Identification of phase II in vivo metabolites of alkyl-anthracenes in rainbow trout (Oncorhynchus mykiss).

Metabolism of xenobiotics is a two-step process that increases the polarity of compounds to facilitate their excretion. In previous work, the major in vitro phase I metabolites of alkyl-anthracenes by rainbow trout (Oncorhynchus mykiss) CYP enzymes were shown to be predominantly ring hydroxylated metabolites. Here, we present the first report on the identification of in vivo phase II metabolites of alkyl-anthracenes in juvenile rainbow trout. Bile was collected from trout injected with individual alkyl-anthracenes with, in some cases, a co-injection of ß-naphthoflavone (BNF). Some samples were digested with the ß-glucuronidase enzyme to confirm the presence of glucuronide conjugates. The metabolites were separated using a water-acetonitrile gradient on a HPLC system equipped with a C(18) column and a UV-diode array detector. Trout with endogenous and BNF-induced enzymes produced the same metabolites, but higher concentrations of metabolites were detected after enzyme induction. Alkyl-anthracenes were metabolized predominantly on the rings as evidenced by the UV spectral analysis. Likewise, mass spectrometry and UV spectral analysis confirmed a predominance of glucuronide conjugates for all systems investigated.

Measuring the toxicity of alkyl-phenanthrenes to early life stages of medaka (Oryzias latipes) using partition-controlled delivery.

Alkyl-phenanthrenes are a class of compounds present in crude oil and toxic to developing fish. Most research on alkyl-phenanthrenes has focused on retene (7-isopropyl-1-methyl-phenanthrene), but little is known about the chronic toxicity of related congeners to the early life stages of fish. This project is the first to describe the chronic toxicity of a series of alkyl-phenanthrenes to the embryos of Japanese medaka (Oryzias latipes) using the partition-controlled delivery (PCD) method of exposure and is the first to establish a relationship between toxicity of alkyl-phenanthrenes and log P. With PCD, test concentrations were maintained by equilibrium partitioning of test chemicals from polydimethylsiloxane (PDMS) films containing various concentrations of C1 to C4 phenanthrenes. Log film:solution partition constants (log K(fs)) and aqueous solubility limits were determined for each alkyl-phenanthrene. The prevalence of abnormalities in fish embryos increased in an exposure-dependent manner, with median effective concentration (EC50) values lower than experimental solubility limits of the compounds, and typical of environmental concentrations. Alkyl-phenanthrenes were more toxic to medaka embryos than unsubstituted phenanthrene, with effects resembling those of dioxin and indicating a specific receptor-based mechanism of toxicity. These results extend conclusions for the Exxon Valdez oil spill, suggest a specific mechanism of toxicity for alkyl-phenanthrenes, and provide a model for assessing the risks of mixture toxicity.

Variation in toxicity response of Ceriodaphnia dubia to Athabasca oil sands coke leachates.

Coke from the Athabasca (Alberta, Canada) oil sands operations may someday be integrated into reclamation landscapes. It is hypothesized that the metals associated with the solid coke may leach into the surrounding environment. Therefore, the main objectives of this study were to characterize the toxicity and chemistry of coke leachates collected from two field lysimeters (i.e. shallow lysimeter and deep lysimeter) over a period of 20months, as well as from other oil sands coke storage sites. In addition, a batch renewal leaching of coke was conducted to examine the rate of metals release. Chronic toxicity of key metals (e.g. Al, Mn, Ni and V) found in lysimeter coke leachate was evaluated separately. Toxicity test results revealed that whole coke leachates (100% v/v) were acutely toxic to Ceriodaphnia dubia; the 7-day LC50 values were always <25% v/v coke leachate. The deep lysimeter leachate was generally more toxic than the shallow lysimeter leachate, likely because of significantly higher concentrations of vanadium (V) found in the deep lysimeter leachate at all sampling times. Vanadium concentrations were higher than all other metals found in the leachate from both lysimeters, and in the batch renewal leaching study. Furthermore, V found in leachates collected from other oil sands field sites showed a concentration-response relationship with C. dubia survival. Mass balance calculations indicated that 94-98% of potentially leachable V fraction was still present in the coke from two field lysimeters. Evidence gathered from these assessments, including toxic unit (TU) calculations for the elements of concern, suggests that V was the likely cause of toxicity of the deep lysimeter leachate, whereas in the shallow lysimeter leachate both Ni and V could be responsible for the observed toxicity.