GREEN SYNTHESIS OF ZINC OXIDE NANOPARTICLES (ZNO NPS) USING FRUIT AND SEED EXTRACT OF PAPAYA (CARICA PAPAYA) AND ASSESSMENT OF ITS ANTIINFLAMMATORY ACTIVITY ON ZEBRAFISH LARVAE (DANIO RERIO)

• Main Author: Putri Syahbaniati, Alia
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/76645
• Institution: Institut Teknologi Bandung

Zinc oxide nanoparticles (ZnO NP) are metal oxide nanomaterials that are widely used in various industrial fields such as biomedicine. Due to the various material properties of ZnO NP such as biocompatibility, low toxicity, and anti-inflammatory activity. Biosynthesis methods (green synthesis) using plant extracts are considered more environmentally friendly than physical and chemical methods. Papaya (Carica papaya Linn) seeds and fruits contain various bioactive compounds that can be used for the green synthesis of ZnO NPs. This study aims to biosynthesise zinc oxide nanoparticles (ZnO NP) using papaya seed and fruit extracts and to determine the anti-inflammatory activity of the synthesised nanoparticles in vivo on zebrafish (Danio rerio) larvae. Papaya seed and fruit extracts were extracted using water and methanol solvents, then the phytochemical content was characterised by qualitative methods and GC-MS analysis. The papaya seed aqueous extract (BIPA), papaya seed methanolic extract (BIPM), papaya fruit aqueous extract (BUPA) and papaya fruit methanolic extract (BUPM) were used for the green synthesis of ZnO NPs and the properties of the synthesised nanoparticles were characterised. The synthesised ZnO NPs were subjected to acute toxicity test on zebrafish embryos to determine the LC50 value and the LC25 value which were used as the treatment concentration for the anti-inflammatory activity test. The anti-inflammatory activity of the synthesised ZnO NP was determined in vivo on 24 hpf zebrafish larvae exposed to the synthesised ZnO NP sample for 6 hours. Then, 4 days post fertilisation (dpf) zebrafish larvae were induced to inflammatory response by the caudal fin amputation method, then macrophage migration is observed by Neutral Red (NR) staining and Sudan Black (SB) staining to observe neutrophil migration in zebrafish larvae at 4 hours and 48 hours after amputation under stereomicroscope. Data analysis of neutrophil migration, macrophage migration, and caudal fin regeneration area after treatment was performed using one-way ANOVA test and post hoc Dunnett test. Based on the results of the study, it is known that all the synthesised nanoparticles have ZnO NP characteristics, with the absorption value of ZnO NP wavelength in the range of 363—374 nm measured by UV-Vis spectrophotometer DRS. Based on the X-ray fluorescence spectroscopy (XRF) results, it was observed that all the nanoparticles contained ZnO oxide elements. The particle size observed by scanning electron microscopy (SEM) had a size range of 113—479 nm and morphology of flakes and nanoflowers. The hydrodynamic size of the particles (dissolved in water) is in the range of 144—355 nm according to the Particle Size Analyzer (PSA) measurements, with a negative surface charge in the range of -26.37 mV and -39.2 mV according to the Zeta Potential Analyzer measurements. The results of X-ray diffraction (XRD) spectra showed ZnO NPs in the form of hexagonal wurtzite crystals with an average crystallite size of 12—14 nm, and the functional groups of phytochemical compounds of extracts involved in the synthesis of ZnO NPs were identified in Fourier transform infrared (FTIR) spectra. The LC50 values from the ZnO NP toxicity test results are ZnO BIPA 26,08 mg/L; ZnO BIPM 10,83 mg/L; ZnO BUPA 8,25 mg/L; and ZnO BUPM 6,57 mg/L which are classified as toxic and slightly toxic. The results of the anti-inflammatory activity test of the ZnO NP treatment group at 4 hpa and 48 hpa were observed from the migration of neutrophils and macrophages, and the area of regeneration of zebrafish larval caudal fins had a significant difference compared to the negative control (p? 0.05). Based on the results of the research conducted, it can be concluded that ZnO NPs can be biologically synthesised using papaya seed and fruit extracts with the resulting nanoparticles having low toxicity and potential anti-inflammatory activity.