eng Oriental Scientific Publishing Company Biomedical and Pharmacology Journal 0974-6242 2025-12-30 18 4 3057 3069 10.13005/bpj/3318 69391 article I Made Sukadana 1 Sri Rahayu Santi 1 Ni Nyoman Rupiasih 2 Luh Gede Sri Surya Heryani 3 Lilik Parwati 1 Nadya Pamesthi Annisa Ramadhani 1 Chemistry Department, Faculty Mathematics and Natural Sciences Udayana University, Badung, Indonesia Physic Department, Faculty Mathematics and Natural Sciences Udayana University, Badung, Indonesia Faculty of Veterinary Medicine Udayana University, Badung, Indonesia Preclinical studies indicate that gayam seeds (Inocarpus fagiferus Fosb) at a dose of 50 mg/kg body weight exhibit anti- atherosclerotic effects, as evidenced by changes in biomarkers including SOD activity, lipid profile, blood MDA, SOD-3, SOD-2, ICAM-1, IL-6, and TNF-α in aortic endothelial cells. While optimized solid and granule formulations have been developed, the formation of nanoparticles (1–100 nanometers in size) may further enhance the bioactivity of the extract. Nanoparticles provide improved solubility, bioavailability, stability, targeted delivery, increased extract efficacy, reduced dosages, and minimized side effects. This study aimed to prepare and characterize ethanol extract nanoparticles of gayam seeds for anti-atherosclerotic drug development. Nanoparticles were synthesized using the quality by design approach (Fractional Factorial Design, FFD), with the optimal formulation comprising 1.5 mL of 0.1% extract, 10 mL of 1.754% chitosan, and 5 mL of 0.1% NaTPP, stirred at 500 rpm for 15 minutes. The resulting nanoparticles demonstrated 100% encapsulation efficiency, 44.89% transmittance, a polydispersity index (PDI) of 0.3489, a zeta potential of 34.07 mV, and a particle size of 250.63 nm, meeting established nanoparticle criteria. https://biomedpharmajournal.org/vol18no4/formulation-and-characterization-of-ethanol-extract-nanoparticles-of-gayam-inocarpus-fagifer-parkinson-fosberg-seeds-for-anti-atherosclerotic-applications/ Antioxidant Atherosclerosis Drug formulation Inocarpus fagiferus Fosb Nanoparticles