This study aimed to investigate the changes in skin barrier function in human subjects, following long-term topical application of Hippophae rhamnoides oil-in-water (O/W) emulsion whereas effects were measred using non-invasive probes like tewameter and corneometer. For this purpose, two stable oil-in-water (O/W) emulsions were formulated one with 5% Hippophae rhamnoides extract and other without extracts. Thirteen healthy, male subjects with a mean age 27 ± 4.8 years were enrolled after their informed consents. The subjects were instructed to apply either the active formulation or the base formulation over 84 days while they were not known with the contents of either formulation. Biometrological measurements of skin hydration and transepidermal water loss (TEWL) were performed on both sides of the face in each volunteer at baseline and on day 07, 14, 21, 28, 42, 56, 70 and 84. The statistical analysis revealed formulation with 5% plant extract was superior compared to placebo (base formulation) as formulation with extract have shown extremely significant improvements in skin hydration (p=0.0003) and TEWL (p=0.0087) throughout treatment course. Moreover, a significant (p<0.05) correlation between the active formulation and the improvement of the skin barrier functions was observed. The active formulation found to be superior to that of placebo. Results affirmed that future studies are necessary to clinically evaluate the active formulation hence it can be proposed that Hippophae rhamnoides emulsion could be an alternative pharmacological tool in treating barrier compromised conditions of skin.