Depletion of isoeugenol residues from the fillet tissue of AQUI-S™ exposed rainbow trout (Oncorhynchus mykiss)

There is a critical need in U.S. public aquaculture and fishery management for an approved sedative that allows for the immediate release of fish after sedation. AQUI-S^TM is a fish anesthetic/sedative approved for use in several countries and until recently was being developed in the U.S. as a sedative for immediate release of fish after sedation. The U.S. National Toxicology Program reported that isoeugenol (the active ingredient in AQUI-S^TM) exposed male mice showed clear evidence of carcinogenicity, therefore efforts within the U.S. Department of Interior to develop AQUI-S^TM as a sedative that allows for immediate release ceased. Despite the ruling, AQUI-S^TM still has the potential to be approved as an anesthetic with a short withdrawal time. Among the data required to gain approval for use in the U.S. are data describing the composition and depletion of all AQUI-S^TM residues from fish fillet tissue. A total residue depletion study for AQUI-S^TM was conducted by exposing market-sized rainbow trout, Oncorhynchus mykiss (mean weight, 502.7 ± 54 g; s.d.) to 8.9 mg/L of ¹⁴C-[URL]-isoeugenol for 60 min in 17 °C water. The ¹⁴C-[URL]-isoeugenol was mixed with a surfactant resulting in a mixture that mimicked AQUI-S^TM. Groups of fish (n = 6) were sampled immediately after the exposure (0-h sample group) and at 0.5, 1, 2, and 4 h thereafter. Total isoeugenol-equivalent residue concentrations in the fillet tissue were determined by oxidizing triplicate subsamples of homogenized skin-on fillet tissue from each fish to ¹⁴CO₂ and enumerating the radioactivity by static liquid scintillation counting. Isoeugenol concentrations in fillet tissue were determined by extracting homogenized fillet tissue with solvents and determining the isoeugenol concentrations in the extracts with high performance liquid chromatography techniques. The mean total isoeugenol-equivalent residue concentrations in the 0, 0.5, 1, 2, and 4-h sample groups were 55.4, 32.0, 19.8, 11.3, and 4.9 µg/g, respectively. The primary chemical residue in fillet tissue from all exposed fish was isoeugenol. The mean isoeugenol concentrations in the 0, 0.5, 1, 2, and 4-h sample groups were 48.9, 26.5, 15.3, 7.2, and 2.2 µg/g, respectively. The percents of the total radioactivity classified as isoeugenol in the 0, 0.5, 1, 2, and 4-h tissue extracts were 95, 73, 73, 64, and 48%, respectively.