John Coumbaros

Analysis of illicit drug samples traditionally focuses on chemical impurity profiling for identifying drug origin and linking seizures. This method relies on detecting impurities such as bulking agents (e.g. starch), enhancing agents (e.g. procaine hydrochloride), manufacturing precursors (e.g. pseudoephedrine in methamphetamine) and synthesis by-products. However, a trend towards a final product of greater purity has reduced available chemical impurity data, thereby limiting definitive profiling.

An emerging research area explores extracting trace human DNA and environmental DNA (eDNA) from capsule surfaces and illicit drug powders. It may be expected, however, that perpetrators may take greater care to avoid leaving fingerprints or human DNA on drug packaging, highlighting the need for alternative forensic methods. In such circumstances, eDNA presents a promising avenue, identifying bacterial, fungal, plant, and eukaryotic cells from diverse sources like soil, air, and water that may provide clues as to the location of manufacture as well as potential trafficking routes. The research highlights the importance of integrating eDNA analysis with conventional chemical methods in forensic illicit drug investigations.

Drink spiking in social settings is one of the most pervasive forms of drug-facilitated sexual assault (DFSA). There are no current data in Australia on the rates of drink spiking or their associated assaults. There is also little known about the prevalence of different substances involved and how the current substance use trends compare to sexual assault trends. To explore this, a recalculation of sexual assault trends to estimate substance related sexual assault was performed. Data about recent trends of mental health prescriptions and sexual assault were obtained from the Australian Institute of Health and Welfare (AIHW). The analysis of these datasets highlighted that females are the highest consumers of antidepressants and benzodiazepines. Results also indicated a statistically significant positive correlation between females and a higher incidence of sexual assault (r = 0.996, p < .001). This paper demonstrates that females are at most risk of drug-drug interactions (e.g., diazepam and ketamine) with their medications due to the higher rate of prescriptions amongst this population, and therefore more vulnerable to both opportunistic and proactive DFSA. While these findings are preliminary and not causal, they highlight trends in need of further study.

Little is known about the prevalence of incapacitating substances present in drug facilitated sexual assaults (DFSA). Presented here is a literature review conducted to provide background information, such as symptoms, exacerbations, and drug interactions, on drugs typically implicated in DFSA, namely gamma-hydroxybutyrate (GHB), gamma-butyrolactone (GBL), 1,4-butanediol (1,4-BD), ketamine, diazepam, oxycodone, methamphetamine, and alcohol.
Literature found through Scopus and Pubmed was reviewed to determine the current prevalence of these substances in DFSA with a focus on Australian data.
The global literature revealed that there is a wide variety of substances used in DFSA and the prevalence varied by country. For example, it was found that in Northern Ireland, opioids were most prevalent whereas in France, benzodiazepines were most prevalent. In Australia the review revealed a lack of contemporary data with the most recent report in Victoria using data collected during 2011–2013. The literature also revealed there can be an important difference between self-reported substance use and substances discovered via toxicological analysis. This can be due to the challenges of biological detection, reliability of self-reporting, and the possibility of a substance being introduced to a person's food or drink without their knowledge.
This review highlights the need for the collection and analysis of current data pertaining to DFSA reports and the drugs detected, and due to the constantly evolving picture of both licit and illicit drug use an assessment of the role of prescription medications in DFSA due to drug-drug interactions as well as potential to incapacitate is warranted.

Deceased human remains are often buried as a forensic countermeasure or method of disposal by homicide perpetrators. Owing to this, the excavation of clandestine grave sites is a task that forensic crime scene teams may only encounter a few times a year. Not all crime scene units have specialised teams for this task, and even those that do, may not have specific protocols for the optimal recovery of forensic traces retained within grave fill as procedures such as sieving require optimisation for the specific soil conditions of the jurisdiction. This study aimed to define the optimal sieving conditions for a sandy environment when searching for minute traces of paint, glass, hair and fibres. Furthermore, this study justifies the practice of retaining grave fill and examining it under controlled laboratory conditions, rather than in-situ adjacent to the grave site. The results demonstrate that using sieve mesh sizes as fine as 0.1 mm can recover up to 82% of the deposited traces and almost all paint, hair and glass traces. The processing of grave fill in the laboratory lead to increased yield of forensic evidence, which on a case-basis may warrant the increased time needed. These findings merit consideration for clandestine grave crime scenes where evidence is scarce or the case is likely to become cold.

Cold case reviews within police and law enforcement agencies are challenging, not the least owing to the amount of time required to carefully review documentation, forensic exhibit holdings and various other casefile information. Most federal and state agencies are time poor, meaning there are very few dedicated cold case teams fortunate enough to have an abundance of police and expert staff resources. Universities and education organisations, however, have large troves of various expertise, alongside expansive human resources, by way of their academic and student body. In certain circumstances, the academic expertise and course offerings of a university may be well suited to assisting law enforcement in reviewing cold cases.

There is growing desire for university courses to generate job ready graduates. In the field of law enforcement and policing this is difficult, as safety and the security of sensitive material and evidence is paramount. Educators strive to create workplace simulations, and with the correct mix of academic expertise, course offerings and industry linkages, the emerging opportunity for real cold case collaboration is possible. One such example is the Cold Case Review @ Murdoch (CCR) initiative. Since 2020, CCR has worked with the Lower Saxony Police Academy in Germany to develop the novel International Cold Case Analysis Project (ICCAP), now incorporating over 25 member institutions, to assist in solving real cases from both Niedersachsen (Lower Saxony) and federal police jurisdictions. One case, known as “The North Sea Man” has shown great success and demonstrates the power of joining forces between law enforcement and external agencies to help advance cold cases.

Forensic samples for DNA analysis are limited by physical size and volume as DNA extraction methods require small portions placed into 1.5 or 2.0 mL tubes for lysis. Apart from tape lifting techniques, this precludes large sample areas from being harvested of cellular material by any means other than washing or wet vacuuming. In addition, forensically savvy perpetrators may employ clean-up methods to remove DNA evidence by way of wash basins and sinks, although evidence may be retained within the p-trap of plumbing. Both outcomes still leave forensic biologists with samples incompatible with most commercial DNA extraction methods. Fields of microbiology and ecology have long used environmental DNA (eDNA) collection methods to overcome the similar challenges of sampling biota within large volumes of water or liquid samples. This study is the first application of eDNA techniques to capture such dilute traces of human cellular material from large water volumes using a microfiltration method. The technique was able to isolate human DNA from as little as 100 cells in 1 L of water. While future optimization is required to determine ideal filter specifications, this research demonstrates a proof of concept for forensic application in cases with challenging substrates that can be washed or contain water. This research was conducted under the Murdoch University Human Research Ethics Committee approval 2019/025.

Osseous remains provide forensic anthropologists with morphological and osteometric information that can be used in building a biological profile. By conducting a visual and physical examination, an anthropologist can infer information such as the sex and age of the deceased. Traditionally, morphological and osteometric information is gathered by physically handling remains for analysis. With the advancement of digital technology, there has been a shift from direct to indirect methods of analysis by utilizing models generated from three-dimensional (3D) imaging, which includes computed tomography (CT) scanning and 3D photogrammetry. Although CT scanning is more common, photogrammetry has found application in a range of fields such as architecture, geography and road accident reconstruction. The application of modern-day photogrammetry for forensic anthropology purposes, however, has not been discussed extensively. The aim of this research was to validate the accuracy of 3D models generated by photogrammetry by comparing them to both 3D models generated by CT scanning and the actual physical models. In this study, six 3D models were created using photogrammetry (n = 3) and CT scanning (n = 3). The 3D models were generated from three different Bone Clone® human skulls. A mobile phone camera was used to capture images, which were then processed in Agisoft Metashape®. Intrarater, interrater, and intermethod reliability tests gave correlation coefficients of at least 0.9980, 0.9871, and 0.9862, respectively; rTEM results ranged from 0.250 to 6.55%; and an analysis of variance (ANOVA) yielded P values under 0.05 for all measurements except one. Statistical tests therefore showed photogrammetry to be a reliable and accurate alternative to more expensive CT scanning approaches.

This paper describes the involvement of our laboratory in a Western Australian ‘cold-case’ investigation that spanned 24 years. The investigation was widely considered to be the largest in Australian history. During the investigative phase, our laboratory was tasked with the collection of trace evidence of all types in connection with a sexual assault and two homicides that were suspected to be related. Textile fibres represented the vast majority of trace evidence recovered. A much greater quantity of fibres (>10,800) was collected than would be typical for a routine case, as fibres of any colour or type were potentially of investigative value. The investigation was unprecedented in its scale, and presented numerous challenges in terms of evidence recovery, analysis, interpretation, reporting, and provision of testimony. A textile fibre microspectrophotometric (MSP) database was developed specifically for the interpretation of data in connection with the investigation. The database currently contains over 25,000 normalised and first derivative spectra of casework, validation and reference textile fibres. A fibre comparison strategy was devised, involving the identification of preliminary fibre groups on the basis of corresponding/similar MSP spectra, and verification of these groups via brightfield and fluorescence comparison microscopy. A potential link to an automotive source was identified for one of the homicide victims during the investigative phase. After identification of a suspect, a total of 98 fibres recovered from victims and from a seized motor vehicle were found to correspond in properties to six different fibre types from known textile sources in connection with the suspect. A highly publicised criminal trial was held, and textile fibre evidence provided a major contribution to the trial findings, in which the accused was found guilty of two homicides.

Chemical, biological, radiological, nuclear and explosive (CBRNE) response teams are responsible for mitigating and investigating events such as the distribution of anthrax letters throughout the US Postal Service in 2001. One consideration will be the examination of contaminated physical evidence to identify those responsible. Decontamination agents are often used to inactivate or neutralize chemical and/or biological warfare agents so that once-contaminated objects can be handled safely. The objective of this project was to determine the impact of different decontamination agents on physical evidence including DNA, documents, fingermark and footwear evidence. In Phase 1, vaporous hydrogen peroxide (VHP), gamma irradiation, dry fogging, chlorine dioxide, MODEC MDF-500, Bioxy-S, ozone and formaldehyde were tested. Promising decontamination agents were selected for Phase 2 when physical evidence was exposed to Bacillus thuringiensis (Bt), an anthrax surrogate, prior to decontamination. VHP, gamma irradiation, dry fogging, formaldehyde and chlorine dioxide were tested in Phase 2. Results indicate that several forensic analyses were impacted. VHP and gamma irradiation were used across different laboratories with inconsistent results observed for some forensic protocols. Further investigation indicated that these results could be explained by variation in contamination protocols between laboratories.

Les équipes d'intervention chimique, biologique, radiologique, nucléaire et explosive (CBRNE) sont chargées d'atténuer et d'enquêter sur des événements tels que la distribution de lettres d'anthrax dans l'ensemble du service postal américain en 2001. Une considération sera l'examen des preuves physiques contaminées pour identifier les responsables. Les agents de décontamination sont souvent utilisés pour inactiver ou neutraliser les agents de guerre chimiques et/ou biologiques afin que des objets autrefois contaminés puissent être manipulés en toute sécurité. L'objectif de ce projet était de déterminer l'impact de différents agents de décontamination sur les preuves physiques, y compris l'ADN, les documents, les traces de doigt et les chaussures. En phase 1, le peroxyde d'hydrogène vaporeux (VHP), l'irradiation gamma, la brumisation sèche, le dioxyde de chlore, le MODEC MDF-500, le Bioxy-S, l'ozone et le formaldéhyde ont été testés. Des agents de décontamination prometteurs ont été sélectionnés pour la phase 2 lorsque des preuves physiques ont été exposées à Bacillus thuringiensis (Bt), un substitut de l'anthrax, avant la décontamination. Le VHP, l'irradiation gamma, la brumisation sèche, le formaldéhyde et le dioxyde de chlore ont été testés à la phase 2. Les résultats indiquent que plusieurs analyses médico-légales ont été touchées. VHP et l'irradiation gamma ont été utilisée dans différents laboratoires avec des résultats incohérents observés pour certains protocoles médico-légaux. Une enquête plus approfondie a indiqué que ces résultats pourraient s'expliquer par la variation des protocoles de contamination entre les laboratoires.

Purpose
Dermorphin, growth hormone releasing peptide (GHRP), TB-500 and their analogues have been used illegally in the horse racing industry to improve the performance of the horses. This study aims to present dispersive solid phase extraction (dSPE) as an alternative to solid phase extraction (SPE) for the clean-up of equine urine samples prior to liquid chromatography combined with tandem mass spectrometry (LC–MS/MS) screening of 43 illegal performance enhancing peptides.

Methods
Sorbent types and mass, washing and eluting solvents were tested to obtain the optimal clean-up conditions for these peptides in horse urine matrices.

Results
The resulting dSPE clean-up method gave optimal recovery and reproducibility of 43 target peptides; for the first time dSPE is proven as a viable alternative to SPE and achieves limits of detection (LOD) that are sufficient for the screening of these peptides. The LODs for all dermorphin, TB-500 and GHRP peptides were 1 ng/mL. Recoveries of the 43 target analytes extracted from 3 spiked urine samples ranged from 8.9 to 58.8%. The intra-day and inter-day precision for all target analytes ranged from 0.6 to 24.1% and 1.4 to 27.8% respectively.

Conclusions
Using dSPE as a clean-up method, 43 peptide analytes of interest were successfully screened by LC–MS/MS.