Nanotechnology is a multidisciplinary approach to scientific research that successfully collaborates with other science disciplines like agriculture, pharmaceuticals, biotechnology, and engineering. The synthesis of nanoparticle methods has also progressed from the classic chemical approach to the green synthesis method. The improved procedure of plant-mediated nanoparticle synthesis has been verified to be less hazardous to the environment, and its inexpensive cost of production adds to its attributes. With the world executing every function at the nanoscale, studying the role, mode of action and impacts of phytotherapeutic preparations combined with nanotechnology is critical. The Cissus quadrangularis stem is also employed in Ayurveda as an alternative, anthelmintic, dyspeptic, digestive, tonic, and analgesic in eye and ear illnesses and irregular menstruation, asthma, and back and spine problems. The current research attempted the green production and characterization of zinc oxide nanoparticles from the plant Cissus quadrangularis L stem. The GC-MS analysis identified the bioactive compounds in the stem of the plant extract. Long-chain fatty acids, phenols, sterols like stigmasterol, alkylating agents, triterpenoids like lupeol, amines, esters and unsaturated hydrocarbons were found in abundance in the zinc oxide nanoparticles extract of C. quadrangularis. The FTIR analysis revealed the presence of several functional groups like alcohol, esters, amines, halo compounds and carboxylic acid. The SEM analysis portrayed the size and shape of the synthesis zinc oxide nanoparticles in the range of 88 nm to 182 nm and was found to be spherical.