Abstract— Hibiscus rostellatus belonging to family, malvaceae,is a herb ,used in traditional medicine for the treatment of inflammation and renal diseases. Methanolic extract of Hibiscus rostellatus was fractionated using different solvents of increasing polarity like petroleum ether, ethylacetate, methanol and water. The preliminary phytochemical screening showed the presence of carbohydrates, steroids, flavonoids, tannin. Methanolic extract of hibiscus rostellatus roots were screened for their antimicrobial, antioxidant activity. The test microorganisms included bacteria of Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa. It was concluded that Hibiscus rostellatus root extract was well effective against all these microorganisms. HPTLC-DPPH assays were used to identify the antioxidant activity of plant extract. This activity was estimated by using the mobile phase Chloroform:EthylAcetate:Acetic Acid (4.6:3.1:2.3 v/v/v). There was a successful chromatogram developed and detection of a band in the sample was observed. Later, the band showing antioxidant characteristic property was identified by DPPH Assay.