Identification of compounds in heavy fuel oil 7102 that are chronically toxic to rainbow trout (Oncorhynchus mykiss) embryos
rainbow trout , embryos , ecotoxicology , PAH , heavy fuel oil
Spilled heavy fuel oil (HFO) sinks within the water column and accumulates in sediments, affecting aquatic organisms that are not typically exposed to oils that float. Previously, the 3-4 ring alkyl substituted polycyclic aromatic hydrocarbons (PAHs) have been identified as the major toxic components in crude oil. Since HFO is comprised of higher concentrations of 3-4 ringed alkyl PAH and an abundance of 5-6 ringed PAH relative to crude oil, it is predicted to be more toxic to the early life stages of fish. An effects-driven chemical fractionation (EDCF) of HFO 7102 was undertaken to establish the toxicity relative to crude oil, and to identify the compounds that are bioavailable and chronically toxic to the early life stages of fish. In this EDCF, the complex HFO 7102 mixture was separated by low temperature vacuum distillation into three distinct fractions, 2, 3 and 4. Each fraction was assessed using a chronic bioassay to determine whether it contained components that caused toxicity to rainbow trout embryos similar to that of the whole oil. Acute bioassays with juvenile trout demonstrated the presence of compounds that induce cytochrome P450 enzymes, an indicator of exposure to PAH. Fraction 3, the fraction more toxic than the parent mixture, was further separated by cold acetone extraction into fraction 3-1 (PAH-rich extract) and fraction 3-2 (wax residue), and assessed with the same bioassays. Simultaneous chemical analysis with bioassays guided the fractionation, and identified compounds abundant and consistently present in toxic fractions. Due to resistance to dispersion of HFO, a chemical dispersant was used with vigorous mixing to drive the maximum amount of oil into solution to minimize the potential for false negatives and the volume of test material used. The potency of HFO 7102 and its fractions were also measured using water accommodated fractions (WAFs) produced by a continuous flow system of water flowing through oil coated gravel. Both exposure methods traced the toxicity from whole oil into fractions containing higher concentrations of 3-4 ring alkyl PAH, similar to crude oil. This research is the first toxicological assessment of HFO 7102, which is essential for determining the risk of spills of HFO to fish, and whether the risk of oils can be predicted from their alkyl PAH composition.