Skin simulants for wound ballistic investigation - an experimental study.

Gunshot wound analysis is an important part of medicolegal practice, in both autopsies and examinations of living persons. Well-established and studied simulants exist that exhibit both physical and biomechanical properties of soft-tissues and bones. Current research literature on ballistic wounds focuses on the biomechanical properties of skin simulants. In our extensive experimental study, we tested numerous synthetic and natural materials, regarding their macromorphological bullet impact characteristics, and compared these data with those from real bullet injuries gathered from medicolegal practice. Over thirty varieties of potential skin simulants were shot perpendicularly, and at 45°, at a distance of 10 m and 0.3 m, using full metal jacket (FMJ) projectiles (9 × 19 mm Luger). Simulants included ballistic gelatine at various concentrations, dental silicones with several degrees of hardness, alginates, latex, chamois leather, suture trainers for medical training purposes and various material compound models. In addition to complying to the general requirements for a synthetic simulant, results obtained from dental silicones shore hardness 70 (backed with 20 % by mass gelatine), were especially highly comparable to gunshot entry wounds in skin from real cases. Based on these results, particularly focusing on the macroscopically detectable criteria, we can strongly recommend dental silicone shore hardness 70 as a skin simulant for wound ballistics examinations.

RNA Analysis in Forensic Molecular Biology.

BACKGROUND: Different types of RNA take on multiple crucial functions in living cells and tissues. Messenger RNA (mRNA) is a temporary molecular carrier of genetic information. Analysis of the composition of all mRNA contained in a cell at a given moment, the so-called transcriptome, enables the determination of the type of cell and its condition, e.g., in pathologically altered states. METHODS: This review is based on pertinent publications retrieved by a selective literature search. RESULTS: The analysis of differential gene expression has already been used in forensic molecular biology to determine the type of tissue contained in biological specimens. It is also being used in criminal investigations to determine the composition of mixed traces of various bodily fluids and/or organ tissues. The method is limited by degradation of the mRNA molecules through environmental influences. The use of newly developed molecular biological methods such as massive parallel sequencing can expand the information obtainable by this investigative method. Current research also addresses the forensic potential of deriving relevant information about the crime-e.g., its timing, or the condition of the involved persons-from the totality of mRNA species present in the specimens. CONCLUSION: Forensic RNA analysis can yield a great deal of relevant information. It is likely to be applicable in a much wider variety of forensic situations in the near future.

Phase I-metabolism studies of the synthetic cannabinoids PX-1 and PX-2 using three different in vitro models.

PURPOSE: Synthetic cannabinoids (SCs), highly metabolized substances, are rarely found unmodified in urine samples. Urine screening relies on SC metabolite detection, requiring metabolism knowledge. Metabolism data can be acquired via in vitro assays, e.g., human hepatocytes, pooled human liver microsomes (pHLM), cytochrome P450 isoforms and a fungal model; or in vivo by screening, e.g., authentic human samples or rat urine. This work describes the comprehensive study of PX-1 and PX-2 in vitro metabolism using three in vitro models. 5F-APP-PICA (PX-1) and 5F-APP-PINACA (PX-2) were studied as they share structural similarity with AM-2201, THJ-2201 and 5F-AB-PINACA, the metabolism of which was described in the literature. METHODS: For SC incubation, pHLM, cytochrome P450 isoenzymes and the fungal model Cunninghamella elegans LENDNER (C. elegans) were used. PX-1 and PX-2 in vitro metabolites were revealed comprehensively by liquid chromatography-high-resolution mass spectrometry measurements. RESULTS: In total, 30 metabolites for PX 1 and 15 for PX-2 were detected. The main metabolites for PX-1 and PX-2 were the amide hydrolyzed metabolites, along with an indole monohydroxylated (for PX-1) and a defluorinated pentyl-monohydroxylated metabolite (for PX-2). CONCLUSIONS: CYP isoforms along with fungal incubation results were in good agreement to those obtained with pHLM incubation. CYP2E1 was responsible for many of the metabolic pathways; particularly for PX-1. This study shows that all three in vitro assays are suitable for predicting metabolic pathways of synthetic cannabinoids. To establish completeness of the PX-1 and PX-2 metabolic pathways, it is not only recommended but also necessary to use different assays.

Detectability of oxandrolone, metandienone, clostebol and dehydrochloromethyltestosterone in urine after transdermal application.

Situations of both, intentional and inadvertent or accidental doping, necessitate consideration in today's doping controls, especially in the light of the substantial consequences that athletes are facing in case of so-called adverse analytical findings. The aim of this study was to investigate, whether a transdermal uptake of doping substances would be possible. In addition to the period of detectability of the particular substances or respective characteristic metabolites, the possibility of deducing the route of administration by metabolite patterns was also assessed. Twelve male subjects were included in the study. Four common anabolic androgenic steroids (AAS) were dissolved in dimethylsulfoxide to facilitate transdermal administration on different skin regions. One half of the test persons received only oxandrolone (17α-methyl-2-oxa-4,5α-dihydrotestosterone), and the other half were applied a mixture of oxandrolone, metandienone (17ß-hydroxy-17α-methylandrosta-1,4-dien-3-one), clostebol (4-chlorotestosterone-17ß-acetate) and dehydrochloromethyltestosterone (DHCMT). Urine samples were collected 1 h, 6 h and one sample per day for the next 14 consecutive days. Measurements were conducted on a tandem-gas chromatography-mass spectrometry (GC-MS/MS) or tandem-liquid chromatography-MS/MS (LC-MS/MS) system. Substance findings were obtained at least 1 day after application on nearly all skin locations. The results indicated inter-individual variability in detection windows, also varying between the different analytes and possible impact of skin location and skin thickness, respectively. Nevertheless, a rapid and rather long detectability of all substances (or respective metabolites) was given, in some cases within hours after administration and for up to 10-14 days. Hence, the transdermal application or exposure to the investigated AAS is a plausible scenario that warrants consideration in anti-doping.

Fixed-Wing Motorized Aircraft Accidents: Analysis of Injury Severity and Concomitant Factors (2000-2019).

BACKGROUND: There is a paucity of research on general aviation accidents in Germany. The authorities investigate only a fraction of all national accidents. The current study analyzes existing accident reports and aims to identify injury severity in regard to concomitant risk factors.METHODS: Data of flight accidents was analyzed for aircraft of <5700 kg maximum takeoff weight (MTOW) over a 20-yr period. Besides descriptive data, concomitant factors (type and category of aircraft, date, occupants and outcome, flight phase, etc.) were analyzed. Statistical analysis was performed using the Chi-squared test.RESULTS: The authorities list 1595 aircraft accidents between 2000 and 2019, but only 17.9% of these were analyzed in detail. Accidents of aircraft of <2000 kg MTOW were over-represented between May and September and between Friday and Sunday. The fraction of fatal accidents was highest during cruise. During landing, significantly more mishaps of larger aircraft occurred. The number of seriously injured or deceased occupants was significantly higher for accidents involving private pilots. An occupancy rate of more than three persons on board correlated significantly with fewer number of deaths.CONCLUSIONS: The annual count of aircraft accidents has almost halved during the previous 20 yr. Unfortunately, only a small number of mishaps were further investigated by authorities, which leads to a lack of evaluable data needed for in-depth investigations. The accumulation of larger aircraft mishaps in winter and the superior outcome of professional pilots in terms of safety, as well as the fewer number of mishaps in larger aircraft, should be further investigated.Liebold F, Hippler C, Schmitz J, Yücetepe S, Rothschild M, Hinkelbein J. Fixed-wing motorized aircraft accidents: analysis of injury severity and concomitant factors (2000-2019). Aerosp Med Hum Perform. 2022; 93(5):442-449.

Variant interpretation in molecular autopsy: a useful dilemma.

Sudden cardiac death (SCD) in adolescents and young adults may be the first manifestation of an inherited arrhythmic syndrome. Thus identification of a genetic origin in sudden death cases deemed inconclusive after a comprehensive autopsy and may help to reduce the risk of lethal episodes in the remaining family. Using next-generation sequencing (NGS), a large number of variants of unknown significance (VUS) are detected. In the majority of cases, there is insufficient evidence of pathogenicity, representing a huge dilemma in current genetic investigations. Misinterpretation of such variants may lead to inaccurate genetic diagnoses and/or the adoption of unnecessary and/or inappropriate therapeutic approaches. In our study, we applied current (ACMG) recommendations for variant classification in post-mortem genetic screening of a cohort of 56 SCD victims. We identified a total 53 rare protein-altering variants (MAF < 0.2%) classified as VUS or worse. Twelve percent of the cases exhibited a clinically actionable variant (pathogenic, likely pathogenic or VUS - potentially pathogenic) that would warrant cascade genetic screening in relatives. Most of the variants detected by means of the post-mortem genetic investigations were VUS. Thus, genetic testing by itself might be fairly meaningless without supporting background data. This data reinforces the need for an experienced multidisciplinary team for obtaining reliable and accountable interpretations of variant significance for elucidating potential causes for SCDs in the young. This enables the early identification of relatives at risk or excludes family members as genetic carriers. Also, development of adequate forensic guidelines to enable appropriate interpretation of rare genetic variants is fundamental.

Anisotropic and age-dependent elastic material behavior of the human costal cartilage.

Compared to articular cartilage, the biomechanical properties of costal cartilage have not yet been extensively explored. The research presented addresses this problem by studying for the first time the anisotropic elastic behavior of human costal cartilage. Samples were taken from 12 male and female cadavers and unconfined compression and indentation tests were performed in mediolateral and dorsoventral direction to determine Young's Moduli EC for compression and Ei5%, Ei10% and Eimax at 5%, 10% and maximum strain for indentation. Furthermore, the crack direction of the unconfined compression samples was determined and histological samples of the cartilage tissue were examined with the picrosirius-polarization staining method. The tests revealed mean Young's Moduli of EC = 32.9 ± 17.9 MPa (N = 10), Ei5% = 11.1 ± 5.6 MPa (N = 12), Ei10% = 13.3 ± 6.3 MPa (N = 12) and Eimax = 14.6 ± 6.6 MPa (N = 12). We found that the Young's Moduli in the indentation test are clearly anisotropic with significant higher results in the mediolateral direction (all P = 0.002). In addition, a dependence of the crack direction of the compressed specimens on the load orientation was observed. Those findings were supported by the orientation of the structure of the collagen fibers determined in the histological examination. Also, a significant age-related elastic behavior of human costal cartilage could be shown with the unconfined compression test (P = 0.009) and the indentation test (P = 0.004), but no sex effect could be detected. Those results are helpful in the field of autologous grafts for rhinoplastic surgery and for the refinement of material parameters in Finite Element models e.g., for accident analyses with traumatic impact on the thorax.

Insights to enhance the examination of tool marks in human cartilage.

This work deals with the examination of tool marks in human cartilage. We compared the effectiveness of several cleaning methods on cut marks in porcine cartilage. The method cleaning by multiple casts achieved the significantly highest scores (P = 0.02). Furthermore, we examined the grain-like elevations (dots) located on casts of cut cartilage. The results of this study suggest that the casting material forms these dots when penetrating cartilage cavities, which are areas where the strong collagen fibres leave space for the chondrocytes. We performed fixation experiments to avoid this, without success. In addition, 31 casting materials were compared regarding contrast under light-microscope and 3D tool marks scanner. Under the light-microscope, brown materials achieved significantly higher values than grey (P = 0.02) or black (P = 0.00) whereas under the 3D scanner, black materials reached higher contrast values than grey (P = 0.04) or brown (P = 0.047). To compare the accuracy and reproducibility of 6 test materials for cartilage, we used 10 knives to create cut marks that were subsequently scanned. During the alignment of the individual signals of each mark, the cross-correlation coefficients (Xmax) and lags (LXmax) were calculated. The signals of the marks in agarose were aligned with significantly fewer lags and achieved significantly higher cross-correlation coefficients compared to all tested materials (both P = 0.00). Moreover, we determined the cross-correlation coefficients (XC) for known-matches (KM) per material. Agarose achieved significantly higher values than AccuTrans®, Clear Ballistics™, and gelatine (all P = 0.00). The results of this work provide valuable insights for the forensic investigation of marks in human costal cartilage.

Anatomical In Vitro Investigations of the Pediatric Larynx: A Call for Manufacturer Redesign of Tracheal Tube Cuff Location and Perhaps a Call to Reconsider the Use of Uncuffed Tracheal Tubes.

BACKGROUND: Some in vivo studies question the traditional "funnel-shaped" infant larynx; further anatomic examinations were warranted. Examination of fixative free fresh autopsy laryngeal and upper tracheal specimens and multiple measurements was needed to determine consistency between current tracheal tube designs and anatomic observations. METHODS: Larynges from 19 males and 11 females (Caucasian term newborn to 126 months) were examined by the same forensic pathologist. Measurements included anterior/posterior (A/P) and transverse (T) diameters of the cricoid outlet (CO), interarytenoid diameter (IAD), cricothyroid membrane (CTM), distance from the vocal cords (VC) to CO (VC-CO), and calibration of the larynx lumen with uncuffed tracheal tubes as measuring rods. Assessment of "safe tracheal tube placement" was assessed using manufacturer recommended cuffed Microcuff (Kimberly-Clark, Koblenz, Germany) tubes. RESULTS: In 77% (95% confidence interval [CI], 58-90) of specimens, the proximal end of the cuff was within the CO and in 23% even with or close to the CO. The VC-CO varied from 9.1 to 13.17 mm in infants, 11.55 to 15.17 mm in toddlers, and 13.19 to 18.34 mm in children. The A-P/T ratio of the CO was nearly 0.99 in most larynges; the IAD was greater than CO in all specimens. The CTM could be minimally distended in all specimens. CONCLUSIONS: First, despite being marketed as a safer tracheal tube design, the proximal end of the Microcuff cuff rested within or close to the cricoid cartilage theoretically increasing potential cuff-induced injury when using the VC markings for positioning. Our data suggest that the optimal cuff free distance (VC-CO) would be ~13.5 mm for a Microcuff internal diameter (ID) size 3.0, ~15 mm for size 3.5, and ~16 to 19 mm for greater sizes.Second, the CO was virtually circular in all specimens, suggesting that appropriately sized uncuffed tubes should provide an adequate seal in most neonates and toddlers, thus avoiding the potential for cuff-related necrosis injury.Third, the IAD was always greater than CO confirming that the narrowest point of the infant larynx is the nondistensible cricoid cartilage and not the easily distended glottis.Fourth, appropriately sized Microcuff tubes with the cuff deflated completely filled the lumen of the CO and proximal trachea in all specimens. Our data suggest the need for all manufacturers to further evaluate tracheal tube cuff locations and lengths in relation to the VC safe insertion markings, particularly for neonates and toddlers.Fifth, the CTM is minimally distensible, thus having important implications for emergency surgical airway access with most currently available emergency airway devices.

Evidence for the transfer of methadone and EDDP by sweat to children's hair.

In cases where there is a question as to whether children have come into contact with drugs, examinations of their scalp hair are frequently carried out. Positive test results are often discussed in the forensic community due to the various possible modes via which drugs and their metabolites can be incorporated into the hair. These include drug uptake by the child (e.g. oral ingestion or inhalation), but also contamination of hair via contact with the sweat from drug users. In this study, the possibility of methadone and its metabolite EDDP being incorporated into children's hair by contact with sweat from persons undergoing opiate maintenance therapy (methadone) was examined. The transfer of methadone and EDDP via sweat from methadone patients (n = 15) to children's hair was simulated by close skin contact of drug-free children's hair, encased in mesh-pouches, for 5 days. Sweat-collecting patches (hereafter referred to as 'sweat patches') were applied to the test persons' skin. One strand of hair and one sweat patch were collected daily from each patient. Analyses were performed using GC-MS/MS (hair) and LC-MS/MS (serum, sweat patches). After 4 days of skin contact, methadone was detectable in the formerly drug-free hair strands in all 15 study participants. EDDP was detectable in 34 of 75 hair strands, with the maximum number of positive results (11 EDDP-positive hair strands) being detected after 5 days. These results show that transfer of methadone and EDDP to drug-free hair is possible through close skin contact with individuals taking part in methadone substitution programmes. A correlation between serum concentration, sweat concentration and substance concentration in hair strands could not be demonstrated, but a tendency towards higher concentrations due to longer contact time is clearly evident.

Sharp force trauma with two katana swords: identifying the murder weapon by comparing tool marks on the skull bone.

This paper describes the variety of information that a tool mark analysis on human tissue can provide based on a case of multiple sharp violence. The perpetrator attacked the victim with a sharp-edged weapon against the head, leaving several deep wounds on the back of the skull bone. Three of those marks on the skull bone could be used for a forensic tool mark examination. Silicone casts of the marks were compared by light microscopy with casts of test marks of Japanese katana swords found at the crime scene. One of the swords could be identified as the one responsible for the marks. In addition, the marks and the test marks were scanned in 3D and examined in a visual on-screen comparison confirming the results from the light microscopic examination. Furthermore, a mathematical approach in which the signatures of the marks from the skull bone and the test marks from the sword were compared by cross correlation confirms those findings. In addition, the aforementioned results were used to determine the orientation of the sword in relation to the cranial bone at the time of the respective impact.

Sudden death of a young adult with coronary artery vasculitis, coronary aneurysms, parvovirus B19 infection and Kawasaki disease.

We present a case of a 20-year-old man who suffered from Kawasaki disease (KD) associated with a florid parvovirus infection, and who died suddenly from thrombotic occlusion of the coronary arteries. The autopsy revealed several aneurysms of the coronary arteries, a chronic vasculitis and a myofibroblast proliferation leading to focal luminal narrowing. The inflammatory response as well as the detection of the viral particles by PCR in blood and in the lesional tissue demonstrated a possible cause by Parvovirus infection. The expression of endoglin on endothelial cells of neoangiogenesis indicates the involvement of the TGF-beta pathway, necessary for maintaining chronic inflammation. In addition, a possible connection between the intake of methylphenidate, arteritis and a possible pre-existing heart disease must be discussed. Furthermore, KD must also be considered as a cause of sudden death in the adult population.

Studies on acceptance, evaluation and impact of the Cologne program "Research and Medical Studies".

Introduction: The curricular implementation of events (or programs) for science-related training in human medicine has been on the agenda of the medical faculties since the publication of the Federal-State Working Group [1]. The Medical Faculty of the University of Cologne developed and established a systematic, longitudinal science curriculum together with the start of the model curriculum in human medicine in 2003. Here, we investigate the questions of whether the described (para-) curricular elements are accepted by students and lecturers and how they are evaluated, especially by students. In addition, we investigate whether selected parameters can be used to demonstrate changes in the students' scientific activities. Project description: The program "Research and Medical Studies" (RaMS) consists of several components: these elements of the mandatory curricular (Scientific Projects, SP) and optional components (Research in Medical Studies (RiMS), Research Track (RT), Research Fair Cologne (RFC)) are described here. Results were recorded at various levels: Likert Scale evaluation of the event's elements were collected as satisfaction parameters from the studentsProcess data on participation in the voluntary events were collected and evaluated as absolute and relational figures (WS 12/13-SS 17). Data on the outcome of the RaMS program were collected: Type of scientific projects in the academic years 2011/12-2014/15), number and type of available projects offered at the RFC (in the years 2011-18) and number of student research funding applications in a comparison of the periods 2010-13 vs. 2014-17). Results: The students' acceptance of mandatory and paracurricular courses of the RaMS program is pleasingly high, which is not surprising, at least in the case of the voluntary courses. The participation of students in RiMS, RT and RFC is satisfactory for voluntary courses. In the case of the RT, with certified participation of approximately 47% of all registrations (corresponding to 10% of the total cohort), this is comparable to similar programs. It can be shown that the number of experimental science projects has more than doubled over time in parallel with the development of RaMS. The average number of provided projects according to the RFC is 42 (which corresponds to a placement rate of approx. 1:4). The number of successful student applications for a research support grant during the period the measures were implemented has doubled. Discussion and conclusion: The RaMS program shows a route for the implementation of the SP required by the next licensing regulations in medical education, which was initially supported and expanded solitarily, later by further elements (RiMS), also in the sense of a science-based career development (RT, RFC). The student acceptance and the measured success, in the form of successful participation in the Research Track, increased choice of experimental projects, significant increase of submitted as well as approved research grants and the high project placement rate of the Research Fair, encourage the further development of the program, which is indicated in the conclusion.

Fatalities associated with NPS stimulants in the Greater Cologne area.

This study centres on the prevalence of new psychoactive substances (NPS) stimulant use, and its relevance as a cause of death amongst individuals between the ages of 12 and 35 in the greater Cologne area. An automated solid-phase extraction-liquid chromatography-tandem mass spectrometry method was developed for the determination of 97 stimulants in urine (including conventional stimulants, e.g. amphetamine and MDMA), of which 68 analytes were fully validated for quantification. Samples of urine or kidney tissue (in cases where urine was unavailable) of 268 deceased were collected, during autopsy, between January 2011 and May 2017 and analyzed. Blood (if available) was also investigated in cases where urine/kidney samples were tested positive for NPS. An intake of stimulants (including NPS stimulants) was proven in 50 cases. In 33 cases, only conventional stimulants were detected. A total of 17 cases were tested positive for NPS. Of the 17 NPS-positive cases, 13 were also tested positive for other conventional drugs of abuse (mostly amphetamine and MDMA). In six NPS-positive cases, at least three different NPS were proven to be ingested. Due to the determined blood concentrations, NPS was assigned as the leading cause of death, or of toxicological relevance, in the cause of death in only 5 cases. In two of the cases, NPS was judged to be a component of a multidrug poisoning, but of minor relevance.

Organ distribution of diclazepam, pyrazolam and 3-fluorophenmetrazine.

The organ distribution of 3-fluorophenmetrazine (3-FPM), pyrazolam, diclazepam as well as its main metabolites delorazepam, lormetazepam and lorazepam, was investigated. A solid phase extraction (SPE) and a QuEChERS (acronym for quick, easy, cheap, effective, rugged and safe) - approach were used for the extraction of the analytes from human tissues, body fluids and stomach contents. The detection was performed on a liquid chromatography-tandem mass spectrometry system (LCMS/MS). The analytes of interest were detected in all body fluids and tissues. Results showed femoral blood concentrations of 10 µg/L for 3-FPM, 28 µg/L for pyrazolam, 1 µg/L for diclazepam, 100 µg/L for delorazepam, 6 µg/L for lormetazepam, and 22 µg/L for lorazepam. Tissues (muscle, kidney and liver) and bile exhibited higher concentrations of the mentioned analytes than in blood. Additional positive findings in femoral blood were for 2-fluoroamphetamine (2-FA, approx. 89 µg/L), 2-flourometamphetamine (2-FMA, hint), methiopropamine (approx. 2.2 µg/L), amphetamine (approx. 21 µg/L) and caffeine (positive). Delorazepam showed the highest ratio of heart (C) and femoral blood (P) concentration (C/P ratio = 2.5), supported by the concentrations detected in psoas muscle (430 µg/kg) and stomach content (approx. 210 µg/L, absolute 84 µg). The C/P ratio indicates that delorazepam displays susceptibility for post-mortem redistribution (PMR), supported by the findings in muscle tissue. 3-FPM, pyrazolam, diclazepam, lorazepam and lormetazepam did apparently not exhibit any PMR. The cause of death, in conjunction with autopsy findings was concluded as a positional asphyxia promoted by poly-drug intoxication by arising from designer benzodiazepines and the presence of synthetic stimulants.

Death after misuse of anabolic substances (clenbuterol, stanozolol and metandienone).

A 34-year old male was found breathless and panting at home by his girlfriend three hours after a gym workout. Minutes later, he collapsed and died. Autopsy, histological and chemical analyses were conducted. The examination of the heart showed left ventricular hypertrophy, while the right coronary artery showed only a small vascular lumen (3 mm in diameter), due to its anatomical structure. In femoral blood concentrations of approx. 1 µg/L clenbuterol, approx. 56 µg/L stanozolol and approx. 8 µg/L metandienone, with trenbolone (

Distribution of bone injuries in cases of non-natural death: A retrospective, encompassing study from 2008 to 2017.

For this study all autopsy reports of non-natural deaths performed at the Institute of Legal Medicine at the University of Cologne from a 10-year period (2008-2017) were examined in regards to the manner of non-natural death and their frequency and distribution of bone injuries (BI). 2026 cases (220 homicides, 421 suicides, 760 accidents, 393 possible suicides, 232 possible accidents) were reviewed. The highest frequency of BI was registered within homicides with 89.1%, second being accidents (79.3%), while suicides and poss. suicides were at 53.4% and 43.3%. Gunshots/explosions split between 52% for suicides and 33.6% for homicides, sharp force was to 56% found within homicides and with 29.8% in suicides. 63.7% of all blunt force was found in the accident category. The highest participation of BI and injuries across all body regions was found with gunshot homicides (100% BI, over every region). Intoxication showed the lowest BI over all with only 19.9% of all cases. Albeit several distinctive patterns were found, e.g. distinction between gunshots in suicides and homicides or high amount of BI in decayed corpses in homicides, a close examination on case by case basis, utilizing the forensic toolkit to its fullest, should always be attempted when examining decomposed or skeletal remains.

Simultaneous quantification of THC-COOH, OH-THC, and further cannabinoids in human hair by gas chromatography-tandem mass spectrometry with electron ionization applying automated sample preparation.

The detection of Δ9 -tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN) in hair, for the purpose of identifying cannabis consumption, is conducted in many forensic laboratories. Since external contamination of hair with these cannabis components cannot be excluded, even after hair decontamination, only the detection of THC metabolites such as 11-nor-9-carboxy-Δ9 -tetrahydrocannabinol (THC-COOH) or 11-hydroxy-Δ9 -tetrahydrocannabinol (OH-THC), is considered to prove cannabis consumption. At present, testing for THC metabolites is not standard practice due to its analytical complexity. For these reasons, we developed a novel method for the detection of THC-COOH and OH-THC as well as THC, CBD, and CBN in one single analytical run using gas chromatography-tandem mass spectrometry (GC-MS/MS) with electron ionization. After manual hair washing and grinding, sample preparation was fully automated, by means of a robotic autosampler. The hair extraction took place by digestion with sodium hydroxide. A solid-phase extraction (SPE) was chosen for sample clean-up, using a mixed-mode anion exchange sorbent. Derivatization of all analytes was by silylation. The method has been fully validated according to guidelines of the Society of Toxicological and Forensic Chemistry (GTFCh), with a limit of detection (LOD) of 0.2 pg/mg for THC-COOH and OH-THC and 2 pg/mg for THC, CBD and CBN, respectively, thus fulfilling the Society of Hair Testing (SoHT) recommendations. The validated method has been successfully applied to our routine forensic case work and a summary of data from authentic hair samples is given, as well as data from proficiency tests.