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The Effect of the Burial Environment on Adipocere Formation Shari L Forensic Science International 154 (2005) 24–34 www.elsevier.com/locate/forsciint The effect of the burial environment on adipocere formation Shari L. Forbesa,*, Barbara H. Stuartb, Boyd B. Dentc aCentre for Forensic Science, University of Western Australia, 35 Stirling Highway, Crawley 6009, Australia bDepartment of Chemistry, Materials and Forensic Science, University of Technology, PO Box 123, Broadway 2007, Sydney, Australia cDepartment of Environmental Sciences, University of Technology, PO Box 123, Broadway 2007, Sydney, Australia Received 4 November 2003; accepted 15 September 2004 Available online 10 November 2004 Abstract Adipocere is a decomposition product comprising predominantly of saturated fatty acids which results from the hydrolysis and hydrogenation of neutral fats in the body. Adipocere formation may occur in various decomposition environments but is chiefly dependent on the surrounding conditions. In a soil burial environment these conditions may include such factors as soil pH, temperature, moisture and the oxygen content within the grave site. This study was conducted to investigate the effect of these particular burial factors on the rate and extent of adipocere formation. Controlled laboratory experiments were conducted in an attempt to form adipocere from pig adipose tissue in model burial environments. Infrared spectroscopy, inductively coupled plasma–mass spectrometry, and gas chromatography–mass spectrometry were employed to determine the lipid profile and fatty acid composition of the adipocere product which formed in the burial environments. The results suggest that adipocere can form under a variety of burial conditions. Several burial factors were identified as enhancing adipocere formation whilst others clearly inhibited its formation. This study acts as a preliminary investigation into the effect of the burial environment on the resultant preservation of decomposing tissue via adipocere formation. # 2004 Elsevier Ireland Ltd. All rights reserved. Keywords: Adipocere; Fatty acids; Burial environment; Soil pH; Temperature; Moisture; Oxygen content 1. Introduction formation refers to the process of conversion of body fat into a solid white substance [4]. The process of decay of human remains commences Adipocere is a postmortem decomposition product which almost immediately following death and will usually pass forms from the fatty tissue in a body. Adipose tissue through a number of stages including autolysis and putre- comprises mainly triglycerides which during decomposition faction. Putrefaction can either proceed until the soft tissue undergo hydrolysis to yield free fatty acids [4]. Hydrogena- has liquefied and the cadaver becomes skeletonised [1],orit tion of the free fatty acids will yield saturated fatty acids can become arrested by later postmortem changes such as which, under the right conditions, form into adipocere [5]. mummification or adipocere formation [2]. Mummification Adipocere is comprised chiefly of the saturated fatty acids, refers to the desiccation and preservation of soft tissue and myristic, palmitic and stearic acids. Unsaturated fatty acids usually occurs in hot, dry environments [3]. Adipocere [6,7], salts of fatty acids [8] and hydroxy- and oxo-fatty acids [6,9] have all been identified as constituents of adipocere. * Corresponding author. Tel.: +61 8 6488 3408; Adipocere formation is of interest to forensic scientists as fax: +61 8 6488 7285. it has the ability to preserve the remains it encases by E-mail address: [email protected] (S.L. Forbes). inhibiting decomposition. The degree of decomposition 0379-0738/$ – see front matter # 2004 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.forsciint.2004.09.107 S.L. Forbes et al. / Forensic Science International 154 (2005) 24–34 25 and differential preservation observed depends on the sur- 2. Materials and methods rounding environment [10]. In a burial environment, a number of factors may inhibit bacterial growth, thus retard- 2.1. Adipocere formation ing decomposition and setting up favourable preservation conditions. The nature of the soil and sediments has been In order to determine the effect of particular burial investigated [2,10] and its affect on adipocere formation conditions on adipocere formation, experiments were reported previously by the authors [11]. Hence, whilst soil conducted in a laboratory environment so that the indi- type is an important burial factor, it is not considered in this vidual variables could be adequately controlled. The study. experiments were conducted in large decomposition con- The soil pH is important, as a highly acidic or highly tainers which included a tap for draining purposes and an alkaline environment will inhibit bacterial growth and thus, airlock seal for the release of gases present within the decomposition. The effect of pH on adipocere formation has container. Due to ethical restrictions within Australia, the been studied by Mant but a contradictory result was use of human adipose tissue was not a viable option for obtained. In Mant’s laboratory experiments, the pH of the this study. Current considerations suggest that decompos- fluid in which the fat decomposed fell rapidly to produce an ing pigs are the next most reliable model [16,17] and this acidic environment in which adipocere formed [4].How- study therefore utilised domestic pigs which were reared ever, his field studies identified adipocere formation on for commercial use. Pig adipose tissue collected from the bodies in sealed coffins demonstrating a strong ammoniacal abdominal region and still containing muscle and skin was odour and presumably an alkaline environment [2]. used for each experiment. To ensure adipocere formation The temperature of the burial environment can have a occurred, each experiment included those factors known significant effect on decomposition as most bacteria thrive at to enhance adipocere formation, namely sufficient adipose an optimal temperature of approximately 37 8C [12,13]. Soil tissue, moisture, bacteria, and a relatively anaerobic envir- temperatures are typically much lower than this and bacterial onment. A control environment was created whereby the growth may be inhibited. Putrefactive activity is greatly tissue was buried within a damp soil containing a loamy inhibited at temperatures below 10 8C or above 40 8C sand composition with pH approximately 5.2. Additional [12]. Such variations in temperature may be experienced environments were prepared in the same manner with the in a shallow burial environment and will ultimately affect the individual variable of interest altered accordingly. The extent of decomposition and preservation which occurs. experiment was conducted in a temperate environment The oxygen content of the burial environment is likely to which averaged approximately 22 8C over a 12-month affect the rate of decomposition as the aerobic bacteria and period. fungi often found on decomposing remains require sufficient The factors investigated in this study were pH (including oxygen to survive and proliferate. An environment with the presence of lime), temperature, moisture and oxygen minimal air circulation tends to delay decomposition and content. The pH of the burial environment was altered to promote preservation [12,14]. Several studies have pre- investigate both acidic and alkaline environments. A highly viously reported the requirement of an anaerobic environ- acidic environment (pH approximately 2.4) was created ment for the successful formation of adipocere [5,15]. through the addition of 10 M hydrochloric acid (HCl) to The formation of adipocere requires moisture for the the soil. A mildly alkaline environment (pH approximately hydrolytic conversion of triglycerides to free fatty acids. It 8.5) was produced through the addition of 5 M sodium was previously believed that adipocere formation could only hydroxide (NaOH) to the soil whilst the highly alkaline occur in large bodies of water or water-logged environments. environment (pH approximately 12.6) was created by cover- Studies have since shown that adipocere can form by ing the pig tissue in lime. The latter environment was utilising the moisture present within the body’s tissues included due to its occurrence in forensic situations whereby [2,4,10]. Hence, the presence of adipocere in a burial lime is often used to hide the odour and alter the rate of environment is not unusual even in a dry, soil environment. decomposition [18]. The pH values were determined using The factors described represent some of the more com- calibrated meters for measuring pH. mon conditions present in a burial environment which may A variation in temperature was explored by investigating affect decomposition and preservation of human remains. the extremes of cold and warm environments. A cold Whilst the effect of these conditions on decomposition in environment was achieved by placing the decomposition soil has been studied in detail, their effect on adipocere containers in a refrigerated environment held at 4 8C. A formation in a burial environment is mostly based on warm environment was created by placing the containers in a observation. This study was conducted to experimentally large water bath maintained at 40 8C. Both environments investigate the burial factors of soil pH, temperature, moist- mimicked extremes in temperature variation when compared ure and oxygen content, with regard to their effect on to the mild temperature of the control environment. The adipocere formation. The results are useful in
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