Suggested Citation:
Murawski, Steven. 2016. PAH Analysis: Red Snapper Bile PAH Metabolite Concentrations, Northern Gulf of Mexico, 2012. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7T43R3T
Data Parameters and Units:
Red Snapper Station, ID, Data file, Nap Area, Nap Conc (ug/g), Nap Mean (ug/g), Nap STDEV, Nap %CV, Phen Area, Phen Conc (ug/g), Phen Mean (ug/g), Phen STDEV, Phen %CV, BaP Area, BaP Conc (ng/g), BaP Mean (ng/g), BaP STDEV, BaP %CV. Red Snapper Stations-- Latitude (decimal degrees), Longitude (decimal degrees), Napth, BaP, station.
Methods:
Frozen bile samples were stored at -20°C until analysis. A total of 15 samples from 2012 (red snapper) were analyzed for metabolites of PAHs using a high-performance liquid chromatography/fluorescence (HPLC-F) method (Krahn et al. 1984; Krahn et al. 2005). This method results in the determination of the concentrations of classes of PAH metabolites fluorescing in the regions typified by NPH, PHN and benzo[a]pyrene (BaP). Bile was injected directly onto a Waters © high-performance liquid chromatography/fluorescence system equipped with a C-18 reverse-phase column (Phenomenex Synergi Hydro©). The PAH metabolites were eluted with a linear gradient from 100% water (containing a trace amount of acetic acid) to 100% methanol at a flow of 1.0 mL/min. Chromatograms were recorded at the following wavelength pairs: (1) 292/335 nm where many 2-3 benzene ring aromatic compounds (e.g., NPH) fluoresce, (2) 260/380 nm where several 3-4 ring compounds (e.g., PHN) fluoresce and (3) 380/430 nm where 4-5 ring compounds (e.g., BaP) fluoresce. Peaks eluting after nine minutes were integrated and the areas of these peaks were summed. The concentrations of fluorescent PAHs in the bile samples of the marine fish were determined using NPH, PHN, BaP as external standards and converting the fluorescence response of bile to PHN (ng PHN equivalents g-1 bile), NHP (ng NPH equivalents g-1 bile) or BaP (ng BaP equivalents g-1 bile) equivalents. To ensure that the HPLC/fluorescence system was operating properly, a NPH/PHN/BaP calibration standard was analyzed five times to obtain a relative standard deviation < 15% for each PAH. Methods blanks revealed no traces of PAH metabolites, and the calibration standard was reproduced within the standard deviation of the source compound. Bile samples taken in 2012 were analyzed at the Mote Marine Laboratory using identical methods as described above. We re-reanalyzed 2011 samples at Mote Marine laboratory, resulting in identical results as those obtained by the NWFSC. Methods blanks were also run periodically to assure that no contamination of samples was skewing contaminant assessment results. Sampling in 2012 occurred in August, using the R/V Weatherbird II. For selected specimens we determined PAH levels in muscle and liver tissues and PAH metabolites in bile using standard methods. At each station, eight km of 3.2 mm galvanized steel main line was deployed, with 322-500 baited hooks. We used 91 kg test leaders, 3.7 m long attached to #13 circle hooks, with alternating cut fish and squid as bait. At the beginning and end of the main line we deployed Star:Oddi© CDST Centi temperature/time/depth recorders to record actual bottom time, as well as bottom temperature and depth fished. The recording interval of these instruments was 5 min. At set-out and haul-back we recorded latitude and longitude, time, depth from the vessel’s depth finder, the unique numbers of the TD instruments deployed at either end of the string, and local weather conditions. Once the longline was deployed, usually the vessel steamed back to the start buoy and began haul-back. The average soak time was 2 hours 1 minute. Large sharks (e.g., ~2 m and greater) were photographed for species identification at the rail and released alive. Each fish obtained was inspected for a variety of externally-symptomatic diseases and other samples obtained. Each fish was examined for the following: (1) presence of external skin lesions (e.g., ulcers, or other external eruptions of the integument or skin irritation unrelated to mechanical damage, (2) the presence of fin rot disease, (3) gills examined for the presence of parasites (data not reported here) and tumors, (4) body inspected for evidence of recent mechanical damage, thought to occur through trauma of the catching process or due to predators, and (4) inspection of the skin and internal organs for the presence of obvious tumors and tumor-like growths. Photographs were taken of skin lesions and other pathologies and the status of each lesion was evaluated (e.g., open bloody ulcer, closed skin contusions, healing or old injury). To obtain bile samples we dissected and punctured the gall bladder into a clear 15x45 mm vial. A foil liner was inserted between the cap and vial to prevent hydrocarbon contamination. The vial was wrapped in foil to prevent photo oxidation, and placed in a plastic bag. Bile samples were kept on ice or frozen at sea until return to the laboratory where they were maintained at -20°C. Data for skin lesions for these samples can be found in dataset R1.x135.120:0002. Methods blanks and standard exposure material (Atlantic salmon tissue) were routinely done for these samples (see published paper for methods) and results are reported in the spreadsheet.Methods described in: Krahn, M.M., M.S. Myers, D.G. Burrows, and D.C Malins. 1984. Determination of xenobiotics in bile of fish from polluted waterways. Xenobiotica 14: 633-646. Krahn, M.M., G.M. Ylitalo, J. Buzitis, J.L. Bolton, C.A. Wigren, S.-L. Chan, and U. Varanasi. 1993. Analyses of petroleum-related contaminants in marine fish and sediments following the Gulf oil spill. Marine Pollution Bulletin 27: 285-292. Krahn, M.M., G.M. Ylitalo, and T.K. Collier. 2005. Analysis of bile of fish collected in coastal waters of the Gulf of Mexico potentially affected by Hurricane Katrina to determine recent exposure to polycyclic aromatic compounds (PACs). M.S. NOAA, National Marine Fisheries Service, Northwest Fisheries Science Center, Seattle, Washington.
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