American Society of Hematology

Fig. 3.

$Fig. 3. Separation and identification of OH-PUFA isomers in stage-separated parasitized RBCs and hemozoin. / Rings, trophozoites, and hemozoin were separated from synchronized cultures, and the lipids were extracted. The solvents were removed, the remaining lipids were hydrolyzed under alkaline conditions, and OH-PUFA was analyzed by RP-HPLC (see “Materials and methods” for details). Aliquots of the OH-PUFA fraction were dried under vacuum, the remaining lipids were reconstituted in N-hexane containing 0.1% acetic acid, and aliquots were analyzed by SP-HPLC as described in “Materials and methods.” The chemical structure of the major OH-PUFAs (12-HETE, 13-HODE, 9-HODE) was confirmed from cochromatography with authentic standards in reverse phase–, normal phase–, and chiral phase–HPLC, from UV-spectroscopy and gas chromatography/mass spectroscopy.$

Separation and identification of OH-PUFA isomers in stage-separated parasitized RBCs and hemozoin.

Rings, trophozoites, and hemozoin were separated from synchronized cultures, and the lipids were extracted. The solvents were removed, the remaining lipids were hydrolyzed under alkaline conditions, and OH-PUFA was analyzed by RP-HPLC (see “Materials and methods” for details). Aliquots of the OH-PUFA fraction were dried under vacuum, the remaining lipids were reconstituted in N-hexane containing 0.1% acetic acid, and aliquots were analyzed by SP-HPLC as described in “Materials and methods.” The chemical structure of the major OH-PUFAs (12-HETE, 13-HODE, 9-HODE) was confirmed from cochromatography with authentic standards in reverse phase–, normal phase–, and chiral phase–HPLC, from UV-spectroscopy and gas chromatography/mass spectroscopy.

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