A seizure containing 37 items was analyzed with laboratory identification scheme for drugs and chemicals in seized products applying accredited methods based on gas chromatography mass spectrometry (GC-MS) and liquid chromatography with photodiode-array detection (UPLC-PDA), a category A technique combined with category B and C techniques. No identification was obtained for 1 item [analogue of lysergic acid diethylamide (LSD)] and none detected or identified by GC-MS and/or UPLC-PDA for 8 items. All items were subsequently analyzed using ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-Tof) to investigate whether it is appropriate to include this technique in laboratory identification scheme for seized materials. The seizure consisted of individual packages containing blotter papers, capsules and powders, respectively. The powders were present in sealed alu-bags labeled 'not for human consumption' and Falcon® tubes, presumably purchased as 'research chemical'. Methanolic solutions were analyzed. The GC-MS method was described elsewere (Coopman V. Toxicol Anal Clin 2018;30:75–79). Automated library search was performed against NIST, SWGDRUG and CAYMAN mass spectral library and mass spectra published in scientific literature were consulted. The UPLC-PDA analysis was performed with a Waters Acquity UPLC H-Class system. The compounds were separeted on an Acquity UPLC C18 column using gradient elution with a mobile phase consisting of 0.1% (v/v) formic acid aqueous solution and 0.1% (v/v) formic acid in acetonitrile, and photodiode-arra y detection. Automated library search was performed with Empower™ 3 software against in-house and Waters library. A Waters Acquity UPLC I-Class with Xevo G2-XS QTof was used, equipped with UNIFI® scientific information system and Forensic Toxicology Screening Application from Waters with data acquisition parameters for accurate mass analysis in positive and negative mode. Analogues of LSD (1P-LSD, ETH-LAD, AL-LAD) and a designer benzodiazepine (clonazolam) respectively, were identified in blotter paper by UPLC-PDA, GC-MS and confirmed by UPLC-Q-Tof. The unknown analogue of LSD was identified as ALD-52 by UPLC-Q-Tof. 4-fluoromethylphenidate (4F-MPH), N-ethyldeschloroketamine (O-PCE) and mirtagynine were identified in powder by UPLC-PDA, GC-MS and UPLC-Q-Tof. O-PCE was not present in the Forensic Toxicology Screening Application library and was therefore not automatically identified. Salicylic acid, caffeine, trazodone, pseudoephedrine, melatonin and ETH-LAD respectively, were identified in tablet or capsule with routine laboratory identification scheme and confirmed by UPLC-Q-Tof. In a capsule found to contain pseudoephedrine and unknown, a library hit was found for the designer drug N-methyl-2-aminoindane (NM-2AI) by UPLC-Q-Tof. Laboratory identification scheme for the identification of drugs and chemicals in seizures is based on GC-MS and UPLC-PDA, a category A technique and category B, C techniques (SWGDRUG recommendations). Not all drugs and chemicals were detected with both techniques. When identification is obtained with one or any of the applied techniques, a more extensive analysis is recommended. UPLC-Q-Tof is a sensitive and powerful technique, appropriate to include in laboratory identification scheme for seized materials when identification is obtained with one or any of the applied techniques or if the presence of low concentration, a NPS or designer drugs is suspected, allowing confirmation or identification.

In the rapidly evolving ‘legal highs’ market, synthetic cannabinoids play a predominant role. These potent synthetic cannabinoid receptor agonists produce effects similiar to those of delta-9-tetrahydrocannabinol and are usually added to herbal smoking mixtures by soaking or spraying. Typical marketed as so-called herbal incense (potpourri), they are widely and openly sold on the web with the synthetic cannabinoids used in the mixtures continuously being substituted by ‘legal’ alternatives, in pace with new control measures. A seizure of ‘spice-like’ herbal incense was submitted to the laboratory for toxicological analysis. A methanolic sample solution was analyzed using liquid chromatography with diode array detector and gas chromatography-mass spectrometry. The μ-opioid receptor agonist U-47700 was found to be the only psychoactive substance present in the herbal mixture. Although considerable inter-batch differences in the concentration can be expected, a quantitative determination was performed in order to obtain an indication regarding the potency of the seized herbal mixture. In light of the emerging threat of synthetic opioids fatalities, the presence of U-47700 in ‘spice-like’ herbal incense raises great concerns, since the abuse may pose serious risk, as users will likely not be aware that they are using a potent synthetic opioid. This case illustrates once again that new psychoactive active substances openly sold on the Internet as ‘legal highs’ pose a risk for public health.

This paper reports the structure elucidation and full characterization of 4-chloromethamphetamine (4-CMA), a compound which was never reported previously outside of laboratory settings in seized drug samples, or samples actively being used at large dance festivals. Identification of 4-CMA was obtained by liquid chromatography with diode array detector (HPLC-PDA) and gas chromatography mass spectrometry (GC-MS). Further structure elucidation was performed by fragment pattern analysis of the trimethylsilyl and trifluoroacetyl derivatives with GC-MS. The region-isomeric form was confirmed by 1H nuclear magnetic resonance spectroscopy (1H NMR). HPLC-PDA was used for quantitation of 4-CMA in the seized tablet to obtain an indication of the potency. A literature review of the toxic effects of 4-CMA was performed, and mechanisms for serotonin neurotoxicity were proposed and discussed. Finally the risks for potential widespread harm to the public in events where similar substances or tablets start appearing and circulating on a larger scale in the general population is discussed.

A brown powder and different product packages of "spice-like" herbal incenses were analyzed using a systematic identification approach based on liquid chromatography with diode array detector (HPLC-PDA) and gas chromatography mass spectrometry (GC-MS) with computer based library search against spectral libraries. However, the most predominant compound in the methanolic sample solutions could not be identified. In order to elucidate the chemical structure, a more extensive analysis of the material was initiated using liquid chromatography mass spectrometry (LC-MS), electrospray high resolution mass spectrometry (ESI-HRMS) and H-1, C-13 and F-19 nuclear magnetic resonance spectroscopy (NMR), which allowed the identification and characterization of the major compound as methyl 2-[[1-(5-fluoropentyl) indole-3-carbonyl] amino]-3,3-dimethyl-butanoate (5F-MDMB-PICA).The goal of this study is to provide analytical information for the identification of this new tert-leucinate class synthetic cannabinoid by various analytical methods. (C) 2017 Elsevier B.V. All rights reserved.

A brown powder and different product packages of “spice-like” herbal incenses were analyzed using a systematic identification approach based on liquid chromatography with diode array detector (HPLC-PDA) and gas chromatography mass spectrometry (GC-MS) with computer based library search against spectral libraries. However, the most predominant compound in the methanolic sample solutions could not be identified. In order to elucidate the chemical structure, a more extensive analysis of the material was initiated using liquid chromatography mass spectrometry (LC-MS), electrospray high resolution mass spectrometry (ESI-HRMS) and 1H, 13C and 19F nuclear magnetic resonance spectroscopy (NMR), which allowed the identification and characterization of the major compound as methyl 2-[[1-(5-fluoropentyl)indole-3-carbonyl]amino]-3,3-dimethyl-butanoate (5F-MDMBPICA). The goal of this study is to provide analytical information for the identification of this new tert-leucinate class synthetic cannabinoid by various analytical methods.

This paper describes the first reported death involving ocfentanil, a potent synthetic opioid and structure analogue of fentanyl abused as a new psychoactive substance in the recreational drug scene. A 17-year-old man with a history of illegal substance abuse was found dead in his home after snorting a brown powder purchased over the internet with bitcoins. Acetaminophen, caffeine and ocfentanil were identified in the powder by gas chromatography mass spectrometry and reversed-phase liquid chromatography with diode array detector. Quantitation of ocfentanil in biological samples was performed using a target analysis based on liquid-liquid extraction and ultra performance liquid chromatography tandem mass spectrometry. In the femoral blood taken at the external body examination, the following concentrations were measured: ocfentanil 15.3μg/L, acetaminophen 45mg/L and caffeine 0.23mg/L. Tissues sampled at autopsy were analyzed to study the distribution of ocfentanil. The comprehensive systematic toxicological analysis on the post-mortem blood and tissue samples was negative for other compounds. Based on circumstantial evidence, autopsy findings and the results of the toxicological analysis, the medical examiner concluded that the cause of death was an acute intoxication with ocfentanil. The manner of death was assumed to be accidental after snorting the powder.

A 30-year old man was found dead in his home after inhaling fumes of a powder burned on aluminum foil. Blood and urine were taken by the medical examiner during the external body examination and submitted to the laboratory for a comprehensive systematic toxicological analysis. A toxic fentanyl level of 10.9μg/L was measured in the subclavian blood. Police investigation revealed that the man searched the internet for information on new psychotropic substances, among others including U-47700. A powder found in the victims' home was transferred to the laboratory for analysis, in which trace amounts of fentanyl (0.0035%, m/m) and U-47700 (0.0012%, m/m) were identified by gas chromatography mass spectrometry. U-47700 is an opioid analgesic drug, considered to have a potency of approximately 7.5 times that of morphine. A target analysis on U-47700 was performed using liquid-liquid extraction and ultra performance liquid chromatography tandem mass spectrometry operating in multiple reaction monitoring mode. The method validation was based on the Scientific Working Group of Forensic Toxicology document 'Standard Practices of Method Validation in Forensic Toxicology'. In blood and urine the U-47700 concentration was 13.8 and 71.0μg/L, respectively. To the author's knowledge, this is the first case report of a fatal intoxication involving U-47700 abused as a new psychotropic substance.

The detection of autochthonous aquatic bacteria in tissue samples from drowning cases is increasingly considered as an alternative approach to assist the medico-legal diagnosis of death by drowning. Bacteria belonging to the genus Aeromonas may be suitable candidates for this application as they are ubiquitous in natural aquatic environments but are generally not part of the human microbiota. The research aims of this study were (i) to develop a sensitive, specific and rapid screening and confirmation method for Aeromonas species in tissue samples and (ii) to evaluate aseptic sternal puncture as a post-mortem sample technique and bone marrow as an alternative matrix to provide evidence of death by drowning. The presence of Aeromonas in tissue samples was verified by cultivation using the selective media Ampicillin Dextrin Agar (ADA) and Ryan's Aeromonas Medium. The use of ADA medium was found most optimal for the sensitive, inexpensive and quick detection of aeromonads in human tissue samples. Positive culture plates were confirmed by harvesting all colonies for DNA extraction and subsequent PCR amplification using Aeromonas genus-specific primers. Aeromonads were detected in lung swab, blood and bone marrow of drowned bodies (n=3), but were negative in these three matrices for all negative controls (n=90) tested. Bone marrow proved to be a suitable alternative matrix and can be sampled post-mortem by an aseptic sternal puncture. In conclusion, this study confirms previous indications that aeromonads in cultures from blood of water bodies can be considered a potential marker for drowning. Given the fact that the number of immersed bodies (drowned and non-drowned) included in this study is statistically not significant, however, more tissue samples need to be investigated to confirm the validity of these methods to aid the diagnosis of death by wet drowning.

A powder was submitted to the author's laboratory for analysis on the presence of illegal drugs. Based on the structure elucidation performed by nuclear nuclear magnetic resonance spectroscopy and analysis of the trimethylsi- lyl and trifluoroacetyl derivatives with gas chromatography mass spectrometry, the powder was found to contain the designer drugs 4-methylamphetamine, besides amphetamine and caffeine. The electron impact mass spectrum of 4-methylamphetamine obtained from the analysis of a methanolic sample solution showed a base ion of m/ z5 6 rather than the expected m/z 44, therefore no hit was found with an electron impact mass spectrum present in the mass spectral library and literature. The increase of 12 atomic mass units was explained by formaldehyde-adduct formation.

A powder was submitted to the author’s laboratory for analysis on the presence of illegal drugs. Based on the structure elucidation performed by nuclear nuclear magnetic resonance spectroscopy and analysis of the trimethylsilyl and trifluoroacetyl derivatives with gas chromatography mass spectrometry, the powder was found to contain the designer drugs 4-methylamphetamine, besides amphetamine and caffeine. The electron impact mass spectrum of 4-methylamphetamine obtained from the analysis of a methanolic sample solution showed a base ion of m/z 56 rather than the expected m/z 44, therefore no hit was found with an electron impact mass spectrum present in the mass spectral library and literature. The increase of 12 atomic mass units was explained by formaldehyde-adduct formation.

Objectives: The aim of this study was to identify active ingredients present in counterfeit drugs and pharmaceutical preparations seized from the black market among bodybuilders. Preparations in closed or sealed packaging with different batch numbers (n = 74) and two preparations without labels were analyzed. Methods: The identification scheme was based on reversed-phase liquid chromatography with a photodiode array UV detector and gas chromatography-mass spectrometry. Methanolic sample extracts of tablets, capsules, oral jellies and water–based injectables were chromatographed on a ChromSpher 5 C18 column using a gradient program of triehylammonium phosphate and acetonitrile as mobile phases and were monitored at the wavelengths 220 nm and 254 nm. Oil-based injectables (n = 41) were extracted with hexane and methanol. The methanolic sample extracts and diluted supernatant layers were injected into an Agilent 6890 N gas chromatograph with a fused silica CPSIL 8CB low-bleed capillary column and Agilent 5973 inert mass selective detector operated in full-scan mode. The identified compound(s) were checked against the active ingredient(s) declared on the label. Conclusion: Packaging from counterfeits produced in underground labs cannot be visually distinguished from packaging originating from legitimate pharmaceutical companies. Out of 74 labeled products, the content of 25 (33.8%) did not match the label. Anabolic androgenic steroids are commonly found on the black market among bodybuilders, predominately esterified in oil-based injectables. Out of 40 steroidal oily solutions, the active ingredients of 21 (52.5%) did not match the label: one preparation did not contain an active compound, 20 contained other or more active ingredients. The additional compounds were often present in smaller quantities, which could be indicative of contamination. Active ingredients such as methandrostenolone, sildenafil, tamoxifen, quinine, clomiphene, dehydroepiandrosterone, anastrazole, clenbuterol, stanozolol, oxandrolone, liothyronine, finasteride and melatonin were identified in tablets, oral jellies and water-based injectables.

Objectives: The aim of this study was to identify active ingredients present in counterfeit drugs and phar- maceutical preparations seized from the black market among bodybuilders. Preparations in closed or sealed packaging with different batch numbers (n = 74) and two preparations without labels were analyzed. Methods: The identi- fication scheme was based on reversed-phase liquid chromatography with a photodiode array UV detector and gas chromatography-mass spectrometry. Methanolic sample extracts of tablets, capsules, oral jellies and water-based in- jectables were chromatographed on a ChromSpher 5 C18 column using a gradient program of triehylammonium phos- phate and acetonitrile as mobile phases and were monitored at the wavelengths 220 nm and 254 nm. Oil-based in- jectables (n = 41) were extracted with hexane and methanol. The methanolic sample extracts and diluted supernatant layers were injected into an Agilent 6890 N gas chromatograph with a fused silica CPSIL 8CB low-bleed capillary column and Agilent 5973 inert mass selective detector operated in full-scan mode. The identified compound(s) were checked against the active ingredient(s) declared on the label. Conclusion: Packaging from counterfeits produced in underground labs cannot be visually distinguished from packaging originating from legitimate pharmaceutical compa- nies. Out of 74 labeled products, the content of 25 (33.8%) did not match the label. Anabolic androgenic steroids are commonly found on the black market among bodybuilders, predominately esterified in oil-based injectables. Out of 40 steroidal oily solutions, the active ingredients of 21 (52.5%) did not match the label: one preparation did not con- tain an active compound, 20 contained other or more active ingredients. The additional compounds were often present in smaller quantities, which could be indicative of contamination. Active ingredients such as methandrostenolone, sildenafil, tamoxifen, quinine, clomiphene, dehydroepiandrosterone, anastrazole, clenbuterol, stanozolol, oxandrolone, liothyronine, finasteride and melatonin were identified in tablets, oral jellies and water-based injectables.

Two adult dogs with the same owner were intoxicated by ingestion of fertilizer composed of residual plant material of the castor bean plant (Ricinus communis L.). Both dogs died within 2 and 3 days, respectively, after the first signs of vomiting and abundant hemorrhagic diarrhea. Toxicologic and histopathologic examinations were performed on different organs. Histopathologic examination of the kidneys revealed tubular degeneration and necrosis and membranous glomerulonephritis. Additionally, myocardial degeneration with localized inflammation, lymphoid necrosis, and depletion in the spleen and mesenteric lymph nodes and hemorrhagic ulcerative gastroenteritis were found. The 2 cases could be used to elucidate the lethal dose of ricin and the histopathologic lesions in dogs.

The castor bean plant (Ricinus communis L.) or wonder tree is cultivated in many countries as an ornamental annual plant in gardens. The highest concentration of the lectin ricin is present in the seeds and pods. Ricin is considered as one of the most toxic natural poisons. Ricinine is a piperidine alkaloidal toxin present in castor bean and is described as a biomarker for the exposure to ricin. A case report is presented of a 49-year-old man who committed suicide by intravenous and subcutaneous injection of a castor bean extract. He was brought to the emergency department 24 h after injecting himself. On admission, the patient was conscious and he presented with a history of nausea, vomiting, diarrhoea, dyspnoea, vertigo and muscular pain. Despite symptomatic and supportive intensive care, the man died 9 h after admission due to multiorgan failure. A body external examination was performed. Blood, urine, vitreous humour and the castor bean extract were submitted to the laboratory for toxicological analysis. The identification of ricinine in the extract was performed by solid phase extraction in combination with full-scan gas chromatography/mass spectrometry, high-performance liquid chromatography with photodiode array detection and liquid chromatography/mass spectrometry operated in the full-scan mode, respectively. An extraction procedure with Oasis HLB solid phase extraction cartridges was applied. Chromatography was achieved using a Symmetry C18 column using a gradient mode with 0.15% formic acid and 0.15% formic acid in acetonitrile as mobile phase. Exposure to the castor bean extract was confirmed by identification of the biomarker ricinine in blood, urine and vitreous humour using solid phase extraction and liquid chromatography tandem mass spectrometry with electro spray source in positive ionization mode. Multiple reaction monitoring was used for specific detection. To the authors' knowledge, it is the first time that ricinine has been identified in vitreous humour in a case of castor bean poisoning. Based on the clinical symptoms and the results of the toxicological analysis, we concluded that death was caused by intoxication with plant toxins originated from R. communis L.

Poisoning may also lead to both coma and multiple organ failure, also in youngsters without a known major medical history. As not all toxic agents are routinely screened when a poisoning is suspected, it is useful to consider less frequently encountered poisons in certain cases. We describe the occurrence of asystole and multiple organ failure which occurred in a young man after a suspected tramadol overdose. The tramadol concentration on admission in the ICU was indeed 8 microg/ml (mg/l), far above the therapeutic range. Subsequently, the patient developed severe acute liver failure, finally leading to death. Post-mortem toxicology did not reveal any other poison responsible for this unfavourable course as only very high serum and tissue tramadol and desmethyltramadol concentrations were found. Only a few fatal poisonings attributable to tramadol alone, as observed in our case, have been reported. An overview of these cases is presented.

In this manuscript, a newly identified compound, 3,4-methylenedioxy-N,N-dimethylamphetamine (MDDM or also called MDDA), was quantified. The substance was identified in the biological specimens of a 31-year-old man who died following a massive 3,4-methylenedioxymethamphetamine (MDMA) overdose. In addition, the postmortem distribution of the identified substance in various body fluids and tissues was evaluated. For MDDM quantitation, a formerly reported and validated liquid chromatography tandem mass spectrometry (LC-MS/MS) method was adapted. The following quantitative results of the MDDM quantitation were obtained: Femoral blood, aorta ascendens, and right atrial blood contained 2.5, 21.7, and 11.6 ng MDDM/ml, respectively. In left and right pleural fluid and pericardial fluid, concentrations of 47.0, 21.7, and 31.9 ng/ml, respectively, were found. MDDM levels in urine, bile, and stomach contents were 42.4, 1,101, and 1,113 ng/ml, respectively. MDDM concentrations in lungs, liver, kidney, and left cardiac muscle ranged from 12.8 to 39.8 ng/g, whereas these levels were below the limit of quantitation (< LOQ) in right cardiac and iliopsoas muscle. In conclusion, for the first time, MDDM was unambiguously identified in a fatal MDMA overdose. MDDM was probably present as a synthesis by-product or impurity in the MDMA tablets, which were taken in a huge amount by the victim, or MDDM was ingested separately and prior to the MDMA overdose. A third option, i.e., the eventual formation of MDDM as a result of postmortem methylation of MDMA by formaldehyde, produced by putrefaction processes or during storage under frozen conditions, is also discussed. The MDDM levels, substantiated in various body fluids and tissues, are in line with the distribution established for other amphetamine derivatives and confirm that peripheral blood sampling, such as that of femoral blood, remains the "golden standard".

Fentanyl is a potent synthetic narcotic analgesic administered in the form of a transdermal patch for the management of chronic pain. A 78-year-old woman with a history of cancer was found dead in bed. She was lying on her back. The external examination revealed 10 Durogesic® transdermal therapeutic systems (100μg/h fentanyl) on the body.Liquid–liquid extraction and liquid chromatography tandem mass spectrometry with electrospray source in positive ionization mode was applied for the quantitation of fentanyl and its major metabolite norfentanyl in the post-mortem samples. Fentanyl-d5 and norfentanyl-d5 were used as internal standards. Multiple reaction monitoring was used for specific detection. Calibration was performed by addition of standard solutions to drug-free matrix (blood, urine and liver) prior to extraction. The method showed good linearity for fentanyl and norfentanyl over a concentration range of 5–150μg/L in reconstituted extracts with coefficients of determination equal or greater than 0.998. Percent mean within-day precision and accuracy of 0.9–1.0% and 99.4–101.1% for fentanyl and 2.0–4.5% and 93.1–101.0% for norfentanyl were obtained.Mean extraction recoveries varied between 95.5% and 100.3% for fentanyl and 39.2–57.4% for norfentanyl. The following fentanyl (norfentanyl) concentration in the post-mortem samples were measured; 28.6μg/L (3.0μg/L) in right and 28.2μg/L (3.5μg/L) in left subclavian blood, 21.3μg/L (<2μg/L) in right and 20.9μg/L (<2μg/L) in left femoral blood, 37.6μg/L (4.2μg/L) in right and 33.9μg/L (4.4μg/L) in left ventricular blood, 282.9μg/L (121.2μg/L) in urine, 688.2μg/L in stomach contents, 122.5μg/L (25.4μg/L) in bile, 19.5μg/L (< 2μg/L) in vitreous humour, 203.0μg/kg (26.6μg/kg) in liver and 78.6μg/kg (46.3μg/kg) in kidney. We concluded that the woman's death was caused by acute intoxication with fentanyl. The manner of death was presumed to be suicide due to excessive administered Durogesic® transdermal therapeutic systems.