Quantitation and prediction of sorptive losses during toxicity testing of polycyclic aromatic hydrocarbon (PAH) and nitrated PAH (NPAH) using polystyrene 96-well plates

Neurotoxicol Teratol. 2016 Sep-Oct:57:30-38. doi: 10.1016/j.ntt.2016.05.001. Epub 2016 May 8.


Developing zebrafish are increasingly being used for rapid assessments of chemical toxicity, and these assays are frequently conducted in multi-well plastic plates. This study investigated the sorptive behavior of polycyclic aromatic hydrocarbons (PAHs) and nitrated PAHs (NPAHs) to uncoated 96-well polystyrene plates typically used for zebrafish (Danio rerio) testing. We measured the percent sorption in the presence and absence of zebrafish embryos, at two exposure concentrations, as well as using two different procedures (addition of embryos to polystyrene plates either before analyte addition, or allowing 24h of equilibrium between analyte addition and embryo addition to the polystyrene plates). Following exposure, the plates were extracted with hexane and analyzed using gas chromatography coupled with mass spectrometry (GC/MS). Allowing 24h of pre-incubation between the addition of analytes and embryos did not significantly impact the percent sorption. The percent sorption was higher for both PAHs and NPAHs at the lower exposure concentration, and sorption was lower in the presence of zebrafish embryos. A mass balance model was developed to predict the sorption to polystyrene plates, based on the PAH and NPAH mass distribution ratios between polystyrene and water. While PAH sorption was significantly correlated with subcooled liquid solubility, NPAH sorption did not correlate with any of the physical-chemical properties investigated. This indicates the need to better understand the sorptive behavior of hydrophobic analytes to plastics, and to better account for sorptive losses during toxicity testing in polystyrene plates.

Keywords: 96-Well plate; Nitrated polycyclic aromatic hydrocarbon (NPAH); Polycyclic aromatic hydrocarbon (PAH); Polystyrene; Sorption.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Gas Chromatography-Mass Spectrometry
  • Nitro Compounds / chemistry
  • Polycyclic Aromatic Hydrocarbons / chemistry*
  • Polystyrenes / chemistry*
  • Toxicity Tests / instrumentation*
  • Toxicity Tests / methods*
  • Zebrafish


  • Nitro Compounds
  • Polycyclic Aromatic Hydrocarbons
  • Polystyrenes