Study of acute lethality, teratogenesis, and metabolomic changes of N-(2’-hydroxyphenyl)-2-propylpentanamide (HO-AAVPA) on Artemia franciscana

The brine shrimp lethality test (Artemia spp.) is a classical model for assessing the toxicity of bioactive compounds. This study evaluated the toxicity and metabolomic changes induced by N-(2'- hydroxyphenyl)-2-propylpentanamide (HO-AAVPA), a derivative of valproic acid (VPA), in Artemia franciscana larvae using untargeted metabolomics through liquid chromatography-mass spectrometry (LC-MS/MS). The lethal concentration 50 (LC₅₀) was determined by acute toxicity tests at 24 and 48 h, and teratogenic effects were assessed by measuring the larvae body length. Larval metabolomic changes were examined following 24- and 48-hour exposures to sublethal concentrations of HO-AAVPA (LC₁ = 0.04 mM, LC₁₀ = 0.2 mM) and VPA (LC₁ = 1.79 mM, LC₁₀ = 8.95 mM). After 48 h, HO-AAVPA had an LC₅₀ of 0.32 mM, while VPA had 18.7 mM. VPA induced teratogenic effects at 9.6 mM; in contrast, HO-AAVPA only significantly affected the body length at 0.56 mM. Metabolomic analysis revealed that sublethal concentrations of HO-AAVPA affected the sphingolipid and glycerophospholipid metabolism, while VPA impacted alanine, aspartate, and glutamate metabolism. These findings suggest HO-AAVPA has high toxicity, but lower teratogenicity compared to VPA. In conclusion, the present study indicates that alterations in lipid and amino acid metabolism could be critical points in the mode of action of these compounds in A. franciscana.