A variety of plants are available by nature such as ornamental and medicinal. Medicinal plants contain many biologically active ingredients such as alkaloids, saponins, poly phenol, anthracyanin, terpenoids and flavonoids. These potent bio active components have therapeutic properties and have the ability to treat different health issues including stomach ulcers, inflammation, gastrointestinal issues, skin diseases, respiratory problems and various pathogenic diseases etc. Ratanjot (Alkanna tinctoria) is a herbaceous flowering plant. It contains many important phytochemicals and biological active compounds. It shows many pharmacological activities such as antioxidant, anti-proliferation, antimicrobial and anti-inflammatory. Ratanjot majorly contains two important naphthoquinone derivatives namely alkannins and shikonins. They are powerful components and show anti-inflammatory, antimicrobial and wound healing properties. The main objective of this study is to identify the important biological active natural compounds from the roots of Alkanna tinctoria by different extraction methods. Plant extracts will be prepared by the conventional and modern extraction methods. Phytochemical profiling will be carried out by UV-vis spectroscopy and Fourier transform infrared spectroscopy (FTIR). After characterization the extracts will be used to study the anti-cancer activity of hepatic carcinoma cell lines (Huh7 and HepG2) and will evaluate the anti-microbial activity. This study will be helpful for the treatment of liver cancer and infectious diseases. Antioxidant activity will also be performed for different roots extracts.