The data presented in this thesis has been organized in three parts: First part included the development and validation of a quantitative HPLC-DAD analytical method of mephedrone after extraction from spiked whole blood and serum samples, alone and with methcathinone. The second part included in vitro metabolism of mephedrone and other NPS, which are methoxetamine and methcathinone, using an in-house prepared in vitro metabolic system, namely liver microsomes, followed by performing analysis for the drugs and their proposed metabolites utilizing LC-MS. Third part included in vitro studies of selected NPS using purchased HepaRG and hepatocytes. In vitro study included in vitro cytotoxicity studies of 4-fluoromethamphetamine, mephedrone, methoxetamine and methcathinone, and analytical studies of these drugs of abuse and their potentially produced metabolites using GC-MS. In the first part of this thesis, a HPLC method for the analysis of mephedrone after LLE from blood matrix was developed and validated and shown to be linear with R2> 0.995, precise with intraday and interday RSD values of 4.36 and 4.77% respectively and LOD and LOQ of 0.025 and 0.082 μg/mL respectively. Recovery percentages were low and ranged between 28-37%. Emulsion formation was the major problem effaced which negatively affected recovery and precision values. The previously developed method was optimised and fully validated for the simultaneous analysis of mephedrone and methcathinone after liquid-liquid extraction (LLE) from whole blood and serum samples. The LLE method was optimised through selection of extraction solvent and adjustment of pH values achieving the best validation parameters and minimal emulsion formation. The LLE protocol involved extraction with a mixture of dichloromethane: n-butanol (80:20 v: v) after buffering the sample with borate buffer pH=9.2 and using aniline as internal standard. The HPLC-DAD method was optimized, using reverse mode chromatography and buffered mobile phase of (acetate buffer pH 4.1: ACN – 85:15) for qualitative and quantitative analysis of these drugs in less than 10 minutes under isocratic elution and ambient temperature. The method was fully validated for both drugs and showed to be linear over the specified range of 0.1-10 μg/mL with R2 > 0.99 for both drugs. The accuracy was assessed by calculating percentage recovery at different concentrations for xii both drugs, and retained recovery percent between 84-110%. For repeatability and intermediate precision tests, RSD values were ≤ 6.73%. Specificity was assessed by good resolution between the peaks and by checking peak purities. Limit of detection and limit of quantification, calculated mathematically for both drugs either extracted from whole blood or serum samples, were 0.010- 0.013 μg/ml and 0.032 - 0.043 μg/mL, respectively. In the second part, in vitro studies on the metabolism of the selected NPS using pig liver microsomes and liquid chromatography-mass spectrometry (LC-MS) analysis were performed. Microsomes were prepared by a conventional ultracentrifugation method. In brief, pig liver was brought freshly from local abattoir, sliced into small pieces, homogenised and ultra-centrifuged to produce microsomes and S9 fractions. Produced microsomes were incubated with the drugs of interest under optimised conditions and followed by analysis utilizing LC-MS for the detection of the drugs and the potentially produced metabolites. It was possible to detect two metabolites of the drug mephedrone, hydroxytolyl-mephedrone and nor-dihydro mephedrone. For MXE, one metabolite produced by the O-demethylation was detected and it identity confirmed by MS/MS study to be o-desmethyl-MXE. Another metabolite was detected is suggestively produced by the reduction of the ketone moiety to produce dihydro-MXE or by two steps of O-demethylation and hydroxylation to produce O-desmethyl –hydroxy-MXE. However, due to low intensity signal recorded, MS/MS study was not conclusive for the identity of the molecule In the third part, two types of hepatocytes were used for the study of the metabolism and cytotoxicity of the selected NPS - Mephedrone, Methoxetamine, Methcathinone and 4-Fluoromethamphetamine. Studying the metabolism of selected NPS followed utilizing HepaRG™ followed by GC-MS analysis, it was possible to detect new peaks in the chromatograms of mephedrone and methcathinone which is suggestively the product of N-demethylation. However, it was not possible to detect any new peaks in the chromatograms of methoxetamine nor 4-flouromethamphetamine. The cytotoxicity study utilizing HepaRG cell line showed that these drugs have cytotoxic effects causing in vitro cell death, within the specified range of 4.0x10-2-1.6x101 mM. These drugs were able to cause 43-83% ii cell death, and EC50 values were 0.2323-0.6297 mM. The most potent drug was 4-fluoromethamphetamine, while mephedrone showed the least biological effect to produce.

Short tandem repeat typing is the primary method of DNA identification used in the field of forensic science. Over the past several years the need to improve on this method has moved to the forefront of research. Due to the increasing number of criminal cases and the substantial backlogs most laboratories are facing, it is vital to evaluate methods which can produce quality DNA profiles in a fast and reliable manner. Direct amplification, also referred to as direct PCR, is one alternative method that has been proposed to address this issue. Direct amplification allows for the generating of DNA profiles without using the DNA isolation process. While direct PCR would reduce processing time and resources, it is unknown if this technique would be able to generate a robust full or partial profile from samples which could be collected from scenes of crime. Often crime scene personnel must use visualization techniques, either in powder or chemical form, in order to see and collect biological evidence for submission to a crime laboratory. In order to evaluate if direct PCR is a feasible solution a comparative study between a direct PCR kit and standard DNA profiling practices was undertaken using mock crime scene type samples. Samples of this nature include surfaces which have been exposed to fingerprint powders and whole blood which has been chemically enhanced for visualization. PowerPlex® 18D, a direct amplification system, and PowerPlex® 16HS, an extraction-based method, were used to produce the profiles. An assessment of the kits aimed to critically evaluate and compare how the direct amplification kit performs on samples which have been exposed to powder and chemical processing for visual enhancement. This will be done by reviewing two types of samples; epithelial cells which have been exposed the fingerprint powders (black, magnetic and white) and whole blood which has been exposed to chemicals (luecocrystal violet, amido black and ninhydrin). Samples subjected to direct amplification using PowerPlex® 18D generated DNA profiles with greater peak heights when compared to the extraction- based method. The peak balances for heterozygous loci were also higher and more full profiles were generated with direct amplification than with the extraction method. The amount of DNA retrieved from each substrate also varied even though the same amounts of starting material were deposited, proving that the type of substrate can affect the retrieval of DNA. Epithelial cell samples were most successful when processed with white powder. Magnetic powder samples also yielded a positive result when using direct amplification which was not expected as in previous data magnetic powder samples have not been successful. Whole blood samples which were processed with amido black produced profiles with lower overall peak heights when compared to the two other chemical processes. This could be attributed to the rinse step which is required when working with amido black. Ninhydrin was the most successful of the chemicals in generating full, good quality profiles.

The ability to discriminate between different inks and to determine the length of time an ink has been on a substrate can provide important scientific evidence, especially in cases involving document fraud. Many techniques have been used to analyse inks for ink dating including chromatography and spectroscopy, but the results are unreliable as a result of factors affecting the aging process such as light. This study utilises established techniques in Forensic Document Examination, including filtered light examination but also novel techniques for ink analysis; Atmospheric Pressure Chemical Ionisation (APCI) to analyse inks and dyes with the aim of discriminating between samples based on their degradation products. APCI-MS was used for the first time to study nineteen ballpoint pens from a range of manufacturers by investigating the chemical processes that occur and the products that are formed following the deposition of ink onto a substrate and in solution. Monitoring the degradation process as an ink ages and fades enables the identification of components present in the inks. Using molecular mass data, accurate ink component identifications could be made over a period of two years on samples subjected to a range of external influences. Light, hydrogen peroxide and sodium hypochlorite bleach were used to simulate natural and deliberate fading of inks and dye solutions. Benzophenone and phenol molecules were identified as degradation products but their presence differed for each of the different conditions tested such as no phenol products when bleach was used. This novel approach to ink analysis utilises existing equipment commonly used by document examiner to analyse inks that are old or faded in some way, in order to discriminate between the inks or determine method of alteration.

A series of seven published peer-reviewed papers and reports are presented. The body of work to be considered is concerned with the reexamination of previously excavated Earlier Neolithic human remains from Southern Britain. These form the subject of a Synoptic Overview. The Synoptic Overview places the papers within the context of academic debate and knowledge as it stood in the mid 1980s and 1990s. The background events surrounding the writing of the papers and the extent of the author's involvement in collaborative papers are detailed. The papers present new information concerning Ecirlier Neolithic mortuary assemblages and their formation and subsequent taphonomic histories, new information concerning the extent of interpersonal violence in the Earlier Neolithic and new chronological data and interpretations. The contribution to knowledge of each of the papers is summarised and critically reflected on.

The thesis concerns an examination of victim engagement with the police investigation of domestic abuse. Notwithstanding the huge efforts being made in tackling the problem by police forces across the UK, national inspections still find that the services provided to victims are “not good enough” (HMIC, 2014, p.6). Subsequently, the thesis argues that in order to build an approach around empowering victims of Intimate Partner Violence (IPV), there first needs to be further research into victim engagement with the police investigation (Birdsall et al., 2016; Hoyle & Sanders, 2000). Using the rationale, the research examined 540 cases of IPV to determine which factors were significantly associated with victim engagement. It controlled for suspect charging, cross validated the results with qualitative case file information and brought together the findings through an analysis of their co-occurrence. The process resulted in distinct themes and an overall model of victim engagement. The thesis concludes that the current risk assessment used routinely by the police to identify victim vulnerability does not take into account victim engagement. The thesis therefore proposes that the factors, themes and model of victim engagement developed throughout the thesis, as well as other means of assessing victim engagement, would need to precede the DASH risk assessment to provide a more effective evaluation of victim vulnerability. Doing so would allow the police to critically communicate and provide suitable support that is applicable to all victims of IPV. Crucially, the early indication of victim withdrawal would allow the police to identify some of the most vulnerable victims of abuse who would otherwise disengage from professional support and place themselves at greater risk of harm, injury and abuse.

In the study a novel method for synthesis of a dark magnetic flake powder for detecting latent fingerprint has been developed. Even though, flake powders ,of aluminium, brass and highly reflective magnetic iron flake are already in use in crime scenes , it is highly desirable to develop a suitable darker variety of magnetic flake powder for print development on light background surfaces. In order to achieve rapid production of dark metal flakes, a new high energy prototype vibratory mill has been designed, manufactured and develo ed. The design concepts were developed following a comprehensive review of the various commercially available milling devices and undertaking some initial experimentation. The mechanical milling process which results in changing the particle morphology of starting atomised iron powder to flaky shaped powder was investigated in terms of different milling phases like micro forging, fracture and agglomeration. The effect of milling process parameters on the flake quality was investigated. It was shown that the amount of stearic acid content, ball packing fraction and weight loading were important parameters in determining the final flake qualities. The quality of latent fingerprint development with the dark flake powder was investigated. Some of the flake powders produced excellent ridge quality details with good adherence quality on a range of background surfaces. The present study has been able to establish the relationship between the visual characteristics ( dark) and adherence property of the flake fingerprint powder with respect to the particle dimensions and surface characteristics of the flakes. It was also found that finer flakes are darker in colour and the low weight percentage of stearic acid has produced the best adherence quality for the dark flake powder. Some unique relationship between the flake dimensions with the nature of the print deposit was also established as it was found that slightly coarser flakes are suitable for heavy print deposit whereas finer flakes are more sensitive to aged prints. Further, the role of process variables in influencing the milling process were established such that optimum conditions by vibration milling can be determined to obtain dark flakes of desired quality. While the weight loading significantly influenced the milling behaviour of the powder, the role of stearic acid as an additive was found to influence the surface quality of flakes, besides restricting particle welding. Increasing the pre determined theoretical ball size does not result in faster milling, but increasing the ball packing fraction from 50% to 70% has proved to be more effective. Some relation with the oxidising behaviour of the powder with vibration milling has also been established. Moreover, the study has demonstrated that high energy milling by a vibration mill can be utilised for consistent production of a novel dark magnetic flake powder.

Model prison, Lucknow

Thesis (M.S.)--Oklahoma State University, 2009. / Vita. Includes bibliographical references.

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Thesis (M.S)--Oklahoma State University, 2007. / Vita. Includes bibliographical references.