The method described here allows noninvasive quantification of reducing LMWA or the lipid hydroperoxide present on the surface of the skin. Quantification of reducing antioxidants can be achieved because they are secreted from the skin surface into a well containing an extraction solution. Analysis of the reducing equivalents released indicates the presence of uric acid and ascorbic acid. Other LMWA released from the skin are as yet unidentified. The secretion of the LMWA reaches a plateau following 20-30 min of incubation. Therefore, a 30-min incubation period was chosen as the optimal time for the extraction solution to be present in the well and in contact with the skin. This extraction procedure can be repeated after 24 hr. This period of time is needed for regeneration of the LMWA to their initial levels. Direct measurement allows continuous determination of the release of LMWA and their interaction with the iron chelate. The reaction is completed after 25-35 min, at which time the final potential can be recorded. When organic peroxides on the surface of the skin are determined, it is important that the glassy carbon electrode be in close contact with the skin, since the reaction occurs on the surface of the electrode and the bound peroxide on the outer layer of the skin. Furthermore, close contact is needed to avoid interference of reducing equivalents secreted from the skin into the well.
Bibliographical noteFunding Information:
This work was supported in part by the Foundation for Research and Development,b y the Bergman Foundation, and by the David R. Bloom Center of Pharmacy.