Interpretative problems due to the presence of chloromethcathinone isomers in the biological material from postmortem cases.

While many new psychoactive substances often disappear from the drug market rather quickly, some, such as synthetic cathinones (SCs), still remain due to their popularity among users. The current knowledge of SC concentrations in blood samples is based mainly on the published case reports of intoxications or fatalities caused by SC intake. The aim of the present study was to present and interpret the obtained toxicological analysis results of these cases, in which it was possible to determine or detect the presence of one of the isomers of chloromethcathinone (CMC) along with its intake biomarker-dihydro-CMC. These cases include 27 deaths reported at the Department of Forensic Medicine in Kraków in 2016-2022. CMC constitutes a major toxicological opinion challenge, in terms of toxicological evaluation of poisonings. As presented in this paper, a significant problem is its stability in the biological material and practices in the reporting of the obtained data. It is therefore important to monitor potential intake biomarkers that may show greater stability in the biological material than the parent drug. In the case of CMC isomers, the good biomarker of intake is the dihydro-CMC metabolite, which was detected in the blood sample in every case presented, even with the absence of the parent substance. Interpretation of the results obtained for CMC in terms of assessing their toxicity and possible cause of death is difficult. However, it should be taken into account that in cases of new psychoactive substance poisoning, an in-depth risk assessment is mandatory and the opinion of the unpredictability of the effects is taken as a principle.

The Stability of Synthetic Cathinones and the Study of Potential Intake Biomarkers in the Biological Material from a Case of 3-CMC Poisoning.

The objective of the present study was to identify the metabolites of synthetic cathinone (SC), 3-chloromethcathinone (3-CMC), and to select a potential intake biomarker for this compound. The basis of the experiment was the analysis of blood and urine samples from a case of fatal poisoning with this substance. We also evaluated the stability of 3-CMC and the selected potential biomarker, the dihydro-3-CMC metabolite, depending on the time elapsed since the autopsy as well as the storage conditions of the biological material. Studies have shown that 3-CMC shows low stability in the biological material. However, acidification of the biological material or storage at low temperatures has a great impact on maintaining its stability. It is crucial to monitor the presence of the dihydro-3-CMC metabolite in the biological material during analysis, which showed high stability under all of the given storage conditions. SCs might not be detected in the biological material and, if they are, the determined concentrations at the time of testing may not correspond to the actual concentrations at the time of the event or death. Consequently, the interpretation of the results obtained for 3-CMC and dihydro-3-CMC in terms of assessing their toxicity and possible cause of death is difficult. The area of research into the search for other intake biomarkers of unstable halogen derivatives of SCs remains open.

Post-aggression suicide under the influence of new psychoactive substances AMB-FUBINACA and U-47700.

The objective of the present study was the development and validation of the method for determining AMB-FUBINACA and its metabolite - AMB-FUBINACA O-desmethyl acid - in blood samples, followed by verification of the method in toxicological judicial and forensic medicine practice employing the example of post-aggression suicide. Most likely in consequence of development of adverse effects resulting in psychotic symptoms, a male being under the influence of the synthetic cannabinoid AMB-FUBINACA and the new synthetic opioid U-47700, mortally wounded his female partner and subsequently committed suicide. Identification and determination of the afore-mentioned xenobiotics in blood samples collected from the male and female victims were performed employing high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The analytes were isolated from blood samples using the solid phase extraction (SPE) method. The blood samples collected from the male and female demonstrated respectively 110 and 196 ng/mL of AMB-FUBINACA O-desmethyl acid metabolite, 1935 and 357 ng/mL of U-47700, 250 and 200 ng/mL of N-desmethyl-U-47700, as well as 410 and 200 ng/mL of N,N-didesmethyl-U-47700. The concentration values of new psychoactive substances (NPS's) in blood samples originating from the male and female were within the ranges encountered in cases of poisoning, including these resulting in death. Nevertheless, the evident signs of exsanguination proof that the woman was alive when she sustained lethal injuries. The presented cases illustrate the difficult to be anticipated effect exerted on the users by NPS's.

Forensic-psychiatric relationships in the context of forensic medical examination of new psychoactive substance-related deaths.

INTRODUCTION: The aim of this study was to find causal relationships between the use of new psychoactive substances (NPS), the so-called "legal highs", and the cause of death, taking into account information indicating the victim's mental disorders before death. MATERIAL AND METHODS: The research material includes the results of toxicological analyses of blood samples collected during autopsies of 40 deceased persons whose cause of death was associated with the ingestion of new psychoactive substances (NPS). The blood samples were subjected to solid phase extraction (SPE), and then analyzed by liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) for quantification. Information on individual cases was taken from case files provided by the prosecutor's office for research purposes. RESULTS: As a result of the analyses, 24 types of NPS were detected, comprising synthetic cathinones - 15 cases, synthetic cannabinoids - 10 cases, synthetic opioids - 13 cases, other - 2 cases. In the study group there were 5 women (12.5%) and 35 men (87.5%). Suicide was demonstrated in 10 cases (25%), while the remaining 30 cases (75%) were accidental deaths, including those in which the actual motivation of drug use could not be determined. Drugs used to treat mental disorders were detected in 6 cases, while analgesics were demonstrated in 5 cases. CONCLUSIONS: In the majority of cases, including suicides and accidental deaths where the manner of death was unknown, NPS use and consequent death from various causes are associated with psychiatric disorders.

Increased Ethanol Consumption and Locomotion Develop upon Ethanol Deprivation in Rats Overexpressing the Adenosine (A)2A Receptor.

Preclinical data indicate that ethanol produces behavioral effects that can be regulated by many neurotransmitters and neuromodulators like adenosine (A). The most important receptors with respect to the rewarding effects of ethanol seem to be the A2A receptors. This study used a transgenic strategy, specifically rats overexpressing the A2A receptor, to characterize the neurobiological mechanisms of ethanol consumption as measured by intermittent access to 20% ethanol in a two-bottle choice paradigm. In this model, no change in ethanol consumption was observed in transgenic animals compared to wild type controls during the acquisition/maintenance phase. Following alcohol deprivation, only transgenic rats overexpressing the A2A receptor exhibited escalation of ethanol consumption and drank more (by ca. 90%), but not significantly, ethanol than did the wild type rats. During ethanol withdrawal, the immobility time of rats overexpressing the A2A receptor in the forced swim test was lower than that of wild type rats. Moreover, transgenic rats withdrawn from ethanol, compared to the drug-naive transgenic animals, exhibited an increase above 70% in locomotion. The results indicated that the overexpression of A2A receptors may be a risk factor for the escalation of ethanol consumption despite the reduction in depression-like signs of ethanol withdrawal.

Aspects related to analysis and medico-legal assessment in the context of the presence of synthetic cannabinoids in biological material based on the example of AB-CHMINACA.

AIM OF THE STUDY: The aim of this study was to develop and validate a method for the determination of the synthetic cannabinoid AB-CHMINACA in blood, followed by its verification in forensic toxicological practice. MATERIAL AND METHODS: The case of a 41-year-old man admitted to hospital because of scheduled cardiac surgery was discussed. The man died after 12 hours of hospitalization. Based on collected evidence, AB-CHMINACA poisoning was suspected. The identification and determination of AB-CHMINACA in the man's blood was performed by high-performance liquid chromatography coupled with tandem mass spectrometry using electrospray ionization (HPLC-ESI-MS-MS), after prior solid phase extraction. RESULTS: The concentration of AB-CHMINACA determined in the man's blood sample was 0.5 ng/ml. CONCLUSIONS: In the interpretation of the case, it was concluded that AB-CHMINACA had no direct effect on the patient's death, the cause of which was ascertained as chronic heart failure secondary to aortic valve disease, decompensated by pneumonia. However, an indirect impact of side effects resulting from the use of synthetic cannabinoids cannot be ruled out. They might have exacerbated the man's disease process leading to sudden cardiac arrest caused by asystole.

A new challenge in forensic toxicology exemplified by a case of murder under the influence of a synthetic cannabinoid - AM-2201.

Among new psychoactive substances (NPS) available on the narcotic market, a significant number consists of synthetic cannabinoids commonly known as smokable herbal "spice" and "K2", and which are legally treated as a legal alternative to marijuana. The dearth of information on the pharmacology of these intoxicants as they are introduced into the market has created the urgent need among healthcare providers for case studies on the substances belonging to this group, both in terms of the consequences of using such intoxicants, and in methods of detection. The subject of the present report is a multi-parameter analysis of a criminal case of an 18-year-old male who was charged with murder of his female relative and attempted murder of two other victims by stabbing. The defendant pleaded guilty, but he claimed that he had been acting without volition, because he was under the influence of the synthetic cannabinoid AM-2201, which had been purchased from a dealer as a 10g package labelled "Mr Green - No bad trip". Analytical methods including gas chromatography - electron ionization - quadrupole ion trap mass spectrometry (GC-EI-QIT/MS) and liquid chromatography, electrospray ionization, tandem mass spectrometry (LC-ESI-MS-MS) were developed to determine the presence of AM-2201 in the Mr Green - No Bad Trip, and in the blood of the perpetrator, respectively. Toxicological findings are discussed in the context of psychoactive and adverse physical effects resulting from the presence of AM-2201 in the human body; the observations were also analyzed in conjunction with data from the literature.

New psychoactive substances in substantive evidence in expert practice of the Department of Forensic Medicine, UJCM in the years 2010-2015.

AIM OF THE STUDY:: Aim of the study was to analyse of 2075 evidences containing new psychoactive substances (NPS). MATERIAL AND METHODS: The prepared samples were identified employing an analytical procedure where the analytes were investigated by gas chromatography-electron impact mass spectrometry (GC-EI-MS) using a created library of mass spectra. RESULTS: The analysis revealed the following substances in the investigated products: piperazine derivatives (including BZP, MPMP, TFMPP), cathinone derivatives (including: pentedrone, 3-MMC, butylone, 4-MEC), pyrovalerone derivatives (MDPV, naphyrone, α-PVP, α-PVT), synthetic cannabinoids (such as AM-2201, UR-144, XLR-11, JWH073, JWH081, PB-22, AB-CHMINACA). Research conducted in 2010-2015 made it possible to track changes in the composition of investigated preparations. CONCLUSIONS: The following relationships has been shown: number of components decreased and in the end of 2015 dominated preparations with single component; introduction of amendments Act on Preventing Drug Addictionn affect the elimination from the market of one compounds and replacing them by their derivatives; since 2011 on the market of 'legal highs' we did not observe occurrence of compounds of piperazine group.

Medicolegal aspects of PMA-related deaths.

Unlike amphetamine, amphetamine-like substances accessible on the drug market are less expensive and more easily available; they also produce hallucinogenic effects expected by the users. Such properties render them more attractive as compared to amphetamine. On the other hand, the knowledge of the toxicity of these compounds is very limited, what in consequence generates problems that create ever-expanding research areas, including analytical, clinical and medicolegal issues, thus leading to development of systemic databases. An example here is paramethoxyamphetamine (PMA), which appeared on the drug market in recent years as a result of creative inventiveness of producers of psychoactive substances, who aimed at PMA replacing the popular ecstasy (MDMA) as a less expensive and more available product. It is more potent than MDMA, but has a slower onset of action, which encourages users to take more. The problem is illustrated in the present paper by three fatal cases involving PMA, which were comprehensively investigated taking into consideration case histories, pathological and toxicological findings obtained with the use of LC-MS-MS method. In blood samples taken from all the three victims, very high concentrations of PMA were found (in the range of 10-27mg/L) and thus the cause of deaths was determined as overdoses of PMA with the underlying mechanism of acute cardiorespiratory failure.

New psychoactive substance α-PVP in a traffic accident case.

The problems of new psychoactive substances (NPSs), especially related to drivers, constitute an open research area. In this case report, we present a traffic accident case, in which two passengers of five individuals died instantly, while the other three persons survived the accident with minor injuries only. From the blood samples of the driver and the passengers, α-pyrrolidinovalerophenone (α-PVP), an NPS belonging to the category of cathinone derivatives, was disclosed. Therefore, we established a detailed procedure for analysis of α-PVP in blood samples by liquid chromatography-tandem mass spectrometry. After careful validation tests of this method, α-PVP concentration in blood samples from the surviving driver and passengers, and from the two deceased, were measured. The concentrations varied from 20 to 650 ng/mL. Access to detailed information originating from the court files and from explanations provided by the driver and eye witnesses revealed extremely valuable illustrative details addressing the symptoms and pharmacological effects of α-PVP on the human organism, thus contributing to enriching the body of knowledge of α-PVP abuse.

Opium alcaloids in toxicological medico-legal practice of Department of Forensic Medicine, Jagiellonian University Medical College / Alkaloidy opium w toksykologicznej praktyce medyczno-sadowej Zakladu Medycyny Sadowej UJ CM.

Most likely, opium was the first narcotic substance discovered at the dawn of mankind. Contemporary drug abuse predominantly poses a social and clinical problem and encompasses among other aspects emergency procedures in cases of intoxication and treatment of addictions. On the other hand, this is also a problem of the judicial system, which implements the rule of apt punishment in criminal cases (rapes, robberies, drivers, production and trade in narcotic substances) and of the necessity of monitoring drug-associates deaths. In all drug-associated cases, investigative capabilities have increased with the introduction of extremely sensitive and specific analytical methods (GC-MS, LC/MS, HPLC/DAD) allowing for detection and identification of multi-component mixtures of xenobiotics found at low concentration levels in complex biological matrices. The history of the Krakow Department of Forensic Medicine dates back to the year 1877, since archival materials have been kept since that time. Isolated deaths resulting from morphine poisoning, mostly involving individuals employed in the health care sector, constituted the subject of medico-legal expert opinions starting at the beginning of the 20th century, but only the eighties did bring the need for multidirectional toxicological examinations of opiates and their metabolites in diversified biological and non-biological materials. The present report, in addition to the historical background of opiate addiction, discusses selected problems derived from published by Department reports on opiates, including cases of fatal intoxication, hair analysis of drug addicts in its various aspects, interactions in cases of poisoning and others.

"Legal highs"--toxicity in the clinical and medico-legal aspect as exemplified by suicide with bk-MBDB administration.

The easily available "legal highs", which are products containing psychoactive substances, such as cathinones, piperazines and synthetic cannabinoids, are abused by adolescents in Poland and in the world as alternatives to classic drugs, such as amphetamines or marijuana. The majority of these potentially dangerous substances are still legal and they are associated with a risk of severe poisoning or even death, and provide new challenges in clinical and forensic toxicological practice. Investigations in the field of "designer drugs" may be well illustrated by the case of a suicide of a 21-year old male who ingested a specified dose of a preparation called "Amphi-bi-a" that contains bk-MBDB, chemically 2-methylamino-1-(3,4-methylenedioxyphenyl) butan-1-one, which belongs to the cathinone group, as a synthetic euphoric empathogen and psychoactive stimulant that is chemically similar to MDMA. It is one of more common components of "legal highs" examined in Poland and other countries. The documentation of the case includes a clinical assessment of the patient's health status performed during his almost 4-h hospitalization before death, autopsy and histological examinations supported by toxicological findings revealing bk-MBDB at extremely high concentrations (at 20 mg/l in the blood and 33 mg/kg in the liver); hence, this body of evidence contributes to knowledge in the field of "designer drugs". Inventions of designers of new psychoactive xenobiotics, which are much in demand, especially in view of the dynamic Internet marketing, which drums up narcobusiness, must be balanced by a national strategy developed by medical, legal and educational circles in the modern civilized world in order to prevent the spreading of the phenomenon.

[Analysis of taxus baccata alkaloids in biological samples with the use of chromatographic methods]. / Analiza alkaloidów cisa pospolitego w materiale biologicznym z zastosowaniem metod chromatograficznych.

The aim of the experiment was to develop a method of the identification of taxines (taxine B and isotaxine B) from Taxus leaves and its application to the analysis of biological samples collected from the deceased girl. The object of the investigation was the analysis of taxines in post-mortem samples collected from a 15-year-old girl, who consumed leaves of Taxus baccata to commit suicide. She died after several hours of hospitalization. Taxus alkaloids, especially taxine B and isotaxine B, are not commercially available. Due to this fact, the authors worked out a preliminary method of isolating taxines from Taxus leaves and their extraction using thin layer chromatography (TLC) and solid-phase extraction (SPE). Identification of the presence of taxine B and isotaxine B in the extract from Taxus leaves was performed by high performance liquid chromatography coupled to tandem mass spectrometry with electrospray ionization (HPLC-ESI-MS-MS). The developed method was applied to determination of taxine B and isotaxine B in the biological samples collected from the girl's body. The samples were prepared using solid-phase extraction (SPE) and analyzed by HPLC-ESI-MS-MS. The results demonstrated that all the samples contained taxine B and isotaxine B. The results of toxicological investigations were confirmed by macroscopic and microscopic histological examinations.