From: Larry Roberts (enclosure)

Illegal trafficking of pharmaceutical products by criminal organisations is a global threat for public health. Drugs for erectile dysfunction such as phosphodiesterase type 5 inhibitors are the most commonly counterfeited medicines in Europe. The search of possible toxic chemical substances in seized products is needed to provide early warning for public health. Furthermore, the elemental profile of the seized products can be useful in criminal investigations. For the first time an ion beam analysis (IBA) procedure to characterise authentic Viagra® tablets and sildenafil-based illegal products is described. Moreover, results are compared with the ones obtained by instrumental neutron activation analysis (INAA) on authentic Viagra® tablets in two reactors. IBA results showed that a combination of particle-induced X-ray emission (PIXE) and secondary ion mass spectrometry using primary ions with energies in the range of several MeV (MeV-SIMS) is a powerful tool to characterise different products in a straightforward manner, allowing discrimination between legal and illegal products. INAA allowed accurate elemental quantification and also showed a great potential for the future implementation of an inter- laboratory classification system

A Probable Fatal Case of Oleander (Nerium oleander) Poisoning on a Cattle Farm: A New Method of Detection and Quantification of the Oleandrin Toxin in Rumen

Oleander (Nerium oleander) is an ornamental plant common in tropical and sub-tropical regions that is becoming increasingly widespread, even in temperate regions. Oleander poisoning may occur in animals and humans. The main active components contained in the plant are cardiac glycosides belonging to the class of cardenolides that are toxic to many species, from human to insects. This work describes a case of oleander poisoning that occurred on a small cattle farm and resulted in the fatality of all six resident animals. Furthermore, the investigation of the poisonous agent is described, with particular focus on the characterization of the oleandrin toxin that was recovered from the forage and rumen contents. The innovation of this study is the first description of the detection and quantification of the oleandrin toxin by liquid chromatography-high resolution mass spectrometry (LC-HRMS) in rumen

LC-HRMS characterization of the skin pigmentation and sexualenhancers melanotan II and bremelanotide sold on the blackmarket of performance and image enhancing drugs

The spread of performance and image enhancing drugs (PIEDs) often requires foren-sic toxicology laboratories to identify unknown compounds without reference stan-dards. We characterized the PIEDs melanotan II and bremelanotide, not legallymarketed, in eight unknown samples confiscated by police together with anabolicsteroids, hormone modulators, sexual enhancers and stimulants, intended for theblack market of bodybuilders, using liquid chromatography–high resolution/highaccuracy Orbitrap mass spectrometry (LC-HRMS). The characterization was carriedout by the accurate mass measurements of MH+ionic species, the study of theirisotopic patterns and the associated relative isotopic abundance (RIA) values, as wellas the accurate mass measurements of collision-induced product ions obtained infragmentation experiments. LC-HRMS confirmed itself as a powerful analytical toolto elucidate the elemental composition and structural characteristics of unknowncompounds

APPLICATION OF ULTRASOUND-ASSISTED LIQUID-LIQUID MICRO EXTRACTION COUPLED WITH GAS CHROMATOGRAPHY-MASS SPECTROMETRY FOR THE RAPID DETERMINATION OF SYNTHETIC CANNABINOIDS AND METABOLITES IN BIOLOGICAL SAMPLES.

The constant emergence of New Psychoactive Substances is a challenge to clinical and forensic toxicologists that need to constantly update analytical techniques to detecting them. A large portion of these substances are synthetic cannabinoids. The aim of this study was to develop a rapid and simple method for the determination of synthetic cannabinoids and their metabolites in urine and blood using GC-MS. The method involves an Ultrasound-Assisted Dispersive Liquid-Liquid Microextraction which implies a rapid procedure, giving excellent extraction efficiencies with minimal use of toxic solvents. This is followed by silylation and analysis with GC-MS. The chromatographic method allows for the separation and identification of 29 selected synthetic cannabinoids and some metabolites. The method was validated on urine and blood samples with the ability to detect and quantify all analytes with satisfactory limits of detection (from 1 to 5 ng/mL), limits of quantification (5 ng/mL), selectivity and linearity (in the range 5 - 200 ng/mL). The developed assay is highly applicable to laboratories with limited instrumental availability, thanks to the use of efficient and low-cost sample preparation and instrumental equipment. The latter may contribute to enhance the detection of NPS in clinical and forensic toxicology laboratories

Metabolism Study of N-Methyl 2-amino Indane (NM2AI) and Determination of Metabolites in Biological Samples by LC-HRMS

Since the widespread diffusion of new psychoactive substances, forensic laboratories are often required to identify new drugs and their metabolites for which information or reference standards are lacking. We performed a study on N-methyl-2-aminoindane (NM2AI) metabolism in silico and in vivo, in order to identify the main metabolites to be screened in the different biological samples. We performed the in silico metabolism prediction of NM2AI using MetaSiteTM software, and subsequently verified the presence of metabolites in the blood, urine and hair of mice after NM2AI administration. The samples were analyzed by liquid chromatography-high-resolution mass spectrometry (LC-HRMS) with a benchtop Orbitrap Exactive mass detector. This allowed the evaluation of the agreement between software prediction and experimental results in biological samples. LC-HRMS analysis identified seven main metabolites in the urine. They were identified, by their accurate masses and fragmentation patterns, as 2-aminoindane (2AI), two hydroxy-2AI and four hydroxy-NM2AI; one of the hydroxy-NM2AI and one of the hydroxy-2AI underwent also to conjugation. NM2AI and 2AI were also detected by LC-HRMS in the hair and blood. Based on these findings, we developed an LC-HRMS method for the screening of NM2AI and metabolites in urine, blood and hair samples. This can be of primary effectiveness to uncover the abuse of NM2AI and related possible intoxications

Behavioral and Pharmacokinetics Studies of <i>N</i>-Methyl-2-Aminoindane (NM2AI) in Mice: An Aminoindane Briefly Used in the Illicit Drug Market

Drug forums are considered as the main platform sources that have contributed to the increase in NPS popularity, especially for those not yet known to law enforcement and therefore not yet illegal. An example is the new synthetic stimulant NM2AI, which has a very short history of human use and abuse. Little is known regarding this compound, but some information from internet forums and the scientific literature indicates NM2AI as a structural derivate of MDAI, which is known for its entactogenic activity. Indeed, the purpose of this study is to evaluate, for the first time, the in vivo acute effect induced by the intraperitoneal injection of NM2AI (1–10–30–100 mg/kg) in mice. We demonstrate the sensory (by visual placing and object tests) and physiological (core temperature measurement) function variations, nociceptor (by tail pinch test) and strength (grip test) alterations, and sensorimotor (time on rod and mobility) decrease. Moreover, we verify the mild hallucinogenic effect of NM2AI (by startle/prepulse inhibition test). Lastly, we perform a pharmacokinetic study on mice blood samples, highlighting that the main active metabolite of NM2AI is 2-aminoindane (2AI). Taken together, our data confirm the suspected entactogenic activity of NM2AI; however, these in vivo effects appear atypical and less intense with respect to those induced by the classic stimulants, in surprising analogy with what is reported by networked users

Scopolamine fatal outcome in an inmate after buscopan\uae smoking

Scopolamine is an alkaloid which acts as competitive antagonists to acetylcholine at central and peripheral muscarinic receptors. We report the case of a 41-year-old male convict with a 27-year history of cannabis abuse who suddenly died in the bed of his cell after having smoked buscopan\uae tablets. Since both abuse of substances and recent physical assaults had been reported, we opted for a comprehensive approach (post-mortem computed tomography CT (PMCT), full forensic autopsy, and toxicology testing) to determine which was the cause of the death. Virtopsy found significant cerebral edema and lungs edema that were confirmed at the autopsy and at the histopathological examination. Scopolamine was detected in peripheral blood at the toxic concentration of 14 ng/mL in blood and at 263 ng/mL in urine, and scopolamine butyl bromide at 17 ng/mL in blood and 90 ng/mL in urine. Quetiapine, mirtazapine, lorazepam, diazepam, and metabolites and valproate were also detected (at therapeutic concentrations). Inmates, especially when they have a history of drug abuse, are at risk to use any substance they can find for recreational purposes. In prisons, active surveillance on the management and assumption of prescribed drugs could avoid fatal acute intoxication

Ion beam analysis (IBA) and instrumental neutron activation analysis (INAA) for forensic characterisation of authentic Viagra® and of sildenafil-based illegal products

Illegal trafficking of pharmaceutical products by criminal organisations is a global threat for public health. Drugs for erectile dysfunction such as phosphodiesterase type 5 inhibitors are the most commonly counterfeited medicines in Europe. The search of possible toxic chemical substances in seized products is needed to provide early warning for public health. Furthermore, the elemental profile of the seized products can be useful in criminal investigations. For the first time an ion beam analysis (IBA) procedure to characterise authentic Viagra® tablets and sildenafil-based illegal products is described. Moreover, results are compared with the ones obtained by instrumental neutron activation analysis (INAA) on authentic Viagra® tablets in two reactors. IBA results showed that a combination of particle-induced X-ray emission (PIXE) and secondary ion mass spectrometry using primary ions with energies in the range of several MeV (MeV-SIMS) is a powerful tool to characterise different products in a straightforward manner, allowing discrimination between legal and illegal products. INAA allowed accurate elemental quantification and also showed a great potential for the future implementation of an inter-laboratory classification system.</p

Method development for the identification of methoxpropamine, 2-fluoro-deschloroketamine and deschloroketamine and their main metabolites in blood and hair and forensic application

The constant increase of new psychoactive substances, often available on the illicit drug market as 'research chemicals', poses a concern for public health and a significant analytical and legislative challenge. \u392-keto-arylcyclohexamines represent a class of dissociative anesthetics recently introduced on the market of New Psychoactive Substances (NPS). There is still a lack of information about the pharmacological activity of many of such substances, usually depending on the potential chemical modifications introduced to circumvent the law. Furthermore, their intake may not be fully intentional, since consumers do not always have knowledge of the content of online purchases. The present study describes the characterization by liquid chromatography-high resolution mass spectrometry (LC-HRMS), using a benchtop Orbitrap instrument, of the novel ketamine analogues methoxpropamine, 2-fluoro-deschloroketamine and deschloroketamine, found in the post-mortem blood and hair samples from a forensic case of suicide by fall from height, and of some of their metabolites. This allowed the development of analytical methods for the determination of both the \u3b2-keto-arylcyclohexamines and the metabolites in LC-HRMS and in LC-MS/MS, providing a starting point for studying their toxicokinetics

Ion beam analysis (IBA) and instrumental neutron activation analysis (INAA) for forensic characterisation of authentic Viagra® and of sildenafil-based illegal products

Illegal trafficking of pharmaceutical products by criminal organisations is a global threat for public health. Drugs for erectile dysfunction such as phosphodiesterase type 5 inhibitors are the most commonly counterfeited medicines in Europe. The search of possible toxic chemical substances in seized products is needed to provide early warning for public health. Furthermore, the elemental profile of the seized products can be useful in criminal investigations. For the first time an ion beam analysis (IBA) procedure to characterise authentic Viagra® tablets and sildenafil-based illegal products is described. Moreover, results are compared with the ones obtained by instrumental neutron activation analysis (INAA) on authentic Viagra® tablets in two reactors. IBA results showed that a combination of particle-induced X-ray emission (PIXE) and secondary ion mass spectrometry using primary ions with energies in the range of several MeV (MeV-SIMS) is a powerful tool to characterise different products in a straightforward manner, allowing discrimination between legal and illegal products. INAA allowed accurate elemental quantification and also showed a great potential for the future implementation of an inter-laboratory classification system.Accepted Author ManuscriptTechnici Poo