Forensic Analysis of Explosives Using Isotope Ratio Mass Spectrometry (IRMS)—Part 1: Instrument Validation of the DELTAplusXP IRMS for Bulk Nitrogen Isotope Ratio Measurements

Abstract: A significant amount of research has been conducted into the use of stable isotopes to assist in determining the origin of various materials. The research conducted in the forensic field shows the potential of isotope ratio mass spectrometry (IRMS) to provide a level of discrimination not achievable utilizing traditional forensic techniques. Despite the research there have been few, if any, publications addressing the validation and measurement uncertainty of the technique for forensic applications. This study, the first in a planned series, presents validation data for the measurement of bulk nitrogen isotope ratios in ammonium nitrate (AN) using the DELTA^plusXP (Thermo Finnigan) IRMS instrument equipped with a ConFlo III interface and FlashEA? 1112 elemental analyzer (EA). Appropriate laboratory standards, analytical methods and correction calculations were developed and evaluated. A validation protocol was developed in line with the guidelines provided by the National Association of Testing Authorities, Australia (NATA). Performance characteristics including: accuracy, precision/repeatability, reproducibility/ruggedness, robustness, linear range, and measurement uncertainty were evaluated for the measurement of nitrogen isotope ratios in AN. AN (99.5%) and ammonium thiocyanate (99.99+%) were determined to be the most suitable laboratory standards and were calibrated against international standards (certified reference materials). All performance characteristics were within an acceptable range when potential uncertainties, including the manufacturer’s uncertainty of the technique and standards, were taken into account. The experiments described in this article could be used as a model for validation of other instruments for similar purposes. Later studies in this series will address the more general issue of demonstrating that the IRMS technique is scientifically sound and fit‐for‐purpose in the forensic explosives analysis field.     

The kinetics of colour change in textiles and fibres treated with detergent solutions: Part II — Spectrophotometric measurements

The aim of this study was to assess colour variations that occur in several types of textiles and their constituent fibres, resulting from the long-term influence of various laundry detergents. A 14-day experiment was conducted using blue, red and grey/black cotton, wool, acrylic and polyester textiles. The spectrophotometric measurement of colour changes in fabric samples and test solutions, as well as the microspectrophotometric analysis of colour changes in single fibres were described. An evaluation of the observed colour changes from a forensic fibre analysis expert's point of view, as well as that of an average user/consumer of the textiles and laundry detergents is also provided.     

Forensic science does not start in the lab: the concept of diagnostic field tests

An attempt to improve an analytical system can focus either on the actual processing or on the input. In forensic science, much emphasis has been placed on improving laboratory procedures, as though the input is already the best that can be obtained. Means of improving the basic input have gained much less attention. Yet, it must be agreed that even the best laboratory cannot gain from an item more than has been contained in it when it arrived from the field. The detection of latent materials at the crime scene by physical or chemical techniques and the diagnostic examination of material already discovered belong to the concept of diagnostic field tests. This group also includes "mapping" for the presence of certain materials, such as latent fingerprints through the distribution of amino acids on the surface. These tests are conducted outside the laboratory, without sophisticated instrumentation, at the crime scene, the suspect's home, or elsewhere. A significant advantage of the use of diagnostic field tests is the ability to deal with "dissipating evidence" such as gunshot residue or explosive traces on the hands of suspects. If time is lost, there is a risk of losing such evidence, which is liable to deteriorate rapidly. In my presentation, I will discuss older and some newly developed forensic field tests, with specific emphasis on the Israeli experience.     

DNA条形码技术在法医昆虫学中的研究进展

嗜尸性昆虫的准确鉴定是应用昆虫判断死亡时间的先决条件,但是昆虫的鉴定只有相关类群的昆虫专家能够鉴定。DNA条形码（DNA Barcoding）技术是利用一个或少数几个DNA片段对地球上现有物种进行识别和鉴定的一项新技术。这一技术给生物分类研究带来了空前的繁荣,同时也给法医昆虫学中各昆虫种类的鉴别研究带来新的动力。     

Nuclear and mitochondrial DNA quantification of various forensic materials

Due to the different types and quality of forensic evidence materials, their DNA content can vary substantially, and particularly low quantities can impact the results in an identification analysis. In this study, the quantity of mitochondrial and nuclear DNA was determined in a variety of materials using a previously described real-time PCR method. DNA quantification in the roots and distal sections of plucked and shed head hairs revealed large variations in DNA content particularly between the root and the shaft of plucked hairs. Also large intra- and inter-individual variations were found among hairs. In addition, DNA content was estimated in samples collected from fingerprints and accessories. The quantification of DNA on various items also displayed large variations, with some materials containing large amounts of nuclear DNA while no detectable nuclear DNA and only limited amounts of mitochondrial DNA were seen in others. Using this sensitive real-time PCR quantification assay, a better understanding was obtained regarding DNA content and variation in commonly analysed forensic evidence materials and this may guide the forensic scientist as to the best molecular biology approach for analysing various forensic evidence materials.     

Validation studies of an immunochromatographic 1-step test for the forensic identification of human blood.

An immunochromatographic 1-step test for the detection of fecal occult blood was evaluated for applicability for the forensic identification of human blood in stained material. The following experiments were conducted: 1) determination of the sensitivity and specificity of the assay; 2) evaluation of different extraction media for bloodstains (sterile water, Tris buffer pH 7.5 provided in the test kit, 5% ammonia); 3) analysis of biological samples subjected to a variety of environmental insults; and 4) evaluation of casework samples. This immunochromatographic 1-step occult blood test is specific for human (primate) hemoglobin and is at least an order of magnitude more sensitive than previous methods for detecting human hemoglobin in bloodstains. The antigen is insensitive to a variety of environmental insults, except for exposure to certain detergents and household bleaches and prolonged exposure to certain preparations of luminol. The entire assay can be conducted in field testing conditions within minutes. When in the laboratory the supernatant from a DNA extraction is used for the assay, there is essentially no consumption of DNA for determining the presence of human hemoglobin in a forensic sample. The data demonstrate that this test is robust and suitable for forensic analyses.     

The Use of an Alternate Light Source for Detecting Bones Underwater

When searching underwater crime scenes or disaster scenes for fragmentary human remains, it may be advantageous for forensic divers to be able to detect the presence of bones and teeth among other marine materials (such as shells and rocks). In terrestrial environments, this can typically be accomplished by visual and instrumental methods, but underwater conditions make it difficult to employ detection and sorting techniques in these environments. This study investigates fluorescence of bones and teeth and other marine materials using a submersible alternate light source (ALS) and concludes that an ALS can be a useful tool for detecting bones and teeth in underwater searches as well in terrestrial searches and laboratory environments. The results could impact the methods and equipment used by forensic divers and forensic anthropologists when searching for skeletal remains, potentially increasing the quantity and efficiency of forensic evidence recovered.     

Optimization and validation of 10 mitochondrial DNA SNPs using SNaPshot Kit

In some forensic cases, nuclear DNA is degraded and cannot be analyzed. In such a case mitochondrial DNA (mtDNA) is usually used in forensic cases for identification because of its special features as high number of copies per cell, maternal inheritance and high mutation rate. Single nucleotide polymorphisms (SNPs) represent the most abundant class of human polymorphisms. The aim of this study was optimization of 10 mtDNA SNPs by using SNaPshot minisequencing technique on ABI310 genetic analyser in forensic molecular genetics laboratory. At the end of this study, the optimization of minisequencing technique was done by changing some assay parameters. Also, during the optimization of 10 mtSNP loci in our laboratory.     

Forensic entomology: application, education and research in Western Australia.

Forensic entomology as a science and a tool for investigation has had slow beginnings in Australia. A number of small animal decomposition trials have been recorded in the literature but mostly from an ecological rather than a forensic entomology perspective. In the last 20 years, a number of more forensically orientated field trials on small pigs and some fly developmental trials in the laboratory have been conducted but lack any replication. The following article was presented at an international seminar to detail the current research in forensic entomology, the applications of forensic entomology in scene of crime (SOC) and homicide investigations and the education of police and judiciary in the discipline of forensic entomology in Western Australia over the last 10 years.     

Forensic Confirmation Bias: Do Jurors Discount Examiners Who Were Exposed to Task-Irrelevant Information?*†

Knowledge of task-irrelevant information influences judgments of forensic science evidence and thereby undermines their probative value (i.e., forensic confirmation bias). The current studies tested whether laypeople discount the opinion of a forensic examiner who had a priori knowledge of biasing information (i.e., a defendant's confession) that could have influenced his opinion. In three experiments, laypeople (N = 765) read and evaluated a trial summary which, for some, included testimony from a forensic examiner who was either unaware or aware of the defendant's confession, and either denied or admitted that it could have impacted his opinion. When the examiner admitted that the confession could have influenced his opinion, laypeople generally discounted his testimony, as evidenced by their verdicts and other ratings. However, when the examiner denied being vulnerable to bias, laypeople tended to believe him—and they weighted his testimony as strongly as that of the confession-unaware examiner. In short, laypeople generally failed to recognize the superiority of forensic science judgments made by context-blind examiners, and they instead trusted examiners who claimed to be impervious to bias. As such, our findings highlight the value of implementing context management procedures in forensic laboratories so as not to mislead fact-finders.     

Forensic DNA laboratory automation – Principles and guidelines

There is a lack of clear guidelines for project managers, laboratory managers and forensic scientists on strategies for the automation of forensic DNA laboratory processes and operational implementation of new technologies. This is reflected in the failure rate of projects in the forensic DNA testing environment. We present a set of guidelines and concepts important for forensic laboratory automation. Some case studies from past projects are presented. These consist of partial (or modular) automation (n = 2) and full automated robotically integrated systems (n = 2).Technology Management principles and concepts are crucial to prevent failure of projects, e.g. early adoption of untried technologies, and organizational factors. The future of laboratory automation is modular until such time as new discontinuous technologies will replace the need of the traditional manual laboratory configuration in totality.     

Estimation of postmortem interval in real cases based on experimentally obtained entomological evidence

Using the entomological evidence obtained in several forensic cases analyzed in our laboratory for comparison, we evaluated the results of an experimental study carried out in a semiurban setting to determine the structure of the sarcosaprophagous fauna from a Mediterranean region of SE Spain. In all, 18 orders of arthropods were collected. The summarized experimental results refer to the most important taxa for estimating the postmortem interval. Thus, the seasonal character of certain Diptera species, such as Phaenicia sericata, Calliphora vicina, Chrysomya albiceps and Musca domestica, is cited. Among the Coleoptera, the role of Dermestidae as a necrophagous species, and of Staphylinidae and Histeridae as necrophilous, or Cleridae, Tenebrionidae and Nitidulidae as omnivorous, as well as their appearance on the corpse, is described. Among the Hymenoptera, Formicidae were as the most abundant group, acting as omnivores and not apparently related to any particular decomposition stage. The real cases are discussed using data from the literature and the experimentally obtained results. In every case, the most relevant factors for estimating PMI are briefly discussed, mentioning, when possible, the relation with the experimental results. We confirm the significance of the experimental results because they seem to be applicable to actual forensic cases, the details of which enlarge our very little knowledge on the subject in the Iberian Peninsula. The importance of regional faunistic studies of the sarcosaprophagous arthropod community, the results of which may be applied to forensic practice, is mentioned.     

Cyclic Pentanone Peroxide: Sensitiveness and Suitability as a Model for Triacetone Triperoxide

Research to counter the threat of organic peroxides such as triacetone triperoxide (TATP) is at times hampered by their inherent extreme sensitiveness and volatility. This work describes an approach to lowering some risks associated with the handling of TATP in the laboratory through the use of an analog species, tripentanone triperoxide (TPTP). Sensitiveness has been tested via standard methods. GCMS analysis has shown that TPTP degrades via similar mechanisms to TATP under a range of conditions. Slight differences in product composition were traced to side reactions which may also affect impurities present in homemade TATP synthesis. A pilot field trial was conducted to evaluate TPTP as a substitute for TATP in explosive detection dog (EDD) scent training. The degradation studies have yielded insights into the complexities of the acidic degradation of cyclic peroxides with potential forensic application, and TPTP's inadequacy as a TATP pseudoscent is a valuable example of the limitations of such training aids.     

A New Robust Epigenetic Model for Forensic Age Prediction

Forensic DNA phenotyping refers to an emerging field of forensic sciences aimed at the prediction of externally visible characteristics of unknown sample donors directly from biological materials. The aging process significantly affects most of the above characteristics making the development of a reliable method of age prediction very important. Today, the so-called “epigenetic clocks” represent the most accurate models for age prediction. Since they are technically not achievable in a typical forensic laboratory, forensic DNA technology has triggered efforts toward the simplification of these models. The present study aimed to build an epigenetic clock using a set of methylation markers of five different genes in a sample of the Italian population of different ages covering the whole span of adult life. In a sample of 330 subjects, 42 selected markers were analyzed with a machine learning approach for building a prediction model for age prediction. A ridge linear regression model including eight of the proposed markers was identified as the best performing model across a plethora of candidates. This model was tested on an independent sample of 83 subjects providing a median error of 4.5 years. In the present study, an epigenetic model for age prediction was validated in a sample of the Italian population. However, its applicability to advanced ages still represents the main limitation in forensic caseworks.     

A NATIONAL SURVEY OF US POLICE ON PREPARATION AND CONDUCT OF IDENTIFICATION LINEUPS

A great deal of research has been aimed at identifying the factors that produce errors in eyewitness identification. However, most of this work has been conducted in laboratory environments using undergraduates and naive lay persons as research participants. Little information is available on what police officers do in the course of their identification activities. The present research investigated the procedures that police officers report employing when constructing and administering eyewitness identification lineups. We developed a survey that addresses a number of these issues, including experience, sources of lineup foils, lineup formation and display, lineup fairness evaluation, presence of suspect's attorney, witness instructions, historical records, and legal challenges. Surveys were sent to 500 US police jurisdictions; 220 were returned. Several survey items showed results consistent with previous laboratory research, however police officers reported using some procedures that are different from those established as most effective in the research literature. This paper summarizes the results of our survey and discusses the implications for future research and forensic procedures.     

Forensic Analysis of a Contact Lens in a Murder Case

Contact lenses have had rare relevance in trials and/or investigations. After 5 years of burial, orbital remnants were retrieved from an exhumed body and subsequently identified as a key piece of material evidence in a murder trial. The exhumed case materials were evaluated under laboratory conditions and were determined to be contact lens remnants. Contact lens fracture and burial simulation studies were performed to provide additional corroboration of the physical findings of the exhumed contact lens remnants. This material evidence was instrumental in providing factual proof refuting the defendant's testimony in the murder trial. A brief history of contact lens composition and use is provided for understanding the methods and observational results. This forensic case study represents the first published documentation of a contact lens from an exhumed body being used in a murder investigation and establishes an operational procedure for future forensic contact lens examinations.     

Organizational and Human Factors Affecting Forensic Decision-Making: Workplace Stress and Feedback

Although forensic examiners operate in a stressful environment, there is a lack of understanding about workplace stress and feedback. These organizational and human factors can potentially impact forensic science judgments. In this study, 150 practicing forensic examiners from one laboratory were surveyed about their experiences of workplace stress, and the explicit and implicit feedback they receive. Forensic examiners reported that their high stress levels originated more from workplace-related factors (management and/or supervision, backlogs, and the pressure to do many cases) than from personal related factors (family, medical, and/or financial). The findings showed that a few (8%) of the forensic examiners sometimes felt strong implicit feedback about what conclusions were expected from them and that some (14%) also strongly felt that they were more appreciated when they helped to solve a case (e.g., by reaching a “match” as opposed to an “inconclusive” conclusion). Differences were found when comparing workplace stress and feedback levels across three core forensic science fields (forensic biology, chemistry, and latent prints) and across career stages (early, mid, and late). Gaining insights into the stress factors within a workplace and explicit and implicit feedback has implications for developing policies to improve the well-being, motivation, and performance of forensic examiners.     

Estimation of stature and determination of sex from radial and ulnar bone lengths in a Turkish corpse sample

Determination of sex and estimation of stature are important aspects of forensic identification of an unknown individual. In the absence of pelvis the sex is assessed from long bones and cranium as they both provide high accuracy in sexing. The present study is an attempt to assess sex and stature from long bones of the forearm using recently deceased forensic cases in Istanbul, Turkey. The sample is composed of 80 males and 47 females with an average age of 36 and 30 years, respectively. Length measurements from the radius and ulna were obtained by exposing the epiphyseal ends of the long bones in a fashion similar to dry long bones. Discriminant function statistics showed a sex determination accuracy as high as 96%. Regression analysis was used in stature estimation from these two bones. Ideally osteological remains are necessary to make standards for osteological identification. These materials are not always easy to obtain and those available seem to be less ideal when they do not represent a current population. Forensic anthropologists therefore should develop techniques that utilize autopsy remains.     

Ultraviolet illumination as an adjunctive aid in dental inspection

Tooth-colored resin fillings have become increasingly popular as restorative materials. Their presence in the dentition presents a challenge to the clinician and the forensic odontologist, as detection of the fillings can be difficult both visually and radiographically. As they necessarily form part of the unique dentition of an individual, recognition of the resins is important for forensic identification. Alternative light sources have been used with success in various fields of forensic science. In recent years small LED flashlights emitting at specific wavelengths in the ultraviolet light (UV) range have been developed. Their low cost, small size, and ready availability makes their use practical in both forensic dental inspection and clinical settings. UV inspection is of interest because enamel, dentin and dental materials all have differing fluorescent properties when illuminated by UV light. It was one goal of this research to quantitatively assess the fluorescence properties of modern restorative resins in order to predict their behavior during inspection using UV illumination. The second goal was to demonstrate practical use of UV in dental inspection with examples of how different materials fluoresce. Quantitative measurements were obtained for optical emission wavelength and intensity for 15 modern resins using a spectrophotometer. Results indicated that resin brands fluoresce at different wavelengths and with varying intensities. Practical use and comparison of the flashlights revealed that the most useful excitation wavelengths for resin detection were in the UVA range (365 and 380 nm). Porcelain restorations and composite resin fillings exhibited different responses to these two wavelengths and thus use of both is recommended for forensic dental inspection.