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With the advancement of nanotechnology, the nano-sized particles make an imprint on our daily lives.The present investigation revealed that biomolecules present in seed exudates of Vigna radiata are responsible for the synthesis of AuNPs, confirmed by the routine characterization techniques. Anticancer efficacy showed by AuNPs might be due to the release of phytochemicals in the exudate which is being adsorbed on the surface of AuNPs referencing their anticancer efficacy against the tested breast cancer cell lines. Inhibition of clonogenicity and cell cycle arrest at G2/M phase then apoptosis of AuNPs was also observed, but found nontoxic to the human PBMC cells which further confirms its biocompatible property Among the various physicochemical study, present AuNPs shows unique information, they show photoluminescent property which may be used for bioimaging purposes. However, further molecular analysis needs to be explored to understand the underlying mechanism for therapeutic and biomedical application.