Chitosan nanoparticles functionalized with plant extracts for the inhibition of the toxic effects of aflatoxin B1 and Ochratoxin A

M.Sc. (Nanoscience)Ochratoxin A and Aflatoxin B1 are important food contaminates as they are known to be mutagenic, genotoxic, nephrotoxic, hepatotoxic, immunosuppressive and teratogenic to both animals and humans. These mycotoxins are associated with the contamination of food stuff such as grapes, maize, red pepper, meat, milk, beans and processed products from contaminated raw material. Current physical, biological and chemical methods employed to improve the safety of food often compromise the nutritional value and result in huge losses. The alternative to these treatments are addition of supplements with protective properties to reduce the toxicity of mycotoxins or prevent their formation. The work presented in this dissertation reports an attempt to develop such materials to prevent damage caused by ochratoxin A and aflatoxin B1. This was done through the synthesis; characterisation and cytotoxicity study of chitosan nanoparticles with methanolic plant extracts (L. leonurus, M. longifolia and A. montanus). Inhibition of cellular damage due to mycotoxins for possible application in prevention of cellular damage by mycotoxins also presented. Chitosan nanoparticles were synthesised using an ionic gelation method with sodium triphosphate as the cross linker. The methanolic medicinal plants extracts were incorporated into the chitosan solution before synthesising nanoparticles, and nanoparticle synthesis initiated by the addition of sodium triphosphate solution. The synthesised products were characterised using zetasizer, transmission electron microscopy, x-ray diffraction and Fourier-transform infrared spectroscopy. The extracts’ antioxidant ability was evaluated before incorporation into chitosan using 2, 2-diphenyl- 1-picrylhydrazy (DPPH) radical scavenging assay. This assay was performed using UVvis spectroscopy. The cytotoxicity of the synthesised nanoparticles was assessed using a Vero cell line and by evaluating the cell viability with an MTS assay. The nanoparticles were successfully synthesised and showed the presence of different functional groups as expected. Plain chitosan nanoparticles were roughly spherical shaped and had smooth surfaces, nanoparticles containing extracts similarly were spherical in shape as well but had rougher surfaces when visualised under TEM. All nanoparticles had positive zeta potentials between 26 – 28 mV. The average particle sizes ranged between 31 – 65 nm as measured using TEM and average particle sizes obtained using zetasiser was 78 – 190 nm. The cytotoxicity studies of plain nanoparticles and nanoparticles with extract showed that the synthesised nanomaterials were not toxic even at concentration of 500 μg/ml and less than 20% of the Vero cells were affected under these conditions