title: Antimalarial and safety evaluation of extracts from Toddalia asiatica (L) Lam. (Rutaceae).
abstract: BACKGROUND
  Toddalia asiatica (L) Lam. (Rutaceae) is a medicinal plant traditionally used in Kenya by many communities for the treatment of malaria and other ailments. All parts of the plant are claimed to have medicinal value, but the root bark in particular is believed to be more potent. Decoctions or infusions of the roots are taken orally to treat malaria, fever and stomach ache.
  
  
  OBJECTIVE
  To evaluate antimalarial activity of aqueous and organic extracts prepared from Toddalia asiatica and determine in vitro and in vivo safety of the extracts.
  
  
  METHODS
  Aqueous, ethyl acetate, hexane and methanol extracts were obtained from Toddalia asiatica root bark, fruits and leaves. In vitro antiplasmodial activity was done using chloroquine-sensitive (D6) and chloroquine-resistant (W2) Plasmodium falciparum strains and the concentration causing 50% inhibition of radioisotope incorporation (IC(50)) was determined. In vivo assay was done by administering mice infected with Plasmodium berghei four consecutive daily doses of the extracts through oral route following Peters 4-Day suppressive test. The percentage suppression of parasitaemia was calculated for each dose level by comparing the parasitaemia in untreated control with those of treated mice. Quinine hydrochloride was used as positive control while double distilled water or 20% Tween-80 was used as a negative control. In vivo acute toxicity was determined in mice using standard procedures. In vitro cytotoxicity assay was carried out using actively dividing sub-confluent Vero cells.
  
  
  RESULTS
  Inhibitory concentrations of ethyl acetate extract of Toddalia asiatica fruits showed high activity against chloroquine resistant (W2) strains of Plasmodium falciparum (IC(50)=1.87 μg/ml), followed by root bark aqueous extract (IC(50)=2.43 μg/ml). Tested in vivo against Plasmodium berghei, the fruit ethyl acetate extract (500 mg/kg) and root bark aqueous extract (250 mg/kg) reduced malaria parasitaemia by 81.34% and 56.8% respectively. Higher doses were found to be less effective in vivo. Acute toxicity and cytotoxictiy of the tested extracts, with the exception of hexane extract from the roots, showed LD(50)>1000 mg/kg and CC(50)>100 μg/ml respectively.
  
  
  CONCLUSIONS
  The results obtained contribute to the validation of traditional use of Toddalia asiatica and provides in vivo and safety data of the plant extracts tested for the first time. Ethyl acetate extract of the fruits was active against chloroquine resistant Plasmodium falciparum as well as against Plasmodium berghei. These findings confirm the suitability of Toddalia asiatica as a good candidate for further tests to obtain a prototype for antimalarial medicine.
authors.affiliation: Kenya Medical Research Institute, Center for Traditional Medicine and Drug Research, P.O. Box 54840-00200, Nairobi, Kenya. jorwa@kemri.org
authors.forename: J A
authors.initials: JA
authors.lastname: Orwa
authors.forename: L
authors.initials: L
authors.lastname: Ngeny
authors.forename: N M
authors.initials: NM
authors.lastname: Mwikwabe
authors.forename: J
authors.initials: J
authors.lastname: Ondicho
authors.forename: I J O
authors.initials: IJ
authors.lastname: Jondiko
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D005638:Fruit; D008288:Malaria / Q000188:drug therapy* / Q000469:parasitology; D051379:Mice; D008517:Phytotherapy; D024301:Plant Bark; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D018515:Plant Leaves; D010963:Plasmodium falciparum / Q000187:drug effects; D027821:Rutaceae
chemical_list: D000962:Antimalarials; D010936:Plant Extracts