Fisher, Russell S.

Freimuth, Henry C.

Volatile, Michael T.

English

Northwestern University Illinois, School of Law: Scholarly Commons

Journal of Criminal Law and CriminologyVolume 42Issue 4 November-December Article 9Winter 1951Results of the Studies on the Determination ofEthyl Alcohol in TissuesHenry C. FreimuthMichael T. VolatileRussell S. FisherFollow this and additional works at: https://scholarlycommons.law.northwestern.edu/jclcPart of the Criminal Law Commons, Criminology Commons, and the Criminology and CriminalJustice CommonsThis Criminology is brought to you for free and open access by Northwestern University School of Law Scholarly Commons. It has been accepted forinclusion in Journal of Criminal Law and Criminology by an authorized editor of Northwestern University School of Law Scholarly Commons.Recommended CitationHenry C. Freimuth, Michael T. Volatile, Russell S. Fisher, Results of the Studies on the Determination of Ethyl Alcohol in Tissues, 42 J.Crim. L. Criminology & Police Sci. 529 (1951-1952)THE RESULTS OF STUDIES ON THE DETERMINATION OFETHYL ALCOHOL IN TISSUESHenry C. Freimuth, Michael T. Volatile, and Russell S. FisherHenry C. Freimuth, Ph. D., is Toxicologist, Office of the Chief Medical Examiner,State of Maryland, and an associate in Legal Medicine, University of MarylandMedical School. In addition to these duties he has served since 1941 as an asso-ciate instructor in the Maryland State Police Training School.Michael T. Volatile formerly served on the staff of the Office of the Chief MedicalExaminer, State of Maryland, as a junior chemist and is at present a student indentistry at the University of Maryland.Russell S. Fisher, M. D., is Chief Medical Examiner, State of Maryland, andHead of the Department of Legal Medicine, University of Maryland MedicalSchool. He is also a lecturer in Forensic Pathology at Johns Hopkins Universityand prior to his appointment as Chief Medical Examiner held a research fellow-ship in the Department of Legal Medicine, Harvard Medical School.This paper on the determination of ethyl alcohol in tissues was read before theorganizational meeting (1950) of the Academy of Forensic Sciences.-EDiTor.There are many methods for the quantitative estimation of ethylalcohol in tissues, some of which are quite specific (1), (2), (3), whileothers are based upon the relatively non-specific reaction of tissue dis-tillates with potassium dichromate (4), (5), (6). The former methodsare very time consuming so that one must sacrifice some specificity indaily routine analyses of a large number of cases such as are encounteredin a Medical Examiner's Office. As a result, recourse is usually hadto one of the modifications of the original Nicloux procedure, and theconcentration of reducing substances in the tissue distillate is reportedas ethyl alcohol after methyl alcohol has been eliminated by qualita-tive tests.The authors have adopted the procedure of Muehlberger (7) forroutine quantitative analysis of tissues. In this procedure, 50 gramsof minced tissue are taken, tartaric acid and mineral oil are added,and the whole is steam distilled. 100 ml. of distillate are collected. To5 ml. of distillate there are added 5 ml. of standard potassium dichro-mate solution and 5 ml. of concentrated sulphuric acid. This mixtureis heated in a boiling water bath for fifteen minutes, cooled, and dilutedto approximately 200 ml. with distilled water. 3 grams of potassiumiodide are added, and the liberated iodine is titrated with standardsodium thiosulfate.There were several points involved in this procedure which, it wasfelt, required investigation and clarification.I. Information was lacking as to the percentage of alcohol re-covered by the method employed. Hence, known amounts of ethylalcohol were added to 50 gram samples of brain which were thensubjected to analysis. The results obtained are recorded in Table 1.H. FREIMUTH, M. VOLATILE AND R. FISHERTable 1.RECOVERY OF ETHYL ALCOHOLMg. Alcohol Mg. Alcohol Per centSample No. Added Recovered Recovery1 51.5 52.0 1012 51.5 52.0 1013 103.0 101.0 984 103.0 102.3 99.45 154.5 157.0 101.56 154.5 152.0 98.47 206.0 200.0 97.28 206.0 198.5 96.59 257.5 251.0 97.410 257.5 246.5 95.811 309.0 296.5 96.012 309.0 296.5 96.0Average 98.1It may be seen that the percentage recovery varied from 95.8 to101.5 with an average recovery of 98.1%. This means a maximumvariation of 0.01% in reported concentrations at levels of 0.40% orless and a maximum variation of 0.03% at the higher levels.II. Distribution of ethyl alcohol in the various parts of the brain.Inasmuch as the procedure involves the use of 50 gram samples oftissue, it was deemed advisable to determine whether it was necessaryto grind the entire organ, mix, and then remove a 50 gram aliquotor whether 50 grams could be removed from the intact organ for useas a sample. Accordingly, the brains were obtained from five differentcases, and 50 gram samples were taken for analysis from the left frontallobe, the right frontal lobe, the cerebellum, the left occipital lobe, andthe right occipital lobe. The results of these analyses are shown inTable 2.Table 2.DISTRIBUTION OF ETHYL ALCOHOL IN BRAINLeft Right Left RightFrontal Frontal Occipital OccipitalCase No. Lobe - Lobe Cerebellum Lobe Lobe1 0.20% 0.20% 0.23% 0.21% 0.21%2 0.07% 0.08% 0.07% 0.08% 0.08%3 0.27% 0.28% 0.27% 0.28% 0.27%4 0.22% 0.21% 0.21% 0.21% 0.21%5 0.23% 0.24% 0.26% 0.25% 0.24%The maximum variation was shown in cases 1 and 5 in each of whicha difference of 0.03 % was shown between the concentration of alcohol[Vol. 42RESULTS OF STUDIES OF ALCOHOL IN. TISSUESTable 3REDUCING SUBSTANCES (As ALCoHOL) FoumIED By PUTREFACTION"InitialConc. 2 days 4 days 6 days 8 days 10 days 13 daysSample No.1[Brain 0.00% 0.07% 0.16% 0.29% 0.29% 0.29% 0.35%A* Liver 0.00% 0.01% 0.02% 0.02% 0.11% 0.09% 0.06%[Blood 0.00% 0.01% 0.04% 0.05% 0.06% 0.05% 0.06%rBrain 0.00%. 0.01% 0.04% 0.03% 0.03% 0.02% 0.02%B* Liver 0.00% 0.01% 0.01%" 0.03% 0.02% 0.02% 0.01%IBlood 0.00% 0.01% 0.02% 0.01%q 0.01%Sample No. 2(Brain 0.00% 0.06% 0.12% 0.19% 0.27% 0.31% 0.31%A* Liver 0.00% 0.04% 0.03% 0.04% 0.08% 0.07% 0.05%IBlood 0.00% 0.00% 0.02% 0.02% 0.01% 0.01% 0.03%(Brain 0.00% 0.00% 0.01% 0.02% 0.02% 0.02% 0.02%B* Liver 0.00% D.01% 0.01% 0.02% 0.07% 0.11% 0.14%IBlood 0.00% 0.01% 0.01% 0.00% 0.01%Sample No. 3fBrain 0.00% 0.21% 0.26% 0.34% 0.37% 0.42% 0.34%A* Liver 0.00% 0.05% 0.20% 0.20% 0.14% 0.21% 0.11%[Blood 0.00% 0.04% 0.03% 0.10% 0.13% 0.16% 0.21%(Brain 0.00% 0.01% 0.00% 0.01% 0.01% 0.01% 0.01%B* Liver 0.00% 0.01% 0.01% 0.01% 0.01% 0.01% 0.01%IBlood 0.00% 0.01% 0.01% 0.01% 0.02% 0.01% 0.00%*Group A represents the results on specimens kept at room temperatures, 200 to 26°C;group B the results on specimens kept in the refrigerator at 5C.in the left frontal lobe and the cerebellum. The concentration in thecerebellum was highest in both of these cases. The variations in thedifferent parts of the brain were not deemed to be significant enoughto necessitate the taking of aliquots from the entire minced organ.III. Inasmuch as the work of the Medical Examiner's Office fre-quently involves the examination of bodies in varying stages of putre-faction, a study was undertaken to determine the effects of putrefactionupon the concentration of alcohol as determined by this method.Analyses were conducted on fresh samples of brain, liver, and blood.With the exception of the tissues of sample 5 (Table 4), 50 gramportions of each of the aforementioned tissues were placed in tightlycapped glass jars, half of which were allowed to remain at room tem-peratures of 200C to 26 0 C while the other half were maintained atrefrigerator temperature of 50C. At various intervals of time afterthe original analysis, these specimens were re-analyzed.1951]H. FREIMUTH, M. FOLI TILE AND R. FISHERTable 4REDUCING SUBSTANCES (AS ALCOHOL) FORMED By PUTREFACTIONInitialCone. 2 days 4 days 7 days 9 days 11 days 14 daysSample No. 4[Brain 0.16% 0.39% 0.49% 0.42% 0.41% 0.46% 0.47%A* Liver 0.13% 0.15% 0.14% 0.02% 0.02% 0.11% 0.03%(Blood 0.21% 0.22% 0.22% 0.31% 0.21% 0.16% 0.13%fBrain 0.16% 0.16% 0.15% 0.15% 0.16% 0.14% 0.11%B*{ Liver 0.13% 0.13% 0.12% 0.12% 0.11% 0.12% 0.12%LBlood 0.21% 0.21% 0.19% 0.16% 0.21% 0.20% 0.20%InitialCone. 2 days 5 days 7 days 9 days 12 days 14 daysSample No. 5rBrain 0.22% 0.40% 0.58% 0.57% 0.56% 0.55% 0.47%A*- Liver 0.18% 0.24% 0.19% 0.21% 0.21% 0.18% 0.12%Blood 0.24% 0.20% 0.26% 0.27% 0.19% 0.14% 0.02%rBrain 0.22% 0.22% 0.23% 0.23% 0.23% 0.21% 0.12%B* Liver 0.18% 0.18% 0.20% 0.18% 0.18% 0.18% 0.17%tBlood 0.24% 0.22% 0.21% 0.23% 0.24% 0.23% 0.21%*Group A represents the results on specimens kept at room temperatures, 200 to 26C;group B the results on specimens kept in the refrigerator at 5°C.In the case of sample 5, each of the tissues was minced and dividedinto two portions which were placed in tightly covered jars. One por-tion was then kept at room temperature while the other was kept atrefrigerator temperature. At the indicated intervals of time after theoriginal analysis, 50 gram samples were withdrawn from the con-tainers for analysis.The results of these analyses are shown in Tables 3 and 4. It maybe seen that in all of the tissues kept at room temperatures, a definiteincrease in reducing substances (calculated as ethyl alcohol) occurs,with the increase being most marked with brain tissue. This is truewhether or not the original tissue contained any alcohol.Surprisingly, although the liver shows some increase in reducing sub-stances, it is never as great as that shown by the brain. Consideringthe relatively large carbohydrate stores in the liver, one would expectthat fermentation would result in a higher alcohol production in thattissue than in brain or blood. However, it is known that the livercontains an enzyme which converts ethyl alcohol to acetic acid (8).If this reaction proceeds at a rate greater than the rate of alcoholproduction, it might account for the lower results in the liver.Except for the liver of sample 2 and the brain of sample 5, specimens[Vol. 42RESULTS OF STUDIES OF ALCOHOL IN TISSUESkept under refrigeration showed no appreciable change ih alcohol con-centration even after fourteen days.Much remains to be done in the study of the effects of putrefactionon alcohol production, and it is our intention to investigate further todetermine whether the reducing substances found are, in fact, ethylalcohol. Also, it is hoped to determine whether the same order ofresults would ensue with putrefaction of the intact animal as occurswith tissues removed from the body.In conclusion, the work reported here is a preliminary report of astudy of one of the modifications of the Nicloux procedure for the de-termination of ethyl alcohol in tissues. To summarize the results:(1) The percentage recovery of ethyl alcohol averages 98.1%, thusmaking it unnecessary to make any correction for this factor.(2) There is no significant variation in alcohol distribution in thedifferent areas of the brain.(3) In putrefaction, apparent alcohol production is so great, evenafter two days, as to make the results of an analysis under thesecircumstances completely unreliable.REFERENCES1. Gettler, A. 0. and Tiber, A.: Arch. Path. 3, 218 (1927).2. Gettler, A. 0. and Niederl, J. B., and Benedetti-Pichler, A. A.: J.A.C.S. 54, 1476(1932).3. Gettler, A. O.'and Umberger, C. J.: J. Biol. Chem. 143, 633 (1942).4. Nicloux, M.: Compt. rend. Soc. de Biol. 48, 841 (1896).S. Harger, R. N.: J. Lab. & Clin. Med. 20, 746 (1935).6. Widmark, E.: Biochem. Ztschr. 131, 473 (1922).7. Muehlberger, C. W.: p. 648 in Toxicology by W. D. McNally, Industrial Medicine,Chicago (1937).8. Leloir, L. F. and Munoz, J. M.: Biochem. J. 32, 299 (1938).19511

Results of the Studies on the Determination of Ethyl Alcohol in Tissues

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Results of the Studies on the Determination of Ethyl Alcohol in Tissues

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