Human skin consists of the stratum corneum, which is composed of corneocytes and intercellular lipids, including ceramides, cholesterol, and free fatty acids. These intercellular lipids form a multi-lamellar structure that serves as the skin barrier, protecting the body from external hazards. However, this barrier also poses challenges for the delivery of active ingredients and drugs. Consequently, research on drug delivery systems (DDS) is being actively conducted to address this issue. In this study, we experimentally verified the effectiveness of multi-lamellar emulsion (MLE), which is structurally similar to intercellular lipids, as a carrier for active ingredients. First, MLE and non-MLE formulations were manufactured in the laboratory, and structural differences were confirmed through polarization microscopy and small-angle x-ray scattering (SAXS) analysis. Subsequently, we conducted a skin absorption test (in vitro method, OECD TG 428) using human or animal skin with an active ingredient in MLE. The results of this study confirm the potential of MLE as an excellent carrier for delivering active ingredients through the skin.