Organic impurity profiling of fentanyl samples associated with recent clandestine laboratory methods

Nearly a decade ago, fentanyl reappeared in the United States illicit drug market. In the years since, overdose deaths have continued to rise as well as the amount of fentanyl seized by law enforcement agencies. Research surrounding fentanyl production has been beneficial to regulatory actions and understanding illicit fentanyl production. In 2017, the Drug Enforcement Administration (DEA) began collecting seized fentanyl samples from throughout the United States to track purity, adulteration trends, and synthetic impurity profiles for intelligence purposes. The appearance of a specific organic impurity, phenethyl-4-anilino-N-phenethylpiperidine (phenethyl-4-ANPP) indicates a shift in fentanyl production from the traditional Siegfried and Janssen routes to the Gupta-patent route. Through a collaboration between the DEA and the US Army's Combat Capabilities Development Command Chemical Biological Center (DEVCOM CBC), the synthesis of fentanyl was investigated via six synthetic routes, and the impurity profiles were compared to those of seized samples. The synthetic impurity phenethyl-4-ANPP was reliably observed in the Gupta-patent route published in 2013, and its structure was confirmed through isolation and structure elucidation. Organic impurity profiling results for illicit fentanyl samples seized in late 2021 have indicated yet another change in processing with the appearance of the impurity ethyl-4-anilino-N-phenethylpiperidine (ethyl-4-ANPP). Through altering reagents traditionally used in the Gupta-patent route, the formation of this impurity was determined to occur through a modification of the route as originally described in the Gupta patent.     

Fatal intoxication involving 2-methyl AP-237

Novel synthetic opioids contribute considerably to the opioid epidemic, especially with the frequent emergence of structurally similar compounds. This case report describes a fatal intoxication involving 2-methyl AP-237. A 54-year-old Caucasian male was found deceased from an apparent drug overdose. A plastic container labeled “2MAP” and a cut straw were found in the decedent's backpack at the scene. A white substance found in the container tested positive for fentanyl by field testing. According to his medical history, the decedent was treated for a drug overdose 3 years prior to his death. With no diagnostic findings at autopsy, the case was submitted for toxicological analysis. An unknown substance was detected in peripheral blood and urine using gas chromatography with nitrogen phosphorous detection (GC-NPD). Further testing was conducted using gas chromatography–mass spectrometry (GC–MS) and liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS) which confirmed the presence of 2-methyl AP-237 and potential metabolites in blood and urine. Quantitation by GC-NPD revealed concentrations of 2-methyl AP-237 in blood and urine at 480 ng/mL and 4200 ng/mL, respectively. The toxicological analysis also identified and quantitated alprazolam in the blood at 55 ng/mL. Additionally, the metabolism of 2-methyl AP-237 was investigated and three hydroxylated metabolites were identified in peripheral blood and urine. Limited literature is available for the detection and quantitation of 2-methyl AP-237 in postmortem specimens. Given the toxicological findings with unremarkable autopsy findings, this case is an example of a fatal intoxication involving 2-methyl AP-237.     

2,4,6-Tri(2′-pyridyl)-1,3,5-triazine for determination of iron(II), Iron(III), and total iron contents on human palms

When iron-based tools, such as knives or guns, are held, traces of iron can transfer to the skin. However, no previous studies have been published regarding the effect of the elapsed contact time on the transfer of iron species with different valences to the palm. Compared with 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine (PDT), 2,4,6-tri(2′-pyridyl)-1,3,5-triazine (TPTZ) was found to exhibit a higher sensitivity to iron(II) spectrophotometrically. This work employed 2,4,6-tri(2′-pyridyl)-1,3,5-triazine (TPTZ) and UV spectrophotometry to determine the amounts of iron(II), iron(III), and total iron transferred to human palms from iron tools. It was found that the palmar moisture level was an important factor in determining the amounts of total iron, including iron(II), transferred to the palm. For identical contact times, the amounts of total iron transferred to the palm was proportional to the palmar moisture, and the difference between the maximum and minimum amounts was 12 μg per hand. However, the amounts of iron(II) transferred to the palm gradually decreased over time for low palmar moisture levels, but steadily increased over time for high palmar moisture levels. Additionally, for average levels of palm moisture, the amounts of iron(II) and iron(III) transferred to the palm gradually decreased and increased, respectively, with longer contact times. Notably, this research could serve as a theoretical basis and guide for the detection of trace iron species with different valences on human palms for criminal investigations.     

Ballistic impact of hollow-point ammunition on porcine bone

Identifying failure mechanisms in skeletal tissue allows a deeper understanding of the effects of specific projectile impacts on bone. While ballistic trauma in flat bones is largely researched, knowledge of how long bones react to gunshot impacts is limited in the literature. The impacts of deforming ammunition appear to produce higher levels of fragmentation; however, these have not been studied in depth. This study compares the damage to femora bone by HP 0.357 and 9 mm projectiles constructed with both full and semi-metal jackets. Impact experiments were undertaken on a single-stage light gas gun involving the use of a high-speed video camera and full reconstruction of the bones to ascertain fracture patterns occurring in the femora. Higher degrees of fragmentation are likened to the presence of semi-jacketed HP projectiles than jacketed HP projectiles. The observations of external facing beveled edges are believed to be associated with the increased separation of the jacket and lead core of projectiles. Additionally, experimentation has shown that the amount of kinetic energy lost postimpact is likely related to the presence or the absence of a metal jacket on an HP projectile. The observed data, therefore, suggest that the composition, rather than the configuration, of a projectile affects the type and extent of the damage.     

A reference database of Windows artifacts for file-wiping tool execution analysis

Anti-forensic technology can play an effective role in protecting information, but it can make forensic investigations difficult. Specifically, file-wiping permanently erases evidence, making it challenging for investigators to determine whether a file ever existed and prolonging the investigation process. To address this issue, forensic researchers have studied anti-forensic techniques that detect file-wiping activities. Many previous studies have focused on the effects of file-wiping tools on $MFT, $LogFile, and $DATA, rather than on Windows artifacts. Additionally, previous studies that have examined Windows artifacts have considered different artifacts, making it difficult to study them in a comprehensive manner. To address this, we focused on analyzing traces in 13 Windows artifacts of 10 file-wiping tools' operations in the Windows operating system comprehensively. For our experiments, we installed each file-wiping tool on separate virtual machines and checked the traces that the tools left behind in each artifact. We then organized the results in a database format. Our analysis revealed that most of the tools left traces on other artifacts, except for JumpList, Open&SavePidlMRU, and lnk. There were also some cases where traces remained on the other three artifacts. Based on our research, forensic investigators can quickly identify whether a file-wiping tool has been used, and it can assist in decision-making for evidence collection and forensic triage.     

Pulp volume quantification methods in cone-beam computed tomography for age estimation: A critical review and meta-analysis

The secondary dentin deposition that occurs after the tooth formation process results in a decrease in pulp cavity volume in adolescents and adults. The purpose of this critical review was to correlate pulpal and/or dental volume on cone-beam computed tomography (CBCT) with chronological age approximation. A subobjective was to investigate which methodology and CBCT technical parameters would be most appropriate to evaluate this correlation. This critical review followed the PRISMA guidelines, and it was conducted by a search through PubMed, Embase, SciELO, Scopus, Web of Science, Cochrane Library databases, as well as gray literature. Primary studies that used pulp volume, or pulp chamber to tooth volume ratio measured using CBCT were included. Seven hundred and eight indexed and 31 non-indexed records were identified. A qualitative analysis was performed including 25 selected studies with a total of 5100 individuals, age ranging from 8 to 87 years without sex predilection. The most used method was pulp volume/tooth volume. CBCT voxel size ranged between 0.09 and 0.5. Manual segmentation associated with threshold algorithms was used in most of the studies. Correlation between the pulp volume/tooth volume ratio was moderate: −0.66 for the upper central incisors, −0.59 for upper canines and −0.56 for lower canines. High heterogeneity was observed among the studies. It is concluded that pulp volume should be used with caution in age estimation. Evidence supports the preferable use of upper incisors with pulp volume/tooth volume ratio for age estimation. There is not enough evidence that voxel size interferes in age estimation by pulp volume.     

Internal validation of reduced PCR reaction volume of the Qiagen Investigator® Argus X-12 QS Kit from blood samples on FTA cards

The onus of proof in criminal cases is beyond any reasonable doubt, and the issue on the lack of complete internal validation data can be manipulated when it comes to justifying the validity and reliability of the X-chromosomal short tandem repeats analysis for court representation. Therefore, this research evaluated the efficiency of the optimized 60% reduced volumes for polymerase chain reaction (PCR) amplification using the Qiagen Investigator® Argus X-12 QS Kit, as well as the capillary electrophoresis (CE) sample preparation for blood samples on Flinder's Technology Associates (FTA) cards. Good-quality DNA profile (3000–12,000 RFU) from the purified blood sample on FTA card (1.2 mm) were obtained using the optimized PCR (10.0 μL of PCR reaction volume and 21 cycles) and CE (9.0 μL Hi-Di™ Formamide and 0.3 μL DNA Size Standard 550 [BTO] and 27 s injection time) conditions. The analytical and stochastic thresholds were 100 and 200 RFU, respectively. Hence, the internal validation data supported the use of the optimized 60% reduced PCR amplification reaction volume of the Qiagen Investigator® Argus X-12 QS Kit as well as the CE sample preparation for producing reliable DNA profiles that comply with the quality assurance standards for forensic DNA testing laboratories, while optimizing the analytical cost.     

Characterization of an odor permeable membrane device for the storage of explosives and use as canine training aids

The storage and use of explosives is regulated at the state and federal level, with a particular focus on physical security and rigorous accounting of the explosive inventory. For those working with explosives for the training and testing of explosive-detecting canines, cross-contamination is an important concern. Hence, explosives intended for use with canine teams must be placed into secondary storage containers that are new, clean, and airtight. A variety of containers meet these requirements and include screw-top glass jars (e.g., mason jars). However, an additional need from the explosive-detecting canine community is secondary containers that can also be used as training aids whereby the volatiles emitted by explosives are emitted in a predictable and stable manner. Currently, a generally accepted method for the storage of explosives and controlled emission of explosive vapor for canine detection does not exist. Ideally, such containers should allow odor to escape from the training aid but block external contaminates such as particulates or other volatiles. One method in use places the explosive inside a permeable cotton bag when in use for training and then stores the cotton bag inside an impermeable nylon bag for long-term storage. This paper describes the testing of an odor permeable membrane device (OPMD) as a new way to store and deploy training aids. We measured the evaporation rate and flux of various liquid explosives and volatile compounds that have been identified in the headspace of actual explosives. OPMDs were used in addition to traditional storage containers to monitor the contamination and degradation of 14 explosives used as canine training aids. Explosives were stored individually using traditional storage bags or inside an OPMD at two locations, one of which actively used the training aids. Samples from each storage type at both locations were collected at 0, 3, 6, and 9 months and analyzed using Fourier Transform Infrared (FTIR) Spectroscopy and Gas Chromatography–Mass Spectrometry (GC–MS) with Solid-Phase Microextraction (SPME). FTIR analyses showed no signs of degradation. GC–MS identified cross-contamination from ethylene glycol dinitrate (EGDN) and/or 2,3-dimethyl-2,3-dinitrobutane (DMNB) across almost all samples regardless of storage condition. The contamination was found to be higher among training aids that were stored in traditional ways and that were in active use by canine teams.     

Study on the standard components of digital forensics laboratory

Recently, digital forensics has become increasingly important as it is used by investigation agencies, corporate, and private sector. To supplement the limitations of evidence capacity and be recognized in court, it is essential to establish an environment that ensures the integrity of the entire process ranging from collecting and analyzing to submitting digital evidence to court. In this study, common elements were extracted by comparing and analyzing ISO/IEC 17025, 27001 standards and Interpol and Council of Europe (CoE) guidelines to derive the necessary components for building a digital forensic laboratory. Subsequently, based on 21 digital forensic experts in the field, Delphi survey and verifications were conducted in three rounds. As a result, 40 components from seven areas were derived. The research results are based on the establishment, operation, management, and authentication of a digital forensics laboratory suitable for the domestic environment, with added credibility through collection of the opinions of 21 experts in the field of digital forensics in Korea. This study can be referred to in establishing digital forensic laboratories in national, public, and private digital forensic organizations as well as for employing as competency measurement criteria in courts to evaluate the reliability of the analysis results.     

Overview of the seized and examined pharmaceutical in the state of Minas Gerais-Brazil by the Civil Police from 2017 to 2022: A fundamental analysis of the recent impact over the drugs market

This work presents a data survey regarding the qualitative chemical analysis of drugs seized by the Police in the state of Minas Gerais between July 2017 and June 2022, including an evaluation of labeling of 265 samples of anabolic androgenic steroids (AAS) seized in 2020. The Active Pharmaceutical Ingredients (API) present in the samples were identified through chemical analysis and classified by system Anatomical Therapeutic Chemical (ATC) methods. Analysis of the labeling information for 265 samples of AAS followed the guidance of legislation RDC 71 (2009) from ANVISA. For this study 6355 seized pharmaceuticals underwent qualitative chemical analysis that corresponded to 7739 APIs successfully identified and classified. Among the components studied AAS, psychostimulants, anesthetics, and analgesics were the most commonly examined. AAS seized and tested increased by over 100% and for the majority of the samples analyzed were found to not match the labeling on the packaging. In the meantime, anti-obesity drugs presented a prominent increase of 400% from 2020/1 to 2021/2, during covid-19 quarantine. Seized pharmaceuticals and tests can support information in the planning of public health and safety policies.