Isoquinoline alkaloids of the Amaryllidaceae have proven to be of significant value in drug discovery and development. Crossyne guttata is frequently used in traditional medicine as an antiseptic and wound cleaner as well as to treat alcoholism. The aim of the study was to identify the variation in alkaloid content of the species in relation to published works on the genus Boophone. We isolated Distichamine 1 from an ethanolic fraction of Crossyne guttata using column chromatography. Spectroscopic analysis yielded three alkaloids, distichamine 1, crinamine 2 and epibuphanisine 3. Distichamine 1 is a rare β-crinane alkaloid previously isolated only from the genus Boophone. The presence of distichamine 1 in the genus Crossyne suggests that it may be more widespread in tribe Amaryllideae than originally thought. © 2020 SAAB