International Journal of Food Safety and Public Health

International Journal of Food Safety and Public Health ISSN 2756-3693 Vol. 10 (10), pp. 001-015, October, 2023. © International Scholars Journals

Full Length Research Paper

Cytotoxicity Assessment of Aflatoxin B1 (AFB1) after High Voltage Atmospheric Cold Plasma (HVACP) Generated Reactive Gas Species (RGS) Treatment Using Artemia salina and HepG2 Mammalian Cell MTT Bioassays

Michael M. Ndengele^{1 *}, Jackson F. Cummings¹

¹NanoGuard Technologies, Inc. 1100 Corporate Square Drive St. Louis, Missouri 63132, USA. *Corresponding author Email:[email protected]

Accepted October 14, 2023

Abstract 

Mycotoxins are toxic, heat-resistant metabolites of mold and fungi prevalent in food and animal feeds. Recent studies have shown that air passed through high voltage atmospheric cold plasma (HVACP) becomes temporarily ionized, forming reactive gas species (RGS) capable of destroying mycotoxins before reverting to normal air. This study evaluated aflatoxin B1 (AFB1) cytotoxicity with and without RGS treatment using two commonly applied toxicity evaluation model systems: Artemia salina [Brine Shrimp Test (BST)] and HepG23-(4-5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide salt (HepG2-MTT) in vitro bioassays. The BST dose response assay tested 0.25-1.13 µg AFB1/mL. Results obtained showed 11-73% mortality after 72h and 50% mortality (LD₅₀) at 0.6 µg AFB1/mL. Untreated AFB1 (1.7 µg/mL) caused 92% mortality of Artemia while RGS-treated AFB1 caused significantly less, 12% (p<.0001). The HepG2-MTT dose response assay tested 0.2-2.8 AFB1 µg/mL, causing 10-32% cell death and 13-57% for 24h and 48h respectively. Untreated AFB1 (0.8 µg/mL) caused 22% and 29% cell death after 24h and 48h while RGS-treated AFB1 caused significantly less, 3% and 2% (p<.05). Overall, this study confirms HVACP generated RGS significantly reduces AFB1 and the AFB1 byproducts formed are less toxic to cells and living organisms. 

Keywords: Aflatoxin. High voltage atmospheric cold plasma (HVACP). Reactive gas species (RGS) treatment. Cytotoxicity assessment.  HepG2 cell MTT bioassay. Artemia salina brine shrimp bioassay.