Forensic examination of textile fibres using Raman imaging and multivariate analysis.

Vibrational spectroscopic techniques have shown to be highly suitable for the identification and comparison of textile fibres and clothing fabrics. On the other hand, new chemical imaging modes based on these spectroscopic techniques are becoming useful in multiple fields. This is particularly important to, for instance, chemically visualize and screen different samples including forensic evidence (crime scene investigation), chemical and food products (quality control), biological tissues and living beings (medical imaging), among others. This study explores the forensic examination and selective chemical visualization of textile fibres and clothing fabrics using Raman imaging. Four experiments were performed, which were focused on the screening of (i) white different materials made of 100 % cotton (gauze, cotton wool, t-shirt, and swab), (ii) polyester and cotton fabrics evidence of the same colour, (iii) five different coloured cotton fabrics, and (iv) textile fibres of different materials (acrylic, cotton, nylon, polyester, and silk). Several methods of multivariate chemometric analysis including principal component analysis (PCA), multivariate analysis of variance (MANOVA), and multivariate curve resolution (MCR) were applied to enhance the limited visual comparison of the spectra accomplished with the unaided eye. The results evidenced the suitability of Raman imaging to statistically discriminate textile fibres and fabrics due to the chemical composition of both the clothing material and the dyestuff.

Prevalence study of drugs and new psychoactive substances in hair of ketamine consumers using a methanolic direct extraction prior to high-resolution mass spectrometry.

Few studies have reported the prevalence or incidence about the consumption of new psychoactive substances (NPS). The hair analysis can be useful for this purpose. At the present, ketamine is the most consumed arylcyclohexylamine associated to young consumers and polyconsumption profiles. For this reason, ketamine consumer cases become very interesting to provide information on NPS prevalence. In this work, ten former cases of the National Institute of Toxicology and Forensic Science (INTCF) of Madrid Department (INTCFM), all of them belonging to defendants accused of crimes against public health and who had been found positive to ketamine, were reassessed. At the first toxicological analysis of those hair samples, a positive consume in ketamine had been determined by gas chromatography coupled to mass spectrometry (GC-MS). In this work, the same hair samples were reanalyzed by high-resolution mass spectrometry ( UHPLC-HRMS/MS) using an incubation methanolic extraction combined with a single, simpler, non-selective and direct sample pre-treatment. After corroborating the GC-MS results previously obtained for the same samples, the detection of additional NPS using this new methodology evidenced its benefits and opened the possibility to perform a NPS prevalence study. In brief, in those cases with a positive consumption in ketamine, a polyconsumption of other drugs and NPS was found, including different arylcyclohexylamines as deschloroketamine, 3-MeO-PCP and methoxetamine; and cathinones as methylmetcathinone and N-ethyl-pentylone.

Evaluation of an Ozone Chamber as a Routine Method to Decontaminate Firefighters' PPE.

Ozone chambers have emerged as an alternative method to decontaminate firefighters' Personal Protective Equipment (PPE) from toxic fire residues. This work evaluated the efficiency of using an ozone chamber to clean firefighters' PPE. This was achieved by studying the degradation of pyrene and 9-methylanthracene polycyclic aromatic hydrocarbons (PAHs). The following experiments were performed: (i) insufflating ozone into PAH solutions (homogeneous setup), and (ii) exposing pieces of PPE impregnated with the PAHs to an ozone atmosphere for up to one hour (heterogeneous setup). The ozonolysis products were assessed by Fourier Transform Infrared Spectroscopy (FTIR), Thin-Layer Chromatography (TLC), and Mass Spectrometry (MS) analysis. In the homogeneous experiments, compounds of a higher molecular weight were produced due to the incorporation of oxygen into the PAH structures. Some of these new compounds included 4-oxapyren-5-one (m/z 220) and phenanthrene-4,5-dicarboxaldehyde (m/z 234) from pyrene; or 9-anthracenecarboxaldehyde (m/z 207) and hydroxy-9,10-anthracenedione (m/z 225) from 9-methylanthracene. In the heterogeneous experiments, a lower oxidation was revealed, since no byproducts were detected using FTIR and TLC, but only using MS. However, in both experiments, significant amounts of the original PAHs were still present even after one hour of ozone treatment. Thus, although some partial chemical degradation was observed, the remaining PAH and the new oxygenated-PAH compounds (equally or more toxic than the initial molecules) alerted us of the risks to firefighters' health when using an ozone chamber as a unique decontamination method. These results do not prove the ozone-advertised efficiency of the ozone chambers for decontaminating (degrading the toxic combustion residues into innocuous compounds) firefighters' PPE.

Identification of 2C-B in Hair by UHPLC-HRMS/MS. A Real Forensic Case.

The analysis of drugs of abuse in hair and other biological matrices of forensic interest requires great selectivity and sensitivity. This has been traditionally achieved through target analysis, using one or more analytical methods that include different preanalytical stages, and more complex procedures followed by toxicological laboratories. There is no exception with 2C-series drugs, such as 2C-B, a new psychoactive substance (NPS), which use has emerged and significantly increased, year by year, in the last decades. Continuously new analytical methods are required to selectively detect and identify these new marketed substances at very low concentrations. In this case report, one former case of a polydrug consumer (charged of a crime against public health in Spain) was reanalyzed in hair matrix. In this reanalysis, 2C-B has been positively detected and identified using liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS/MS). The most selective analytical UHPLC-HRMS/MS method alongside a universal and simpler pretreatment methodology has opened up more possibilities for the detection of substances of different chemical structure and optimization of different HRMS/MS detection approaches allowing the identification of 2-CB in the hair of a real forensic case.

Human ultra-weak photon emission as non-invasive spectroscopic tool for diagnosis of internal states - A review.

In the knowledge that human ultra-weak photon emission (UPE) is mainly due to the metabolic oxidative stress processes that the skin cells undergo in the presence of reactive oxygen species (ROS), external stressors (like UV radiation), but also internal stressors (like diseases or brain activity) might strongly influence the UPE. This manuscript revises the scientific advances focused on the influence of internal factors on the human UPE. According to literature, the UPE seems to be influenced by some diseases (including diabetes, hemiparesis, protoporphyria, or a typical cold), and even by the cerebral intention/relaxation (brain activity/meditation). These allow to consider UPE as a natural and promising non-invasive spectroscopic tool for helping during the diagnosis of a variety of illnesses or stress- / mood-state disorders. Nonetheless, further research is required for answering some still unresolved controversial points.

Introducing ATR-FTIR Spectroscopy through Analysis of Acetaminophen Drugs: Practical Lessons for Interdisciplinary and Progressive Learning for Undergraduate Students.

Infrared (IR) spectroscopy is a vibrational spectroscopic technique useful in chemical, pharmaceutical, and forensic sciences. It is essential to identify chemicals for reasons spanning from scientific research and academic practices to quality control in companies. However, in some university degrees, graduate students do not get the proficiency to optimize the experimental parameters to obtain the best IR spectra; to correlate the IR spectral bands with the molecular vibrations (chemical elucidation); to have some criteria for any substance identification (especially relevant in quality control to recognize counterfeit); and to apply chemometrics for comparing, visualizing, and classifying the IR spectra. This work presents an experimental laboratory practice for an introductory teaching of the IR instrumental conditions in the identification of substances based on visual spectra comparison and statistical analysis and matching. Then, the selected IR conditions are applied to different commercial drugs, in the solid state or in solution, mostly composed of acetaminophen. Finally, the students apply chemometrics analysis to the IR data. This practice was designed for the training in a chemistry subject for undergraduate students of the chemistry, pharmacy, or forensics degrees, among others related to science, medical, food, or technological sciences.

Chemical Classification of Explosives.

This work comprehensively reviews some fundamental concepts about explosives and their two commonly used classifications based on either their velocity of detonation or their application. These classifications are highly useful in the military/legal field, but completely useless for the chemical determination of explosives. Because of this reason, a classification of explosives based on their chemical composition is comprehensively revised, discussed and updated. This classification seeks to merge those dispersed chemical classifications of explosives found in literature into a unique general classification, which might be useful for every researcher dealing with the analytical chemical identification of explosives. In the knowledge of the chemical composition of explosives, the most adequate analytical techniques to determine them are finally discussed.

Comparison between computed tomography and silicone-casting methods to determine gunshot cavities in ballistic soap.

Current methods used in terminal ballistics to determine the volume of temporary cavities created by projectiles in soft tissue simulants (such as ballistic soap) usually involve silicone-casting to obtain the cavity moulds. However, these methods have important drawbacks including their little sensitivity and precision, besides the fact that they are destructive. Imaging techniques such as computed tomography (CT) might not only overcome those limitations but also offer useful tools for digitally reporting the scientific results. This work accomplished the 3D digital reconstruction of the cavities created by different projectiles in ballistic soap blocks. This way, the total volume of the cavities, the projectile penetration depths, and other measurements were determined, rendering better capabilities when compared to the current silicone method. All these features were achieved through the CT analysis and 3D Slicer imaging software. In addition, it is worth mentioning that the method can preserve the evidence by digitally obtaining, signing, and storing the infographic videos displaying the 3D-reconstructed cavities. Graphical abstract.

Occurrence and identification of microplastics along a beach in the Biosphere Reserve of Lanzarote.

This work studied the accumulation of plastic debris in a remote beach located in La Graciosa island (Chinijo archipelago, Canary Islands). Microplastics were sampled in the 1-5 mm mesh opening range. An average plastic density of 36.3 g/m2 was obtained with a large variability along the 90 m of the beach (from 8.5 g/m2 to 103.4 g/m2). Microplastic particles preferentially accumulated in the part of the beach protected by rocks. A total number of 9149 plastic particles were collected, recorded and measured, 87% of which corresponded to fragments. Clear colours and microscopic evidence of weathering corresponded to aged plastics wind-driven by the surface Canary Current. The chemical composition of plastics particles corresponded to PE (63%), PP (32%) and PS (3%). Higher PE/PP ratios were recorded in the more protected parts of the beach, suggesting preferential accumulation of more aged fragments.

Study of the adhesion of explosive residues to the finger and transfer to clothing and luggage.

It is important to understand the extent of transfer of explosive particles to different surfaces in order to better evaluate potential cross-contamination by explosives in crowded security controls such as those at airports. This work investigated the transfer of nine explosive residues (ANFO, dynamite, black powder, TNT, HMTD, PETN, NH4NO3, KNO3, NaClO3) through fingerprints from one surface to another. First, the extent of adhesion of explosive residues from different surfaces to the bare finger, nitrile and latex gloves was studied. Then, the transfer of explosive residues from one surface to another through fingerprints was investigated. Cotton fabric (hereinafter referred to as cotton) as clothing material and polycarbonate plastic (hereinafter referred to as polycarbonate) as luggage material were chosen for the experiments. These surfaces containing explosive particles were imaged using a reflex camera before and after the particles were transferred. Afterwards the images were processed in MATLAB where pixels corresponding to explosive residues were quantified. Results demonstrated that transfer of explosive residues frequently occurred with certain differences among materials. Generally, the amount of explosive particles adhered to the finger decreased in the following order: skin>latex>nitrile, while the transfer of particles from the finger to another surface was the opposite. The adhesion of explosive residues from polycarbonate to the finger was found to be better compared to cotton, while the amount of particles transferred to cotton was higher.

Selective Monitoring of Oxyanion Mixtures by a Flow System with Raman Detection.

Raman spectroscopy is a selective detection system scarcely applied for the flow analysis of solutions with the aim of detecting several compounds at once without a previous separation step. This work explores the potential of a portable Raman system in a flow system for the selective detection of a mixture of seven oxyanions (carbonate, sulphate, nitrate, phosphate, chlorate, perchlorate, and thiosulphate). The specific bands of these compounds (symmetric stretching Raman active vibrations of carbonate at 1068 cm−1, nitrate at 1049 cm−1, thiosulphate at 998 cm−1, phosphate at 989 cm−1, sulphate at 982 cm−1, perchlorate at 935 cm−1, and chlorate at 932 cm−1) enabled their simultaneous detection in mixtures. Although the oxyanions’ limit of detection (LOD) was rather poor (in the millimolar range), this extremely simple system is very useful for the single-measurement detection of most of the oxyanions in mixtures, without requiring a previous separation step. In addition, quantitative determination of the desired oxyanion can be performed by means of the corresponding calibration line. These are important advantages for controlling in-line processes in industries like those manufacturing fertilizers, pharmaceuticals, chemicals, or food, among others.

Interpreting the near infrared region of explosives.

The NIR spectra from 1000 to 2500 nm of 18 different explosives, propellant powders and energetic salts were collected and interpreted. NIR spectroscopy is known to provide information about the combination bands and overtones of highly anharmonic vibrations as those occurring in XH bonds (CH, NH and OH). Particularly intense and complex were the bands corresponding to the first combination region (2500-1900 nm) and first overtone stretching mode (2ν) of CH and NH bonds (1750-1450 nm). Inorganic oxidizing salts including sodium/potassium nitrate, sodium/potassium chlorate, and sodium/potassium perchlorate displayed low intense or no NIR bands.

Simple multispectral imaging approach for determining the transfer of explosive residues in consecutive fingerprints.

This novel investigation focused on studying the transfer of explosive residues (TNT, HMTD, PETN, ANFO, dynamite, black powder, NH4NO3, KNO3, NaClO3) in ten consecutive fingerprints to two different surfaces - cotton fabric and polycarbonate plastic - by using multispectral imaging (MSI). Imaging was performed employing a reflex camera in a purpose-built photo studio. Images were processed in MATLAB to select the most discriminating frame - the one that provided the sharpest contrast between the explosive and the material in the red-green-blue (RGB) visible region. The amount of explosive residues transferred in each fingerprint was determined as the number of pixels containing explosive particles. First, the pattern of PETN transfer by ten different persons in successive fingerprints was studied. No significant differences in the pattern of transfer of PETN between subjects were observed, which was also confirmed by multivariate analysis of variance (MANOVA). Then, the transfer of traces of the nine above explosives in ten consecutive fingerprints to cotton fabric and polycarbonate plastic was investigated. The obtained results demonstrated that the amount of explosive residues deposited on successive fingerprints tended to undergo a power or exponential decrease, with the exception of inorganic salts (NH4NO3, KNO3, NaClO3) and ANFO (consists of 90% NH4NO3).

The discrimination of 72 nitrate, chlorate and perchlorate salts using IR and Raman spectroscopy.

Inorganic oxidizing energetic salts including nitrates, chlorates and perchlorates are widely used in the manufacture of not only licit pyrotechnic compositions, but also illicit homemade explosive mixtures. Their identification in forensic laboratories is usually accomplished by either capillary electrophoresis or ion chromatography, with the disadvantage of dissociating the salt into its ions. On the contrary, vibrational spectroscopy, including IR and Raman, enables the non-invasive identification of the salt, i.e. avoiding its dissociation. This study focuses on the discrimination of all nitrate, chlorate and perchlorate salts that are commercially available, using both Raman and IR spectroscopy, with the aim of testing whether every salt can be unequivocally identified. Besides the visual spectra comparison by assigning every band with the corresponding molecular vibrational mode, a statistical analysis based on Pearson correlation was performed to ensure an objective identification, either using Raman, IR or both. Positively, 25 salts (out of 72) were unequivocally identified using Raman, 30 salts when using IR and 44 when combining both techniques. Negatively, some salts were undistinguishable even using both techniques demonstrating there are some salts that provide very similar Raman and IR spectra.

Multi-spectral imaging for the estimation of shooting distances.

Multispectral images of clothing targets shot at seven different distances (from 10 to 220cm) were recorded at 18 specific wavelengths in the 400-1000nm range to visualize the gunshot residue (GSR) pattern. Principal component analysis (PCA) showed that the use of violet-blue wavelengths (430, 450 and 470nm) provided the largest contrast between the GSR particles and the white cotton fabric. Then, the correlation between the amount of GSR particles on clothing targets and the shooting distance was studied. By selecting the blue frame of multispectral images (i.e. the blue frame in the red-green-blue (RGB) system which falls at 470nm), the amount of pixels containing GSR particles was accounted based on the intensity of pixels in that frame. Results demonstrated that the number of pixels containing GSR exponentially decreases with the shooting distance from 30 to 220cm following a particular exponential equation. However, the targets shot at the shortest distance (10cm) did not satisfy the above equation, probably due to the noticeable differences of the GSR-pattern of these targets (e.g. high presence of soot). Then, the equation was applied to validation samples to estimate the shooting distances, obtaining results with an error below 10%.

Statistical approach for ATR-FTIR screening of semen in sexual evidence.

Genetic identification has revolutionized the Forensic Sciences, especially in sexual aggression cases. For the successful extraction of the genetic information of a criminal, a crucial step is the prior detection of bodily fluids on evidence. In this article, a method for non-destructive screening of semen samples is reported. Using chemometric tools, bodily fluids can be detected and differentiated without damaging the sample, by correlating the infrared spectra of sexual evidence with previously recorded spectra from undamaged stains of individual bodily fluids. In modern hospitals/laboratories, the proposed method would not require additional equipment/material nor specialized personnel. Furthermore, the method provides qualitative and reliable results, without requiring human interpretation. Therefore, the proposed method opens a door for a low-cost, fully automated and efficient system for non-destructive screening of semen, which could be easily and massively implemented.

Analysis of different materials subjected to open-air explosions in search of explosive traces by Raman microscopy.

Post-explosion scenes offer such chaos and destruction that evidence recovery and detection of post-blast residues from the explosive in the surrounding materials is highly challenging and difficult. The suitability of materials to retain explosives residues and their subsequent analysis has been scarcely investigated. Particularly, the use of explosive mixtures containing inorganic oxidizing salts to make improvised explosive devices (IEDs) is a current security concern due to their wide availability and lax control. In this work, a wide variety of materials such as glass, steel, plywood, plastic bag, brick, cardboard or cotton subjected to open-air explosions were examined using confocal Raman microscopy, aiming to detect the inorganic oxidizing salts contained in explosives as black powder, chloratite, dynamite, ammonium nitrate fuel oil and ammonal. Post-blast residues were detected through microscopic examination of materials surfaces. In general, the more homogeneous and smoother the surface was, the less difficulties and better results in terms of identification were obtained. However, those highly irregular surfaces were the most unsuitable collectors for the posterior identification of explosive traces by Raman microscopy. The findings, difficulties and some recommendations related to the identification of post-blast particles in the different materials studied are thoroughly discussed.

Revealing the location of semen, vaginal fluid and urine in stained evidence through near infrared chemical imaging.

Crime scene investigation (CSI) requires the ultimate available technology for a rapid, non-destructive, and accurate detection of a wide variety of evidence including invisible stains of bodily fluids. Particularly crucial is the discrimination of semen in stained evidence from sexual abuse cases. This is because those evidence have high odds of containing the DNA from the aggressor. To this aim, we demonstrated the potential of near infrared hyperspectral imaging (NIR-HSI) to make visible stains of semen, vaginal fluid, and urine on fabrics, which lays the bases to face the challenging visualization and discrimination of semen within bodily fluids mixtures. Combining the NIR-HSI data and simple chemometric techniques such as principal component analysis and classical least squares regression, we have revealed the location of semen, vaginal fluid and urine in bodily fluids stained evidence.

Analysis of human bodily fluids on superabsorbent pads by ATR-FTIR.

Superabsorbent pads are composed of different layers with different grades of absorbent capacity, being the lower one the most absorbent layer. Due to their complexity, the analysis of bodily fluids on superabsorbent pads is certainly difficult. In this study, semen, vaginal fluid and urine stains placed on superabsorbent pads including sanitary napkins, panty-liners and diapers were non-destructively detected by Attenuated Total Reflectance (ATR) Fourier Transform Infrared spectroscopy (FTIR). In spite of the higher absorbent capacity of the lower layers, this technique was able to detect the three fluids on the upper layer of all pads, showing that bodily fluids are distributed within all layers. Additionally, mixtures of these bodily fluids prepared on superabsorbent pads and cotton were studied, since real forensic investigations involving sexual abuse cases usually deal with mixtures of these fluids. Due to their IR marked protein region (1800-1480cm-1), semen and vaginal fluid were easily distinguished from urine. However, since semen and vaginal fluid have both a high protein composition, that region of their IR signatures were quite similar, except for slight visual differences, that should be further analysed. Therefore, we propose ATR-FTIR as a suitable, presumptive, non-destructive and rapid approach to detect stains of human bodily fluids on the upper layer of superabsorbent pads from sexual crimes.