The current study focused on the green synthesis of zinc oxide nanoparticle (ZnO) from the Dodonaea viscosa roots and evaluation of its anti-inflammatory properties. The presence of phytochemicals in the root extract of D. viscosa root extract was analyzed with different solvents including methanol, chloroform, aqueous and petroleum ether. The methanolic extract had higher activity among the four solvents. UV-Visible and FT-IR spectrophotometry was used to validate the synthesized ZnO. The FT-IR bands revealed the existence of several functional groups, and the UV-Vis spectrum showed the absorption peak of 371 nm for ZnO nanoparticles. The green synthesized ZnO nanoparticles showed an anti-inflammatory activity that was confirmed by albumin denaturation and membrane stabilization. Maximum inhibition of 70% was observed from ZnO nanoparticle when compared with aspirin. IC50 value of plant extract was 36.34 μg/ml and for aspirin was found to be 36.26 μg/ml. The in vitro RBCs membrane stabilization study emphasized the anti-inflammatory activity of ZnO nanoparticles. The IC50 valve of aspirin was determined to be 21.19, whereas the plant extract was 23.49. At varying doses, the methanolic extract of D. viscosa showed efficacy in preventing heat-induced hemolysis. The three-dimensional data of the protein structure is used to predict the structural levels of proteins. Proteins of D. viscosa were analyzed using Expasy (structural prediction), Protparam server (functional characterization), and (3-D structure) SWISS MODEL, and homology modeling was used to compare them with known plant biomaterials. Bioinformation on useful proteins was also analyzed and reported. Thus, the synthesized nanoparticles possess strong anti-inflammatory activity that could be utilized in numerous pharmacological formulations. © 2024 Elsevier B.V., All rights reserved.