The synthesis of metal nanoparticles of different sizes, shapes, chemical composition and controlled monodispersity is an important area of research in nanotechnology because of their interesting physical properties and technological applications. Present work describes an eco-friendly method for the synthesis of spherical gold nanoparticles using aqueous extract of Macrotyloma uniflorum. The effects of quantity of extract, temperature and pH on the formation of nanoparticles are studied. The nanoparticles are characterized by UV-visible spectroscopy, transmission electron microscopy (TEM), X-ray diffraction (XRD) and FTIR analysis. The high crystallinity of nanoparticles with fcc phase is evident from HRTEM images, SAED and XRD patterns. Synthesized nanoparticles have size in the range 14-17nm. FTIR spectrum indicates the presence of different functional groups present in the bio-molecule capping the nanoparticles. The possible mechanism leading to the formation of gold nanoparticles is suggested.