Quantification of OH-PUFA in stage-separated parasitized RBCs and hemozoin.

The OH-PUFA/PUFA ratio is expressed as percentage of OH-PUFA ± SD from 4 separate experiments related to total PUFAs in nonparasitized RBCs, rings, trophozoites, and hemozoin. The ring and trophozoite data were corrected to 100% parasitemia. Rings and trophozoites were harvested 12 and 36 hours after reinfection from synchronized cultures (parasitemia, 23%). Control RBCs kept in culture for the same time periods were treated identically. Lipids were extracted from the intact cells or hemozoin and analyzed for OH-PUFA and PUFA by HPLC. The levels of extracted PUFAs from 4 separate experiments were 713 ± 49 mg/L RBCs in nonparasitized RBCs, 706 ± 56 mg/L RBCs in rings, 848 ± 119 mg/L RBCs in trophozoites, and 2383 ± 405 mg/L RBC equivalents in hemozoin. For details, see “Materials and methods.”