Chromatography and isolation of the Kav 1.0 pheromone of female Nippostrongylus brasiliensis (Nematoda).

The Kav 1.0 pheromone from incubation of mouse-adapted females of Nippostrongylus brasiliensis was soluble in organic solvents, as shown by bioassay of males and presence of a peak at Kav 1.0 during gel filtration. Recovery of the biological activity from the females was enhanced when the Kav 1.0 component was purified by organic extraction-gel filtration, C-18 cartridge separation with 50% methanol, C-18 cartridge-gel filtration, and C-18 cartridge-TLC. The activity recovered from organic extraction, TLC, or gel filtration of solutions in which females had been incubated was less than the above combined techniques. Reverse-phase cartridges retained activity of 20,000 female-hr of incubation without loss and enabled storage of the pheromone component for 3 days at 4 C. The HPLC separation of incubate from females revealed an active peak(s) with a 4- to 5-min retention time. This peak(s) was present also in regions containing activity from other purification procedures.