The prospect of depletion of natural energy resources on the Earth forces researchers to seek and explore new and alternative energy sources. Biomass is a composite resource that can be used in many ways leading to diversity of products. Therefore, microalgal biomass offers great potential. The main aim of this study is to find the best physical method of microalgal biomass pretreatment that guarantees efficient lipid extraction. These studies identifies biochemical composition of microalgal biomass as source for biodisel production. The influence of drying at different temperatures and lyophilization was investigated. In addition, wet and untreated biomass was examined. Cell disruption (sonication and microwave) techniques were used to improve lipid extraction from wet biomass. Additionally, two different extraction methods were carried out to select the best method of crude oil extraction. The results of this study show that wet biomass after sonication is the most suitable for extraction. The fatty acid composition of microalgal biomass includes linoleic acid (C18:2), palmitic acid (C16:0), oleic acid (C18:1), linolenic acid (C18:3), and stearic acid (C18:0), which play a key role in biodiesel production.