Latent print evidence collected from crime scenes are analyzed and compared by examiners using the ACE-V protocol (Analysis, Comparison, Evaluation-Verification) to identify potential suspects. However, smudged and poor-quality prints, usually deemed inconclusive, could be utilized for source association or discrimination based on the chemistry of the oils and sweat that comprises the latent print residue (LPR). For this research, LPRs were collected from various places on the face and analyzed via two-dimensional gas chromatography-mass spectrometry (2D GC-MS), one-dimensional (1D) GC-MS, and direct analysis in real time-high resolution mass spectrometry (DART-HRMS). LPR recovery with two common sample preparation methods, derivatization and non-derivatization, was examined when collected on a porous and non-porous substrate. The method that provided high correlation and low relative standard deviation for each substrate was used for subsequent studies. Then, the LPRs were investigated to observe if association to a source can be achieved in either 1D or 2D GC. Comparison methods of Pearson correlation coefficient (PCC), principal component analysis (PCA), hierarchical cluster analysis (HCA), and receiving operating characteristic (ROC) curves were used to test source association. An aging study was performed to analyze the change in latent print chemistry when simulating evidence storage for up to three months via 1D GC and DART-HRMS. During this time, a longevity study was conducted by collecting LPRs every two weeks to test the intra- and inter-variability of the recovered chemistry. Correlation and similarity metrics such as PCC, Spearman's rank correlation, and Euclidean distance were used to compare the monthly and weekly changes of the LPRs.

The ability to associate a smokeless powder, smokeless powder residue, or organic gunshot residue (OGSR) to one another may be helpful in determining the origin of a suspected sample and aid in linking a suspect to a crime scene. In this study, smokeless powders were extracted and analyzed via gas chromatography – mass spectrometry (GC-MS) and direct analysis in real time – high resolution mass spectrometry (DART-HRMS). Subsequently, group definition was performed using hierarchical cluster analysis and principal component analysis followed by internally validated classification models. Then, smokeless powder residues were generated in-lab and extracted. Resulting residue data from each instrument was classified within the respective smokeless powder model using linear discriminant analysis (LDA) with external test sets. Residue groupings and classification models were also generated. Ammunition was loaded with known smokeless powder, then fired to collect OGSR from cloth targets. The OGSR was extracted and analyzed via DART-HRMS and GC-MS, then tested against the smokeless powder and residue models to determine the association of OGSR to its intact smokeless powder as well as to lab generated residues. Reference classes for the OGSR samples in the LDA prediction were determined via flow charts for informed analyst determination of class in smokeless powder and residue models. Standards of common smokeless powder components were pyrolyzed and an expected pyrolysis products profile was created for each sample based on the intact composition. Similarity and correlation metrics including Pearson's correlation, Sørensen-Dice similarity coefficient, and Concordance correlation were evaluated in the comparison of smokeless powder to residue and residue to expected pyrolysis products. Pearson's correlation was used in the comparison of smokeless powder to OGSR and smokeless powder residue to OGSR.

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.

Currently, DNA profiling is the gold standard to identify an individual. However, determining body fluid origin is important in criminal investigations, offering additional information surrounding the circumstances of a crime. However, crime labs can only definitively identify blood and semen and presumptively saliva using techniques that consume time and sample and do not simultaneously identify all forensically relevant body fluids. This causes many crime labs to want to bypass body fluid identification altogether. Therefore, advances into more definitive molecular-based body fluid methods are necessary. One such technique is mRNA profiling because it provides a highly sensitive and specific approach to definitively identifying all relevant body fluids in parallel. Although advancements have been made, improvements to mRNA profiling methodologies still need to be researched such as 1) possible mRNA recovery from established DNA workflows and 2) possible integration of mRNA profiling into an upfront male DNA screening assay for triaging sexual-assault evidence likely to contain male DNA and reduce/eliminate a significant bottleneck in the standard DNA workflow of microscopic sperm identification. This study was designed to address these two issues by evaluating a novel way to recover RNA, for body fluid identification, from the waste fractions of a PrepFiler™ DNA extraction, and from the DNA extracts directly. Next, this study aimed to provide a relatively quick molecular-based approach for screening sexual-assault evidence. It involves extraction of RNA using the Dynabeads™ mRNA DIRECT™ Kit, while saving the extraction waste fractions for downstream male-DNA quantitation and STR profiling. The RNA is then used in a rapid and sensitive 1-step combined reverse transcription-HRM assay to positively detect the presence of sperm. Both non-conventional co-extraction methods successfully addressed current body fluid identification challenges and allowed for easy integration into existing workflows when single sourced, mixture and mock casework samples were analyzed.