Recent forensic applications of enhanced chromatographic separation methods.

This study reviews the recent applications of enhanced separation methods employed in forensic analysis utilizing gas chromatography, liquid chromatography, and supercritical fluid chromatography published between 2015 to 2020, except papers previously covered in relevant review articles. Applications of enhanced chromatographic separation methods to arson investigations, environmental forensics, sexual assault investigations, drug analysis, and toxicology are discussed. Future directions for enhanced chromatographic separation methods in forensic science are also explored.

Capillary zone electrophoresis separation and collection of spermatozoa for the forensic analysis of sexual assault evidence.

The processing of sexual assault kits (SAKs) relies on the genetic analysis of material extracted from swabs collected from the assault victim. A vital step in producing an identifiable DNA profile of the perpetrator is the effective separation of perpetrator (sperm) and victim (epithelial) DNA that have been isolated from the collected evidence. We report the use of capillary zone electrophoresis for the separation of intact sperm from whole and lysed epithelial cells in SAKs. The separated components are deposited into wells of a microtiter plate using a computer-controlled fraction collector, and quantitative PCR is used to verify the collection of sperm cells by targeted amplification of male DNA. We present results from simulated sexual assault samples that have been aged for up to 18 months, as well as vaginal swabs from authentic forensic kits. Components extracted from the vaginal swabs from the SAK comigrated with an aged semen sample at 6.25 ± 0.25 min. Epithelial cells migrated from 10-12 min, producing baseline resolution of the components. Sperm cells were collected in a microtiter plate for downstream analysis.

Exploitation of microbial forensics and nanotechnology for the monitoring of emerging pathogens.

Emerging infectious diseases remain among the leading causes of global mortality. Traditional laboratory diagnostic approaches designed to detect and track infectious disease agents provide a framework for surveillance of bio threats. However, surveillance and outbreak investigations using such time-consuming approaches for early detection of pathogens remain the major pitfall. Hence, reasonable real-time surveillance systems to anticipate threats to public health and environment are critical for identifying specific aetiologies and preventing the global spread of infectious disease. The current review discusses the growing need for monitoring and surveillance of pathogens with the same zeal and approach as adopted by microbial forensics laboratories, and further strengthening it by integrating with the innovative nanotechnology for rapid detection of microbial pathogens. Such innovative diagnostics platforms will help to track pathogens from high risk areas and environment by pre-emptive approach that will minimize damages. The various scenarios with the examples are discussed where the high risk associated human pathogens in particular were successfully detected using various nanotechnology approaches with potential future prospects in the field of microbial forensics.

[Development of Lung Compression Degree Measurement Software of Pneumothorax and Its Application in Forensic Medicine].

OBJECTIVES: To develop a measurement software of lung compression degree to calculate the lung compression ratio in pneumothorax patients accurately and quickly, and then provide an objective assessment of damage degree in forensic clinical identification. METHODS: A volume calculation software was established according to the working principle of the CT instrument. CT data of 15 pneumothorax patients were selected as research objects. The lung compression ratio of pneumothorax patient was calculated by the lung compression volume calculation software of the CT instrument. Meanwhile, the lung compression ratio was also calculated by the developed volume calculation software. The lung compression ratio and operation time calculated by the two methods were analyzed statistically. Scatter plot graphs were draw based on related data, and the developed volume calculation software was verified. RESULTS: The difference between the lung compression ratios calculated by the two methods was not statistically significant, but showed a linear correlation （P<0.05）. The operation time of the developed volume calculation software was obviously shorter. CONCLUSIONS: The volume calculation software developed in this study can calculate the lung compression degree of pneumothorax more conveniently and rapidly with easy accessibility, which shows an application value in the forensic practice.

Current applications of high-resolution mass spectrometry for the analysis of new psychoactive substances: a critical review.

The proliferation of new psychoactive substances (NPS) in recent years has resulted in the development of numerous analytical methods for the detection and identification of known and unknown NPS derivatives. High-resolution mass spectrometry (HRMS) has been identified as the method of choice for broad screening of NPS in a wide range of analytical contexts because of its ability to measure accurate masses using data-independent acquisition (DIA) techniques. Additionally, it has shown promise for non-targeted screening strategies that have been developed in order to detect and identify novel analogues without the need for certified reference materials (CRMs) or comprehensive mass spectral libraries. This paper reviews the applications of HRMS for the analysis of NPS in forensic drug chemistry and analytical toxicology. It provides an overview of the sample preparation procedures in addition to data acquisition, instrumental analysis, and data processing techniques. Furthermore, it gives an overview of the current state of non-targeted screening strategies with discussion on future directions and perspectives of this technique. Graphical Abstract Missing the bullseye - a graphical respresentation of non-targeted screening. Image courtesy of Christian Alonzo.

Medico-Legal Aspects of Pulmonary Thromboembolism.

Pulmonary ThromboEmolism (PTE) is an important disease for legal medicine. Because of their sudden lethal onset, generally medicolegal autopsies show few clinical information when PTE is the cause of death. During medicolegal autopsies, the autopsy operator must answer to important questions. For example, autopsy operator can need to assess the casual relationship between PTE and recent accident, such as trauma or long air travel. Furthermore, the autopsy operator needs to investigate the pathology of PTE as a cause of sudden cardiovascular death. It is relatively simple to confirm a fatal massive thromboembolus in the initial stage of thoracic investigations, but sometimes it might be difficult to distinguish this from postmortem clot. In such cases histopathological examination can help in the differentiation. Histological examination is also required for observation of chronological changes of the thrombi. Chronological evaluation is an important factor especially to determine whether the death coincides with the date of a specific accident/event or instead there is an earlier onset of PTE. In addition, histological sections sometimes show additional information, such as tumor fragments in cases of malignancy or small fragments of bone marrow in cases of active resuscitation, that can be useful in a medicolegal scenario. Furthermore, new diagnostic tools are arising, which they can be very helpful in the individuation of this frequently underdiagnosed disease. The goal of our work is to investigate these aspects through the review of the recent literature.

FORENSIC RADIOLOGY AND IMAGING FOR VETERINARY RADIOLOGISTS.

Imaging studies are often of evidentiary value in medicolegal investigations involving animals and the role of the veterinary radiologist is to interpret those images for courts as an expert or opinion witness. With progressing interest in prosecuting animal crimes and strengthening of penalties for crimes against animals, the participation of veterinary radiologists in medicolegal investigations is expected to increase. Veterinary radiologists who are aware of radiographic and imaging signs that result in animal suffering, abuse, or neglect; knowledgeable in ways radiology and imaging may support cause of death determinations; conversant in postmortem imaging; comfortable discussing mechanisms and timing of blunt or sharp force and projectile trauma in imaging; and prepared to identify mimics of abuse can assist court participants in understanding imaging evidence. The goal of this commentary review is to familiarize veterinary radiologists with the forensic radiology and imaging literature and with the advantages and disadvantages of various imaging modalities utilized in forensic investigations. Another goal is to provide background information for future research studies in veterinary forensic radiology and imaging.

Surface-assisted laser desorption ionization mass spectrometry techniques for application in forensics.

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) is an excellent analytical technique for the rapid and sensitive analysis of macromolecules (>700 Da), such as peptides, proteins, nucleic acids, and synthetic polymers. However, the detection of smaller organic molecules with masses below 700 Da using MALDI-MS is challenging due to the appearance of matrix adducts and matrix fragment peaks in the same spectral range. Recently, nanostructured substrates have been developed that facilitate matrix-free laser desorption ionization (LDI), contributing to an emerging analytical paradigm referred to as surface-assisted laser desorption ionization (SALDI) MS. Since SALDI enables the detection of small organic molecules, it is rapidly growing in popularity, including in the field of forensics. At the same time, SALDI also holds significant potential as a high throughput analytical tool in roadside, work place and athlete drug testing. In this review, we discuss recent advances in SALDI techniques such as desorption ionization on porous silicon (DIOS), nano-initiator mass spectrometry (NIMS) and nano assisted laser desorption ionization (NALDI™) and compare their strengths and weaknesses with particular focus on forensic applications. These include the detection of illicit drug molecules and their metabolites in biological matrices and small molecule detection from forensic samples including banknotes and fingerprints. Finally, the review highlights recent advances in mass spectrometry imaging (MSI) using SALDI techniques.

A Raman "spectroscopic clock" for bloodstain age determination: the first week after deposition.

Knowing the time since deposition (TSD) of an evidentiary bloodstain is highly desired in forensics, yet it can be extremely complicated to accurately determine in practice. Although there have been numerous attempts to solve this problem using a variety of different techniques, currently, no established, well-accepted method exists. Here, a Raman spectroscopic approach was developed for determining the age of bloodstains up to 1 week old. Raman spectroscopy, along with two-dimensional correlation spectroscopy (2D CoS) and statistical modeling, was used to analyze fresh bloodstains at ten time points under ambient conditions. The 2D CoS results indicate a high correlation between several Raman bands and the age of a bloodstain. A regression model was built to provide quantitative predictions of the TSD, with cross-validated root mean squared error and R (2) values of 0.13 and 0.97, respectively. It was determined that a "new" (1 h) bloodstain could be easily distinguished from older bloodstains, which is very important for forensic science in helping to establish the relevant association of multiple bloodstains. Additionally, all bloodstains were confirmatively identified as blood by comparing the experimentally measured spectra to multidimensional body fluid spectroscopic signatures of blood, saliva, semen, sweat, and vaginal fluid. These results demonstrate that Raman spectroscopy can be used as a nondestructive analytical tool for discriminating between bloodstains on the scale of hours to days. This approach shows promise for immediate practical use in the field to predict the TSD with a high degree of accuracy. Graphical Abstract Bloodstain aging over time illustrating naturally ocurring processes.

Guilty by dissociation-development of gas chromatography-mass spectrometry (GC-MS) and other rapid screening methods for the analysis of 13 diphenidine-derived new psychoactive substances (NPSs).

The prevalence of new psychoactive substances (NPSs) in forensic casework has increased prominently in recent years. This has given rise to significant legal and analytical challenges in the identification of these substances. The requirement for validated, robust and rapid testing methodologies for these compounds is obvious. This study details the analysis of 13 synthesised diphenidine derivatives encountered in casework using presumptive testing, thin layer chromatography and gas chromatography-mass spectrometry (GC-MS). Specifically, the validated GC-MS method provides, for the first time, both a general screening method and quantification of the active components for seized solid samples, both in their pure form and in the presence of common adulterants. Graphical Abstract Chemical synthesis and forensic analysis of 13 diphenidine-derived new psychoactive substance(s).

Computer-assisted virtual autopsy using surgical navigation techniques.

UNLABELLED: OBJECTIVE; Virtual autopsy methods, such as postmortem CT and MRI, are increasingly being used in forensic medicine. Forensic investigators with little to no training in diagnostic radiology and medical laypeople such as state's attorneys often find it difficult to understand the anatomic orientation of axial postmortem CT images. We present a computer-assisted system that permits postmortem CT datasets to be quickly and intuitively resliced in real time at the body to narrow the gap between radiologic imaging and autopsy. CONCLUSION: Our system is a potentially valuable tool for planning autopsies, showing findings to medical laypeople, and teaching CT anatomy, thus further closing the gap between radiology and forensic pathology.

Postmortem computed tomography (PMCT) and disaster victim identification.

Radiography has been used for identification since 1927, and established a role in mass fatality investigations in 1949. More recently, postmortem computed tomography (PMCT) has been used for disaster victim identification (DVI). PMCT offers several advantages compared with fluoroscopy, plain film and dental X-rays, including: speed, reducing the number of on-site personnel and imaging modalities required, making it potentially more efficient. However, there are limitations that inhibit the international adoption of PMCT into routine practice. One particular problem is that due to the fact that forensic radiology is a relatively new sub-speciality, there are no internationally established standards for image acquisition, image interpretation and archiving. This is reflected by the current INTERPOL DVI form, which does not contain a PMCT section. The DVI working group of the International Society of Forensic Radiology and Imaging supports the use of imaging in mass fatality response and has published positional statements in this area. This review will discuss forensic radiology, PMCT, and its role in disaster victim identification.

Simple New Method of Detecting Lies By Identifying Invisible Unique Physiological Reflex Response Appearing Often Less Than 10-15 Seconds on the Specific Parts of Face of Lying Person; Quick Screening of Potential Murderers & Problematic Persons.

Frequently, we cannot find any significant visible changes when somebody lies, but we found there are significant invisible changes appearing in specific areas of the face when somebody lies and their location often depends on whether the lie is serious with or without physical violence involvement. These abnormalities were detected non-invasively at areas: 1) lobules and c) a small round area of each upper lateral side of forehead; 2) the skin between the base of the 2 orifices of the nose and the upper end of upper lip and 3) Alae of both sides of nose. These invisible significant changes usually last less than 15 seconds after telling a lie. In these areas, Bi-Digital O-Ring Test (BDORT), which received a U.S. Patent in 1993, became significantly weak with an abnormal value of (-)7 and TXB2, measured non-invasively, was increased from 0.125-0.5ng to 12.5-15ng (within the first 5 seconds) and then went back down to less than 1ng (after 15 seconds). These unique changes can be documented semi-permanently by taking photographs of the face of people who tell a lie, within as short as 10 seconds after saying a lying statement. These abnormal responses appear in one or more of the above-mentioned 3 areas 1), 2) & 3). At least one abnormal pupil with BDORT of (-)8-(-)12 & marked reduction in Acetylcholine and abnormal increase in any of 3 Alzheimer's disease associated factors Apolipoprotein (Apo) E4, ß-Amyloid (1-42), Tau protein, viral and bacterial infections were detected in both pupils and forehead of murderers and people who often have problems with others. Analysis of well-known typical examples of recent mass murderers was presented as examples. Using these findings, potential murderers and people who are very likely to develop problems with others can be screened within 5-10 minutes by examining their facial photographs and signatures before school admission or employment.

Mass spectrometry imaging under ambient conditions.

Mass spectrometry imaging (MSI) has emerged as an important tool in the last decade and it is beginning to show potential to provide new information in many fields owing to its unique ability to acquire molecularly specific images and to provide multiplexed information, without the need for labeling or staining. In MSI, the chemical identity of molecules present on a surface is investigated as a function of spatial distribution. In addition to now standard methods involving MSI in vacuum, recently developed ambient ionization techniques allow MSI to be performed under atmospheric pressure on untreated samples outside the mass spectrometer. Here we review recent developments and applications of MSI emphasizing the ambient ionization techniques of desorption electrospray ionization (DESI), laser ablation electrospray ionization (LAESI), probe electrospray ionization (PESI), desorption atmospheric pressure photoionization (DAPPI), femtosecond laser desorption ionization (fs-LDI), laser electrospray mass spectrometry (LEMS), infrared laser ablation metastable-induced chemical ionization (IR-LAMICI), liquid microjunction surface sampling probe mass spectrometry (LMJ-SSP MS), nanospray desorption electrospray ionization (nano-DESI), and plasma sources such as the low temperature plasma (LTP) probe and laser ablation coupled to flowing atmospheric-pressure afterglow (LA-FAPA). Included are discussions of some of the features of ambient MSI for example the ability to implement chemical reactions with the goal of providing high abundance ions characteristic of specific compounds of interest and the use of tandem mass spectrometry to either map the distribution of targeted molecules with high specificity or to provide additional MS information on the structural identification of compounds. We also describe the role of bioinformatics in acquiring and interpreting the chemical and spatial information obtained through MSI, especially in biological applications for tissue diagnostic purposes. Finally, we discuss the challenges in ambient MSI and include perspectives on the future of the field.

Google Glass for documentation of medical findings: evaluation in forensic medicine.

BACKGROUND: Google Glass is a promising premarket device that includes an optical head-mounted display. Several proof of concept reports exist, but there is little scientific evidence regarding its use in a medical setting. OBJECTIVE: The objective of this study was to empirically determine the feasibility of deploying Glass in a forensics setting. METHODS: Glass was used in combination with a self-developed app that allowed for hands-free operation during autopsy and postmortem examinations of 4 decedents performed by 2 physicians. A digital single-lens reflex (DSLR) camera was used for image comparison. In addition, 6 forensic examiners (3 male, 3 female; age range 23-48 years, age mean 32.8 years, SD 9.6; mean work experience 6.2 years, SD 8.5) were asked to evaluate 159 images for image quality on a 5-point Likert scale, specifically color discrimination, brightness, sharpness, and their satisfaction with the acquired region of interest. Statistical evaluations were performed to determine how Glass compares with conventionally acquired digital images. RESULTS: All images received good (median 4) and very good ratings (median 5) for all 4 categories. Autopsy images taken by Glass (n=32) received significantly lower ratings than those acquired by DSLR camera (n=17) (region of interest: z=-5.154, P<.001; sharpness: z=-7.898, P<.001; color: z=-4.407, P<.001, brightness: z=-3.187, P=.001). For 110 images of postmortem examinations (Glass: n=54, DSLR camera: n=56), ratings for region of interest (z=-8.390, P<.001) and brightness (z=-540, P=.007) were significantly lower. For interrater reliability, intraclass correlation (ICC) values were good for autopsy (ICC=.723, 95% CI .667-.771, P<.001) and postmortem examination (ICC=.758, 95% CI .727-.787, P<.001). Postmortem examinations performed using Glass took 42.6 seconds longer than those done with the DSLR camera (z=-2.100, P=.04 using Wilcoxon signed rank test). The battery charge of Glass quickly decreased; an average 5.5% (SD 1.85) of its battery capacity was spent per postmortem examination (0.81% per minute or 0.79% per picture). CONCLUSIONS: Glass was efficient for acquiring images for documentation in forensic medicine, but the image quality was inferior compared to a DSLR camera. Images taken with Glass received significantly lower ratings for all 4 categories in an autopsy setting and for region of interest and brightness in postmortem examination. The effort necessary for achieving the objectives was higher when using the device compared to the DSLR camera thus extending the postmortem examination duration. Its relative high power consumption and low battery capacity is also a disadvantage. At the current stage of development, Glass may be an adequate tool for education. For deployment in clinical care, issues such as hygiene, data protection, and privacy need to be addressed and are currently limiting chances for professional use.

[The determination of the natural content of chemical elements in human biological objects (liver, kidney, stomach) by mass spectrometry with inductively coupled plasma].

The objective of the present work was to continue the studies of the sites of concentration of the chemical elements corresponding to normal homeostasis in human biological objects by mass spectrometry with inductively coupled plasma. The study yielded the data on the natural content of 27 elements in the cadaveric liver, kidney, and stomach. It is recommended to use these findings as the reference parameters corresponding to normal homeostasis.

Forensic drug analysis and microfluidics.

The analysis of drugs of abuse in microfluidic devices has the potential to provide solutions to today's on-site analysis challenges. The use of such devices has not been limited to miniaturising conventional analytical methods used routinely in forensic laboratories; new and interesting approaches have been implemented in microfluidics and benefit from the ability to control minute amounts of liquids in the small channels. The microfluidic platforms developed so far have been used successfully to carry out single or multiple analytical processes and offer a great opportunity for new technologies for on-site drug testing.

Tunable filter-based multispectral imaging for detection of blood stains on construction material substrates part 2: realization of rapid blood stain detection.

Based on the blood stain detection method and criteria established in part 1 of this article, we combine and organize all necessary tasks to realize the multispectral imaging-based rapid blood stain detection system. To rapidly detect blood stains on the test surface, the developed system automatically captures the spectral images, extracts their spectral data, determines the positions of blood stains, and accurately highlights the positions of blood stains on the display. To achieve such a system, several tasks are newly introduced, including adjustment of camera exposure times to prevent image saturation or excessive darkness, the search for the sampled clean positions of the substrate to determine the substrate reflectance spectrum, and suitable detection procedures and proper arrangement of criteria to eliminate unnecessary calculations. Parallel processes between image capturing and blood stain identification help shorten the time for blood stain identifications despite a large amount of spectral data to be processed. The developed system can identify blood against several other reddish brown stains on several substrates. The measured average identification times on different test surfaces range from only 23.3 to 28.7 s, including the image capturing process.

Intra- and inter-rater reliability of digital image analysis for skin color measurement.

BACKGROUND: We determined the intra- and inter-rater reliability of data from digital image color analysis between an expert and novice analyst. METHODS: Following training, the expert and novice independently analyzed 210 randomly ordered images. Both analysts used Adobe(®) Photoshop lasso or color sampler tools based on the type of image file. After color correction with Pictocolor(®) in camera software, they recorded L*a*b* (L*=light/dark; a*=red/green; b*=yellow/blue) color values for all skin sites. We computed intra-rater and inter-rater agreement within anatomical region, color value (L*, a*, b*), and technique (lasso, color sampler) using a series of one-way intra-class correlation coefficients (ICCs). RESULTS: Results of ICCs for intra-rater agreement showed high levels of internal consistency reliability within each rater for the lasso technique (ICC ≥ 0.99) and somewhat lower, yet acceptable, level of agreement for the color sampler technique (ICC = 0.91 for expert, ICC = 0.81 for novice). Skin L*, skin b*, and labia L* values reached the highest level of agreement (ICC ≥ 0.92) and skin a*, labia b*, and vaginal wall b* were the lowest (ICC ≥ 0.64). CONCLUSION: Data from novice analysts can achieve high levels of agreement with data from expert analysts with training and the use of a detailed, standard protocol.

Material differentiation in forensic radiology with single-source dual-energy computed tomography.

The goal of this study was to investigate the use of dual-energy computed tomography (CT) in differentiating frequently encountered foreign material on CT images using a standard single-source CT scanner. We scanned 20 different, forensically relevant materials at two X-Ray energy levels (80 and 130 kVp) on CT. CT values were measured in each object at both energy levels. Intraclass correlation coefficient (ICC) was used to determine intra-reader reliability. Analysis of variance (ANOVA) was performed to assess significance levels between X-Ray attenuation at 80 and 130 kVp. T test was used to investigate significance levels between mean HU values of individual object pairings at single energy levels of 80 and 130 kVp, respectively. ANOVA revealed that the difference in attenuation between beam energies of 80 kVp compared to 130 kVp was statistically significant (p < 0.005) for all materials except brass and lead. ICC was excellent at 80 kVp (0.999, p < 0.001) and at 130 kVp (0.998, p < 0.001). T test showed that using single energy levels of 80 and 130 kVp respectively 181/190 objects pairs could be differentiated from one another based on HU measurements. Using the combined information from both energy levels, 189/190 object pairs could be differentiated. Scanning with different energy levels is a simple way to apply dual-energy technique on a regular single-energy CT and improves the ability to differentiate foreign bodies with CT, based on their attenuation values.