of 1383 chemicals. The model predicts that the PAMPA measured permeability can be highly uncertain for hydrophobic chemicals because of considerable retention by the artificial membrane and for volatile chemicals because of substantial volatilization to the headspace. Notably, the permeabilities of hydrophobic chemicals are remarkably sensitive to specific experimental conditions, for example, the frequency of stirring and incubation time, challenging the comparison between measurements under different conditions. For hydrophobic chemicals, the PAMPA measured permeability may largely indicate the permeability of the unstirred water layer over the membrane, instead of the “intrinsic” permeability of the membrane, and therefore, may not be of interest for environmental exposure and risk assessments. The model also predicts that the time for mass transfer of highly hydrophobic chemicals to reach the steady state likely exceeds the incubation time, which violates the steady-state assumption used in calculating permeability from measured concentrations. Overall, our theoretical analysis underscores the importance to consider chemical properties when applying the current design of PAMPA to chemicals of environmental concern.