Synthesis of 26-methyl cholestane and identification of cryostanes in mid-Neoproterozoic sediments

The biomarker distributions of mid-Neoproterozoic sediments (800–717 million yr, Ma) from various locations are characterized by the occurrence, besides cholestane isomers, of a novel series of C28 steranes, whereas classical C-24 alkylated steranes are absent. These unusual C28 steranes, termed cryostanes, seem to be restricted to pre-Snowball Earth sediments. We report their conclusive identification as 26-methyl cholestanes based on comparison of gas chromatographic (GC) behavior and mass spectrum of the last eluting cryostane isomer with those of a 26-methyl 5alpha(H),14alpha(H),17alpha(H),20R-cholestane standard obtained by way of synthesis. Sterols methylated at C-26 seem to be restricted to some demosponges and, based on molecular clock estimates, demosponges may have emerged in the time interval 800–700 Ma. A sponge origin of cryostanes is thereby conceivable, making cryostanes the oldest molecular markers for animals. However, other biological sources need to be explored, including the wide variety of eukaryotic protists