The in vitro antidiabetic efficacy of ethanolic extract Boerhavia diffusa (B.diffusa) synthesized silver nanoparticles (AgNPs) was investigated by inhibition of α-amylase, α-glucosidase, protein glycation assay, non-enzymatic glycosylation of hemoglobin, glucose uptake by yeast cells and glucose diffusion at varying concentrations (10 to 100µg/ml). The alpha-amylase assay shows that the acarbose (standard) and B. diffusa had IC50 values of 46.2 µg/ml and 55.4 µg/ml, whereas alpha-glucosidase inhibitory activity was found to be 63.4 µg/ml and 93.0 µg/ml respectively. Further, non-enzymatic glycosylation analysis showed IC50 value of metformin (standard) as 28.6 µg/ml and B. diffusa as 63.9 µg/ml. The protein glycation activity was inhibited in non-enzymatic glycosylation of hemoglobin. The glycosylation was induced using pioglitazone (standard) which gives IC50 value of 616.4 µg/ml by which B. diffusa showed 756.3 µg/ml. The uptake of glucose by yeast cells was analyzed and the result shows that the glucose concentration increased steadily from 5mM to 25mM (maximum absorption) of both metronidazole (standard) and B. diffusa. From 30 to 180 minutes, the glucose diffusion experiment revealed that the concentration of the metformin and B. diffusa extract was positively correlated with the time. The ethanolic extract of synthesized AgNPs and the reference medication employed in all experiments both benefit their curative potential for the treatment of insulin resistance. The generated silver nanoparticles can be used for industrial and therapeutic purposes and can be released into the environment without harm. More in vivo study can be reviewed, however the green synthesized ethanolic extract of B. diffusa exhibits promising affect for the treatment of diabetes mellitus.