α-Tocopherol, the major biologically active form of vitamin E, represents a frequently added lipophilic compound of skin care products. Despite its emerging use in rinse-off formulations, little is known on its efficacy with respect to its deposition or its antioxidant potential in human skin. The objective of this study was to investigate whether the single use of an α-tocopherol-enriched rinse-off product provides effective deposition of α-tocopherol on human stratum corneum. To test this, forearm skin of 13 volunteers was washed either with an α-tocopherol-enriched rinse-off product (test product, TP) or with an α-tocopherol-free vehicle control (control product, CP) (contralateral arm) using a standardized wash protocol. Thereafter, skin surface lipids were extracted with pure ethanol after the wash procedure as well as after 24 h. Additionally, one group of volunteers was subjected to irradiation of their forearms with low-dose UVA (8 J/cm2) prior to lipid extraction. Skin lipid extracts were analyzed by high performance liquid chromatography using electrochemical detection for vitamin E and UV detection for squalene (SQ) and squalene monohydroperoxide. The results of this in vivo study demonstrated that (1) while CP treatment lowers, TP treatment strongly increases α-tocopherol levels of skin barrier lipids; (2) increased vitamin E deposition levels were maintained for a period of at least 24 h, and (3) TP treatment significantly inhibited photooxidation of SQ. In conclusion, the use of α-tocopherol-enriched rinse-off products may help to maintain the integrity of the skin barrier by providing protection against photooxidative stress at the level of skin surface lipids.