The evaluation of the skin health is an essential step in the skincare industry in order to assess the safety and the efficacy of active ingredients. 12R-LOX (gene ALOX12B) is a lipoxygenase expressed in keratinocytes and is implicated in the oxygenation of the esterified omega-hydroxyacyl-sphingosine (EOS) ceramides, a required process to their covalent linkage to proteins of the cornified envelope. It is an important step in establishing the water barrier by preventing unnecessary evaporation through epithelial cells. HSP70 family members are among the most abundant HSPs in the skin expressed constitutively within keratinocytes. HSP70 expression is elevated in both epidermis and dermis for cytoprotection after skin samples are heat shocked or after stressors like UVB. Traditionally transepidermal water loss (TEWL) has been widely used as a way to evaluate skin barrier function and tape stripping as a way to evaluate biological markers (biomarkers). Our aim was to understand how TEWL, 12R-LOX and HSP70 could be correlated clinically. Subjects were treated with a hydrating cream for 3 weeks on their lower inner forearm. After treatment, TEWL was measured and tape strips were collected from the treatment area and untreated site. 12R-LOX enzyme activity was measured with a fluorescence-based assay and HSP70 by ELISA using the protein samples isolated from the tape stripes. The 12R-LOX enzyme activity had a negative correlation with TEWL as previously described and had a positive correlation with HSP70. Moreover, HSP70 also had a negative correlation with TEWL confirming the correlation between the two biomarkers 12R-LOX and HSP70 with barrier function. The study of these two biomarkers is a new interesting and non-invasive approach for studying skin under different conditions.