Chlorella sp. was cultivated with Conwy medium, BG-11 medium and MN medium at microalgae laboratory. The pure strain with different media and pH were observed within one month. There are various methods for extracting the oil from freshwater microalgae, such as physical factors, mechanical pressing and solvent extraction etc. Solvent extraction method was used for extracting of crude oil from algae biomass by using n-hexane. In this study, crude oil extract of Chlorella sp. was identified and interpreted by Fourier Trans-form Infra-Red spectroscopy (FT-IR) to employ as a fast and reliable analytical technique for the quantification of various fatty acids esters and refined oil spectra in the produced biodiesel. This research has been focused on growing pure algae with high oil content and then extracting the potential of biodiesel from it.