The demand for products and technologies based on plants to control plant pathogens has increased in recent years due to concern about the use of hazardous pesticides. In the present investigation, washings of vermicompost (called biowash) prepared from foliage of Jatropha (Jatropha curcas), Annona (Annona squamosa) and Parthenium (Parthenium hysterophorus) were evaluated against fungal pathogens viz. Fusarium oxysporum f. sp. ciceri (FOC; causes wilt in chickpea), Sclerotium rolfsii (causes collar rot in chickpea) and Macrophomina phaseolina (causes charcoal rot in sorghum). Crude biowash of the botanicals were partitioned against ethyl acetate and the resultant organic and aqueous fractions were tested against the fungi. Similarly, crude biowash was also passed through C18 solid phase extraction cartridges and the resultant adsorbed and non-adsorbed fractions were tested against the fungi. Organic fractions of all the three biowash at 0.5% inhibited the growth of S. rolfsii between 78 and 87%, M. phaseolina between 62 and 65%, whereas only Parthenium was able to effectively inhibit FOC (91%), compared to control. Adsorbed fractions of all the three biowash at 0.5% inhibited the growth of S. rolfsii between 81 and 92%, M. phaseolina between 76 and 77% and FOC between 26 and 49%, compared to control. Both aqueous and non-adsorbed fractions of all the three biowash did not inhibit any of the fungi. Since Jatropha biowash showed consistently higher levels of inhibition (>80%) in both fractionation methods on S. rolfsii, this was selected for further purification of their secondary metabolites. When the organic fraction of Jatropha biowash was further fractionated by C18 open column chromatography with eluent 5, 10, 20, 40, 60, 80 and 100% MeOH fractions, only 80% methanol (MeOH) fraction was found to inhibit S. rolfsii. The active 80% MeOH fraction showed three clear bands when chromatographed on Silica Gel 60 F 254 thin layer chromatography (TLC) plates with Rf values 0.95, 0.90 and 0.70. Hence, it was concluded that one of these three bands could be the active ingredients that inhibited S. rolfsii and can be further exploited as a bio-fungicide.