Studies of the synthesis, environmental occurrence and toxicity of unresolved complex mixtures (UCMs) of hydrocarbons
The occurrence of unresolved complex mixtures of hydrocarbons (UCMs) in the aliphatic fraction of marine sediments and organisms from areas impacted by petroleum hydrocarbons is well documented and widely accepted as an indication of fossil fuel contamination. In contrast, the presence of an aromatic UCM is often ignored and environmental concentrations of aromatic UCM hydrocarbons in marine biota and sediments are rarely reported. The aims of this study were to establish the quantitative significance of aromatic UCMs in environmental samples and to assess the toxicological significance of both aliphatic and aromatic UCMs. A reproducible method was developed and validated for the analysis and quantification of petroleum hydrocarbons in mussel (Mytilus edulis) tissue. Emphasis was placed upon development of a method which minimized losses of more volatile, lower molecular weight, toxicologically significant hydrocarbons, without compromising recovery of higher molecular weight compounds which are useful for source identification in environmental monitoring schemes. Analysis of mussels from a small number of U. K. coastal locations indicated that aromatic hydrocarbon UCMs may form a significant proportion (ca 20 %) of the total hydrocarbon body burden of mussels from areas contaminated with petroleum hydrocarbons. Aromatic UCM hydrocarbons were not observed in mussels from relatively uncontaminated areas but concentrations of 430 μg g'' (dry wt tissue) were measured in mussels from heavily impacted areas. Aliphatic UCM concentrations ranged from 7- 3445 μg gg' (dry wt tissue). For the purposes of toxicological studies, a low molecular weight model aliphatic UCM hydrocarbon, 4-propyloctane (4-PO) was synthesised. Two low molecular weight model aromatic hydrocarbons 7- cyclohexyltetralin and 7-cyclohexyl-l-propyltetralin were also synthesised using a modification of the Haworth synthesis. All three target compounds and synthetic intermediates were characterised by NMR, MSandIR. Exposure of M. edulis to 4-PO caused a significant reduction in mussel ciliary feeding activity indicating that 4-PO was indeed toxic as measured by this bioassay. The demonstrable narcotic activity of 4-PO is presumably related to the greater aqueous solubility of branched hydrocarbons compared with similar straight chain hydrocarbons. Further experiments investigating the effect of 4- PO over exposure periods up to 120h provided a unique and detailed insight into the relationship between concentration of toxicant in the gills of M. edulis and observed feeding rate. The established method of mussel feeding rate determination was modified in light of this relationship to produce an " improved rapid and reproducible screening technique. Both of the model aromatic UCM hydrocarbons were also found to be toxic to mussel ciliary feeding activity. This appears to be the first report of investigations into the toxicity of the aromatic UCM and suggests that previous studies have ignored an environmental burden of toxicological significance. Estimates of the tissue effective concentration (TEC50) for the model UCM hydrocarbons gave comparable values with those reported for the effect of other narcotic hydrocarbons upon mussel feeding rate, providing support for the theory that non-specific narcosis occurs at a relatively constant tissue concentration of toxicant. The demonstrated narcotic activity of each of the three model UCM hydrocarbons has extended the molecular weight range of narcotic hydrocarbons studied to date. The results presented herein suggest that a small proportion of low molecular weight aliphatic UCMs and perhaps a greater proportion of aromatic UCMs are of toxicological significance.