Hypnum cupressiforme Hedw. is the main species for Moss surveys (ICP Vegetation programme) in Southeastern Europe and is widely distributed in the region. In addition to their biomonitoring role, mosses are applied in some countries as a traditional medicine for the treatment of eczema, cuts, burns, eye diseases, etc. Therefore, the chemical and lipid composition of the moss H. cupressiforme is of interest to establish their possible application in different fields. The chemical composition of the moss was examined regarding total lipids, proteins, carbohydrates (i.e., fibres), ash, and moisture content. The main lipid-soluble bioactive components were determined as sterols, tocopherols, phospholipids and fatty acids. The major fatty acids were linoleic (14.9%), oleic (13.8%), palmitic (12.5%) and α-linolenic (11.3%) acids. Unsaturated fatty acids (56.4%) prevailed in the glyceride oil, in which the polyunsaturated ones constituted 32.5%. The lipid indices (atherogenicity, thrombogenicity, hypocholesterolemic/hypercholesterolemic ratio, peroxidability, and oxidation stability index) were also theoretically calculated based on the fatty acid composition of the moss lipids to establish their health benefits and the rate of oxidation. The primary results of this study revealed H. cupressiforme to be a promising alternative source of bioactive compounds that could be implemented in supplements with health-promoting effects.