In recent years, the marked increase in the abuse of fentanyl and its analogues has emphasized the importance of developing highly sensitive and selective analytical methods for their detection in biological matrices. Oral fluid (OF) has emerged as a useful alternative to blood in forensic toxicology, offering a non-invasive and easily accessible matrix for the identification of a recent drug intake. However, its composition requires rigorous sample preparation and robust analytical techniques. A gas chromatography–tandem mass spectrometry (GC-MS/MS) method was developed and validated for the quantification of four opioids and seven fentanyl analogues. A fast and simple solid-phase extraction (SPE) procedure was optimized, enabling the identification and quantification of all analytes in 11 min. The method was validated according to international guidelines, showing a satisfactory degree of linearity (R² ≥ 0.993), precision, accuracy, and sensitivity, with limit of detections (LODs) ranging from 0.10 to 0.20 ng/mL. The method was then successfully applied to n = 10 real OF samples collected during traffic stops set up by police forces which tested negative at the screening tests. Two samples tested positive for codeine and morphine, and one was positive for fentanyl and norfentanyl. The small number of samples currently limits the interpretation of the results. However, our study represents a good starting point for further application of this method to a wider population of real samples.

In Italy, the law on road homicide (Law no. 41/2016) introduced specific provisions for drivers who cause severe injuries or death to a person due to the violation of the Highway Code. The use of alcohol or drugs while driving constitutes an aggravating circumstance of the offence and provides for a tightening of penalties. Our study aims to report on the analysis performed on blood samples collected between January 2018 and December 2024 from drivers convicted of road homicide and who tested positive for alcohol and/or drugs. The majority of the involved subjects were males belonging to the 18–30 and 41–50 age groups. Alcohol, cocaine and cannabinoids were the most detected substances and the most frequent polydrug combination was alcohol and cocaine. We also investigated other influencing factors in road traffic accidents as the day of the week and the time of the day in which fatal road traffic accident occurred, and the time elapsed between the road accident and the collection of biological samples. Our data, in line with the international scenario, strongly support that, in addition to the tightening of penalties, raising awareness plays a key role in preventing alcohol- and drug-related traffic accidents by increasing risk perception and encouraging safer driving behaviors.

Over the past decade, veterinary forensic toxicology has gained increasing relevance due to the recognition of animal poisoning as a criminal, environmental, and public health concern. This review provides an updated overview on current perspectives in veterinary forensic toxicology, focusing on common toxicological scenarios, analytical innovations, and interpretative challenges. Intentional poisoning of companion animals, wildlife intoxication, and environmental contamination are discussed within a One Health framework. Mass spectrometry-based techniques, including targeted gas chromatography–tandem mass spectrometry (GC-MS/MS), liquid chromatography–tandem mass spectrometry (LC–MS/MS) and high-resolution mass spectrometry (HRMS) for non-targeted screening, and emerging ambient mass spectrometry approaches are highlighted as pivotal tools in modern forensic investigations. Major limitations related to post-mortem changes, species-specific toxicokinetic, and the lack of harmonized interpretative criteria are critically examined. The review also identified future priorities such as methodological standardization, expansion of toxicokinetic databases across species, detection of emerging contaminants, and strengthened international cooperation. Overall, veterinary forensic toxicology is an emerging discipline at the intersection of animal welfare, environmental conservation, and public health providing essential evidence for legal, regulatory, and investigative purposes.

Background/Objectives: Antidepressants are widely prescribed medications that may affect psychomotor performance and driving ability depending on their pharmacological profile. This study investigated the prevalence and patterns of antidepressant use among drivers undergoing forensic toxicological evaluation and explored their potential implications for road safety. Methods: An observational study was conducted on n = 6316 drivers undergoing forensic toxicological assessment following licence suspension for driving under the influence (DUI) of alcohol and/or drugs between January 2023 and December 2025. Reported antidepressants were classified into selective serotonin reuptake inhibitors (SSRIs), serotonin–norepinephrine reuptake inhibitors (SNRIs), serotonin antagonist and reuptake inhibitors (SARIs), tricyclic antidepressants (TCAs), norepinephrine–dopamine reuptake inhibitors (NDRIs), noradrenergic and specific serotonergic antidepressants (NaSSAs), and monoamine oxidase inhibitors (MAOIs). Distribution patterns were analysed descriptively and discussed considering the available literature. Results: Antidepressant use was reported by n = 132 participants (2.1%). SSRIs were the most common class (44.0%), followed by SNRIs (26.5%) and SARIs (17.4%). TCAs (6.1%), NDRIs (3.7%), and NaSSAs (2.3%) were less common, while no MAOI use was reported. Among antidepressant users, n = 28 individuals (21.2%) tested positive for other psychoactive substances, including benzodiazepines, cocaine, and cannabinoids. Conclusions: The use of antidepressants was relatively uncommon in the study population. However, clinical evaluation remains important, particularly at the start of treatment and during dose adjustments to determine the potential for an increased risk while driving. Further studies integrating toxicological analyses and clinical data are needed to better define the relationship between antidepressant exposure, polysubstance use, and road safety.

Synthetic cannabinoids (SCs) are an evolving group of new psychoactive substances (NPSs), characterized by structural diversity and continuously emerging analogues. Their detection in biological matrices is challenging because of low concentrations, rapid metabolism, and incomplete coverage by routine methods. Hair analysis offers a detection window for retrospective exposure assessment. This study aimed to develop and validate an ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method for simultaneous identification and quantification of 26 synthetic, semi-synthetic, and related cannabinoids in human hair and to assess its applicability to authentic forensic samples. The method was validated for linearity, limits of detection (LOD) and quantification (LLOQ), accuracy, precision, matrix effect, recovery and process efficiency, with additional robustness assessments under freeze–thaw and processed-extract conditions, and was then applied to n = 100 hair samples collected from subjects tested positive for Δ⁹-tetrahydrocannabinol (Δ⁹-THC) during forensic assessments following driving license suspension. LODs were 1.56–3.13 pg/mg, LLOQs 6.25–12.5 pg/mg, and R² ≥ 0.992; accuracy and precision met the predefined acceptance criteria, while ME, RE, and PE showed generally consistent mean performance with wider level-specific variability for selected analytes. Target cannabinoids were detected in 23/100 samples, with 14 different compounds identified. MDMB-4en-PINACA, 9R-HHC-O, and ADB-BUTINACA were the most frequently detected compounds. The method enabled sensitive and reliable detection of multiple emerging cannabinoid classes in hair, supporting the continuous updating of targeted analytical panels in response to the evolving NPS landscape.

Vipera aspis envenomation represents a potentially life-threatening condition in dogs, particularly in rural or semi-rural areas of Central Italy, where this species is endemic. This case report describes the clinical presentation, diagnostic findings, therapeutic management, and outcome of a dog bitten by Vipera aspis in the Umbria region. At admission, the dog exhibited mild fever, localized swelling, pain at the bite site, and mild systemic clinical signs. Antivenom was not administered, and the dog was managed with supportive and medical therapy aimed at controlling inflammation, preventing secondary infection, and monitoring systemic involvement. The clinical course was favorable, and the dog recovered without complications. As this report describes a single clinical case, no conclusions regarding treatment efficacy can be drawn. However, this case provides additional clinical information on the management and outcome of Vipera aspis envenomation in dogs.