| Summary: | Current artemisinin-based combination therapy (ACT) regimen for the treatment of malaria is effective, but cases of malaria parasite's resistance to existing antimalarial drugs is worrisome. This necessitates the development of new, safe, effective and affordable chemotherapy. We describe protocols for each step involved in the Anti-Plasmodial evaluation of Calotropis procera latex in mice infected with Plasmodium berghei. The protocols include: (1) determination of the chemical/ phytochemical constituents of Calotropis procera latex, (2) determination of the acute toxicity/ median lethal dose (LD50) and therapeutic dose of the plant latex, in vivo, and (3) in vivo determination of the Anti-Plasmodial potential of Calotropis procera latex in mice infected with Plasmodium berghei. We likewise describe our methodology for direct quantitation of percentage yield of the extract of Calotropis procera latex, and the statistical methodology for assessment of toxicity and efficacy in evaluating the Anti-Plasmodial activity of the plant. • Multi-step pipeline for the extraction of the bioactive constituents of the plant latex using 0.2M phosphate buffer (pH 7.0) and cold acetone. • Detailed protocols for the determination of acute toxicity/ median lethal dose (LD50) and calculation of therapeutic dose for intraperitoneal injection to achieve effective dose levels. • Determination of the phytochemical constituents using standard procedures, and in vivo efficacy against Plasmodium berghei using methodology to directly quantify the parasite level after treatment.
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