The sulfur-rich anal sac secretion of the mink,Mustela vison, consisted of immiscible lipid (1.7% sulfur) and aqueous (0.7% sulfur) phases. Light and electron microscopy revealed secretory tissue of two types, sebaceous (holocrine) and apocrine. A major input of sulfur into the sac appeared to be associated with glycoprotein granules present in the apical portions of the apocrine cells as X-ray energy probe microanalysis showed these to contain relatively high levels of sulfur. The lipid of the secretion, presumed to be largely of sebaceous origin, consisted mainly of wax monoesters, while the aqueous phase contained volatile fatty acids, ammonia, and amines, including putrescine (1,4-diaminobutane). The identity of the major headspace volatiles was confirmed by NMR, MS, and Raney nickel desulfuration as being 2,2-dimethylthiacyclobutane and 3,3-dimethyl-1, 2-dithiacyclopentane. These compounds were not detected by GC-MS in the headspace volatiles of the anal sac secretions of eight other mustelid species examined. Other sulfur compounds detected included isomeric dimethylthiacyclobutanes, a number of disulfides and 3-methyl-but-3-enyl methyl sulfide (isopentenyl methyl sulfide). The significance of these findings is discussed.