Carbon monoxide (CO) is a gas that exerts its toxicity on humans, when inhaled, by bonding with haemoglobin (Hb) to produce carboxyhaemoglobin (COHb). This results in tissue hypoxia which can be fatal at high blood saturation levels. Carboxyhaemoglobin may be measured using a Radiometer ABL825 FLEX analyser, a spectrophotometric instrument that applies a technique called CO-oximetry to measure Hb derivatives such as COHb and methaemoglobin (MetHb). The latter is an oxidised form of Hb that can cause or contribute to mortality at high concentrations. Methaemoglobin is notorious for its instability in vitro. This study, therefore, sought to determine handling conditions best suited for the stability of MetHb in post-mortem blood such that the ABL825 FLEX analyser may be used for the simultaneous measurement of COHb and MetHb. To this end, blood samples collected from 15 cases of potential CO poisoning at Salt River Mortuary were aliquoted into red (no additive)-, green (containing lithium heparin)-, and purple (containing ethylenediaminetetraacetic acid)-top tubes. The samples were stored at 4°C or –80°C and retrieved from storage on days 0, 1, 4, 7, 14, and 30, for analysis. While COHb remained stable in all storage conditions over a 30-day period, this was not true for MetHb. When samples were stored at 4°C, a rapid increase followed by a gradual decline was observed for MetHb in all the tube types investigated. The MetHb content was at its lowest after two weeks of storage, which was consistent with COHb levels at the same time period and temperature. At –80°C, continuous increase of MetHb was observed, with the samples stored in green-top tubes showing the least amount of overall change from the reference (day zero) values. The study provided useful data regarding the stability of MetHb under the considered storage conditions, the investigators concluded that refrigerating blood samples in either red-, purple-, or green-top tube was suitable for the accurate simultaneous determination of both COHb and MetHb, if the analytes are measured immediately after collection or after two weeks of storage. Given that the nature of post-mortem forensic casework is such that suitable blood specimens are not always available for toxicological analyses, for the second aim of the study, the researchers sought to investigate the suitability of thoracic cavity fluid as an alternative specimen for the measurement of COHb. For this aim, thoracic cavity fluid collected into green-top tubes from the aforementioned cases was analysed immediately after collection. The results were compared to those obtained from the analysis of blood collected into green-top tubes and analysed on day zero (before storage). The statistical analyses used for this assessment indicated that thoracic cavity fluid would be a suitable alternative to blood for the measurement of COHb using the ABL825 FLEX analyser.

Violence-related injuries are a major cause of mortality in the Western Cape (South Africa). Previous research has demonstrated an association between violent mortalities and drug use. Furthermore, long-term drug use has been shown to alter behaviour that may lead to violence. Hence, this study aimed to investigate the effect of long-term drug use on violence-related mortalities. Due to the drug retention properties of hair, it is the gold standard for demonstrating the historical pattern of drug use. Hair samples were collected from 92 violent death cases admitted to Salt River Mortuary (South Africa). A qualitative toxicological analysis was performed in 90 hair samples using a SCIEX X500R QTOF. Variables pertaining to the colour and length of the hairs were recorded. The majority of the hair samples were black (n=79), while others were black and white (n=5), greyish (n=3), light grey and reddish brown (n=1) and strong brown hair (n=1). Various toxicologically relevant substances were detected in 74 cases (82.2 %) in which a total of 54 different substances were detected in hair samples. Acetaminophen was the most prominent licit substance (47%) detected, followed by caffeine (18.9%) and diphenhydramine (12%). Methamphetamine was the most common illicit detected substance (54%), followed by methaqualone (43%). Segmented hair samples showed historical use in 81.2 % of cases. These results show that hair can be used as a supplementary sample during toxicological investigation in violent fatalities in the local context.

Indiana University-Purdue University Indianapolis (IUPUI) / The use and abuse of fentanyl has risen drastically over the last several decades. The abuse of this substance has created a hazardous situation for law enforcement and first responders because they could arrive at locations and not necessarily know that they will encounter fentanyl or a fentanyl analog. Fentanyl analogs are substances that have a similar structure to fentanyl, and while the analogs may have additional or altered groups on the molecule, the backbone structure remains similar. This work focus on the electrochemical characterization of fentanyl as a stepping stone for the detection of both fentanyl and later fentanyl analogs by electrochemistry. The metabolic reaction of fentanyl is an N-dealkylation to norfentanyl, occurring in the liver, and can be mimicked by electrochemistry through the irreversible oxidation of fentanyl. This electrochemical reaction is hypothesized to generate electroactive metabolites in solution. The combination of the visualization of both the irreversible oxidation with the development of the additional metabolic electrochemical peaks would constitute a unique electrochemical signature for fentanyl and fentanyl analogs towards a universal rapid screening assay. The electrochemical behavior of fentanyl was characterized in depth using multiple electrochemical techniques such as cyclic voltammetry (CV), square wave voltammetry (SWV) and differential pulse voltammetry (DPV). The optimization of the supporting electrolyte, the potential range, and methods to decrease the background current were explored with CV. To work towards a more portable system, screen printed electrodes were used. The observation of the metabolic peaks remained challenging, and different methods were attempted to achieve it. The quantification of fentanyl was successfully demonstrated using the different electrochemical systems proposed in this work. The electrochemical characterization of fentanyl and the optimization of multiple experimental parameters were the first step in developing a universal, rapid, electrochemical sensing method for the detection of fentanyl and fentanyl analogs. / 2023-02-28

Sexual lubricants are used to enable sexual encounters. There are different types of sexual lubricants such as water-based, oil-based, and silicone-based. They come pre-applied to condoms and separately in bottles as personal lubricants. Although sexual lubricants are intended for consensual use, they are also unfortunately used during the commission of sexual assaults. The analysis of sexual lubricants facilitates sexual assault investigations. With the increased usage of condoms in sexual assault cases, the potential of collected DNA evidence in each case is reduced. In the absence of biological evidence, the presence of sexual lubricants after a sexual assault can provide an additional link between a suspect and the crime scene and/or victim. Having the ability to compare known and unknown sexual lubricants may be the only actionable information available for investigators. Current lubricant analysis only classifies samples into lubricant types based on the major component such as glycerol, petrolatum, and polydimethylsiloxane for water-based, oil-based, and silicone-based lubricants respectively. Differentiation within major types has not been explored. Previously, protocols have been developed to detect and categorize personal lubricants using Fourier transform infrared (FTIR) spectroscopy, gas chromatography-mass spectrometry (GC-MS), liquid chromatography mass spectrometry (LC-MS), and pyrolysis GC-MS. FTIR is routinely used as a screening tool to detect peaks of the major lubricant components and the mass spectrometry (MS) techniques are commonly used to confirm the presence of some of the major components, excluding PDMS. This thesis focused on the differentiation of silicone-based personal and condom lubricants because it is a common type of lubricant due to its ability to reduce friction for a longer period of time. Fifty-six (56) silicone personal and condom lubricants were analyzed to identify unique characteristics that can be used to determine individual sub-classes and test those sub-classes. Direct analysis in real time-time of flight mass spectrometry (DART-TOFMS) was utilized because minor and unique molecular ions that could be attributed to different sub-groups can easily be distinguished from the major sample peaks. This is primarily based on the direct mass spectrometry design of the instrumentation that can differentiate minor components from major components that might not be observed using traditional chromatographic separation. The DART source creates molecular ions for individual components in mixed samples under atmospheric conditions in either positive or negative mode. The TOF-MS, which is capable of high resolution and accurate mass analysis, allows more accurate and precise detection of molecular component ions. Additionally, no sample preparation is required to analyze neat samples, which minimizes potential contamination issues. Attenuated total reflectance-FTIR (ATR-FIR) was used to analyze the training set personal lubricants to compare previous methods of analysis to the newly developed DART-TOFMS method of analysis. Principle component analysis (PCA) and cluster analysis were used to identify potential sub-groups and subsequently a classification scheme. Linear discriminant analysis was utilized to conduct leave one out cross validation and to categorize test samples. Eight sub-groups were developed based on the presence and/or absence of PDMS and minor component peaks observed. A classification scheme was developed using the eight sub-groups identified through PCA and cluster analysis. This classification scheme was tested using LDA to classify blind samples. One group includes a scented personal lubricant. Another group includes flavored condom lubricants. The other groups were developed based on the relative intensity of PDMS peaks and minor component peaks. Variation of the intensity of PDMS peaks between and within samples of different lot numbers causes some misclassification of samples. This classification scheme also doesn't take into account real-world factors such as dilution and biodegradation. Although further research is required to create a more stable classification scheme, the identified sub-groups are a good foundation for the creation of a lubricant database and finalized classification scheme.

Ammunition is most often comprised of a lead-based priming mixture which contributes to the traditional characteristics of gunshot residue (GSR). Due to the health risks often associated with lead, lead-free primer alternatives are becoming increasingly more popular. Thus, it is becoming more difficult to determine the presence of GSR based on traditional means, i.e. the presence of lead (Pb), barium (Ba), and antimony (Sb). Eight different ammunitions were purchased which consisted of one lead-based and one lead-free from four different manufacturers, including Winchester, Federal, Liberty, and Sellier & Bellot. Half of the rounds from each manufacturer and chemical composition (i.e. lead-based or lead-free) were disassembled, e.g. the projectile and smokeless powder were removed, leaving the priming cup in place (i.e. primed only). The remaining cartridges were left intact (i.e. full cartridge). Both the full cartridges and the primed only cartridges for each ammunition were fired using a 9mm Glock and the cartridges were collected post-fire, and subsequently deprimed. Five cups and five anvils from each type of ammunition were adhered to aluminum stubs via colloidal graphite. The GSR remaining on the anvils and in the cups was then analyzed using scanning electron microscopy coupled with energy dispersive x-ray spectrometry (SEM-EDX). Three spectra were collected for each anvil and each cup resulting in 30 total spectra per type of ammunition. The primary element peaks were then selected and four different dataset matrices were created for the full cartridge anvils, full cartridge cups, primed only anvils, and primed only cups. The data was processed using unit vector normalization and was then analyzed using principal component analysis (PCA) and linear discriminant analysis (LDA) to determine the characterization between lead-free and lead-based ammunition. The anvils provided better separation and characterization based on the ability to better collect x-rays, and therefore demonstrated the capability of the ammunition to cluster by both primer mixture composition and manufacturer. The lead-based and lead-free primers showed consistencies across samples, such as the presence or absence of K, which allowed for characterization based on primer composition.

Forensic analysts are tasked with determining the presence of ignitable liquid residue in fire debris. Analysis of fire debris allows the analyst to understand how the fire occurred. However, the presence of some substrates can potentially impact the identification of ignitable liquid residue and classification of a sample as positive or negative for the presence of ignitable liquid. Pyrolysis of building materials and furnishings (substrates) lead to background interference within the resulting chromatographic profile. To combat misclassification of a sample as positive for ignitable liquid residue, knowledge of the pyrolysis products from individual substrates is of utmost importance. However, unburned reference samples from a fire scene can be difficult to obtain. The use of a database in conjunction with the analysis of the samples can lead to a more complete analysis of fire debris. Within this research, four different burn methods (modified destruction distillation method, top heat, bottom heat, and tube furnace) were utilized in burning eight different flooring substrates (polyester, nylon, and olefin carpeting, carpet padding, vinyl flooring, laminate flooring, yellow pine, and plywood) to obtain pyrolysis/combustion product profiles. Each burn method was performed at three different burn times for a total of twelve different burns of each substrate. Standard methods, ASTM E1412-12 and ASTM E1618-14, were used in the extraction and interpretation of the laboratory burn products. Principal component analysis (PCA) was used to relate the laboratory burn results to neat ignitable liquid/substrate and large scale burn data sets. Laboratory burn data projected into the PCA space displayed that the laboratory burn data is similar to the data contained within the ILRC and Substrate databases. Differences observed within laboratory burn data projections illustrated the variability of the laboratory burn methods. The composition of the substrate dictated the pyrolysis/combustion products produced. While this research only focuses on flooring substrates, an increase in the number of different types of materials in the Substrate Database can aid analysts in identifying common pyrolysis/combustion products observed in fire debris.

An important forensic application for Y-chromosome short tandem repeats (Y-STRs) is the identification of male DNA. Since the Y-chromosome is non-recombining and most Y-STRs have slippage mutation rates on the order of 1x10-3 or lower, Y-STR commercial multiplex systems do not yet allow personal individualization. This is problematic in criminal investigations where the persons of interest include males of the same paternal line. In order to obtain discrimination among paternal male relatives, it would be necessary to use a larger number of Y-STRs or a selection of Y-loci with higher slippage mutations with the hope of encountering a germline meiotic mutation. In this study, Y-STR loci from three multiplexes were examined: an ultra-high discrimination multiplex of 14 non-core loci; Promega's PowerPlex® Y23; and a rapidly mutating 13-locus panel (mutation rates above 1x10-2). Taking the three multiplexes together provides a unique 40-locus Y-STR panel (the "Masker Set"), a powerful tool to separate closely related males. The loci were examined for discriminative slippage mutations in pairs of paternally related South Brazilian males: 99 grandfather-grandson, 103 uncle-nephew, and 140 brothers. The goal of this study was to analyze the "Masker Set" loci by: describing the characteristics and frequency of germ-line mutations; noting differences in mutation rates between and within loci; determining repeat gain and loss rates; and identifying the most informative loci to differentiate male relatives. Using the "Masker Set", pairs of male relatives were distinguished by at least one mutation 53% of the time for grandfather-grandson, 62% for uncle-nephew, and 54% for brothers. The most discriminating Y-STR loci were: DYF387S1, DYF404S1, DYS526B, DYS389II, DYS449, DYS547, DYF399S1, DYS458, DYS576, DYF403S1A, DYS508, DYS612, DYS403S1B, DYS518, and DYS627.

Pollen evidence has proven to be a powerful forensic tool to trace a suspect or item to a victim or a crime scene. This is possible because it is microscopic, abundant in nature, resistant to degradation and decay; it presents dispersal patterns that can be used to generate a 'fingerprint' within specific areas, and has illustrated a unique morphology that can be used to classify species. While the pollen grain morphology has been extensively used to characterize the specific species, not much has been investigated as pertains to the coating that surrounds the pollen grain aside from it being categorized as waste. This Master thesis focuses on the qualitative and quantitative determination of the elemental composition of this coating surrounding pollen via Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Two methodologies for sample preparation were compared: a complete digestion and analysis of (i) the entire pollen and (ii) the surrounding pollen coating alone removed from the pollen grain by Soxhlet extraction in ethanol. The goal was to discern the elemental composition of the coating and its specific elemental composition in comparison with the whole pollen grain. The results of both F-test and T-test performed for three pollen species indicated that, of the 19 elements investigated, B, Mg, Mn, K, Ti, and Cs resulted in significant differences between the whole grain and the coating alone; while Se, V, Pb, Cr, Al, and Zn can be recognized as being characteristic of the coating surrounding the pollen grain.

In some forensic disciplines various methods of extraction are used to perform analysis. Among these methods, solid phase extraction (SPE) and solid phase microextraction (SPME) are used in fields such as toxicology and explosives analysis. To enhance extraction efficiency in SPE and SPME, molecularly imprinted polymers (MIPs), which are designer polymers, can be more selective for the binding of an analyte or group of analytes that are similarly structured. Separation of analytes from complex mixtures is possible by utilizing these polymers. This may be especially useful in forensic applications where sample sizes may be small and composition may be complex. In this work, MIP solid phase microextraction fibers (MIP-SPME) were fabricated and caffeine was selectively sampled in the presence of theophylline and theobromine. Calibration studies were performed using the MIP-SPME to quantitate the concentration of caffeine in teas and coffees. MIP-SPME fibers were also prepared with 2,4-dinitrotoluene and deuterated 2,6-dinitrotoluene. Less selectivity was obtained for extraction of 2,4-DNT and 2,6-DNT in the presence of other DNT isomers. Fabricated blank polymers extracted analytes at the same response as templated polymers for both caffeine and DNT, despite expected results. MIP-SPE columns were also fabricated using deuterated 2,6-DNT to determine if changing the extraction procedure would increase extraction selectivity. Using different solvents in the extraction procedure changed the extraction performance efficiency of the MIPs due to the change in solvent polarity. All samples were analyzed using gas chromatography mass spectrometry.

NoSQL (Not only SQL) databases have been gaining some popularity over the last few years. Such big companies as Expedia, Shutterfly, MetLife, and Forbes use NoSQL databases to manage data on different projects. These databases can contain a variety of information ranging from nonproprietary data to personally identifiable information like social security numbers. Databases run the risk of cyber intrusion at all times. This paper gives a brief explanation of NoSQL and thoroughly explains a method of Incidence Response with MongoDB, a NoSQL database provider. This method involves an automated process with a new self-built software tool that analyzing MongoDB audit log's and generates an html page with indicators to show possible intrusions and activities on the instance of MongoDB. When dealing with NoSQL databases there is a lot more to consider than with the traditional RDMS's, and since there is not a lot of out of the box support forensics tools can be very helpful.