Many studies investigate the permeation of actives through the skin and ignore the role of excipients. The solvents used in formulations will undoubtedly penetrate the skin where they can have a number of effects. They can extract skin lipids, they can alter the fluidity of the lipids and they can alter the polarity of the skin. The degree to which they do this and the depth into the skin where this occurs will depend on the uptake kinetics. The problem is to distinguish the different effects. Using ATR-FTIR and deuterated materials this can be achieved in vivo. The aim of the present study was to study the higher alkanols (hexanol, octanol, decanol) in vivo using a combination of ATR-FTIR spectroscopy and tape stripping. Studies conducted in vivo using deuterated vehicles confirmed the lipid extraction effects of d-hexanol and d-octanol, whereas d-decanol did not change skin lipid content. The uptake of d-decanol was higher than for the other vehicles consistent with previous observations on mouse skin for alkanols of increasing chain length. In general, solvent uptake was proportional to the induced shift in the C-H stretching frequency. Lipid disorder was induced by all vehicles studied in vivo and was proportional to the amount of vehicle present in the skin.