Cerevisterol was first discovered in 1928 as a component of crude yeast (Saccharomyces cerevisiae) sterols remaining from the manufacture of the related ergosterol. Chemists Edna M. Honeywell and Charles E. Bills purified the compound and reported some of its properties in a 1932 publication. They noted its high melting point (265.3 °C) relative to other sterols, and insolubility in the organic solvent hexane. These characteristics facilitated its purification, and they were able to obtain  of cerevisterol from  of dry yeast. The following year, they determined its chemical formula to be CHO, with two double bonds, and with two of the oxygen molecules occurring in hydroxyl groups.
Its structure was determined in 1954 by comparison with a sample that was chemically synthesized from ergosterol. Purified cerevisterol has the form of a white amorphous solid. When crystallized in ethyl alcohol, it forms elongated prisms, while crystallization in acetone or ethyl acetate produces broad hexagonal prisms. Its UV absorption spectrum shows a maximum at about 248 nm. Cerevisterol is a stable molecule, showing no discoloration or change in melting point even after several weeks of exposure to light and air.