Human skin provides a physical barrier against the external environment. The main constituents of stratum corneum, the outermost layer of skin, are lipids such as ceramides, cholesterol and free fatty acids. Tape stripping is a simple and efficient sampling method for collecting skin lipids. An adhesive tape is used to remove stratum corneum layers subsequently. Since there are several factors that affect the amount of lipids removed by each stripping, a standardized operation procedure is required.

 This study was focused to optimize the tape stripping method in terms of the number of tape strips and the application site. Lipid extracts from ten successive tape strips from the same anatomical site, and lipid extracts from five successive tape strips from four different sites (forehead, forearm, cheek, and neck) were analyzed by nanoflow ultrahigh performance liquid chromatography electrospray ionization tandem mass spectrometry (nUHPLC-ESI-MS/MS). Lipid profiles of skin samples were evaluated by comparing the normalized peak area of individual lipid species. The amount of quantified lipid class and lipid species had a tendency to decrease as the number of tape strips were increased, and it was largely decreased after the 6^th strip. However, lipid composition was largely different between each tape strips and application sites. As a result, it was determined to use pooled lipid extracts from the first five consecutive tape strips which can represent skin lipid composition without losing the number of quantifiable lipids.