Extraction of Hydrophilic Metabolites from Plasmodium falciparum-Infected Erythrocytes for Metabolomic Analysis
Metabolomics is an increasingly common analytical approach for investigating metabolic networks of pathogenic organisms. This may be of particular use in the study of parasitic infections due to the intrinsic metabolic connection between the parasite and its host. In vitro cultures of the malaria parasite Plasmodium falciparum present a valuable platform to elucidate the structure and dynamics of the parasite’s metabolic network and to determine the mechanisms of action of antimalarial drugs and drug resistance mutations. Accurately measuring metabolite levels requires a reproducible method for quantifying intracellular metabolites. Here we present a simple protocol for extracting hydrophilic metabolites from P. falciparum-infected erythrocyte cultures.
Key wordsMalaria Plasmodium falciparum Erythrocyte Red blood cell Metabolomics Mass spectrometry Liquid chromatography LC-MS Nuclear magnetic resonance NMR
This work was funded by the Burroughs Welcome Fund, an NIH Director’s New Innovators award (1DP2OD001315-01) and with support from the Center for Quantitative Biology (P50 GM071508). KO is supported by an NSF Graduate Research Fellowship. We would also like to thank Ian Lewis for stimulating discussion and for critical reading of the manuscript.
- 6.World Health Organization (2008) World Malaria Report 2008Google Scholar
- 8.Dondorp AM et al (2011) Artemisinin resistance: current status and scenarios for containment. Nat Rev Microbiol 8:272–280Google Scholar
- 18.Fernandez V (2008) Enrichment of late-stage infected erythrocytes in 60% Percoll. In Methods in Malaria Research. American Type Culture Collection, Manassas, Virginia, p 25Google Scholar