The importance of green synthesis was revealed with advantages such as: eliminating the use of expensive chemicals; consume less energy; and generate environmentally benign products. With this aim, silver nanoparticles (AgNPs) were synthesized by using isolated eugenol from clove extract. Its antimicrobial potential was determined on three different microorganisms. Clove was extracted and eugenol was isolated from this extract. Green synthesis was performed and an anti-microbial study was performed. All extraction and isolation analyses were performed by high-performance liquid chromatography (HPLC); identification and confirmation were achieved using liquid chromatography-mass spectrometry (LC-MS); and scanning electron microscopy was used for characterization. Both HPLC and LC-MS analyses showed that eugenol obtained purely synthesized AgNPs and 20-25-nm-sized and homogeneous shaped particles seen in images. The antimicrobial effects of AgNPs at eight concentrations were determinated against Staphylococcus aureus, Escherichia coli and Candida albicans, and maximum inhibition zone diameters were found as 2.6 cm, 2.4 cm and 1.5 cm, respectively. The results of the antimicrobial study showed that eugenol as a biological material brought higher antimicrobial effect to AgNPs in comparison to the other materials found in the literature.