Species-dependent clearance of alternariol monomethyl ether, tenuazonic acid and altertoxin II in rat and human primary hepatocytes

Abstract

The ubiquitously occurring food contaminants alternariol monomethyl ether (AME), tenuazonic acid (TeA) and altertoxin II (ATX-II) are recognized as emerging Alternaria mycotoxins, yet substantial data gaps remain regarding their toxicokinetic characteristics. The hepatic metabolism of these three substances was investigated in primary rat (PRH) and human (PHH) hepatocytes by monitoring parent compound depletion and, where applicable, metabolite formation. AME was initially evaluated at 5 {micro}M and subsequently investigated over a concentration range of 0.75-8 {micro}M (0.75, 1.5, 3, and 8 {micro}M), whereas TeA was assessed at 5 {micro}M, and ATX-II was assessed at 0.22 and 1.11 {micro}M, with additional short-term experiments conducted over 30 min for AME and ATX-II, and over 10 min for concentration-dependent AME depletion. For AME, time-dependent clearance was further evaluated in PRHs at two hepatocyte densities, 0.25 and 0.5 million cells/mL. In PRHs, AME metabolism followed Michaelis-Menten kinetics (Vmax = 150.9 pmol{middle dot}min-{superscript 1}{middle dot}10- cells, Km = 1.18 {micro}M), whereas no reliable kinetic model could be established for PHHs. In contrast, TeA exhibited high metabolic stability, with only 9-10% depletion after 4 h, indicating negligible hepatic clearance in both species. ATX-II was also rapidly depleted and became undetectable within 30 min, accompanied by transient formation of altertoxin I (ATX-I), which was more pronounced in PHHs than in PRHs. Substrate depletion revealed pronounced interspecies differences in hepatic clearance capacity and stability. Overall, these findings provide comparative insights in primary human and rat hepatocyte systems, offering a foundation for future studies on their toxicological relevance and impact on human health.