DEVELOPMENT OF ANALYTICAL 



METHODS FOR IDENTIFICATION AND 



QUANTIFICATION OF OIL BREAKDOWN 



PRODUCTS IN TISSUES AND EGGS OF 



DUCKS 



Efforts of chemists involved in this project 

 have been directed toward several goals: 



• The development of methods for the detec- 

 tion and analysis of hydrocarbons in avian 

 tissue 



• The development of methods of quantifi- 

 cation of individual compounds and com- 

 plex mixtures in avian tissues 



• Provision of analytical support for biological 



studies 



A number of problems are encountered in 

 the analysis of petroleum hydrocarbons in avian 

 tissue. By their ubiquitous nature, these hydro- 

 carbons pose continuing contamination prob- 

 lems for sample analysis. The heterogeneity of 

 the mixtures necessitates the use of complex 

 extraction and cleanup procedures. The presence 

 of hundreds of compounds creates a need for 

 high-resolution instruments. 



During our early attempts to analyze sam- 

 ples, very high levels of contamination were 

 detected. To reduce contamination, glass extrac- 

 tion thimbles cleaned in chromic acid were 

 substituted for paper thimbles. Later, in simpli- 

 fying our procedures, we eliminated thimbles 

 altogether, substituting a Polytron tissue homo- 

 genizer/extractor for Soxhlet extraction. The 

 use of nitrogen gas to blow down samples was 

 abandoned in favor of a flash evaporator. Glass- 

 ware was at first soaked in chromic acid; this 

 was an effective but hazardous procedure. 

 Chromic acid was replaced by an equally effec- 

 tive but safer technique of soaking the glass in 

 an ultrasonic bath with 2% 'Micro' solution. 



Briefly, the method of analysis that is being 

 used in our laboratory is as follows. 



The sample is cut into small pieces and me- 

 chanically extracted with a homogenizer. Bases 

 are partitioned into acid from pentane and ana- 

 lyzed by gas chromatography /mass spectrometry. 

 After the extraction of bases, pentane extracts 

 from fat samples are saponified. The saponified 

 mixture is partitioned into hexane. 



The hexane or pentane layer from samples 

 of fat, liver, and kidney (<0.5 g lipid) are 

 cleaned up on Morisil; the aliphatic and aroma- 

 tic hydrocarbons are separated on a silicic acid 



column; and the residues are screened and ana- 

 lyzed by gas chromatography and gas chroma- 

 tography/mass spectrometry/data system. 



Livers, kidneys, and fat from untreated mal- 

 lard drakes and drakes fed 0.4% of the reconsti- 

 tuted aromatic mixture for 6 months were ana- 

 lyzed in our laboratory. Significantly higher resi- 

 dues were found in the tissues of ducks fed the 

 aromatic mixture than in tissues from controls. 



Researchers at the University of New Orleans 

 developed a hydrocarbon analysis procedure 

 similar to the one in use at our laboratory while 

 under contract to the Patuxent Wildlife Re- 

 search Center. Livers and kidneys from mallard 

 ducklings and adult birds fed South Louisiana 

 crude oil were analyzed. There were no signifi- 

 cant differences in hydrocarbon accumulation in 

 tissues between adult males and females. Hydro- 

 carbon concentrations in both ducklings and 

 adults increased from the control group through 

 those fed 2.5% oil in the diet. 



The research group at the University of New- 

 Orleans also analyzed tissues from four birds 

 killed at the Amoco Cadiz oil spill site off the 

 coast of France. One, a shag (cormorant), was 

 heavily contaminated with hydrocarbons; the 

 other three, a herring gull, a razorbill, and a guil- 

 lemot, contained smaller amounts. 



LITERATURE CITED 



Albers, P. H., and R. C. Szaro. 1978. Effects of 

 No. 2 fuel oil on common eider eggs. Mar. 

 Poll. Bull. 9:138-139. 



Coon, N. C, P. H. Albers, and R. C. Szaro. 

 1979. No. 2 fuel oil decreased embryonic 

 survival of great black-backed gulls. Bull. 

 Environ. Contam. Toxicol. 21:152 -156. 



Coon, N. C, and M. P. Dieter. Effects of dietary 

 ingestion of crude oil on condition and sur- 

 vival of mallard ducks. Patuxent Wildlife 

 Research Center Study Plan Progress Report, 

 October 1, 1976-September 30, 1977 

 (unpubl). 



Hoffman, D. J. 1978. Embryotoxic effects of 

 crude oil in mallard ducks and chicks. 

 Toxicol. Appl. Pharmacol. 46:183-190. 



Holmes, W. N., J. Cronshaw, and J. Gorsline. 

 1978. Some effects of ingested petroleum on 

 seawater-adapted ducks (Ancsplatyrh ynchos ). 

 Environ. Res. 17:177-190. 



