Biosynthesis of carnosic acid (CA), one of the most industrially valuable medicinal compounds present in rosemary (Rosmarinus officinalis L.) leaves, is affected by various plant stressors. In this study, effects of silver nanoparticle (AgNP) treatment on the secondary metabolism and CA production of rosemary plants were investigated. AgNP of 0, 25, 50, 100, and 200 ppm were utilized on hydroponically grown plants using foliar spray. Efficient absorbance and translocation of AgNPs to the plant roots were confirmed by XRF (X-ray fluorescence) analysis. The fluctuations of important antioxidant compounds such as CA content, phenolics, flavonoids, and acid ascorbic were analyzed and their correlations evaluated. Results revealed that application of 200 ppm AgNPs for 12 days increased CA level more than 11%, as compared to the control plants. Furthermore, significant positive correlations were observed between total flavonoids and CA content under AgNP treatment, suggesting that AgNP acted as an elicitor and triggered the enhancement of CA accumulation effectively. These data suggest that concentration-dependent AgNP may be used to boost antioxidant activity and phytochemical contents of other medicinal plants.