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EC number: 214-881-6
CAS number: 1205-17-0
incubation of epidermal membranes with the radiolabeled test item, only
67% of the applied dose was accounted for by 48 h. At 24 and 48 h, 42%
and 50%, respectively, of the applied close was recovered in the fluid
retrieved from the receptor chambers. Distribution of remaining
radiolabel in surface wipes, tape strips, remaining epidermis and the
donor chamber surface accounted for an additional 17%. The chemical
nature of the absorbed radiolabel was not characterized (i.e. parent
test item or metabolites). Evaporative loss, estimated from direct
application to PTFE sheets, was approximately 8% and 19% of the applied
dose at 24 and 48 h, respectively.
An in vitro
dermal absorption studies according to the FDA/AAPS guidelines (Skelly
et al., 1987) was performed with full thickness human skin samples. Twenty
µL of a 1% solution in ethanol of radiolabeled test item (0.2 mCi
benzyl-14C. specific activity of 57.26 mCi/mmol;
radiochemical purity of 99.14%) was applied to the surface of the
membrane. Receptor chambers were continuously agitated by submersible
magnetic stirrers and maintained at 37 °C throughout the experiment. The
skin surface temperature was maintained at 32 °C. Samples of the
receptor fluid (50% ethanol/water) were harvested from the receptor
chamber at 2, 8, 24, 36 and 48 h and analysed by liquid scintillation
chromatography. Post-incubation epidermal membranes were wiped with a
cotton bud, and then tape stripped 10 times using adhesive tape. Total
radioactivity was accounted for by extruding and counting test item
equivalents associated with membrane wipes, tape strips, digested
remaining skin, and donor chambers. To assess evaporative loss, 20 µL of
a 1% test item solution was applied to PTFC sheets mounted in diffusion
cells (at 32 °C surface temperature). These sheets were then placed in
chambers for 24 or 48 h. The PTFE
sheet was then removed and washed twice with methanol (10 mL then 5 mL).
The donor chamber was washed with 10 ml methanol. A sample of each wash
solution was submitted to analysis by LSC which allowed the total
remaining radiolabel at each timepoint to be determined. Radiolabel that
was unaccounted for was considered to be associated with evaporative
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