Nanoparticles synthesized by using natural polymers have been applied to various biomedical applications. These procedures are simple, eco-friendly and serve as an alternative to complex chemical methods for the preparation of nanomaterials. In the present study, synthesis of nanoparticles, to examine their antidiabetic potential, toxicity study was focused. Ethanolic extract of Euphorbia thymifolia was used for the synthesis of nanoparticles. The synthesis was monitored and confirmed by Particle size & polydispersibility index, Zeta potential study and Scanning electron microscopy. The particle size of nanoparticle made from extracts of Euphorbia thymifolia was determined. The nanoparticle’s particle size was determined to be 43.2 ± 11 and zeta potential was found to be -28.09. SEM picture of a freeze-dried nanoparticle with a longer cross-linking period, showing a tiny, spherical nanoparticle with a small size. The entrapment efficiency of Euphorbia thymifolia extract loaded nanoparticle was observed 81.12 %, indicating higher drug entrapment efficiency. It was discovered that as the concentration of polymer increased, the yield increased. After 12 weeks, the stability of the nanoparticles of was evaluated by analysing its absorption spectra. There were no significant changes over the storage period, and the nanoparticles did not agglomerate, indicating that they were more stable. The acute oral toxicity in rats indicated that extract of Euphorbia thymifolia was nontoxic at 2000mg/kg body weight. The formulated nanoparticles of extract of Euphorbia thymifolia after administration in Swiss albino mice were studied for antidiabetic activity for 1 month and found to be potent.