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Green River BIOL 211 - The Formation of Ethanol in Postmortem Tissues

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The Formation of Ethanol in Postmortem TissuesRobert D. JohnsonRussell J. LewisMike K. AngierNicole T. VuCivil Aerospace Medical InstituteFederal Aviation AdministrationOklahoma City, OK 73125February 2004Final ReportThis document is available to the publicthrough the National Technical InformationService, Springfield, Virginia 22161. Office of Aerospace MedicineWashington, DC 20591DOT/FAA/AM-04/4NOTICEThis document is disseminated under the sponsorship ofthe U.S. Department of Transportation in the interest ofinformation exchange. The United States Governmentassumes no liability for the contents thereof.iTechnical Report Documentation Page 1. Report No. 2. Government Accession No. 3. Recipient's Catalog No. DOT/FAA/AM-04/4 4. Title and Subtitle 5. Report Date The Formation of Ethanol in Postmortem Tissues February 2004 7. Author(s) 6. Performing Organization Code Johnson RD, Lewis RJ, Angier MK, Vu NT 9. Performing Organization Name and Address 8. Performing Organization Report No. FAA Civil Aerospace Medical Institute P.O. Box 25082 10. Work Unit No. (TRAIS) Oklahoma City, OK 73125 12. Sponsoring Agency name and Address 11. Contract or Grant No. Office of Aerospace Medicine Federal Aviation Administration 13. Type of Report and Period Covered 800 Independence Ave., S.W. Washington, DC 20591 15. Supplemental Notes This work was accomplished under the approved task AM-B-02-TOX-204.14. Sponsoring Agency Code 16. Abstract During the investigation of aviation accidents, postmortem samples obtained from fatal accident victims are submitted to the FAA’s Civil Aerospace Medical Institute for toxicological analysis. During toxicological evaluations, ethanol analysis is performed on all cases. Many species of bacteria, yeast and fungi have the ability to produce ethanol and other volatile organic compounds in postmortem specimens. The potential for postmortem ethanol formation complicates the interpretation of ethanol-positive results from accident victims. Therefore, the prevention of ethanol formation at all steps following specimen collection is a priority. Sodium fluoride is the most commonly used preservative for postmortem specimens. Several studies have been published detailing the effectiveness of sodium fluoride for the prevention of ethanol formation in blood and urine specimens; however, our laboratory receives blood or urine in approximately 70% of cases. Thus, we frequently rely on tissue specimens for ethanol analysis. The postmortem tissue specimens received by our laboratory have generally been subjected to severe trauma and may have been exposed to numerous microbial species capable of ethanol production. With this in mind, we designed an experiment utilizing unadulterated tissue specimens obtained from aviation accident victims to determine the effectiveness of sodium fluoride at various storage temperatures for the prevention of microbial ethanol formation. We found that without preservative, specimens stored at 4°C for 96 h showed an average increase in ethanol concentration of 1470%. At 25°C, these same specimens showed an average ethanol increase of 1432% after 48 h. With the addition of 1.00% sodium fluoride, there was no significant increase in ethanol concentration at either temperature.17. Key Words 18. Distribution Statement Document is available to the public through the National Technical Information Service Postmortem Ethanol, Formation, Microbe, Aircraft Accident Investigation Springfield, Virginia 22161 20. Security Classif. (of this reporte) 20. Security Classif. (of this page) 21. No. of Pages 22. Price Unclassified Unclassified 14 Form DOT F 1700.7 (8-72) Reproduction of completed page authorized1THE FORMATION OF ETHANOL IN POSTMORTEM TISSUESINTRODUCTIONThe Federal Aviation Administration’s (FAA) Civil Aerospace Medical Institute (CAMI) is responsible under Department of Transportation (DOT) orders 8020.11A and 1100.2C, to “conduct toxicologic analysis on speci-mens from … aircraft accident fatalities” and “investigate … general aviation and air carrier accidents and search for biomedical and clinical causes of the accidents, includ-ing evidence of … chemical (use).” Therefore, following an aviation accident, samples are collected at autopsy and sent to CAMI’s Bioaeronautical Sciences Research Laboratory, Forensic Toxicology Research Team where toxicological analysis is conducted on various postmortem fluids and tissues.Ethanol analysis is one of the most common tests per-formed on forensic specimens. The presence of ethanol in aircraft accident victims constitutes an important part of both aircraft accident investigations and litigation. Using modern analytical techniques, there is little doubt as to the quantitative amount of ethanol present in a postmortem specimen. The origin of the ethanol detected, however, is an important variable that must be considered when interpreting ethanol results in postmortem specimens. The microbial formation of ethanol in postmortem specimens is the most significant problem encountered when evaluating ethanol results. The first report dealing with postmortem ethanol production in corpses ap-peared in 1936 (1). At that time, a postmortem increase in ethanol concentration was occasionally observed, but the source of the increase was unknown. The first stud-ies to elucidate the mechanism of postmortem ethanol formation occurred in the early 1970s (2, 3). From these initial studies it became clear that microbes are responsible for postmortem formation of ethanol. Many species of bacteria, yeast and fungi produce ethanol and other volatile organic compounds as a byproduct of their metabolism (4-12). Candida albicans has been identified as the microbe most often responsible for postmortem production of ethanol in humans (9, 13). This species of yeast is commonly found in humans in vivo. However, microbes not normally found in vivo, such as the Mucor species of mold, are commonly found in decaying organic matter and soil (14); they may be introduced from the environment if the circumstances surrounding death are violent, as is often seen in aviation accidents. After death, endogenous and/or exogenous microbes begin rapidly consuming glucose and other nutrients present in the body and produce ethanol and/or other organic vola-tiles as metabolic byproducts (12, 15-17). Under optimal conditions, substantial concentrations of


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Green River BIOL 211 - The Formation of Ethanol in Postmortem Tissues

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