Developing countries, where malaria is one of the most prevalent diseases, still rely on traditional medicine as a source for the treatment of this disease. For the present study, Trigonella foenum-graecum L. (fenugreek) were collected from Coimbatore, Tamilnadu, India. The test plant has been used in India by traditional healers for the treatment of fever as well as other diseases. The active principle was extracted out in different solvent systems to assess the anti-plasmodial potential, with an aim that they can further be utilized to formulate drugs. In vitro anti-plasmodial assay of the extracted fractions of fenugreek leaves was carried out using laboratory adapted chloroquine sensitive and resistant Plasmodium falciparum isolates. Schizont maturation inhibition assay was adopted to analyze the potential of the extracts. Ethanol extract (50%) seemed to possess profound anti-plasmodial activity with IC50 value of 8.75 ± 0.35 µg ml−1 and 10.25 ± 0.35 µg ml−1 against chloroquine sensitive and resistant P. falciparum isolates, respectively. Among the investigated six fractions of the plant extracts, two were found to have significant anti-plasmodial activity with IC50 values <10 µg ml−1, namely ethanol and butanol extracts. Two extracts chloroform and ethyl acetate showed moderate activity with IC50 values ranging from 10 to 20 µg ml−1, and the other two extracts, hexane and water appeared to be inactive with IC50 values >85 µg ml−1. In addition, preliminary phytochemical screening of the various extracts indicated the presence of alkaloids, saponin, tannin like phenolic compounds, flavonoids and steroids.