The present invention discloses a method for effectively isolating and
purifying a physiological active dauer pheromone compound having the
following structural formula I related to aging and stress of C. elegans,
and determination of the structure of such material. First, ethanol
extract is prepared by completely concentrating a liquid medium with
ethanol after culturing C. elegans on a S. basal liquid medium, and then
this ethanol fraction is further extracted with ethyl acetate. Then,
impurities are removed from ethyl acetate extract by a silica-gel
adsorption chromatography and a pheromone extract is isolated and
purified by amine column chromatographic procedures using various
solvents as eluent. Finally, a pheromone compound having structural
formula I is completely isolated and characterized by size-exclusion
HPLC.