Rhynchostylis retusa (Orchidaceae) is an important ethnomedicinal herb in Indian systems of medicine. Tribal healers commonly employ the roots of this epiphytic orchid to treat various ailments. In the present study, the shade-dried root powder of R. retusa was subjected to cold extraction with four solvents, such as n-hexane, acetone, ethyl acetate, and methanol. The crude root extracts were then taken for qualitative phytochemical screening. Furthermore, GC-MS analysis of ethyl acetate and methanol root extracts was carried out. The methanol root extract of R. retusa showed a positive result for all the phytochemicals tested except for saponins. GC-MS analysis of R. retusa methanol root extract revealed the presence of 16 phytoconstituents. Major phytoconstituents such as 9-Hexadecenoic acid, methyl ester, [Z] Heptadecanoic acid, 16-methyl-, methyl ester, Ethanone,1-[4-(4-morpholyl benzylidene amino)phenyl]-, are recorded in the methanol root extract. In the present study, both methanol and ethyl acetate root extracts showed the presence of9-Hexadecenoic acid, methyl ester, [Z] as the major phytocompound. The phytocompounds identified in methanol and ethyl acetate root extracts exhibit various biological activities, including anti-inflammatory, antibacterial, antifungal, antioxidant, and anticancer properties. Based on the findings of the current study, it can be inferred that the roots of R. retusa contain diverse bioactive compounds with medicinal properties. Further investigation of these R. retusa root extracts holds the potential for the discovery and development of innovative pharmaceuticals.