The aim of this study was to investigate the potential use of ELISA and LC-MS-MS in combination and as individual techniques, for the detection of drugs of abuse in biological matrices. Overall the LC-MS-MS method showed good correlation results for opiates compared to the GC-MS method. 6-MAM was however detected in more root segments and segments excluding roots by LC-MS-MS. Morphine was detected in a greater number of root segments by LC-MS-MS compared to GC-MS. However, morphine was detected in a greater number of segments excluding roots by GC-MS. Codeine and dihydrocodeine were also detected in a greater number of root segments and segments excluding roots by GC-MS. The cocaine results showed excellent qualitative correlation between the LC-MS-MS and GC-MS methods for cocaine and benzoylecgonine. The GC-MS method did not however extract greater concentrations of cocaine and its metabolites compared to LC-MS-MS due to the higher recovery of the drug group specific GC-MS method. Cocaethylene and EME were detected in some samples by LC-MS-MS method for opiates and cocaine and its metabolites compared to the GC-MS method; there may be some cases where the GC-MS method would detect the analytes where the LC-MS-MS method would not. This has been demonstrated in 3 samples for morphine and in 6 samples for codeine. The LC-MS-MS method analysed for and detected amphetamines in samples that were not tested for amphetamines by GC-MS. In one sample that was tested by both methods, amphetamine was detected in the root sample by LC-MS-MS where GC-MS failed to detect it. Also a greater concentration of amphetamine was extracted using the LC-MS-MS method in the segment without roots. The LC-MS-MS method was useful for the analysis of 17 drugs of abuse in post-mortem hair samples in forensic toxicology cases. Using this method, it is possible to obtain maximum information from one hair sample which is extremely useful when the sample weight is limited. The ability of the LC-MS-MS method to extract and analyse a greater number of drug groups from one hair sample highlights the advantages of using this method over GC-MS which targets individual drug groups and requires splitting of the sample. This method is particularly applicable for implementation in the forensic toxicology laboratory at the University of Glasgow where currently GC-MS methods that target individual drug groups are used for routine hair screening and confirmation.

Introduction: Care professionals suffering with poor wellbeing is a phenomenon that has been found to have a damaging effect upon individual employees, service users and organisations. Employees working in forensic settings are believed to be at increased risk of damaged wellbeing due to the unique demands of their working environment, including exposure to violence and aggression. This issue was addressed in two ways. Firstly, a systematic review of the literature on the effectiveness of person-centred interventions to improve the wellbeing of forensic professionals was prepared. Following this, an empirical study was completed which examined the ability of demands related to violence and aggression, and resources of two types (cognitive and contextual behavioural) to predict the wellbeing of employees in a high secure forensic mental health (FMH) hospital. Method: For the systematic review, relevant databases were systematically searched and 7 papers that met the inclusion criteria were identified. The included studies were quality assessed to identify strengths and weaknesses. For the empirical study, 142 employees at a high secure FMH hospital completed self-report questionnaires which examined their wellbeing, perceptions of the prevalence of aggression, beliefs about safety, attitudes towards aggression, and psychological flexibility. Results: The reviewed studies included psychological, educational and mixed type interventions. Evidence for the effectiveness of interventions was mixed, and problems with methodological quality common. The results of the empirical paper suggested that exposure to violence and aggression was not a good predictor of wellbeing. However, the beliefs staff held about safety and staff‘s level of psychological flexibility were predictive of wellbeing. Conclusions: The review concluded that the existing evidence for the effectiveness of person-centred wellbeing interventions for forensic professionals was generally of poor quality, and inadequate to provide firm recommendations. Further research to assess the effectiveness of interventions and the underlying mechanisms of wellbeing change in forensic settings was advised. The empirical paper concluded that job demands related to staff‘s cognitive appraisal of safety, and the contextual behavioural resource, psychological flexibility, were predictive of staff wellbeing. It was recommended that future interventions to improve the wellbeing of forensic professionals consider the psychological processes staff encounter in the workplace, with a particular focus on contextual behavioural resources, which have an existing evidence base in broader occupational fields. Further research using contextual behavioural interventions within forensic settings is recommended in order to develop the limited research on forensic professionals‘ workplace wellbeing.

A third of adults in the UK admit to using controlled substances at least once hi their lives and the most commonly abused drug was found to be cannabis, followed by the opiates and ecognine derivatives. Forensic laboratories are continually trying to reduce the turnaround time for samples submitted for analysis, to less than the current time of four days. One of the main aims of this work was to reduce the time taken to prepare, extract and analyse hair for target analytes. A new method combining supercritical fluid extraction and solid phase micro-extraction (SFE/SPME) is introduced. This coupled procedure utilises the advantages of using supercritical carbon dioxide as an extraction solvent and collecting the extract on a selected SPME fibre. Together the combined techniques provide a method that is solvent-free and highly selective, whilst reducing sample preparation to a minimum. Digested hair samples are examined by SEM to find the best method. Alkali hydrolysis and enzyme digestion using proteinase K were found to be the most effective pre-extraction methods. Solid hair samples with the addition of a derivatization reagent (BSTFA) gave the best results using the coupled technique. After optimisation and validation, the extraction from both real and spiked samples resulted in recoveries between 60% to 80% for the target analytes A9 -tetrahydrocannabinol (A9- THC) , cannabinol (CBN) and cannabidiol (CBD). Other forensic applications such as the extraction of polycyclic aromatic hydrocarbons and pesticides from environmental and food samples and the examination of fire debris for the presence of accelerants are also carried out. PAH could easily be extracted from complex matrices to give recoveries of up to 97% for some analytes at the optimized conditions of 90°C at 4500 psi over a 10 minute dynamic extraction period. Pesticide extraction gave recoveries of 60 - 80% while the extraction of accelerants from fire debris clearly showed that accelerants such as diesel can be identified over the coincidental PAH and hydrocarbon peaks found in burnt debris. The coupling of SFE to SPME provides an exciting technique that has a wide range of applications and creates potential for a fully automated on-line system.

This thesis covers a range of important issues in hair analysis and includes 27 scientific works in which the name of the candidate was either listed as the first author or as the major contributor. The work presented in this thesis involved the development of a series of analytical methods to detect trace amounts of drugs in hair and also investigated the mechanisms by which drugs may be incorporated into hair. The major areas covered in this study can be summarized as follows: 1. The methods for the identification and quantification of opiates, amphetamines, ketamine, cannabis, cocaine, benzodiazepines, antidepressants, antipsychotics, and anabolic steroids in hair were developed using gas chromatography–mass spectrometry (GC-MS), liquid chromatography–tandem mass spectrometry (LC-MS/MS) and gas chromatography–tandem mass spectrometry (GC-MS/MS). With GC-MS methods, the limits of detection were 0.1-0.5 ng mg-1 of hair for antidepressants and antipsychotics. For illegal drugs, hair specimens were analyzed by GC-MS with limits of detection of 0.02-2ng mg-1. GC/MS/MS is more sensitive than GC-MS to detect these drugs in hair. The lower limits of detection ranged from 0.001 to 0.020 ng mg-1 for 21 anabolic androgenic steroids and their esters in hair using liquid chromatographic-tandem mass spectrometric method. And the limits of detection ranged from 0.2 to 5 pg mg-1 for benzodiazepines in hair. Tandem mass spectrometry is characterized by its sensitivity, selectivity and specificity, which makes it particularly suitable for the analysis of trace amount of target analytes in hair. 2. Usually, screening for drugs of abuse is the first step in clinical and forensic toxicology. There are a large number of controlled substances and doping agents and novel compounds, which have yet to be characterised. A series of screening methods for drugs of abuse in hair were developed using LC-MS/MS and GC-MS/MS. Using our own library of MRM transitions, the optimum collision energies selected for each transition and retention times were set up. These methods have been applied successfully in forensic casework. 3. Of growing importance to the field of hair analysis is the detection of metabolites related to the parent drugs. Demonstrating the presence of a metabolite of a drug (such as, heroin, amphetamines, cocaine, meperidine, ketamine, triazolam or psychotropic drugs) provides compelling evidence for exposure to the parent drug, and permits distinction between external contamination from ingestion and facilitation of the interpretation of results. The presence of antidepressant and antipsychotic drugs and their metabolites in the hair of psychiatric patients was investigated using GC-MS-EI and GC-MS-PCI. The parent drug and its major metabolite, such as opiates (morphine, 6-acetylmorphine), methamphetamine (methamphetamine, amphetamine), ketamine (ketamine, norketamine), cocaine (cocaine, benzoecognine), meperidine (meperidine, normeperidine), triazolam (triazolam, α-hydroxytriazolam), and clonazepam (clonazepam, 7-aminoclonazepam) were quantified in authentic hair samples simultaneously. The differences were finding in the ratio of parent drug to metabolite. For illegal drugs, the concentrations of parent drugs were higher than that of their metabolites. The results of triazolam and clonazepam were contrary. These data are suitable reference values and are the basis for the interpretation of results. 4. The mechanisms by which drugs are incorporated into hair are not fully understood. Based on experiments with guinea pigs with black, white, or brown hair, the mechanisms of incorporation of cocaine, methamphetamine, ketamine, triazolam and anabolic steroids into hair were investigated. The concentrations of drugs in hair were found to be related to physicochemical properties of drugs. The parent drugs were the predominant analytes in hair. There was an obvious relationship between the concentration of drugs in hair and hair pigmentation. The concentrations of drugs deposited in black hair was found to be higher than that in brown and white hair samples, even when comparing results using hairs on the same multicoloured animal body. This work confirmed that melanin affinity is a governing factor in drug incorporation into hair shafts. These studies on the distribution of drugs in the hair shaft and how their concentration changes along the shaft provide information relevant to the time of ingestion and substance use/abuse. 5. In recent years an increase in drug-facilitated sexual assault (DFSA) has been reported. Segmental hair analysis has proved useful in widening the window of detection, as blood and urine analyses are of limited use, due to the long delays between the actual assaults and obtaining samples from suspects that are frequently encountered in investigations of such crimes. In China, benzodiazepines are the most frequently observed compounds in cases of drug-facilitated crime. In a paper reported here, 14 volunteers ingested a single 1-6 mg estazolam tablet to permit the evaluation of segmental hair analysis after a single drug dosage. Hair was collected one month after administration of the drug. All the proximal segments tested positive for estazolam. With increased dosage, estazolam could be detected in the 2-4 cm segments nearest the hair root in some subject’s hair shafts. In some cases, the 4-6 cm segments also tested positive. Hair analysis was applied to samples from two authentic criminal cases. A significant variation was observed between those obtained from previous studies and the results presented here. The intersubject variability in segmental analysis can be explained mainly due to melanin content and diffusion from sweat or other secretions during formation of the hair shaft. However, more substantial procedural and interpretation guidelines are required to use segmental hair analysis in drug-facilitated crimes. On the other hand, the minimal dosage for detection, which is a critical but previously unknown threshold value of fundamental importance in hair analysis, was determined for triazolam and ketamine in guinea pig hair. 6. Doping with endogenous anabolic steroids is one of the most serious drug issues in sports today. The measurement of anabolic steroid levels in human hair permits the distinction between pharmaceutically produced steroids and naturally occurring steroids. Full-length hair samples were taken at the skin surface from the vertex of 39 males, 30 females and 11 children from China. None of the subjects were professional athletes. Testosterone and dehydroepiandrosterone were detected in all the hair segments. The physiological concentrations of testosterone were in the range 0.8-24.2 pg mg-1, 0.1-16.8 pg mg-1 and 0.2-11.5 pg mg-1 in males, females and children, respectively. However, the mean values of dehydroepiandrosterone were much higher than those for testosterone. This is the first investigation into the physiological concentrations of anabolic steroids in human hair in Chinese subjects. These data provide suitable reference values and form the basis for the interpretation of results from investigations into the abuse of endogenous anabolic steroids. In conclusion, the work presented in this study demonstrates that there was a good correlation between the concentration of drugs in hair and drug dosage. There was an obvious relationship between hair drug concentration and hair colour. Melanin affinity is shown to be a governing factor in determining drug incorporation into hair, and the concentration of drugs deposited in black hair was found to be higher than that in brown and white hair samples. This thesis provides data that will be useful in the application of hair analysis regarding drugs of abuse and in the interpretation of toxicological results.

The purpose of this study was to develop a single method to analyse drugs in hair and to apply this to case samples received from The Security Forces Hospital, Saudi Arabia. First of all, mass fragments and retention times for amfetamines and cannabinoids were identified by GC-MS using derivatization agents, PFPA/ethyl acetate (2:1 v/v) and PFPA / PFPOH (1:0.75 v/v) for amphetamines and cannabinoids, respectively. The results showed good peak shape, good chromatographic resolution and good sensitivity for amfetamine (AF), methamphetamine (MA), 3,4-Methylenedioxyamfetamine (MDA), 3,4-Methylenedioxymethamfetamine (MDMA), 3,4-Methylenedioxyethylamfetamine (MDEA), Δ9-tetrahydrocannabinol (Δ9-THC) and 11-nor-9-carbody- Δ9-tetrahydrocannabinol (Δ9-THC-COOH) compounds. The comparison of the efficiency of four different pre-treatment methods (enzymatic, alkaline, acid and methanol) to extract AF, MA, MDA, MDMA, MDEA, Δ9THC and Δ9-THC-COOH from hair samples obtained from known amfetamines and cannabinoid abusers was investigated. The preliminary results demonstrated difficulty with the cannabinoids recovery and the lower concentration of standards were not detected using any of the pre-treatment methods. As a result of the poor cannabinoids recovery, only the amfetamines were investigated. For the comparison study, only one hair sample positive for amphetamine was available so the pre-treatment comparison study was based on the recovery of AF using the four pre-treatment methods. The positive hair sample was separated into portions and pre-treatment methods, alkaline (1M NaOH), β-Glucuronidase (helix pomatia), methanol (MeOH) and acid (0.1M HCI) were used on these and compared. The best recovery for amphetamine was obtained using the β-glucuronidase pre-treatment method and this extract was also found to be cleaner than the alkaline and methanol pre-treatments. Β-glucuronidase pre-treatment was selected as the method of choice for the extraction of amphetamine content in hair. The method was validated to include linearity, recovery, intra- and inter-day precision, limit of quantitation (LOQ) and limit of detection (LOD) for all five amphetamine compounds. The method was shown to be reliable and robust for these substances. The stability of AF in hair was investigated to assess the validity of analysing hair samples for the presence of AF in victims of drowning. Ten amphetamine positive hair samples were submerged in fresh and sea water for different periods of time. The drug concentrations in the samples were monitored over a period of 8 weeks. Hair samples were analysed using the validated method. The results showed a significant decrease of amphetamine in hair with the time submerged in sea water. Fresh water had a much less significant effect over the study period. The validated method was successfully applied to 16 case samples obtained for living volunteers with a known history of fenethylline (AF precursor) abuse and 6 post-mortem case samples where amfetamines had been detected in the post-mortem blood.

A systematic literature review summarizes what is known about the relationships between attachment, relational security and therapeutic relationships. A paucity of research with forensic populations is noted. Consequently the empirical research paper explores more systematically the relationships between these variables on forensic psychiatric wards. Suggestions for staff and service development are presented. No relationships were found between relational security and service users’ attachment to the service and between relational security and risk events and treatment outcomes. Additional analyses revealed that relational security and ward atmosphere were moderately correlated. A large positive correlation was found between patients’ attachment to the service and ward atmosphere, and a moderate, negative, relationship was found between patients’ attachment to the service and risk incidents Multiple regression revealed that relational security and ward atmosphere significantly predicted patients’ attachment to the service. More robust research is needed to investigate the relationships between attachment, relational security, risk incidents and treatment outcomes on forensic psychiatric wards. An argument is made for promoting stronger therapeutic relationships between staff and service users so that forensic wards can become closer to secure bases which might help clients to overcome past interpersonal traumas and develop safer ways to relate interpersonally.

This study investigated the antecedents and psychological processes inherent in firesetting within an adult intellectually disabled population. A qualitative methodology, grounded theory (GT) was employed to explore the experience of firesetting through verbal self-report. A sample of eight males with mild intellectual disabilities under the care of forensic learning disability services participated in interviews which ranged from 27 minutes 14 seconds to 1 hour 21 minutes 22 seconds. The data produced a preliminary GT which identified eight conceptual categories: Managing internal affect states, the experience of adversity, unsafe others, engagement in multiple offending behaviours, voice entitlement, experiences of fire, and mediators of risk and the function of firesetting. These conceptual categories were linked by an overarching core category of ‘powerlessness’ which was evident in several areas of the participants’ lives and experiences of firesetting. It was concluded that this sense of powerlessness along with the conceptual categories should be considered when risk assessing, delivering firesetting specific treatment and psychological interventions. The clinical implications are discussed and suggestions are made for replication of the current study with larger, more diverse samples and future directions for further research are proposed.

Determining the postmortem interval (PMI) remains one of the most important but challenging factors to establish in a suspicious death investigation. Unfortunately, as time passes current methods lose accuracy and only allow investigators to approximate how long ago death occurred. Bodies interred in clandestine graves prove particularly challenging due to an abundance of variables that need to be taken into consideration. Due to the problems associated with determining the PMI of buried remains this study will utilise macroscopic, microscopic, molecular, chemical and microbiological analyses to systematically document the decompositional changes to human hair and porcine cartilage and bone in a burial environment. The aim was to correlate decompositional changes with time and develop new methods for estimating the PMI of remains found in this context. Whole trotters (from which the cartilage was harvested) exhibited decompositional changes including darkening of the dermis, skin slippage, liquefaction of soft tissues and complete skeletonisation. The decompositional changes to cartilage included a loss of cartilage covering articular facets, changes in colour and texture, formation of orthorhombic crystals, a change in surface pH and colonisation by bacteria. The bacteria found on the cartilage surface were in close proximity to the crystals and when cultured on a B-41 medium were found to precipitate crystals of the same morphology and chemical composition to those found on the cartilage surface. Three species of bacteria (Acinetobacter calcoaceticus, Acinetobacter iwoffii and Grimontia hollisae) were identified based on gas chromatography–mass spectrometry (GC-MS) of their fatty acids and one species (Comamonas sp.) was identified by DNA analysis. Formation of crystals on goat and cow cartilage proved that this was not a porcine specific phenomenon. Human hair exhibited a gradual degradation over time but this was dependent on the characteristics of the burial environment. Decompositional changes included colonisation by fungi, erosions to the cortical surface and formation of tunnels and breaks to the hair shaft. Two fungal species (Aspergillus fumigatus and Penicillium sp.) were identified based on DNA analysis of fungal ribosomal (rDNA) internally transcribed spacer (ITS) regions. The Penicillium sp. was linked with fungal tunnelling of hair. Bone exhibited little modification over time but changes were observed. These included a change in colour of the cortical surface, a change in colour and gradual loss of bone marrow and erosions, cracking and flaking of the cortical bone. Fungi were found to colonise both the bone marrow and bone surface. Whole piglets were buried to document the time period taken to reach skeletonisation. This data was used as a correction factor and combined with the bone results to give an overall time period for the decomposition changes observed. The results of this study suggest that the decompositional changes to cartilage could be used to determine the postmortem interval of buried remains. However, the degradation of hair and bone was too variable to be of use in this context.

When offenders commit crime there is the potential that they may leave behind trace amounts of their DNA, even when there has been no apparent body fluid spill. During the examination of crime scenes, scene investigators try to identify areas that may be sampled to locate these traces. Specialist techniques are then required within the laboratory to enable such small amounts to be analysed to obtain a profile. These techniques are referred to as Low Template DNA analysis (LTDNA), of which Low Copy Number DNA (LCN DNA) is one instance. In 2008, following the Omagh Bombing trial, and comments made by Judge Weir, the UK Forensic Regulator commissioned a review of the science of LTDNA analysis. The subsequent report made specific mention of the fact that there was no available information on the success rate of the use of such DNA techniques and that there seemed to be confusion over what constituted a success. The report went on to state that there was no information on where such trace amounts of DNA were likely to be found, or what factors could influence the likelihood of obtaining a trace DNA profile (Caddy, 2008). This research considered the outcomes of LCN DNA analysis from 3,552 samples to try to establish where trace amounts of DNA could be found, whether some areas sampled were more successful in generating profiles than others, and the likelihood of the profiles obtained being of use to a criminal investigation. Analysis of results identified areas that were more successful in generating profiles of use to an investigation and highlighted significant differences in results across a variety of items from which samples were taken. DNA samples taken from items associated with communication such as mobile phones were much more likely to produce a profile useful to a criminal investigation than those taken from fixed surfaces within premises. The results obtained showed that obtaining a DNA profile did not necessarily correlate with the profile being of use to a criminal investigation. This was due to the fact that a large number of these profiles were anticipated eliminations from legitimate sources. Items that produced high numbers of profiles but were anticipated eliminations, and therefore of no value to an investigation, came from items associated with skin samples and clothing. The research went further to identify key factors that affected the profiling rates. Factors that had a positive influence on the ability to obtain a profile included: any area that had been in close proximity to saliva (direct contact was not required); samples that had been recovered from the inside of premises or vehicles and therefore protected from the elements; those that were dry; items that were of a porous nature; and those that had a rough texture. No differences were found between the actual surface materials (plastic, glass, wood, metal), as all showed a propensity to generate profiles. Other factors that were considered but proved to have no effect on the profiling rates included seasonal differences and whether the area targeted for sampling was clearly defined. Items that had had high contact with a victim, were recovered from outside or had been wet, all proved to be less useful to an nvestigation. A further finding of the research was that swabs that had been recovered and stored frozen appeared to deteriorate in their ability to profile. This was particularly notable if they were submitted later than 5 months after recovery. Items stored in dry conditions did not deteriorate in this way. Overall the research can be used to provide investigators with the knowledge of what areas of crime scenes are most likely to yield trace DNA material, the key factors that can affect the likelihood of obtaining a profile, and those areas that are more likely to produce profiles useful to criminal investigations.

La balistique lésionnelle, carrefour de l’expertise balistique et de la médecine légale, est fréquemment confrontée à l’évaluation et la caractérisation de plaies balistiques crâniennes. De nombreuses recherches ont été effectuées sur les traumatismes de l’extrémité cervico-céphaliques dans une vision d’amélioration de la sécurité mais celles dans le domaine de la balistique sont quasi inexistantesou empreintes de certitudes non démontrées. Le premier objectif de ce travail a été demettre en place un protocole de tirs d’essai sur des voutes crâniennes d’animal dont la structure est comparable à celle de l’homme. Ce setup original, alliant la technologie d’enregistrement vidéo à haute vitesse et l’analyse des dégâts en microtomographie, a permis de mettre en exergue des relations entre vitesse d’impact, vitesse de sortie, énergie dissipée et dommages créés. Ils contredisent souvent certaines théories jusque là proposées et répondant ainsi à notre deuxième objectif. Les nombreuses données acquises par l’expérimentation pourront utilement éclairer l’expert en balistique lésionnelle dans l’argumentation de ses hypothèses quant à l’utilisation d’un couple arme-munition plutôt qu’un autre. Cette étude propose également un nouveau modèle quant à la cinématique complète de l’interaction entre le projectile et la structure osseuse. / Wound ballistics is at the crossing between ballistics and legal medicine. It frequently has to tackle skull gunshot wounds evaluation and characterization. Many researches have been made around head and skull traumas, aiming at an improvement of security devices, but considering the field of ballistics, researches are either inexistent or marred by undemonstrated certainties. This work first aims at setting a firing protocol out, using animal skull bones that are similar with humans’. This original setup allies high speed video recordings and damage analysis with microtomography. It has shown up relationships between impact speed, exit speed, dissipated energy and observed damages. It comes in contradiction with many models formerly exposed, and hence fulfills our second objective. Important data acquired from experiments can show as very useful for wound ballistics expert to support one of his hypothesis about which firearms and ammunition type were used. This work eventually proposes a new model explaining the interaction between projectile and bone structure during damage creation.