Ethanol and diethyl phthalate: vehicle effects in the local lymph node assay

The vehicle in which an allergen is presented to the skin has been recognized to have an effect on the skin-sensitizing potency of the allergen. Typical vehicles used to evaluate the skin sensitization potential of fragrance materials include ethanol, diethyl phthalate, or a combination of the two. The authors conducted a series of studies to evaluate each of these vehicles for their utility in the murine local lymph node assay and to investigate the potential differences in skin sensitization resulting from their use. Four fragrance materials were tested in four different vehicles. The test materials were p-t-butyl-alpha-methylhydrocinnamic aldehyde, geraniol, eugenol, and hydroxycitronellal. The vehicles were diethyl phthalate, 1:3 ethanol:diethyl phthalate, 3:1 ethanol:diethyl phthalate, and ethanol. Each of the fragrance materials was tested at five dose levels ranging from 0.3% to 50% w/v. In all four vehicles, each material tested elicited positive responses, exhibiting weak to moderate skin sensitization potential. Overall, p-t-butyl-alpha-methylhydrocinnamic aldehyde exhibited the most potency, followed by eugenol, geraniol, and hydroxycitronellal. The sensitization potential of both p-t-butyl-alpha-methylhydrocinnamic aldehyde and geraniol was greatest when the vehicle was ethanol. The sensitization potential of eugenol was greatest in 3:1 ethanol:diethyl phthalate, but the sensitization potential for hydroxycitronellal was greatest in 1:3 ethanol:diethyl phthalate. The strength of the sensitization response was observed to vary with the vehicle; however, the results did not show any clear pattern of one vehicle over another regarding skin sensitization.