The quality of the hoof is a significant factor influencing both its market value and the suitability of horses for sports. However, the knowledge is extremely scarce according to the relationship between parameters of quality and composition of the equine hoof. For example, fatty acid profile of bovine claw horn lipids was centred in relationship with lameness in the dairy cow. The current study was conducted to create an approach for analysis of fatty acid composition of equine hoof horn lipids and to relate the results to parameters of hoof horn quality and moisture. Hoof horn samples were collected from totally 24 sport horses. The individual hoof horn quality was evaluated by a professional blacksmith. Finally, 5 subjective categories of horn quality were created (1=excellent quality, 5=insufficient quality) as well as 3 categories for hoof horn moisture (A: moist; B: dry; C: balanced). Fatty acid profile of hoof horn lipids was determined as fatty acid methyl esters by gas chromatographic analysis. Fatty acids were presented as area percentage regarding to total area of 35 detected fatty acids. Due to the high individual variability of fatty acid pattern in Stratum corneum we chose 0.01 ≤ p<0.05 just as trend to significance. Results show a relation between myristic acid (14:0), palmitoleic acid (16:1) and margaric acid (17:0) and hoof horn quality. Furthermore caprylic acid (8:9), heneicosanoic acid (21:0) and undecylic acid (11:0) play a role in hoof horn moisture. Generally results show a very high individual variation of the fatty acid pattern in Stratum corneum.