Plant extracts and their bioactive compounds are considered as the promising options for green synthesis of nanoparticles instead expensive and hazardous materials. Here, Solanum xanthocarpum fruit was used for synthesis of silver nanoparticles (AgNP). The synthesized AgNPs were characterized by using chromatographic and spectroscopic analytical methods. AgNPs were confirmed by UV-visible absorbance at 420-470nm. TEM analysis showed AgNP with 22.45nm average size. X-ray diffraction studies revealed the crystalline and face central cubic nature of AgNPs. FTIR analysis revealed functional group present over AgNPs. The aminodiphenyl acetic acid, clomipramine, and fonisopril from fruit extracts were found to be major capping agents on AgNPs as a result of analysis by HRLC-MS. All clinical isolates showed resistance for ampicilline, amoxyclav, niladixic acid, and sulphafurazole, suggesting multidrug resistance. The results showed that all isolates were sensitive to AgNPs synthesized fruit extracts. On the contrary, all isolates were resistant to whole S. xanthocarpum fruit extracts alone. The antimicrobial activity of AgNP was explored against multidrug-resistant (MDR) Gram-negative clinical isolates including Escherichia coli, Shigella spp., Aeronomonas spp. and Pseudomonas spp. MIC values ranged between 1.25mg/ml and 2.5mg/ml at 8McFarland's standards. Minimum bactericidal concentration was found to be in between 2.5mg/ml to 5mg/ml. Nanoparticles synthesized from fruit extract of S. xanthocarpum containing aminodiphenyl acetic acid, clomipramine, and fonisopril metabolites exhibit promising antimicrobial activity against MDR Gram-negative clinical isolates.