Preliminary quantitative investigation of postmortem adipocere formation

The accurate determination of postmortem interval (PMI) using the formation of adipocere presents a significant challenge to forensic scientists interested in determining the time of death. Several attempts have been made to determine the time since the occurrence of death. However, up to date, this has been difficult because previous approaches have been mainly qualitative, focusing on the later stages of degradation processes. This work presents preliminary results of an experimental model of postmortem adipocere formation using liquid chromatography. Three pig cadavers were submerged in distilled water, chlorinated water, and saline water. Fresh specimens resulting from the degradation in the subcutaneous fat were obtained from the pigs at two-week intervals for a period of ten weeks, and were subjected to chromatographic analysis. By correlating the ratio of the disappearance of hydrolyzed fatty acids with the formation of hydroxystearic and oxostearic acids after death, a simple, quantitative analytical method was developed for the determination of PMI. Experimental observation of the chemistry of adipocere formation indicated that adipocere can be formed only a few hours after an incidence of death and this continues until the saturation of oleic acid degradation after several weeks. Different time courses were obtained for cadavers immersed in distilled, chlorinated, and saline water, respectively. This work has not in any way solved the time since death problem. But it may be an approach to the problem that has not been adequately explored.     

A preliminary investigation of the stages of adipocere formation

Adipocere is a postmortem decomposition product which forms from a body's adipose tissue. This study aimed to chemically demonstrate the process of conversion from adipose tissue to adipocere. Samples of adipocere were collected from pig cadavers that were allowed to decompose for varying intervals. Samples of soil were collected from beneath the cadavers and analyzed to determine the leaching effect of adipocere. Gas chromatography/mass spectrometry (GC/MS) was used to quantify the fatty acid composition of pig adipocere. Fourier transform infrared spectroscopy (FTIR) was used as a confirmatory test and to identify other components such as triglycerides and calcium salts of fatty acids. The study demonstrates the process of adipocere formation and the stages of formation through which the process passes using chemical techniques.     

Waxing grave about adipocere: soft tissue change in an aquatic context

When postmortem environmental conditions are "just right," according to the "Goldilocks Phenomenon," soft tissues (and associated fatty acids) are converted into and preserved as adipocere. To better understand this conversion process and the development of adipocere three human cadavers were immersed in outside, water-filled pits for over 3 months to observe adipocere formation in an underwater context simulating actual field conditions. Recordings of environmental conditions showed that temperatures were between 21 degrees C and 45 degrees C, a range sufficient for the growth of Clostridium perfringens. Chemical analysis of liquid and tissue samples revealed an increase in palmitic acid and decrease in oleic acid. This study tracked the remarkable gross morphological changes that can occur in human bodies subjected to an aquatic postmortem environment. The results support the "Goldilocks Phenomenon" and substantiate previous findings that the presence of bacteria and water is crucial for adipocere to form.     

Marine taphonomy: adipocere formation in a series of bodies recovered from a single shipwreck.

Taphonomy of marine environments has been studied mostly from individual cases. The formation of adipocere, or "grave-wax," is an important indicator of the postmortem interval. In the present paper, the conditions and the timing of adipoceric formation are observed in a series of 15 cadavers recovered at different times, over a period of 433 days, from the same contained environment. Initial foci of adipocere on the subcutaneous tissue of the cadavers were detected as early as 38 days from the time of immersion in cold (10-12 degrees C) sea water. The discrepancies between our findings and previous reports on the correlation between time since death and decomposition stages in marine environments are discussed.     

The formation of early stage adipocere in submerged remains: a preliminary experimental study

In some circumstances, the presence of adipocere may retard decomposition and complicate postmortem interval estimation. This article explores the correlation between Accumulated Degree Days (ADD) and early stage formation of adipocere. Sixty wild rabbit (Oryctolagus cuniculus) carcasses were used in this experiment; a control group (N = 30) deposited directly on the ground surface and an experimental group (N = 30) completely submersed in water in individual buckets. Data (water and inner body temperature, pH, and total body score) were collected every 100 ADD. Results indicated that early stage adipocere is correlated to ADD and that its formation on submersed remains is more likely to occur after 630 ADD. Skin sloughing promoted the formation of adipocere. No adipocere was formed on any of the control group rabbits. This study also highlights the fact that multiple factors influence adipocere formation and it is suggested that further research needs to be conducted into this area.     

Evidence of adipocere in a burial pit from the foot and mouth epidemic of 1967 using gas chromatography-mass spectrometry

Gas-chromatography-mass spectrometry was used to characterise the fatty acids from soils and associated tissues excavated from a 1967 Foot and Mouth burial pit. Subcutaneous fats were mainly comprised of 55-75% palmitic acid, 17-22% stearic acid and 3-16% oleic acid as well as 5-7% myristic acid. The distribution of fatty acids confirmed that the tissues were decayed to adipocere. The loss of oleic acid to <3% in two of the decayed fats suggested advanced stages of adipocere. However, adipocere formation was limited in a third tissue sample recovered from greater depth. Inductively coupled plasma atomic emission spectrometry of the pore waters revealed a decrease in Ca concentration and concurrent increase in Na concentrations this suggested that insoluble calcium salt had formed through displacement of sodium. The use of fatty acid profiles from soils and soil interstitial pore waters provide complementary evidence of adipocere formation in foot and mouth burial pits.     

Arid Climate Adipocere—The Importance of Microenvironment

Skeletal remains were retrieved from a cave in an arid desert environment. The submitted material consisted of dried grass, soil, and malodorous skeletal remains wrapped in leather. After examination, it was revealed that the smell had not been due to recent decomposition but to adipocere. The cause of death was listed as “undetermined” and a suggested date of death as “unknown.” Although adipocere usually requires a damp environment or submersion in water to develop, this case provides further evidence for the rare possibility of adipocere formation in an otherwise very dry environment. Wrapping of the body in leather material had produced an impermeable barrier which had sequestrated and preserved bodily fluids allowing anaerobic decay with fat hydrolysis and adipocere formation. Microenvironments may have potentially significant and quite idiosyncratic effects on decompositional processes which may initially confuse investigations.     

Determination of barbiturates in hair samples by using a validated UHPLC-HRMS method: application in investigation of drug-facilitated sexual assault

In recent years,benzodiazepines and benzodiazepine-like drugs are the most common substances associated with drug-facilitated sexual assaults(DFSA);however,barbiturates are also detected occasionally.Segmental hair analysis provides useful information on the historic pattern of drug use,enabling differentiation between single exposure in DFSA cases and chronic use.However,sensitive and specific methods for barbiturate analysis in hair samples are needed.Herein,we present an ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry(UHPLC-HRMS)method for qualitative and quantitative determination of seven barbiturates in hair samples.Firstly,a hair strand was decontaminated and then freeze-milled in liquid nitrogen.Next,50mg of powdered hair was extracted with methanol in an ultrasonic bath for 10 min in the presence of 10 ng phenobarbital-d5.The supernatant was dried under nitrogen gas,and the pellet was dissolved in 100 μL mobile phase.Afterwards,10 μL of the suspension was injected into the UHPLC-HRMS system.The present method involved two UHPLC conditions for determination of barbiturates(I)and identification of the structural isomers amobarbital and pentobarbital(II).This method showed satisfactory linearity in a range of 0.02–20.00 ng/mg for UHPLC conditions I and II,both with a high determination coefficient(0.9991–0.9999).The selectivity,intra-and interday precision,accuracy and matrix effect of the method were acceptable.Next,the validated method was applied to investigate an authentic DFSA case.Hair samples(black,approximate 25cm long)were collected 3 months after the assault,and the proximal segments(0–5 cm from the root;each segment was 1 cm long)were analysed.Amobarbital was detected at a concentration of相似文献   

Adipocere Formation in Subtropical Climate of Northern India: A Retrospective Study

Adipocere formation depends upon multiple environmental factors. In comparison with temperate countries, it usually develops early in the subtropical climate. We have studied a retrospective data of 31 cases with adipocere formation at Department of Forensic Medicine at All India Institute of Medical Sciences, New Delhi. Most of the cases were recovered during the month of May to October from closed rooms at home, open grounds, open forest areas, various water sources, and riverbanks. The time duration of recovery from the time of death was from 12 h to 7 days 12 h. In 10 cases, adipocere formation was seen within 2 days, and in four male cases among them, the adipocere formed within a day. Most of the bodies showing adipocere formation within 2 days were recovered from land. These facts showed that subtropical climate having hot and humid weather promotes early adipocere formation compared to temperate climate.     

The Initial Changes of Fat Deposits During the Decomposition of Human and Pig Remains

Abstract:  The early stages of adipocere formation in both pig and human adipose tissue in aqueous environments have been investigated. The aims were to determine the short-term changes occurring to fat deposits during decomposition and to ascertain the suitability of pigs as models for human decomposition. Subcutaneous adipose tissue from both species after immersion in distilled water for up to six months was compared using Fourier transform infrared spectroscopy, gas chromatography-mass spectrometry and inductively coupled plasma-mass spectrometry. Changes associated with decomposition were observed, but no adipocere was formed during the initial month of decomposition for either tissue type. Early-stage adipocere formation in pig samples during later months was detected. The variable time courses for adipose tissue decomposition were attributed to differences in the distribution of total fatty acids between species. Variations in the amount of sodium, potassium, calcium, and magnesium were also detected between species. The study shows that differences in total fatty acid composition between species need to be considered when interpreting results from experimental decomposition studies using pigs as human body analogs.     

Postmortem chest computed tomography for the diagnosis of drowning:a feasibility study

It may be difficult to distinguish the cause of death in drowning cases without specific findings. The aim of this study was to explore the forensic value of thoracic postmortem computed tomography (PMCT) using routine images and three-dimensional (3D) image reconstructions. The imaging data of PMCT examinations of six drowning cadavers, aged 21–54 years, were analyzed. Twelve victims of sudden death from coronary artery disease (CAD) were chosen as a control group. After 3D bilateral lung images were reconstructed using image processing software, an interactive medical image control system was used to measure and analyze parameters including lung volume, lung volume ratio, mean CT value of the whole lung, and lung CT value distribution curves. Lung volume and lung volume ratio were used to assess the shape changes of the lung. Lung CT value distribution curves showed the corresponding number of pixels of the different CT values in the lung image. Lung volume was not significantly larger in drowning cases (mean 2 958 cm³) than in controls (mean 2 342 cm³). Lung volume ratio values in the drowning group (mean 0.3156) were greater than those in the control group (mean 0.2763); (P = 0.02). There was no significant difference between the drowning and control group in the mean CT value of the whole lung. There were differences between lung CT value distribution curves in drowning victims and controls, with drowning victims showing a single peak and CAD cases showing a bimodal distribution. Thoracic PMCT is helpful for the forensic medical diagnosis of drowning. Lung volume ratio and lung CT value distribution are potential indicators to distinguish between drowning and CAD.     

The effect of the burial environment on adipocere formation

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.     

The effect of soil type on adipocere formation

Adipocere refers to a postmortem product which forms from body fat in the later stages of decomposition. Factors present in the surrounding decomposition environment will influence adipocere formation and may accelerate or retard the process of conversion. One such factor important in burial environments is the type of grave soil in which the burial has taken place. This study was conducted to investigate the influence of various soil types on the formation of adipocere in grave soils. X-ray diffraction and particle size analysis were used to characterise the soils which were essentially chosen on the basis of grain size. Infrared spectroscopy, inductively coupled plasma-mass spectrometry, and gas chromatography-mass spectrometry were used to investigate the lipid profile and chemical composition of adipocere developed from decomposing tissue. The results suggest that adipocere is able to form in various soil types and that particular soil environments may accelerate its formation.     

大鼠死后肌动蛋白降解与死亡时间的相关性

目的观察大鼠死后不同时间心肌、肝、脾、肺、肾、脑和骨骼肌肌动蛋白（actin）的降解规律,为晚期死亡时间（postmortem interval,PMI）的推断寻找客观指标。方法28只SD大鼠随机分为7组,经机械性窒息致死后．分别置于20℃恒温箱内,于0、24、48、72、96、120和168h后剖取以上器官,应用免疫印迹技术检测各组织内actin的含量,SPSS11．5软件对所得数据进行方差分析。结果大鼠死后上述各器官actin含量均随PMI的延长而逐渐下降,与PMI相关,决定系数（R2）均在0．75以上。但actin在上述各器官内的降解速度有显著差异（P〈O．05）,在脑组织中降解速度最快,其余依次为肺、脾、肝、肾、心肌和骨骼肌。结论尽管大鼠死后心肌、肝、脾、肺、肾、脑和骨骼肌actin的降解规律具有组织差异性,但是actin在上述组织中的含量变化与PMI相关．有助于较晚期PMI的推断。     

Preservation of human tissue immersed for five years in fresh water of known temperature

Two human bodies were recovered from the waters of the Duluth, Minnesota harbor. Extensive adipocere formation resulted in remarkable preservation of gross anatomic features of internal organs. Total time of immersion could be precisely estimated at five years. Water temperature during those five years could also be accurately estimated by direct measurements taken during the year following recovery of the bodies and from information supplied by a local electric power generating company. Immersion occurred at the time of the year when water temperature was highest (70 degrees F [21 degrees C]) facilitating the rapid formation of adipocere. A proposed mechanism for formation of adipocere is described.     

Experimental adipocere formation: implications for adipocere formation on buried bone

Adipocere, or grave wax (adipo = fat, cere = wax), is a distinctive decomposition product composed primarily of fatty acids (FA) and their alkali salts. FA result from the bacterial enzymatic hydrolysis of body fats. Reactions with ammonia and alkali metals originating from body fluids and pore waters of the depositional environment produce alkali salts of FA (soap). Adipocere formation is generally associated with burial of corpses with ample adipose tissue available. No indications that adipocere can form on defleshed remains have been presented in the literature. At the termination of a long-term bone diagenesis experiment, several samples were found to possess growths of an unknown compound. Gas chromatography-mass spectrometry confirmed that the growths are adipocere. The results herein reveal that adipocere can indeed form on defleshed bones under the right conditions and that even residual adipose and lipids in defleshed bones are sufficient to produce adipocere growth on the surfaces of bone.     

Identification of a disinterred grave by molecular and stable isotope analysis

Confirmation of a potential disinterred grave was sought by GC and GC/MS analyses of lipid extracts of whole soils and white particulate matter. Fatty acid profiles and concentrations determined for three of the soils correlated with the deposition of a large amount of exogenous organic matter, most likely adipocere and/or decomposed body fat. Determination of C_16:0 and C_18:0 fatty acid δ¹³C values by GC/C/IRMS revealed the input to be isotopically distinct from common British domesticated animals, plotting closely to values determined for adipose fat obtained from of a murder victim. By considering the difference between δ¹³C values (Δ¹³C_18:0–16:0) a potential isotopic proxy for identifying the source of adipocere (human) and adipose tissue was proposed.     

The effect of body coverings on the formation of adipocere in an aqueous environment

Adipocere is a postmortem decomposition product that consists of a mixture of fatty acids. The rate of formation of adipocere from pig adipose tissue in an aqueous environment has been monitored. The effect of various clothing and carpet material types on the process was investigated. The fatty acid composition of the adipocere was determined at regular intervals using gas chromatography-mass spectrometry. Examination of the changes to fatty acid concentrations allowed the degree of adipocere formation in the different environments to be estimated. The study demonstrated that the rate at which adipocere forms is particularly accelerated by the presence of coverings produced from natural materials. Elemental analysis by inductively coupled plasma-mass spectrometry revealed, for the most part, little change to the cations present in the adipocere formed. However, an increase in Ca concentration was observed for tissue wrapped in acrylic carpet, which was associated with a CaCO(3) additive used in the carpet manufacture.     

Free Fatty Acids Composition in Adipocere of the Kwäday Dän Ts’ìnchí Ancient Remains Found in a Glacier*

Abstract: Adipocere is a postmortem decomposition product consisting of mostly a mixture of free fatty acids (FFAs) that are formed because of the hydrolysis of triglycerides in adipose tissues. This article describes a simple and robust method for the extraction, identification, and quantification of FFA commonly found in adipocere using gas chromatography–mass spectrometry (GC/MS). This method was applied to analyze tissues from Kwäday Dän Ts’ìnchí, ancient remains discovered in a retreating glacier in the Tatshenshini‐Alsek Park, British Columbia, Canada in August 1999. The lyophilized tissues were grinded and extracted with hexane. The trimethylsilyl fatty acid derivatives were analyzed by GC/MS, and the relative abundances of myristic acid, palmitic acid, oleic acid, and stearic acid were determined. Milligram per gram levels of saturated fatty acids were found in the tissues of the ancient remains, while the levels of unsaturated fatty acids, such as palmitoleic acid, were found to be negligible. The results provided further evidence of the existence of adipocere found during forensic examination of the Kwäday Dän Ts’ìnchí ancient remains.     

Identification of decomposition volatile organic compounds from surface-deposited and submerged porcine remains

Cadaver dogs are routinely used internationally by police and civilian search organisations to locate human remains on land and in water, yet little is currently known about the volatile organic compounds (VOCs) that are released by a cadaver underwater; how this compares to those given off by a cadaver deposited on land; and ultimately, how this affects the detection of drowned victims by dogs. The aim of this study was to identify the VOCs released by whole porcine (Sus scrofa domesticus) cadavers deposited on the surface and submerged in water using solid phase microextraction gas chromatography mass spectrometry (SPME GC–MS) to ascertain if there are notable differences in decomposition odour depending on the deposition location.For the first time in the UK, the volatile organic compounds (VOCs) from the headspace of decomposing porcine cadavers deposited in both terrestrial and water environments have been detected and identified using SPME-GCMS, including thirteen new VOCs not previously detected from porcine cadavers. Distinct differences were found between the VOCs emitted by porcine cadavers in terrestrial and water environments. In total, seventy-four VOCs were identified from a variety of different chemical classes; carboxylic acids, alcohols, aromatics, aldehydes, ketones, hydrocarbons, esters, ethers, nitrogen compounds and sulphur compounds. Only forty-one VOCs were detected in the headspace of the submerged pigs with seventy detected in the headspace of the surface-deposited pigs. These deposition-dependent differences have important implications for the training of cadaver dogs in the UK. If dog training does not account for these depositional differences, there is potential for human remains to be missed.Whilst the specific odours that elicit a trained response from cadaver dogs remain unknown, this research means that recommendations can be made for the training of cadaver dogs to incorporate different depositions, to account for odour differences and mitigate the possibility of missed human remains operationally.