Exploring the influence of washing activities on the transfer and persistence of fibres in forensic science.

In forensic science, a robust and sound interpretation and evaluation of transferred fibre evidence requires an understanding of the principles and mechanisms that underpin fibre transfer, yet existing research lacks consistency and repeatability. This study investigates the impact of washing activities on both the release of fibres into wastewater and the transfer of constituent fibres from donor garments to receiver swatches. Using a low-cost friction tester and automated data collection through photography and ImageJ image processing software, controlled conditions were maintained for repeated experiments. Results indicated significant fibre release during wash cycles, with load size and donor garment history playing crucial roles. The donor garments subjected to repetitive washes exhibit a progressive decrease in the number of fibres transferred, independently of the load size. This study underscores the importance of considering a garment's washing history in forensic science contexts, but also for consistency in the way that data are collected.

Trace DNA and its persistence on various surfaces: A long term study investigating the influence of surface type and environmental conditions - Part one, metals.

It is imperative for proper evidence triage that forensic biologists understand what kind of results to expect from certain evidence types submitted for DNA analysis. The persistence of trace DNA has been insufficiently investigated and there is little data available pertaining to the persistence of DNA in different environmental conditions and on different materials. The goal of this study is to increase the available data on this topic which would, in turn, help forensic biologists manage expectations when submitting specific evidence types for DNA testing. The work presented herein is a large-scale persistence project aimed to identify trends in the persistence of trace DNA and indicate how different environmental storage conditions and target surface characteristics influence the persistence of cellular and cell free DNA (cfDNA) over time. To eliminate variation within the experiment we used a proxy DNA deposit consisting of a synthetic fingerprint solution, cellular DNA, and/or cfDNA. Samples were collected and analysed from 7 metals over the course of 1 year (27 time points) under 3 different environmental storage conditions. The results of this experiment show that metal type greatly influences DNA persistence. For instance, copper exhibited an expected poor DNA persistence (up to 4 h) which a purification step did not help increase the DNA yield. Alternatively, DNA can persist for up to a year on lead at levels potentially high enough to allow for forensic DNA testing. Additionally, this study showed that the sample storage environment had no impact on DNA persistence in most cases. When considering DNA type, cfDNA was shown to persist for longer than cellular DNA and persistence as a whole appears to be better when DNA is deposited as mixtures over when deposited alone. Unsurprisingly, it can be expected that DNA recovery rates from trace deposits will decrease over time. However, DNA decay is highly dependent on the metal surface and extremely variable at short time points but slightly less variable as time since deposition increases. This data is intended to add to our understanding of DNA persistence and the factors which affect it.

Log D7.4 and plasma protein binding of synthetic cannabinoid receptor agonists and a comparison of experimental and predicted lipophilicity.

The emergence of new synthetic cannabinoid receptor agonists (SCRAs) onto the illicit drugs market continues to cause harm, and the overall availability of physicochemical and pharmacokinetic data for new psychoactive substances is lacking. The lipophilicity of 23 SCRAs and the plasma protein binding (PPB) of 11 SCRAs was determined. Lipophilicity was determined using a validated chromatographic hydrophobicity index (CHI) log D method; tested SCRAs showed moderate to high lipophilicity, with experimental log D7.4 ranging from 2.48 (AB-FUBINACA) to 4.95 (4F-ABUTINACA). These results were also compared to in silico predictions generated using seven commercially available software packages and online tools (Canvas; ChemDraw; Gastroplus; MoKa; PreADMET; SwissADME; and XlogP). Licenced, dedicated software packages provided more accurate lipophilicity predictions than those which were free or had prediction as a secondary function; however, the latter still provided competitive estimates in most cases. PPB of tested SCRAs, as determined by equilibrium dialysis, was in the upper range of the lipophilicity scale, ranging from 90.8% (ADB-BUTINACA) to 99.9% (BZO-HEXOXIZID). The high PPB of these drugs may contribute to reduced rate of clearance and extended durations of pharmacological effects compared to lesser-bound SCRAs. The presented data improve understanding of the behaviour of these drugs in the body. Ultimately, similar data and predictions may be used in the prediction of the structure and properties of drugs yet to emerge on the illicit market.

In vitro cannabinoid activity profiling of generic ban-evading brominated synthetic cannabinoid receptor agonists and their analogs.

Following the enactment of a generic ban in China in 2021, the synthetic cannabinoid market has been evolving, now encompassing even wider structural diversity. Compounds carrying a brominated core such as ADB-5'Br-BUTINACA (ADMB-B-5Br-INACA) and tail-less analogs, such as ADB-5'Br-INACA (ADMB-5Br-INACA), MDMB-5'Br-INACA, and ADB-INACA (ADMB-INACA), have been detected since late 2021. This study investigated the cannabinoid receptor (CB) activation potential of synthesized (S)-enantiomers of these substances, as well as of two predicted analogs MDMB-5'Br-BUTINACA (MDMB-B-5Br-INACA) and ADB-5'F-BUTINACA (ADMB-B-5F-INACA), using CB1 and CB2 ß-arrestin 2 recruitment assays and a CB1 intracellular calcium release assay. Surprisingly, the tail-less (S)-ADB-5'Br-INACA and (S)-MDMB-5'Br-INACA retained CB activity, albeit with a decreased potency compared to their tailed counterparts (S)-ADB-5'Br-BUTINACA and (S)-MDMB-5'Br-BUTINACA, respectively, which were potent and efficacious CB1 agonists. Also, at CB2 , tail-less analogs showed a lower potency but increased efficacy. Removing the bromine substitution ((S)-ADB-INACA) resulted in a reduced activity at CB1 ; however, this effect was less prominent at CB2 . Looking at tailed analogs, replacing the bromine with a fluorine substitution ((S)-ADB-5'F-BUTINACA) resulted in an increased potency and efficacy at both receptors. Furthermore, as ADB-5'Br-INACA and MDMB-5'Br-INACA have been frequently detected together in Scottish prisons, this study also evaluated the CB1 receptor activation potential of different mixtures of their respective reference standards, showing no unexpected cannabimimetic effect of combining both substances. Lastly, two powders seized by Belgian Customs and confirmed to contain ADB-5'Br-INACA and MDMB-5'Br-INACA, respectively, were assessed for CB activity. Based on the comparison with their reference standards, varying degrees of purity were suspected.

Development of a Dundee Ground Truth imaging protocol for recording indoor crime scenes to facilitate virtual reality reconstruction.

Early and accurate visualisation of a crime scene is highly desirable such that a rapid, agile, and informed decision-making process can be undertaken by an investigative team. We present a new standard operating procedure for imaging an indoor scene using DSLR cameras conventionally used by crime scene investigators and examiners. The standard operating procedure (SOP) enables the systematic photography of indoor spaces in such a way that the Structure from Motion (SfM) photogrammetry technique can be implemented, allowing the scene to be recreated in Virtual Reality (VR). To demonstrate the method's validity, we compare two VR-rendered representations of an example scene using (a) photographs taken by an experienced crime scene examiner using a conventional photographic method and (b) photographs taken by a novice photographer following the developed SOP.

An improved rapid method for DNA recovery from cotton swabs.

We present a novel rapid method for the recovery of cellular and free DNA from cotton swabs based on a simple elution buffer containing a high molecular weight polymer and detergent combined with a short proteinase K digestion to release cellular DNA. This method shows increased yields approaching 80% recovery of the input DNA compared to the QIAamp DNA Mini kit standard extraction protocol for swabs which has a recovery of 20-30%. The buffer components in the described method are compatible with direct PCR analysis of the isolated DNA without further purification. Recovery efficiencies were estimated by qPCR.

Thiolated gamma-cyclodextrin-polymer-functionalized CeFe3O4 magnetic nanocomposite as an intrinsic nanocatalyst for the selective and ultrasensitive colorimetric detection of triacetone triperoxide.

Explosives are powerful destructive weapons used by criminals and terrorists across the globe and their use within military installation sites poses serious environmental health problems. Existing colorimetric sensors for triacetone triperoxide (TATP) relies on detecting its hydrolysed H2O2 form. However, such detection strategy limits the practicability for on-site TATP sensing. In this work, we have developed a novel peroxidase mimic catalytic colorimetric sensor for direct recognition of TATP. Ceria (Ce)-doped Fe3O4 nanoparticles (CeFe3O4) were synthesized via the hot-injection organic synthetic route in the presence of metal precursors and organic ligands. Thereafter, the organic-capped CeFe3O4 nanoparticles were surface-functionalized with amphiphilic polymers (Amp-poly) to render the nanoparticle stable, compact and biocompatible. Thiolated γ-cyclodextrin (γ-CD) was adsorbed on the Amp-poly-CeFe3O4 nanocomposite (NC) surface to form a γ-CD-Amp-poly-CeFe3O4 NC. γ-CD served both as a receptor and as a catalytic enhancer for TATP. Hemin (H), used as a catalytic signal amplifier was adsorbed on the γ-CD-Amp-poly-CeFe3O4 NC surface to form a γ-CD-Amp-poly-CeFe3O4-H NC that served as a functional nanozyme for the enhanced catalytic colorimetric detection of TATP. Under optimum experimental reaction conditions, TATP prepared in BIS-TRIS-Trisma Ac-KAc-NAc buffer, pH 3, was selectively and ultrasensitively detected without the need for acid hydrolysis based on the catalytic oxidation of 3,3',5,5'-tetramethylbenzidine by H2O2 in the presence of the γ-CD-Amp-poly-CeFe3O4-H hybrid nanozyme. The obtained limit of detection of ∼0.05 µg/mL when compared with other published probes demonstrated superior sensitivity. The developed peroxidase mimic γ-CD-Amp-poly-CeFe3O4-H catalytic colorimetric sensor was successfully applied to detect TATP in soil, river water and tap water samples.

Rapid and highly selective colorimetric detection of nitrite based on the catalytic-enhanced reaction of mimetic Au nanoparticle-CeO2 nanoparticle-graphene oxide hybrid nanozyme.

The International Agency for Research cancer (IARC) has classified nitrite in Group 2A of probable carcinogens to human. Herein, we report on the rapid and selective colorimetric detection of nitrite using a chemically modified gold nanoparticle (AuNP)-cerium oxide (CeO2) NP-anchored graphene oxide (GO) hybrid nanozyme in a catalytic colorimetric assay where nitrite acts as the main oxidant/target analyte and 3,3',5,5'-tetramethylbenzidine (TMB) as the substrate. CeO2 NPs and GO were synthesized separately and incorporated in-situ, in a synthetic solution involving the chemical reduction of Au salt to AuNPs. The chemical modification process aided the adsorption of CeO2 NPs and AuNPs on GO nanosheets, yielding a highly catalytic AuNP-CeO2 NP@GO nanohybrid material. Under optimum experimental conditions, a novel colorimetric assay for nitrite recognition was constructed in which AuNP-CeO2 NP@GO hybrid nanozyme catalysed the oxidation of TMB in the presence of nitrite prepared in a 2-(n-morpholino)ethanesulfonic acid-2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)propane-1,3-diol-tris(hydroxymethyl)aminomethane acetate (MES-BIS-TRIS-Trisma Ac)-citric acid buffer solution, pH 2. Nitrite was quantitatively detected in a concentration dependent manner from 100 µM to 5000 µM with a correlation coefficient of 0.9961 and a limit of detection of 4.6 µM. Selective detection of nitrite was confirmed by the generation of a unique green colour reaction upon nitrite interaction in the AuNP-CeO2 NP@GO hybrid nanozyme redox cycle with TMB. None of the several tested metal ions and including H2O2 yielded a positive colour response, thus demonstrating the superior selectivity of the catalytic colorimetric assay for nitrite recognition. The AuNP-CeO2 NP@GO hybrid nanozyme catalytic colorimetric assay was successfully applied in the detection of nitrite in tap water.

Impact of metal ions on PCR inhibition and RT-PCR efficiency.

Inhibition of PCR by metal ions can pose a serious challenge in the process of forensic DNA analysis. Samples contaminated with various types of metal ions encountered at crime scenes include swabs from metal surfaces such as bullets, cartridge casings, weapons (including guns and knives), metal wires and surfaces as well as bone samples which contain calcium. The mechanism behind the impact of metal ions on DNA recovery, extraction and subsequent amplification is not fully understood. In this study, we assessed the inhibitory effects of commonly encountered metals on DNA amplification. Of the nine tested metals, zinc, tin, iron(II) and copper were shown to have the strongest inhibitory properties having IC50 values significantly below 1 mM. In the second part of the study, three commercially available DNA polymerases were tested for their susceptibility to metal inhibition. We found that KOD polymerase was the most resistant to metal inhibition when compared with Q5 and Taq polymerase. We also demonstrate how the calcium chelator ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) can be used as an easy and non-destructive method of reversing calcium-induced inhibition of PCR reactions.

Biomimetic graphene oxide-cationic multi-shaped gold nanoparticle-hemin hybrid nanozyme: Tuning enhanced catalytic activity for the rapid colorimetric apta-biosensing of amphetamine-type stimulants.

Amphetamine-type stimulants are a class of illicit drug that constitutes a worldwide problem to which intelligence agencies, first responders and law enforcement are tasked with identifying them in unknown samples. We report on the development of a graphene oxide (GO)-cationic multi-shaped gold nanoparticle (AuNP)-hemin hybrid nanozyme as a new biomimetic catalytic-induced aptamer-based colorimetric biosensor platform for amphetamine (AMP) and methamphetamine (MAMP). GO was electrostatically bonded to cationic multi-shaped cetyltrimethylammonium bromide (CTAB)-AuNPs to form a GO-CTAB-AuNP hybrid nanozyme exhibiting enhanced catalytic activity in the presence of hemin. The binding of an MNS 4.1 anticocaine DNA aptamer on the GO-CTAB-AuNP-hemin nanozyme assembly and the subsequent catalytic oxidation by 3,3,5,5-tetramethylbenzidine in the presence of H2O2 ensured that the colorimetric reaction was tuned to selectively detect AMP and MAMP with high sensitivity. Under optimum experimental conditions, AMP and MAMP were quantitatively detected within 1 min with a detection limit of 34.1 ng/mL and 28.6 ng/mL respectively. Selected substances and drugs, known to react positively to Marquis and Mandelin reagents (used in AMP and MAMP presumptive testing) and well-known adulterants, were tested for their affinity to react with the aptamer-based GO-CTAB-AuNP-hemin peroxidase mimic biosensor. The deep blue colorimetric reaction, specific to AMP and MAMP detection, was used as the basis to affirm the selectivity of the aptamer-based GO-CTAB-AuNP-hemin peroxidase mimic biosensor. We believe the colorimetric biosensor developed in this work demonstrates a promising new direction in presumptive testing for AMP and MAMP.

Aptamer-based cocaine assay using a nanohybrid composed of ZnS/Ag2Se quantum dots, graphene oxide and gold nanoparticles as a fluorescent probe.

Authors report on a new fluoro-graphene-plasmonic nanohybrid aptamer-based fluorescent nanoprobe for cocaine. To construct the nanoprobe, newly synthesized glutathione-capped ZnS/Ag2Se quantum dots (QDs) were first conjugated to graphene oxide (GO) to form a QD-GO nanocomposite. The binding interaction resulted in a fluorescence turn-ON. Thereafter, cetyltrimethylammonium bromide (CTAB)-gold nanoparticles (AuNPs) were directly adsorbed on the QD-GO nanocomposite to form a novel QD-GO-CTAB-AuNP nanohybrid assembly that resulted in a fluorescence turn-OFF. Streptavidin (strep) was then adsorbed on the QDs-GO-CTAB-AuNP nanohybrid assembly which allowed binding to a biotinylated MNS 4.1 anticocaine DNA aptamer (B) receptor. The addition of cocaine into the strep-B-QDs-GO-CTAB-AuNP aptamer nanoprobe system aided affinity to the aptamer receptor and in turn turned on the fluorescence of the nanoprobe in a concentration-dependent manner. Under optimum experimental conditions, we found the strep-B-QD-GO-CTAB-AuNP to be far superior in its sensitivity to cocaine than the tested strep-B-QDs (no GO and CTAB-AuNPs), strep-B-QD-CTAB-AuNP (no GO) and strep-B-QD-GO (no CTAB-AuNP). In addition, the investigation of localized surface plasmon resonance (LSPR) amplified signal from tested plasmonic NPs shows that CTAB-AuNPs was far superior in amplifying the fluorescence signal of the nanoprobe. A detection limit of 4.6 nM (1.56 ng.mL-1), rapid response time (~2 min) and excellent selectivity against other drugs, substances and cocaine metabolites was achieved. The strep-B-QD-GO-CTAB-AuNP aptamer-based fluorescent nanoprobe was successfully applied for the determination of cocaine in seized adulterated cocaine samples. Graphical abstractSchematic representation of the streptavidin-biotin-quantum dot-graphene oxide-cetyltrimethylammonium bromide-gold nanoparticle aptamer-based fluorescent nanoprobe for cocaine.

On the relationships between applied force, photography technique, and the quantification of bruise appearance.

Bruising is an injury commonly observed within suspect cases of assault or abuse, yet how a blunt impact initiates bruising and influences its severity is not fully understood. Furthermore, the standard method of documenting a bruise with colour photography is known to have limitations which influence the already subjective analysis of a bruise. This research investigated bruising using a standardised blunt impact, delivered to 18 volunteers. The resulting bruise was imaged using colour, cross polarised (CP) and infrared photography. Timelines of the L*a*b* colour space were determined from both colour and CP images for up to 3 weeks. Overall, no single photographic technique out-performed the others, however CP did provide greater contrast than colour photography. L*a*b* colour space timelines were not attributable any physiological characteristics. Whilst impact force negatively correlated with BMI (R2 = 0.321), neither were associated with any measure of bruise appearance. Due to the inter-subject variability in the bruise response to a controlled infliction, none of the methods in the current study could be used to reliably predict the age of a bruise or the severity of force used in creating a bruise. A more comprehensive approach combining impact characteristics, tissue mechanics, enhanced localised physiological measures and improvements in quantifying bruise appearance is likely to be essential in removing subjectivity from their interpretation.

Analysis of chain saw lubricating oils commonly used in Thailand's southern border provinces for forensic science purpose.

In recent years, Thailand's southern border provinces (Malay-Muslim-majority border provinces) have become the scene of violence and insurgency. One of the attack patterns is the blocking of roads with perennial plants followed by planned attacks using improvised explosive devices (IEDs) or weapons on first responders. Containers of viscous dark lubricating oil and traces of lubricants on the felled trees were usually found at the scene. These were suspected to be chain oil lubricant from the chainsaws used to cut down the trees used for the roadblock. This work aimed to differentiate the chromatographic patterns of used lubricating oils available in automobile repair shops from various locations across Thailand's southern border provinces. Lubricating oils were analyzed using gas chromatography/flame ionization detector (GC/FID) every two weeks to study their variation in chemical compositions over time. The results obtained from GC/FID were normalized for differentiation. This included four two-stroke, six four-stroke, and three recycled oils. Two lubricating oils found at an incident scene were also analyzed and the results compared with the chain oil from five seized chainsaws.

Real time quantitative colourimetric test for methamphetamine detection using digital and mobile phone technology.

The Simon presumptive color test was used in combination with the built-in digital camera on a mobile phone to detect methamphetamine. The real-time Red-Green-Blue (RGB) basic color data was obtained using an application installed on the mobile phone and the relationship profile between RGB intensity, including other calculated values, and the colourimetric product was investigated. A wide linear range (0.1-2.5mg mL(-1)) and a low detection limit (0.0110±0.0001-0.044±0.002mg mL(-1)) were achieved. The method also required a small sample size (20µL). The results obtained from the analysis of illicit methamphetamine tablets were comparable to values obtained from gas chromatograph-flame ionization detector (GC-FID) analysis. Method validation indicated good intra- and inter-day precision (2.27-4.49%RSD and 2.65-5.62%RSD, respectively). The results suggest that this is a powerful real-time mobile method with the potential to be applied in field tests.

Using the iPhone as a device for a rapid quantitative analysis of trinitrotoluene in soil.

Mobile 'smart' phones have become almost ubiquitous in society and are typically equipped with a high-resolution digital camera which can be used to produce an image very conveniently. In this study, the built-in digital camera of a smart phone (iPhone) was used to capture the results from a rapid quantitative colorimetric test for trinitrotoluene (TNT) in soil. The results were compared to those from a digital single-lens reflex (DSLR) camera. The colored product from the selective test for TNT was quantified using an innovative application of photography where the relationships between the Red Green Blue (RGB) values and the concentrations of colorimetric product were exploited. The iPhone showed itself to be capable of being used more conveniently than the DSLR while providing similar analytical results with increased sensitivity. The wide linear range and low detection limits achieved were comparable with those from spectrophotometric quantification methods. Low relative errors in the range of 0.4 to 6.3% were achieved in the analysis of control samples and 0.4-6.2% for spiked soil extracts with good precision (2.09-7.43% RSD) for the analysis over 4 days. The results demonstrate that the iPhone provides the potential to be used as an ideal novel platform for the development of a rapid on site semi quantitative field test for the analysis of explosives.

A comparison of enhancement techniques for footwear impressions on dark and patterned fabrics.

The use of chemical enhancement techniques on porous substrates, such as fabrics, poses several challenges predominantly due to the occurrence of background staining and diffusion as well as visualization difficulties. A range of readily available chemical and lighting techniques were utilized to enhance footwear impressions made in blood, soil, and urine on dark and patterned fabrics. Footwear impressions were all prepared at a set force using a specifically built footwear rig. In most cases, results demonstrated that fluorescent chemical techniques were required for visualization as nonfluorescent techniques provided little or no contrast with the background. Occasionally, this contrast was improved by oblique lighting. Successful results were obtained for the enhancement of footwear impressions in blood; however, the enhancement of footwear impressions in urine and soil on dark and patterned fabrics was much more limited. The results demonstrate that visualization and fluorescent enhancement on porous substrates such as fabrics is possible.

Investigation of the reaction impurities associated with methylamphetamine synthesized using the Nagai method.

The synthesis of methylamphetamine hydrochloride from l-ephedrine or d-pseudoephedrine hydrochloride via reduction with hydriodic acid and red phosphorus was investigated. Eighteen batches of methylamphetamine hydrochloride were synthesized in six replicate batches using three different reaction times. This allowed the investigation of the variation of impurities in the final product with reaction time. The results obtained have resolved previously conflicting impurity profile data reported in the literature for this synthesis route. The impurity profile was shown to change with reaction time, and all previously reported impurity components were identified but not in all batches. Additionally, 20 batches of methylamphetamine hydrochloride were synthesized from either from l-ephedrine or d-pseudoephedrine hydrochloride in reactions which were allowed to proceed for 24 h. The impurities present in the resulting batches were investigated, and route-specific impurities present in all batches were identified. Batch-to-batch fluctuations in the resultant chromatographic impurity profile, despite careful synthetic monitoring and control, were also noted.

Multivariate statistical methods for the environmental forensic classification of coal tars from former manufactured gas plants.

Compositional disparity within a set of 23 coal tar samples (obtained from 15 different former manufactured gas plants) was compared and related to differences between historical on-site manufacturing processes. Samples were prepared using accelerated solvent extraction prior to analysis by two-dimensional gas chromatography coupled to time-of-flight mass spectrometry. A suite of statistical techniques, including univariate analysis, hierarchical cluster analysis, two-dimensional cluster analysis, and principal component analysis (PCA), were investigated to determine the optimal method for source identification of coal tars. The results revealed that multivariate statistical analysis (namely, PCA of normalized, preprocessed data) has the greatest potential for environmental forensic source identification of coal tars, including the ability to predict the processes used to create unknown samples.

Chemical enhancement of soil based footwear impressions on fabric.

This study investigates the enhancement of footwear impressions prepared with soils from different locations on a variety of fabric surfaces with different morphology. Preliminary experiments using seventeen techniques were carried out and the best responding reagents were evaluated further. Results indicated that the soils investigated (a cross-section of soils from Scotland) are more likely to respond to reagents that target iron ions rather than calcium, aluminium or phosphorus ions. Furthermore, the concentration of iron and soil pH did not appear to have an effect on the performance of the enhancement techniques. For the techniques tested, colour enhancement was observed on all light coloured substrates while enhancement on dark coloured fabrics, denim and leatherette was limited due to poor contrast with the background. Of the chemical enhancement reagents tested, 2,2'-dipyridil was a suitable replacement for the more common enhancement technique using potassium thiocyanate. The main advantages are the use of less toxic and flammable solvents and improved clarity and sharpness of the enhanced impression. The surface morphology of the fabrics did not have a significant effect on the enhancement ability of the reagents apart from a slight tendency for diffusion to occur on less porous fabrics such as polyester and nylon/lycra blends.