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Studies on percutaneous penetration of chemicals - Impact of storage conditions for excised human skin.

Toxicology in vitro : an international journal published in association with BIBRA (2012-12-12)
Kathrin Dennerlein, Désirée Schneider, Thomas Göen, Karl Heinz Schaller, Hans Drexler, Gintautas Korinth
ABSTRACT

According to international guidelines skin penetration experiments can be carried out using freshly excised or frozen stored skin. However, this recommendation refers to data obtained in experiments with human cadaver skin. In our study, the percutaneous penetration of the occupationally relevant chemicals anisole, cyclohexanone and 1,4-dioxane was investigated for freshly excised as well as for 4 and 30 days at -20°C stored human skin using the diffusion cell technique. As indicator for the impairment of skin barrier by freezing cholesterol dissolution was determined in the solvents in exposure chambers of diffusion cells. Considering the percutaneously penetrated amounts, the following ranking was determined: 1,4-dioxane>anisole>cyclohexanone (decline to a factor of 5.9). The differences of fluxes between freshly excised and frozen stored skin (4 and 30 days) were not significant (p>0.05). Cholesterol dissolved from the skin indicates no significant differences between freshly excised and frozen stored skin. This study shows that freezing of human skin for up to 30 days does not alter the skin barrier function and the permeability of chemicals.

MATERIALS
Product Number
Brand
Product Description

Supelco
Cyclohexanone, Selectophore, ≥99.5%
Sigma-Aldrich
Cyclohexanone, puriss. p.a., ≥99.5% (GC)
Supelco
Cyclohexanone, analytical standard
Supelco
Anisole, analytical standard
Sigma-Aldrich
Anisole, anhydrous, 99.7%
Sigma-Aldrich
Cyclohexanone, ReagentPlus®, 99.8%
Sigma-Aldrich
Cyclohexanone, ACS reagent, ≥99.0%
Sigma-Aldrich
Cyclohexanone, 99.8%
Sigma-Aldrich
Anisole, ≥99%, FCC, FG
Supelco
Anisole, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Anisole, ReagentPlus®, 99%
Supelco
1,4-Dioxane, analytical standard
Sigma-Aldrich
1,4-Dioxane, anhydrous, 99.8%
Sigma-Aldrich
1,4-Dioxane solution, NMR reference standard, 40% in benzene-d6 (99.6 atom % D)
Sigma-Aldrich
1,4-Dioxane, suitable for HPLC, ≥99.5%
Sigma-Aldrich
1,4-Dioxane solution, NMR reference standard, 10 mM in D2O ("100%", 99.96 atom % D), NMR tube size 5 mm × 7 in.
Supelco
1,4-Dioxane solution, NMR reference standard, 40% in benzene-d6 (99.6 atom % D), NMR tube size 5 mm × 8 in.
Sigma-Aldrich
1,4-Dioxane, ACS reagent, ≥99.0%
Sigma-Aldrich
1,4-Dioxane solution, NMR reference standard, 40% in benzene-d6 (99.6 atom % D), chromium(III) acetylacetonate 5 mg/mL, NMR tube size 3 mm × 8 in.
Supelco
1,4-Dioxane solution, NMR reference standard, 40% in benzene-d6 (99.6 atom % D), NMR tube size 3 mm × 8 in.
Sigma-Aldrich
1,4-Dioxane solution, NMR reference standard, 40% in benzene-d6 (99.6 atom % D), chromium(III) acetylacetonate 5 mg/mL, NMR tube size 5 mm × 8 in.
Sigma-Aldrich
1,4-Dioxane solution, NMR reference standard, 40% in benzene-d6 (99.6 atom % D), NMR tube size 10 mm × 8 in.
Supelco
1,4-Dioxane solution, certified reference material, 2000 μg/mL in methanol, ampule of 1 mL
Sigma-Aldrich
1,4-Dioxane, ACS reagent, ≥99.0%