Skin moisture relates to the state of multiple skin constituents and aspects, but unfortunately, a device which could provide comprehensive and in vivo analysis is not available. Nevertheless, several reports have demonstrated accurate estimations of dermal water content using near-infrared spectroscopy (NIRS), and the potential of employing this technique in skin analysis. We aim to investigate whether NIRS could detect changes in skin barrier function through evaluation of skin water uptake in relation to moisturizer application. NIR and capacitance data were collected from 20 volunteers at both forearms, prior to and after seven days of regular moisturizer use. Results indicated lower peak intensities at the 1940-nm minima and higher intensities at the 1450-nm equivalent minima with moisturizer abstinence, while the opposite was true with regular moisturizer application. As the light beam would have traveled deeper into the skin at 1450 nm, it has been concluded that long-term, frequent moisturizer use had limited the penetration of extrinsic water. Partial least squares analysis showed that separation of sample’s scores increased with abstinence of moisturizer use. Thus, NIRS can provide valuable information not only on dermal water contents but also on additional parameters such as skin barrier function.