It is not uncommon for an individual to commit arson as a method of concealing a crime, whether the goal be to destroy a body, DNA evidence, or any other information that may link the suspect to the scene. Fortunately, for investigators, setting a fire to the crime scene does not always destroy all evidence. Some pieces of evidence are more resilient than others. For example, evidence such as ignitable liquids and other accelerants can often be detected after the fire. In the event that an item of evidence like clothing is not completely incinerated, the presence of biological fluids may also be detected through presumptive testing and eventually lead to the identification of an individual through DNA analysis. The purpose of this study was to determine whether or not DNA analysis can be performed effectively after heated passive headspace concentration, without causing irreparable degradation to DNA evidence. Heated passive headspace concentration is a common procedure for extracting ignitable liquids from a substrate to identify and confirm the presence of that substance. This process requires long incubations at fairly high temperatures within a tightly sealed vessel to prevent evaporation. If practitioners can delay DNA analysis steps, move straight into heated passive headspace concentration, and lower the chances of losing a portion of the ignitable liquid to evaporation, it may assist in arson investigations. This study explores DNA quantity and quality in saliva and semen stains after incubation times and temperatures based on the recommended upper and lower temperature and timeframe limits of ASTM Guidelines E1412-19 and E1388-17 entitled “Standard Practice for Separation of Ignitable Liquid Residues from Fire Debris Samples by Passive Headspace Concentration with Activated Charcoal” and “Standard Practice for Static Headspace Sampling of Vapors from Fire Debris Samples,” respectively. Possible DNA analysis inhibitors such as gasoline, open flame, and burnt substrate were also explored. It was determined that while open flame in direct contact with a biological stain caused significant damage in all saliva stains and some semen stains, the presence of gasoline and burnt substrate did not appear to inhibit DNA analysis. Additionally, heated passive headspace concentration conditions did not appear to cause significant degradation or inhibition that would result in an incomplete genetic profile. Further experimentation is necessary given the presence of extraneous factors. For example, the initial amount of DNA deposited onto substrates was unknown. However, it is reasonable to state that performing heated passive headspace concentration before DNA analysis may be a feasible option if desired in a forensic laboratory.

The current methods for identifying ignitable liquid residues in fire debris are heavily based on the holistic, qualitative interpretation of chromatographic patterns with the mass spectral identification of selected peaks. The identification of neat, unweathered ignitable liquids according to ASTM 1618 using these methods is relatively straightforward for the trained analyst. The challenges in fire debris analysis arise with phenomena such as evaporation, substrate interference, and biodegradation. These phenomena result in alterations of chromatographic patterns which can lead to misclassifications or false negatives. The biodegradation of ignitable liquids is generally known to be more complex than evaporation [20], and proceeds in a manner that is dependent on numerous factors such as: composition of the petroleum product/ignitable liquid, structure of the hydrocarbon compound, soil type, bacterial community, the type of microbial metabolism that is occurring, and the environmental conditions surrounding in the sample. While nothing can be done to prevent the biodegradation, continued research on biodegraded ignitable liquids and the characterization of the trends observed may be able to provide insight into how an analyst can identify a biodegraded ignitable liquid residue. This research utilized normalized abundance values of select ions from pre-existing gas chromatography-mass spectrometry (GC-MS) data on samples from three different gasoline and diesel biodegradation studies. A total of 18 ions were selected to indicate the presence of six hydrocarbon classes (three each for alkanes, aromatics, cycloalkanes, naphthalenes, indanes, and adamantanes) based on them being either base peaks or high abundance peaks within the electron impact mass spectra of compounds within that hydrocarbon class. The loss of ion abundance over the degradation periods was assessed by creating scatter plots and performing simple linear regression analyses. Coefficient of determination values, the standard error of the estimate, the slope, and the slope error of the best fit line were assessed to draw conclusions regarding which classes exhibited desirable characteristics, relative to the other classes, such as a linear degradation, low variation in abundance within the sampling days, and a slow rate of abundance loss over the degradation period. Additional analyses included two-way analysis of the variance (ANOVA), to assess the effects of time as well as different soil type on the degradation of the hydrocarbons, stepwise multinomial logistic regressions to identify which classes were the best predictors of the type of ignitable liquid, and one-way ANOVAs to determine where the differences in the ratios of hydrocarbon classes existed within each of the ignitable liquids, as well as between the two liquids. Hydrocarbon classes identified as exhibiting characteristics such as slow and/or reliable rates of abundance loss during biodegradation are thought of as desirable for future validation studies, where specific ranges of hydrocarbon class abundance(s) may be used to identify the presence of a biodegraded ignitable liquid. Classes of hydrocarbons that have experienced biodegradation that maintain an abundance close to that of a neat, non degraded counterpart, or that reliably degrade and have predictable abundance levels given a particular period of degradation, would be instrumental in determining whether or not an unknown sample contains an ignitable liquid residue. It is the hope that these assessments will not only provide helpful information to future researchers in the field of fire debris analysis, but that they will create interest in the quantitative, statistical assessment of ignitable liquid data for detection and identification purposes.

Identification of ignitable liquids in fire debris analysis using pattern recognition is an important step in determining the nature of a suspicious fire. Complex mixtures that make up ignitable liquids are susceptible to microbial degradation when fire debris evidence is presented in the form of soil. Microbial degradation results in a selective metabolism of certain classes of compounds required for identification of an ignitable liquid. Various ignitable liquids that may be used to initiate or propagate a fire contain different classes of organic compounds. These include normal alkanes, branched alkanes, cycloalkanes, aromatics, terpenes, and others. In this work, microbial degradation of nine ignitable liquids in soil was evaluated over a period of twenty-six days. The degradation of aromatic compounds in gasoline was faster with toluene and C2-alkylbenzenes than in C3-alkylbenzenes. However, the overall loss of aromatics made gasoline chromatographically unidentifiable. The complete loss of n-alkanes in medium and petroleum distillates resulted in patterns that resembled naphthenic-paraffinic products. Normal alkanes were more susceptible to microbial degradation than isoalkanes, which was specifically demonstrated in medium and heavy petroleum distillates. In diesel, pristane and phytane remained prominent in comparison to the normally prevalent n-alkanes, which could no longer be detected post-degradation. The degradation of isoalkanes and cycloalkanes was evaluated in a naphthenic-paraffinic product. Isoalkanes were degraded significantly faster than cycloalkanes. The remaining peaks in the naphthenic-paraffinic pattern consisted solely of cycloalkane compounds, and could no longer be classified as a naphthenic-paraffinic product. The terpene compounds in turpentine were also observed to be susceptible to degradation by microorganisms. The loss of !-pinene, limonene, and camphene was significantly noticeable in comparison to other terpene compounds, such as 1,4-cineole. Microbial biodegradation in different soil types was investigated. The difference in soil texture can affect the rate of metabolism of ignitable liquids due to the variance of available oxygen, nutrients and mobility of the microbial population. The degradation of isoalkanes, cycloalkanes, aromatics and heavier normal alkanes was faster in clay, whereas normal alkanes of lower molecular weight were degraded more readily in sand. There has been no explanation of this occurrence within the scientific literature, however it could be hypothesized that the difference in microbial flora and water saturation levels could affect the selective degradation between the two soil types. Fire debris evidence is often stored for long periods of time before analysis due to case backlogs. The storage condition of arson-related soil samples is a sensitive subject. If evidence, containing soil, is stored at room temperature, petroleum compounds in any ignitable liquid residues that are present will be degraded within a week. Therefore, it is important to freeze or refrigerate soil samples. The storage of both refrigerated and frozen soil samples containing gasoline were evaluated over six months. Less than 6% of the aromatic compounds distinctive of gasoline remained when stored at 5 °C, while minimal change was observed in the same compounds when stored at -15 °C. Microbial degradation of petroleum-based ignitable liquids is advantageous from the environmental perspective. However, within the forensic community the effect of microbial action could lead to misclassification or inability to identify the presence of an ignitable liquid in fire debris evidence.

Gasoline is a substance commonly encountered in forensic settings. Unfortunately, gasoline is an easily obtainable ignitable liquid that arsonists commonly use to initiate or expedite the spread of an intentionally set fire. Fires claim the lives of many people each year in addition to causing widespread property damage. Many fire scene investigations result in charges of arson, which has the legal connotation of a committed crime. For this reason, extensive analysis and investigation must be undertaken before any suspected arson scene is deemed an actual case of arson. Although ignitable liquids, including gasoline, may be present at the scene of a fire, it does not necessarily mean they were intentionally used as accelerants. An accelerant is a fuel used to initiate a fire. These realities, in addition to several other factors, demonstrate why a rapid, reliable, gasoline analysis method is crucial to forensic applications. In this thesis, direct analysis in real time – mass spectrometry (DART-MS) is evaluated as a potential method that could better identify, distinguish and classify gasoline brands from one another. Techniques such as DART-MS could enable forensic laboratories to better identify questioned gasoline samples. Many ignitable liquids share similar chemical properties, and forensically relevant evidence is often obtained from a crime scene in less than favorable conditions. Fire debris can encompass various materials, including burnt carpet, flooring, items of furniture and clothing, among others. If gasoline was used as an accelerant, it may be present in trace amounts after the termination of the fire. Materials submitted for laboratory analysis may be substrates with compositions that have components similar to those found in some ignitable liquids. These are just a few of the potential obstacles that could be encountered with analyzing fire debris in a forensic setting. Traditionally, gas chromatography – mass spectrometry (GC-MS) methods are utilized for gasoline analysis in the criminal laboratory setting. While traditional GC-MS methods are sensitive and able to classify samples as gasoline, they are time consuming in terms of both sample preparation and analysis. Additionally, they do not generate differential mass spectral data based on the brand of gasoline. Conversely, gasoline analysis in this research, utilizing the DART-MS method, demonstrated that five different brands of gasoline could be distinguished from one another both by visual examination of mass spectra and with methods of chemometric analysis. Advantageously, the DART-MS method, an ambient ionization technique, requires little sample preparation and a rapid sample analysis time, which could drastically increase the throughput of standard sample analysis with further method development. The goals and objectives of this research were to optimize the DART-MS parameters for gasoline analysis, determine if DART-MS analysis could distinguish gasoline by brand, develop chemometric models to appropriately classify gasoline samples, and finally lay groundwork for future studies that could further develop a more efficient and discriminating DART-MS gasoline analysis method for forensic casework. Each brand of gasoline was observed to have a chemical attribute signature (CAS) consisting of not only low-mass ions, but also a variety of high-mass ions not usually observed with gasoline samples analyzed by GC-MS. Although variables including season, storage time, dilution and age of the gasoline were observed to contribute to the resulting mass spectral data, once the mass spectra are better understood, they could offer even more discriminating power between samples than simple analysis of the gasoline brand. In this research, DART-MS parameters were first optimized for gasoline analysis. Subsequently, the five acquired brands of gasoline: Shell, Sunoco, Irving, Cumberland Farms and Gulf, were analyzed both undiluted (or neat) and diluted utilizing the DART-MS analysis method. GC-MS data was generated and analyzed to show comparisons. After analyzing the data generated by both approaches, it was apparent that the DART-MS method could generate CASs based on the gasoline brand and offer a degree of differentiation that traditional GC-MS does not. Additional chemometric analyses utilizing principle component analysis (PCA) and the construction of models with Analyze IQ Lab software verified that the gasoline brands were distinguishable when samples were analyzed with this ambient ionization method. PCA plots of the neat gasoline demonstrated clustering based on brand. Additionally, models constructed from training samples generated from DART-MS analysis of the various brands were able to accurately classify gasoline samples as "yes" or "no" when a test set of gasoline was compared to all five brands. The lowest associated testing error rate for some of these models was 0%. However, additional analysis with greater sample sizes needs to be further carried out to more accurately evaluate this method of gasoline analysis and classification.

Fire incidents are a major contributor to the number of deaths and property losses within the United States each year. Fire investigations determine the cause of the fire resulting in an assignment of responsibility. Current methods of fire debris analysis are reviewed including the preservation, extraction, detection and characterization of ignitable liquids from fire debris. Leak rates were calculated for the three most common types of fire debris evidence containers. The consequences of leaking containers on the recovery and characterization of ignitable liquids were demonstrated. The interactions of hydrocarbons with activated carbon during the extraction of ignitable liquids from the fire debris were studied. An estimation of available adsorption sites on the activated carbon surface area was calculated based on the number of moles of each hydrocarbon onto the activated carbon. Upon saturation of the surface area, hydrocarbons with weaker interactions with the activated carbon were displaced by more strongly interacting hydrocarbons thus resulting in distortion of the chromatographic profiles used in the interpretation of the GC/MS data. The incorporation of an additional sub-sampling step in the separation of ignitable liquids by passive headspace sampling reduces the concentration of ignitable liquid accessible for adsorption on the activated carbon thus avoiding saturation of the activated carbon. A statistical method of covariance mapping with a coincident measurement to compare GC/MS data sets of two ignitable liquids was able to distinguish ignitable liquids of different classes, sub-classes and states of evaporation. In addition, the method was able to distinguish 10 gasoline samples as having originated from different sources with a known statistical certainty. In a blind test, an unknown gasoline sample was correctly identified from the set of 10 gasoline samples without making a Type II error.

The fire debris analyst is often faced with the complex problem of identifying ignitable liquid residues in the presence of products produced from pyrolysis and incomplete combustion of common building and furnishing materials. The purpose of this research is to investigate a modified destructive distillation methodology provided by the Florida Bureau of Forensic Fire and Explosive Analysis to produce interfering product chromatographic patterns similar to those observed in fire debris case work. The volatile products generated during heating of substrate materials are extracted from the fire debris by passive headspace adsorption and subsequently analyzed by GC-MS. Low density polyethylene (LDPE) is utilized to optimize the modified destructive distillation method to produce the interfering products commonly seen in fire debris. The substrates examined in this research include flooring and construction materials along with a variety of materials commonly analyzed by fire debris analysts. These substrates are also burned in the presence of a variety of ignitable liquids. Comparisons of ignitable liquids, pyrolysis products, and products from pyrolysis in the presence of an ignitable liquid are performed by comparing the summed ion spectra from the GC-MS data. Pearson correlation was used to determine if substrates could be discriminated from one another. A pyrolysis products database and GC-MS database software based on comparison of summed ion spectra are shown to be useful tools for the evaluation of fire debris.

The detection and identification of the oxygenated class of ignitable liquids is a complex issue for the fire debris analyst. The oxygenated compounds are difficult to recover using traditional analytical techniques since their chemical characteristics are vastly different from those of the petroleum products that compose the majority of the ignitable liquid classes. Previous research has demonstrated that the use of zeolite 13X as an adsorbent in heated passive headspace concentration provides increased recovery of oxygenated compounds in comparison to the conventional activated charcoal adsorbent. This hypothesis was further tested in this work using more realistic casework scenarios. Various carpet, carpet padding and wood types were utilized in a number of burn conditions in order to determine if any substrate interferences were present; as well as to monitor the recovery of oxygenated compounds from these substrates using the proposed zeolite extraction method. The substrates explored did not contribute significant background interference to complicate the identification of the oxygenated compounds. In addition, small volumes of the oxygenated ignitable liquids were easily recovered and identified from all burn states using the zeolite method. A dual-mode extraction with both zeolites and activated charcoal strips as adsorbents was utilized with mixtures of oxygenated compounds and petroleum products to determine if a variety of ignitable liquid classes could be detected and identified in the presence of a variety of substrate matrices within a single extraction protocol. The dual-mode extraction showed that both the oxygenated compounds and petroleum products could be detected and identified using a single extraction protocol in the presence of various substrate matrices. Lastly, an experiment was devised to compare the recovery of the oxygenated compounds using various total available surface areas of both zeolites and activated charcoal strips in order to determine which adsorbent exhibits a greater recovery when all other experimental conditions remain constant. When the surface areas were equalized between the zeolites and activated charcoal strips, the activated charcoal exhibited a greater recovery of the oxygenated compounds. However, the cost effectiveness of the zeolites allows for a greater amount of zeolite beads to be used in order to achieve the same recovery as the activated charcoal strips for a much lower price. Therefore, the findings from this work, in combination with previous research, continue to support the use of zeolite 13X as an alternative adsorbent for the recovery of oxygenated ignitable liquids from fire debris evidence.

In Ontario, fire investigators from the Office of the Fire Marshal (OFM) are responsible for determining the origin and cause of suspicious fires. As part of the investigation, fire debris samples are collected from the scene and analyzed by the Centre of Forensic Sciences. The standard practice is to collect items that are porous, highly absorbent or adsorbent with high surface areas as they allow for better retention of the ignitable liquids. The evidence typically collected includes carpets, cardboards, soils, cloths and other items that have not been impinged by flame such as beneath baseboards. These samples are analyzed for the presence of ignitable liquid residues which may be evidence that an accelerant was used at the fire. When a body is recovered from a fire it can provide another source from which to collect samples for analysis. These samples can be especially helpful in instances where the fire generated an intense heat which may cause a loss of ignitable liquid residues from the fire debris. The tissue samples have a greater likelihood of still containing residues as the organs and body fluids can act as a shield protecting the residues from volatilization. The purpose of this study is to validate whether a victim was alive or deceased at the time a fire was intentionally set by detecting presence or absence of gasoline residues within their lungs and heart blood post fire. It was hypothesized that only when a victim was alive and performing respiration would sufficient gasoline vapours enter the airways and bloodstream for detection postmortem. Contamination becomes a significant issue when these samples are collected at autopsy and this study aimed to determine the accuracy with which a gasoline signature can be interpreted following the collection and analysis of lung tissue and heart blood. Pig (Sus domesticus) carcasses were chosen as acceptable analogues for humans in this study. The experiments involved anaesthetizing a pig (with Animal Ethics Approval), exposing the pig to gasoline vapours for 10 minutes, and then euthanizing it. The carcass was clothed with a cotton t-shirt and placed in a house where additional gasoline was poured onto it. The house also contained two additional clothed pig carcasses which did not inhale gasoline vapours; one with gasoline poured directly onto it and the other with no gasoline exposure (negative control). Thermocouples were placed under each carcass and in the centre of each room at ceiling and floor level to record the temperature. The house was set ablaze and monitored by a volunteer fire service. After the fire had reached V flashover and was suppressed, the carcasses were collected and their lungs and heart blood excised at a necropsy. The lungs and heart blood were then placed into glass mason jars following the OFM protocol. The headspace from each sample was analyzed by thermal desorption-gas chromatography-mass spectroscopy to determine the presence or absence of a gasoline signature. Two full scale house fires were conducted in order to obtain three replicates. The results showed that only the lungs and heart blood from the pig that inhaled gasoline contained gasoline residues. This indicates that it is possible to determine a victim’s status at the time of the fire based on the detection of gasoline in the lungs and/or heart blood. It was also concluded that contamination of samples during an autopsy can be minimized by changing gloves before handling the internal tissues. The thermal data showed that the bodies act as an insulator and protects the underside as the temperatures under the carcasses did not exceed 30⁰C while the room reached over 900⁰C at the first full scale house fire. These results will impact the forensic community by demonstrating the importance of analyzing a deceased victim’s internal tissues for ignitable liquid residues post fire as they may provide evidence of an intentionally set fire as well as providing information about the victim’s status when a fire was started. These findings will have a direct impact to the OFM as additional evidence can be obtained by completing internal tissue analysis. This will intern impact the Centre of Forensic Science (CFS) as it confirms the importance of analyzing internal tissues in order to provide results to fire investigators. Finally these findings should be used to implement new protocols at the Coroner’s Office so contamination can be minimized during fire autopsies and accurate samples are collected and sent to the CFS for analysis. / UOIT

In fire debris analysis, substrate contribution refers to compounds present within the material collected that can interfere with the instrumental detection of ignitable liquids or contribute petroleum or alcohol-based compounds, which may complicate the interpretation. The concept of substrate contribution was brought to light by "The petroleum-laced background" by Lentini et al. focusing on commercial products (e.g. tennis shoes, magazines, etc.), the publication successfully illustrated that these products can produce chromatograms similar to those generated by the presence of petroleum-based ignitable liquids (ILs). As a result, Lentini et al. demonstrated that fire debris analysts can identify the presence of ignitable liquids without realizing the compounds in question might be the result of the manufacturing processes, and are inherent to the substrate in question. Therefore, the findings may or may not be probative. Gasoline is easily accessible and is frequently used by arsonists. As such, fire debris analysis focuses primarily on petroleum-based compounds. However, oxygenated solvents, which encompass all oxygen-containing compounds as defined by the American Society for Testing and Materials (ASTM) classification scheme, can also be used in an arson event. Despite the potential to be used as ILs, little is known regarding the recovery of these compounds. Previous thesis projects from the Biomedical Forensic Sciences program at Boston University School of Medicine explored and optimized the use of zeolites in recovering low molecular weight oxygenated ignitable liquids. An isothermal gas chromatography/mass spectrometry (GC/MS) method was also developed to detect these oxygenated ILs. The results from these projects show that zeolites have the potential to be used in forensic casework. Inspired by previous publications and thesis research, the goal of this project was to first develop a reference library on substrate contribution from oxygenates (e.g. ethanol, isopropanol and acetone) present in commercial products using the isothermal GC/MS methods. The development of this reference library included a specific interest in wood treatment products, considering wood is one of the most commonly submitted fire debris materials. The second stage involved an attempt at evaluating extraction efficiencies of activated charcoal strip and zeolites. The results of this project suggested that automotive and food products examined contained only acetone and ethanol respectively, while the variety of oxygenates found in household and personal care products indicated further analysis of additional products in these categories would be beneficial. Moreover, the results also reaffirmed zeolites' role in recovering oxygenated ILs in a controlled testing environment using KimWipes as a non-contributing substrate. However, the instrumental method required some modifications, as there was partial separation between ethanol and acetone. The results from applying products onto wooden blocks suggested that activated charcoal strips recovered more oxygenates than zeolites. This unexpected result prompted an investigation into the existing extraction parameters. The investigation suggested that the wooden blocks themselves were responsible for the unexpected recovery results, and future studies would be needed to understand if this recovery was substrate-specific.

Ignitable liquids consist of either a single organic compound or a complex organic mixture. In regards to fire debris analysis, the analyst is responsible for determining if an ignitable liquid residue is present. However, when extracted from soil-containing fire debris evidence, chemical degradation from microorganisms is observed to result in the loss of compounds based on chemical structure. It can also happen when the evidence container is stored at room temperature before analysis. This can present a challenge to the fire debris analyst when identifying and classifying the ignitable liquid residue based on the criteria established by standard test methods. The purpose of this research was to observe the microbial degradation of fourteen compounds, at room temperature over a period of time, for possible by-product formation that could coincide with compounds normally present in an ignitable liquid. Additionally, a quantitative assessment was performed to observe and record the loss rate of compounds in a representative simple mixture. Finally, the loss rate from the simple mixture was compared to commercially available ignitable liquids. Degradation studies were conducted to observe the microbial degradation of a representative compounds (individually and in a simple mixture, both weathered and unweathered) and seven ignitable liquids of different ASTM E1618 classifications. Potting soil was spiked with 20 µL of a liquid/compound and was allowed to stand at room temperature for a period of time. The simple mixture was evaporated to 50% and 90% using a steady nitrogen gas flow to compare the degradation process to the unweathered mixture. All samples were extracted and analyzed using passive-headspace concentration and gas chromatography-mass spectrometry. The formation of by-products was not observed when degrading the compounds from the simple mixture individually as seen in other research. The simple mixture, unweathered and 50% weathered, resulted in rapid degradation of their oxygenated compounds. The straight-chained alkanes and toluene were observed to be more susceptible to microbial attack than the highly-substituted aromatics and the branched and cyclic alkanes. The 90% weathered mixture followed the same degradation trend as the unweathered and 50% weathered samples, although it only contained two compounds. The loss rates/half-lives for each simple mixture sample (unweathered, 50% weathered, and 90% weathered) were determined to be approximately 3.5, 3.5, and 0.84 days. The unweathered and 50% weathered sample half-lives were similar due to containing compounds with similar susceptibility to degradation, while the 90% weathered sample contained one compound that was more highly susceptible to degradation. When comparing the 3.5 day half-life to the seven different ASTM class liquids, the isoparaffinic product and the naphthenic-paraffinic product had similar rates of degradation while aromatic solvent and normal alkane classes had the shortest half-lives. When observing the degradation of the gasoline, medium petroleum distillate and the miscellaneous, the constituent compounds were seen to exhibit a range of degradation rates that corresponded to half-lives less than and greater than 3.5 days.