Quality assurance testing of an explosives trace analysis laboratory--further improvements

The Forensic Explosives Laboratory (FEL) operates within the Defence Science and Technology Laboratory (DSTL) which is part of the UK Government Ministry of Defence (MOD). The FEL provides support and advice to the Home Office and UK police forces on matters relating to the criminal misuse of explosives. During 1989 the FEL established a weekly quality assurance testing regime in its explosives trace analysis laboratory. The purpose of the regime is to prevent the accumulation of explosives traces within the laboratory at levels that could, if other precautions failed, result in the contamination of samples and controls. Designated areas within the laboratory are swabbed using cotton wool swabs moistened with ethanol water mixture, in equal amounts. The swabs are then extracted, cleaned up and analyzed using Gas Chromatographs with Thermal Energy Analyzer detectors. This paper follows on from a previous published paper describing the regime and summarizing subsequent results from approximately 6 years of tests. Lessons learned and improvements made over the period are also discussed. Monitoring samples taken from surfaces within the trace laboratories and trace vehicle examination bay have, with few exceptions, revealed only low levels of contamination, predominantly of RDX. Analysis of the control swabs, processed alongside the monitoring swabs, has demonstrated that in this environment the risk of forensic sample contamination, assuming all the relevant anti-contamination procedures have been followed, is so small that it is considered to be negligible. The monitoring regime has also been valuable in assessing the process of continuous improvement, allowing sources of contamination transfer into the trace areas to be identified and eliminated.     

Quality assurance testing of an explosives trace analysis laboratory — Further improvements to include peroxide explosives

The Forensic Explosives Laboratory (FEL) operates within the Defence Science and Technology Laboratory (DSTL) which is part of the UK Government Ministry of Defence (MOD). The FEL provides support and advice to the Home Office and UK police forces on matters relating to the criminal misuse of explosives. During 1989 the FEL established a weekly quality assurance testing regime in its explosives trace analysis laboratory. The purpose of the regime is to prevent the accumulation of explosives traces within the laboratory at levels that could, if other precautions failed, result in the contamination of samples and controls. Designated areas within the laboratory are swabbed using cotton wool swabs moistened with ethanol:water mixture, in equal amounts. The swabs are then extracted, cleaned up and analysed using Gas Chromatography with Thermal Energy Analyser detectors or Liquid Chromatography with triple quadrupole Mass Spectrometry. This paper follows on from two previous published papers which described the regime and summarised results from approximately 14 years of tests. This paper presents results from the subsequent 7 years setting them within the context of previous results. It also discusses further improvements made to the systems and procedures and the inclusion of quality assurance sampling for the peroxide explosives TATP and HMTD. Monitoring samples taken from surfaces within the trace laboratories and trace vehicle examination bay have, with few exceptions, revealed only low levels of contamination, predominantly of RDX. Analysis of the control swabs, processed alongside the monitoring swabs, has demonstrated that in this environment the risk of forensic sample contamination, assuming all the relevant anti-contamination procedures have been followed, is so small that it is considered to be negligible. The monitoring regime has also been valuable in assessing the process of continuous improvement, allowing sources of contamination transfer into the trace areas to be identified and eliminated.     

Quality assurance testing of an explosive trace analysis laboratory

During 1989, the Forensic Explosives Laboratory (FEL) established a weekly quality assurance testing regime in its explosives trace analysis laboratory. The purpose of the regime is to prevent the accumulation of explosives traces within the laboratory at levels which could, if other precautions failed, result in the contamination of samples and controls. This paper describes the regime and summarizes the results from approximately eight years of tests. Lessons learned and improvements made over the period are also discussed.     

Report of the blind trial of the Cetus Amplitype HLA DQ alpha forensic deoxyribonucleic acid (DNA) amplification and typing kit

The AmpliType HLA DQ alpha forensic DNA amplification and typing kit is designed for the qualitative analysis of the human leukocyte antigen (HLA) DQ alpha alleles present in deoxyribonucleic acid (DNA) extracted from forensic samples. The AmpliType kit is the first forensic DNA typing product based on the GeneAmp polymerase chain reaction (PCR) process. The kit was evaluated by five forensic science laboratories (test sites) to assess their ability to perform DNA typing using PCR on sample types typically encountered by forensic laboratories. None of the DNA-containing samples was mistyped. Of the 180 DNA-containing samples analyzed, results were reported for 178 (98.9%). Of the 178 samples with results, all were correctly typed. Two sites did not report a result for one sample each. Four of the five laboratories experienced no significant levels of contamination in the DNA-containing samples. At the one site with the highest number of DNA-containing samples with contamination, the typing results were not compromised. This site was able to correct the contamination problem through simple procedural changes and stricter attention to sterile technique. Blank controls were important to monitor contamination. In conclusion, the trial demonstrated that forensic science laboratories are capable of setting up a PCR-based DNA typing laboratory and successfully using the AmpliType HLA DQ alpha forensic DNA amplification and typing kit to analyze forensic samples.     

The evaluation of the extent of transporting or "tracking" an identifiable ignitable liquid (gasoline) throughout fire scenes during the investigative process

Tests have determined that boots or shoes of individuals at a fire scene do not transport sufficient contaminants ("tracking") through the fire scene to produce a positive laboratory result for the presence of gasoline in a fire scene that was not present at the time of the fire. Questions about the validity of forensic laboratory results have been raised on the basis that low-level gasoline residues detected in the laboratory samples could have been the result of transporting the residue by footwear contaminated from the fire scene ("tracking"). The data collected in this study establish that "tracking" does not lead to false-positive laboratory results. Canines trained and experienced in the detection of trace ignitable liquid residues were also utilized in this study. The canine results confirmed that properly trained canines show a higher sensitivity than do standard ASTM laboratory techniques for fire debris analysis. In a few cases, canines responded to contamination, but laboratory testing (which is the definitive indicator) did not produce positive results.     

Hyperspectral imaging in forensic science: An overview of major application areas

Analysis of evidence is a challenge. Crime scene materials are complex, diverse, sometimes of an unknown nature. Forensic science provides the most critical applications for their examination. Chemical tests, analytical methods, and techniques to process the evidence must be carefully selected by the forensic scientist. Ideally, it may be interpreted, analyzed, and judged in the original context of the crime scene. In this sense, hyperspectral imaging (HSI) has been employed as an analytical tool that maintains the integrity of the samples/objects for multiple and sequential analysis and for counter-proof exams. This paper is an overview of forensic science trends for the application of HSI techniques in the last ten years (2011–2021). The examination of documents was the main area of exploration, followed by bloodstain analysis aging process; trace analysis of explosives and gunshot residue. Chemometric tools were also addressed since they are crucial to obtain the most important information from the samples. There are great challenges in applying HSI in forensic science, but there have been clear technological and scientific advances, and a solid foundation has been built for the use of HSI in real-life cases.     

Suicidal deaths using fireworks

The use of commercial explosives is an unusual method of commiting suicide, and only a few cases have been described in the medicolegal literature. In this paper, two cases of suicide are considered that reflect backgrounds of financial problems and psychological illnesses, respectively. Both individuals comitted suicide by detonating an explosive (fireworks). In the first case putting the explosive on his head and in the second case into his mouth. In both cases the cause of death was the destruction of the central nervous system. The following cases emphasize the importance of the forensic pathologist in the recognition of the scene, as well as the systematical collection of trace evidence of the explosion for their subsequent study in the laboratory and their correlation with the autopsy findings.     

Computerized forensic assistance software (FAS 1.0) for training and standardized investigation in distributed and disconnected services

As part of a European union (EU) support program in forensic science for the Palestinian police, a forensic assistance software was designed to respect the quality management of the anti-terrorism laboratory of the Palestinian police by the different forensic teams in the disconnected Palestinian cities. This tool not only proposes up-to-date detection and identification protocols for traces, explosives, drugs, GSR, biological samples, but also appeared to be a convenient support for forensic practice teaching.     

The use of DNA stabilizing solution to enable room temperature storage and transportation of buccal and trace sample swabs

It is proposed that a DNA stabilizing solution (DNA Genotek Inc.) designed to preserve DNA in saliva samples at room temperature can be extrapolated to the storage of swab heads. The aim of this study was to evaluate the effectiveness of the solution for the preservation of reference swabs (buccal) and trace samples (facial swabs). To this end, the solution was used during a twin-site DNA transfer project assessing background levels of carer DNA present in children. Tubes containing 400 μl of solution were used to store and transport swab heads. At the laboratory, samples were extracted using the QIAamp DNA Mini Kit (Qiagen), quantified using the Quantifiler Duo Kit and profiled using the AmpF?STR^® SGM Plus^® PCR Amplification Kit (both Applied Biosystems). Twenty-eight PCR cycles were applied to all samples. Thirty-four cycles or a longer electrophoresis injection time was applied to trace samples where necessary. All Reference swabs produced high quantities of DNA and full DNA profiles after 28 cycles. Profile morphology indicated good quality DNA with no degradation. Of the trace samples, sufficient profiles were achieved to study the transfer of carer DNA making the solution fit for continued use in this project. DNA stabilizing solution enables the storage and transportation of swabs without freezing. This is convenient, reduces transportation costs and enables instant analysis of samples upon arrival at the laboratory. This is a useful alternative for a multi-site research project as well as a reliable storage tool for use in remote areas.     

法医DNA实验室的DNA污染和防范

DNA污染是产生DNA鉴定结论错误的重要因素,法医DNA实验室要努力去解决这一问题。DNA污染有自身污染、交叉污染、PCR污染3种。法医DNA实验室要采取实验室分区、严格检验操作步骤、对试剂及消耗材料进行质量控制等方法防止发生DNA污染,采取设置对照样本、核查DNA结果、建立DNA排查数据库等方法监测和发现DNA污染。     

Optimization and validation of a fully automated silica-coated magnetic beads purification technology in forensics

Automated procedures for forensic DNA analyses are essential not only for large-throughput sample preparation, but are also needed to avoid errors during routine sample preparation. The most critical stage in PCR-based forensic analysis is DNA isolation, which should yield as much highly purified DNA as possible. The extraction method used consists of pre-treatment of stains and samples, cell lysis using chaotropic reagents, binding of the DNA to silica-coated magnetic particles, followed by elution of the DNA. Our work focuses mainly on sample preparation, obtaining the maximum possible amount of biological material from forensic samples, and the following cell lysis, to create a simple standardized lysis protocol suitable for nearly all forensic material. After optimization and validation, the M-48 BioRobot((R)) workstation has been used for more than 20,000 routine lab samples. There has been no evidence of cross contamination. Resulting DNA from as small as three nuclear cells yield reliable complete STR amplification profiles. The DNA remains stable after 2 years of storage.     

Linking ammonium nitrate-aluminum (AN-AL) post-blast residues to pre-blast explosive materials using isotope ratio and trace elemental analysis for source attribution

Forensic science practitioners are often called upon to attribute crimes using trace evidence, such as explosive remnants, with the ultimate goal of associating a crime with a suspect or suspects in order to prevent further attacks. The explosive charge is an attractive component for attribution in crimes involving explosives as there are limited pathways for acquisition. However, there is currently no capability to link an explosive charge to its source via post-blast trace residues using isotope ratios or trace elements. Here, we sought to determine if pre-blast attribution signatures are preserved after detonation and can be subsequently recovered and detected. A field study was conducted to recover samples of post-blast explosives from controlled detonations of ammonium nitrate-aluminum (AN-Al), which were then analyzed via isotope ratio mass spectrometry (IRMS) and inductively coupled plasma-mass spectrometry (ICP-MS) for quantitation and profiling of isotopes ratio and trace element signatures, respectively. Oxygen and nitrogen isotope ratios from AN-Al yielded some of the most promising results with considerable overlap within one standard deviation of the reference between the spreads of pre- and post-blast data. Trace element results from AN-Al support the findings in the isotope ratio data, with 26 elements detected in both pre- and post-blast samples, and several elements including B, Cd, Cr, Ni, Sn, V, and Zn showing considerable overlap. These preliminary results provide a proof-of-concept for the development of forensic examinations that can attribute signatures from post-blast debris to signatures in pre-blast explosive materials for use in future investigations.     

Application of direct PCR in forensic casework

Direct PCR is fast becoming a popular method in forensic science due to the advantages of saving time and money in the lab while increasing the probability of obtaining substantial results has a positive rippling effect. A laboratory is able to reduce the time spent on processing trace DNA samples, which can lead to investigators receiving important information in a timely manner that may not have been possible using standard methods. This study highlights the benefits of direct PCR in forensic casework by analysing trace and touch DNA on a range of substrates and exploring the loss of initial DNA due to extraction.     

Use of a commercially available hydrogel as a novel DNA surface sampling tool

A common requirement in the military, law enforcement, and forensic mission space is the need to collect trace samples from surfaces using a method that not only readily captures the sample but also retains its integrity for downstream identification and characterization. Additionally, collecting samples from three-dimensional objects (e.g., shell casings) is a challenge for which there is currently no validated standardized approach. Recently, hydrogels have been shown to have the potential for surface collection of trace bacterial spores, amino acids, and DNA. To test whether these hydrogels can serve as a viable collection medium for sampling DNA from surfaces, we carried out a series of preliminary tests examining collection efficiency and suitability of hydrogel material to recover samples of diluted, dried human DNA on a smooth polycarbonate surface. The recovery of surface DNA using a commercially available hydrogel was examined, and the efficiency compared to samples collected using a standard foam collection swab. DNA collected using the hydrogel and swab methods was then examined using quantitative polymerase chain reaction (qPCR) and short tandem repeat (STR) analysis to determine whether the collection material was compatible with these downstream processes. The hydrogel material used for this study collected the experimental DNA with comparable efficiency to standard collection swabs. In addition, qPCR and STR analyses demonstrated compatibility with the hydrogel collection and extraction process. These data suggest that hydrogels have the potential to be used as sample collection materials and deserve further characterization to elucidate their utility in collection from irregularly shaped, three-dimensional surfaces/materials.     

Investigations into the hypothesis of transgenic cannabis

The unusual concentration of cannabinoids recently found in marijuana samples submitted to the forensic laboratory for chemical analysis prompted an investigation into whether genetic modifications have been made to the DNA of Cannabis sativa L. to increase its potency. Traditional methods for the detection of genetically modified organisms (GMO) were used to analyze herbal cannabis preparations. Our analyses support the hypothesis that marijuana samples submitted to forensic laboratories and characterized by an abnormal level of Δ(9)-THC are the product of breeding selection rather than of transgenic modifications. Further, this research has shown a risk of false positive results associated with the poor quality of the seized samples and probably due to the contamination by other transgenic vegetable products. On the other hand, based on these data, a conclusive distinction between the hypothesis of GMO plant contamination and the other of genetic modification of cannabis cannot be made requiring further studies on comparative chemical and genetic analyses to find out an explanation for the recently detected increased potency of cannabis.     

Sub-sampling and preparing forensic samples for pollen analysis

The main forensic application of palynology is in providing associative evidence, assisting to prove or disprove a link between people and objects with places or with other people. Although identification and interpretation of pollen is a specialist job, sub-sampling and preparing pollen samples for analysis may be carried out by non-specialists. As few forensic laboratories have residing palynologists, laboratories may wish to reduce the cost of analysis or risk of contamination by doing their own sub-sampling and preparation. Presented is a practical guide for sub-sampling and preparing forensic samples for pollen analysis, providing a complete standard procedure for both the palynologist and non-specialist. Procedures for sub-sampling include a wide variety of materials commonly collected for forensic analysis (soil, clothing and other fabrics, footwear, twine and rope, firearms, granulated materials, plant and animal material, and illicit drugs), many of which palynologists will not be familiar with. Procedures for preparation of samples (pollen concentration) are presented as a detailed, step-by-step method. Minimizing the risks of laboratory and cross-sample contamination during sub-sampling and preparation is emphasized.     

A second survey of high explosives traces in public places

This survey was carried out as a follow-up to a 1994 survey carried out by this laboratory (1) in order to determine the background levels of explosives traces in public places. The first survey concentrated on transport areas and police stations in and around London. This second study examines levels in four of the United Kingdom's major cities: Birmingham, Cardiff, Glasgow, and Manchester. Samples were taken at various transport sites and from hotels, private houses, private vehicles, and clothing. The survey showed that traces of the high explosives nitroglycerine (NG), trinitrotoluene (TNT), pentaerythritol tetranitrate (PETN), and cyclotrimethylene trinitramine (RDX) are rare within the general public environment. Only one low-level trace of RDX was detected. NG, possibly associated with the use of firearms, was detected at low levels in two samples and 2,4-DNT was detected in a separate sample. No PETN was detected in any of the samples. The results of the survey indicate that it is unlikely that persons visiting public areas could become significantly contaminated with explosives. The analytical procedures employed would also have detected ethylene glycol dinitrate (EGDN) if present at levels greater than 2 ng, nitrobenzene (NB) if present at levels greater than 50 ng, mononitrotoluenes if present at levels greater than 50 ng, and the other common isomers of dinitrotoluene if these had been present at levels in excess of 10 ng. None of these were detected. The relatively high volatility of EGDN, NB, and the mononitrotoluenes would, however, cause traces of these compounds to disperse rapidly. A proportion of the samples (approximately 7%) were analyzed for the presence of HMX. No HMX was detected.     

Novel technique for the combined recovery, extraction and clean-up of forensic organic and inorganic trace explosives samples.

This paper describes a simple processing and analysis scheme for explosives trace swab samples which deals both with organic and inorganic materials. Swabs, wetted with ethanol or ethanol/water mixture, were extracted with ethanol/water mixture. The extract was passed directly through a simple column containing an acrylonitrile/styrene copolymer adsorbent. The adsorbent retained common organic explosives, which were recovered with an efficiency of 30-50% as a relatively clean ethyl acetate solution. The concentrated ethyl acetate eluate was analysed using gas chromatography with chemiluminescence or mass spectrometric detection. The unretained inorganic ions and sugars, which were recovered with generally high efficiency as an ethanol/water solution, could be directly analysed using ion chromatography and/or capillary electrophoresis. Minor difficulties encountered in the analysis of sugars, fluoride and phosphate were examined.     

The effect of NucleoSpin® Forensic Filters on DNA recovery from trace DNA swabs

Forensic DNA profiling is a globally accepted method for human identification, however, obtaining full DNA profiles from trace DNA can be challenging. The optimal recovery of DNA from trace DNA swabs is therefore crucial. Methods for extracting DNA from swabs often make use of a spin basket combined with a centrifugation step, to enhance the release of cells from the swab prior to DNA extraction. The NucleoSpin® Forensic Filter (Macherey-Nagel, Düren) is a type of spin basket, but it has not been thoroughly assessed on trace DNA samples. This study aimed to assess if the inclusion of the NucleoSpin® Forensic Filter significantly improved DNA recovery and DNA profiling success from cotton and flocked swabs used to collect trace DNA and buccal cells (control). Buccal cells and trace DNA samples were collected from 25 volunteers using each swab type (cotton and flocked) in duplicate. DNA was extracted from the samples using the NucleoSpin® DNA Forensic kit, one set with, and the other set without, NucleoSpin® Forensic Filters. DNA concentration was assessed using real time PCR, and DNA profiling was done using the PowerPlex® ESX 16 system. The inclusion of the NucleoSpin® Forensic Filters significantly improved DNA concentration for buccal cells that were collected using flocked swabs (p = 0.035). However, no significant differences were noted for trace DNA samples for either swab type. There was also no significant difference in DNA profiling success when NucleoSpin® Forensic Filters were used, regardless of swab and sample type. These results may be helpful for laboratories that are considering the NucleoSpin® Forensic Filters in the DNA extraction workflow, particularly for trace DNA samples.     

Identification of ascorbic acid and its degradation products in black powder substitutes

Low explosives such as smokeless powder, black powder, and black powder substitutes have been used in illicit pipe bombings throughout the United States. Some of the newer black powder substitutes are formulated with ascorbic acid, which gradually decomposes as the powder ages, making it difficult if not impossible for the forensic chemist to identify it by traditional bulk techniques. A sensitive method for the identification of residual levels of ascorbic acid in black powder substitutes is presented. Powder samples are extracted with a mixture of acetonitrile and bis(trimethylsilyl)acetamide (BSA), which converts carboxylic acid and alcohol functional groups to trimethylsilyl esters and ethers, respectively. Samples are then analyzed by gas chromatography-mass spectrometry (GC-MS). Results have shown that trace amounts of ascorbic acid can be identified at detection limits that are well below those for traditional bulk techniques. Degradation products for ascorbic acid (hydroxylated carboxylic acids, furanones, and lactones) can also be detected.