Journal of Analytical Toxicology

Buchweitz, John P; Zyskowski, Justin A; Cajigas, Kevin; Brooks, Jason W

doi: 10.1093/jat/bkag005pmid: 41514164

Rocuronium bromide is a non-depolarizing neuromuscular blocking agent (NMBA) commonly used in anesthesia for its rapid onset and intermediate duration of action. While its therapeutic use has been well-documented in humans and experimentally in dogs, this report describes the first known case of its malicious use in the fatal intoxication of a canine. Herein we describe the case of a six-year-old male Plott Hound found deceased under suspicious circumstances. Postmortem investigation revealed focal hemorrhages near potential injection sites, and toxicological analysis via liquid chromatography–tandem mass spectrometry confirmed the presence of rocuronium in muscle tissue, heart blood, and urine. The dog had no surgical history or indication for anesthesia. However, rocuronium concentrations mirrored those seen in anesthetized human patients under ventilatory support. The owner, a licensed pharmacist, was suspected of drug theft and aggravated animal abuse. This case highlights the lethality of NMBAs when misused outside of the clinical setting and further underscores a collaborative, multi-institutional, multidisciplinary approach to veterinary forensic investigations.

Ballotari, Marco; Truver, Michael T; Hoyer, Jennifer L; Goldberger, Bruce A; Chronister, Chris W

doi: 10.1093/jat/bkag035pmid: 42234862

In the United States (U.S.), fentanyl remains the leading drug associated with opioid-related deaths. Many analogs of fentanyl have been identified and linked to the opioid overdose epidemic. After core-structure scheduling of illicitly manufactured fentanyl (IMF) and related products, para-fluorofentanyl (p-FF) reemerged with varied prevalence in the U.S. Xylazine also emerged as a cutting agent in IMF. The aim of this study was to describe the prevalence and trend use patterns of fentanyl, p-FF, and xylazine in a case series from 2021 to 2023 in the state of Florida. A three-year retrospective analysis of data was conducted. A total of 8121 cases were submitted for analysis over the period of study. Among them, 1530 cases (18.8%) tested positive for fentanyl in one or more of the specimens submitted for these cases. The age of the decedents ranged 16–85 years, with the majority being White males. The concentration range of fentanyl in the blood specimens (n = 1358) analyzed was 2.5–579 ng/mL. p-FF was detected with fentanyl in 164 postmortem blood specimens with a concentration range of 2.5–65 ng/mL. Xylazine was identified in the urine of 391 decedents. The combination of fentanyl, p-FF, and xylazine was detected in 70 cases. When fentanyl co-occurred with both xylazine and p-FF, it was primarily observed with benzodiazepines, cocaine, methamphetamine, and other opioids. This study aimed to provide a better understanding of the fentanyl analog and adulterant prevalence in Florida.

Olson, Aaron

doi: 10.1093/jat/bkag028pmid: 42090454

This letter addresses the article by Deeb et al. describing the analysis of cannabinoids in exhaled breath. While the study represents a technical advancement in breath-based detection, several interpretive and analytical limitations warrant clarification. The manuscript discusses potential impairment-related applications despite the absence of direct measures of impairment. Existing literature, including controlled clinical studies and government reports, indicates that THC concentrations in biological matrices cannot be used as a reliable indicator of impairment. Additionally, key variables influencing cannabinoid exposure, including dose, potency, and inhalation behavior, were not controlled, further limiting interpretability. Analytical concerns are also noted, including subpar chromatographic resolution (Rs ≈ 0.9), which may compromise accurate identification and quantitation of structurally similar cannabinoids. These issues highlight the need for caution in interpreting breath cannabinoid data, particularly in forensic contexts where conclusions regarding impairment may carry significant consequences.

Wood, Rebecca; Moore, Robert

doi: 10.1093/jat/bkag027pmid: 42046253

N-pyrrolidino isotonitazene (Isotonitazepyne) is an extremely potent novel synthetic opioid which has been detected internationally in counterfeit pharmaceutical tablets. Published data from post-mortem cases is extremely limited and no metabolite profiles from authentic case work have been published. This paper describes a number of post-mortem cases in which N-pyrrolidino isotonitazene was thought to play a significant role in the cause of death, as well as describing the detected metabolite profiles. Concentration ranges for N-pyrrolidino isotonitazene was reported to be 1.1–55.9 µg/L across the 4 cases. The presumptive metabolite 5-amino-N-pyrrolidino isotonitazene was also identified in all samples; n-pyrrolidino-4-hydroxy nitazene was present in 3 of the 4 urine samples; and N-pyrrolidino hydroxy isotonitazene was present in 1 sample. This study highlights some of the challenges associated with nitazene interpretation, such as the very wide concentration ranges encountered in potentially fatal cases, as well as providing provisional metabolite data which may aid in detecting future cases.

Vanerio, Sofia; Galante, Nicola; Ravelli, Alessandro; Bergamaschi, Roberta F; Battistini, Alessio; Casati, Sara

doi: 10.1093/jat/bkag014pmid: 41719197

Bone marrow (BM) has emerged as a valuable alternative matrix in postmortem toxicology, when conventional samples such as blood or soft tissues are unavailable, degraded or contaminated. BM, due to its protecting anatomical site within the medullary cavity, slows down decomposition, limits environmental and microbial interference and enables the preservation of xenobiotics over extended postmortem intervals. This review summarizes the updated literature on BM anatomy, sampling site, xenobiotic distribution and stability, as well as the influence of postmortem redistribution (PMR) and putrefaction for toxicological interpretation of the analytical results. Evidence indicates that drug distribution in BM is governed by tissue vascularity and analyte physicochemical properties, with lipophilic compounds often reaching higher concentrations in BM than in blood. Numerous studies demonstrate the long-term stability of drugs such as amphetamines, benzodiazepines, sedative-hypnotics and ethanol in BM, even in severely decomposed, burned, or skeletonized remains. While BM shows reduced susceptibility to PMR and putrefaction compared to blood and soft tissues, post-collection stability depends on proper storage conditions. Findings from human forensic cases assess the detectability of a wide range of illicit and therapeutic substances in BM, supporting its utility in reconstructing ante-mortem drug exposure when traditional matrices are compromised. Despite promising results, data remain limited, and further research is needed to refine interpretative frameworks and establish standardized protocols for BM sampling, analysis, and toxicological evaluation.

Ellefsen, Kayla N; Karschner, Erin L; Mérette, Sandrine A M; Patton, Amy L; Shoff, Elisa N; Smith, Christina R; Truver, Michael T; Mata, Dani C; Papsun, Donna M

doi: 10.1093/jat/bkag026pmid: 42033028

The proliferation and sustained detection of novel psychoactive benzodiazepines in forensic and clinical toxicology, combined with their potential for adverse events, poses an enduring threat to public health and safety with complex characteristics and challenges unique from other subclasses of novel psychoactive substances (NPS). This study aimed to systematically review the published effects, observations, and toxicological data from postmortem and human performance studies [including clinical, driving under the influence of drugs (DUID), and drug-facilitated crimes (DFC)] involving NPS benzodiazepines from 2021 to 2025. The challenges this class of compounds presents to clinicians, toxicologists, and medical professionals were also addressed, including both analytical and interpretative challenges. Literature reviews were performed in PubMed, Google Scholar, Google Search, toxicology journals, and conference abstract proceedings using search terms such as “NPS benzodiazepines,” “designer benzodiazepines,” and compound specific searches; authoritative websites such as NPS Discovery, National Forensic Laboratory Information System, and the European Union Drug Agency were also consulted. A total of 259 NPS benzodiazepine-related studies were identified including 29 postmortem, 15 DUID, 27 clinical, and 5 DFC studies detailed in this review. NPS benzodiazepines were widely pervasive in both postmortem and human performance cases with overlapping toxic and recreational concentrations; often involving polysubstance use with other central nervous system depressants and stimulants. Unique clinical presentations, observed effects, and autopsy findings were also reported for NPS benzodiazepines. This review provides an updated, consolidated resource to support toxicologists and clinicians in interpretation, emerging risk assessment, and evolving challenges associated with NPS benzodiazepines across postmortem and human performance settings.