Oleoresins are secondary metabolites of resinous plants, mainly composed of essential oil and resin acids. The present research work was conducted for the development of methodology for isolation of anticancer compounds from oleoresins of Boswellia and Pinus species. Conventional and advanced extraction techniques such as Hydro-distillation, steam distillation and supercritical fluid extraction (SCFE) method were used for extraction of essential oil from oleoresins. For optimum extraction yield of essential oil, hydro-distillation and steam distillation methods were performed under different temperature conditions. It was observed that essential oil yield in both plants oleoresin was increased on increasing extraction temperatures. These essential oils were further separated in to different fractions and sub fractions on the basis of their boiling points by vacuum fractional distillation method. Moreover, the biological activity of essential oils, fractions and sub fractions were performed under bioactivity guided assays. Antioxidant potential of essential oils, fractions and sub fractions was determined through different antioxidant assays. It was observed that most the essential oils, fractions and sub fractions revealed moderate level of antioxidant activity. Moreover, it was found that all the essential oils, fractions and sub fractions showed excellent antimicrobial and antitumor activity. Cytotoxicity was determined through haemolytic assay and their results expressed that all the tested essential oils, fractions and sub fractions exhibit weak cytotoxic activity. The chemical composition of essential oils was determined through GC-MS. The results showed that α-pinene, β-pinene, 3-carene and longifolene were the major chemical components of Pinus roxburghii oleoresin essential oils. Similarly, the chemical composition of Boswellia serrata oleoresin essential oils showed that α-pinene, β-pinene, pinocarveol and verbenol were the main chemical compounds and these chemical compounds might be responsible for their biological activity. Moreover, it was observed that the chemical composition of essential oils significantly vary with extraction methods.