Enantiomer composition of 12-HETE isolated from trophozoites.

The 12-HETE was prepared from trophozoites by lipid extraction, alkaline hydrolysis, and sequential RP-HPLC and SP-HPLC. The SP-HPLC eluate fractions containing 12-HETE were collected, the solvent was removed under vacuum, the remaining lipids were reconstituted in N-hexane containing 0.1% acetic acid, and aliquots were analyzed by chiral phase HPLC as described in “Materials and methods.”