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In this work, the identification and characterization of the novel synthetic cathinone 5-PPDI found in a suspect drug sample were performed. The suspect sample was analyzed by gas chromatography–mass spectrometry (GC–MS), Fourier-transformed infrared (FTIR) spectroscopy, ultra-high performance liquid chromatography–high-resolution mass spectrometry (HRMS) and nuclear magnetic resonance (NMR) spectroscopy. The fragmentation observed in GC–MS and the identification of functional groups by FTIR was not enough for compound identification. After an exhaustive analysis of the accurate-mass fragmentation observed in HRMS, the compound was tentatively identified as the novel cathinone 5-PPDI. Finally, five different NMR experiments were used for the unequivocal identification and complete characterization of the compound. In addition, the origin of this cathinone was investigated in depth. The analytical data provided in this work will be useful for the identification of 5-PPDI by forensic laboratories. In addition, the origin of this cathinone has been investigated, which could be of interest for the identification of future synthetic cathinones prepared following the similar synthesis route.

Kratom cocktail or the fatal 4 × 100 formula is defined as a mixture of boiled kratom leaves, cola drink, and cough syrup. In the present study, we focused on application of the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) using the anti-mitragynine (MG) monoclonal antibody (anti-MG mAb) to the kratom cocktail. The ic-ELISA is a rapid method for quantification of the major kratom alkaloids including MG, paynantheine, and speciogynine in kratom cocktails. Because some matrices or additives may influence the binding affinity between the alkaloids and the anti-MG mAb, a liquid–liquid extraction method using chloroform was used to clean-up samples and minimize any cross-reactivity with anti-MG mAb. The anti-MG mAb showed slight cross-reactivity to caffeine, codeine, morphine, tramadol, and dextromethorphan (<0.5 %), which are also commonly added to a kratom cocktail. When applied to eight different kratom cocktail samples, the ic-ELISA using the anti-MG mAb allowed the determination of the combined kratom alkaloid content in the range of 0.083–576 mg/L, and these values were in agreement with the results of the high-performance liquid chromatography method (R 2 = 0.9689). To our knowledge, this is the first report for the quantification of total amounts of kratom alkaloids including MG in kratom cocktail by a simple immunoassay. Because of the sharp rise in kratom cocktail abuse in the world, this method will be a useful tool for detection of kratom cocktail consumption.

AMB-FUBINACA is a synthetic cannabinoid receptor agonist (SCRA) which is primarily metabolised by hepatic enzymes producing AMB-FUBINACA carboxylic acid. The metabolising enzymes associated with this biotransformation remain unknown. This study aimed to determine if AMB-FUBINACA metabolism could be reduced in the presence of carboxylesterase (CES) inhibitors and recreational drugs commonly consumed with it. The affinity and activity of the AMB-FUBINACA acid metabolite at the cannabinoid type-1 receptor (CB1) was investigated to determine the activity of the metabolite. The effect of CES1 and CES2 inhibitors, and delta-9-tetrahydrocannabinol (Δ9-THC) on AMB-FUBINACA metabolism were determined using both human liver microsomes (HLM) and recombinant carboxylesterases. Radioligand binding and cAMP assays comparing AMB-FUBINACA and AMB-FUBINACA acid were carried out in HEK293 cells expressing human CB1. AMB-FUBINACA was rapidly metabolised by HLM in the presence and absence of NADPH. Additionally, CES1 and CES2 inhibitors both significantly reduced AMB-FUBINACA metabolism. Furthermore, digitonin (100 µM) significantly inhibited CES1-mediated metabolism of AMB-FUBINACA by ~ 56%, while the effects elicited by Δ9-THC were not statistically significant. AMB-FUBINACA acid produced only 26% radioligand displacement consistent with low affinity binding. In cAMP assays, the potency of AMB-FUBINACA was ~ 3000-fold greater at CB1 as compared to the acid metabolite. CES1A1 was identified as the main hepatic enzyme responsible for the metabolism of AMB-FUBINACA to its less potent carboxylic acid metabolite. This biotransformation was significantly inhibited by digitonin. Since other xenobiotics may also inhibit similar SCRA metabolic pathways, understanding these interactions may elucidate why some users experience high levels of harm following SCRA use.

Animal studies suggested that halogenated hydrocarbons such as 1,1-difluoroethane (DFE) sensitized myocardial tissues to catecholamines and might cause fatal arrhythmia. In this paper, we report a case of a fatality that was associated with DFE abuse, and quantified DFE concentrations in postmortem specimens using gas chromatography–mass spectrometry (GC–MS). Femoral vein blood, cardiac blood, and urine samples were taken from the autopsy for toxicological analysis. We have established a detailed procedure for quantification of DFE in human blood and urine by GC–MS and have presented its validation data. The concentrations of DFE in this case were 481, 591 and 201 µg/mL in femoral vein blood, cardiac blood and urine samples, respectively, which were much higher than those in previous cases measured by gas chromatography–flame ionization detection. Thus, in the absence of other remarkable autopsy findings, the cause of death was determined to be DFE intoxication. To the best of our knowledge, this is the first case report of quantification of DFE in human blood and urine specimens by GC–MS.

One objective was to compare the psychostimulating effects of N,α-diethylphenethylamine (NADEP) with those of methamphetamine (METH) in experiments on behavioral activities of rats. Another objective was to compare concentrations of neurotransmitters dopamine (DA), serotonin (5-HT), and their metabolites such as dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5-HIAA) in brain tissues (striatum and frontal cortex) carried out after administration of NADEP and METH in rats. The animals were treated with NADEP and METH for 6 consecutive days (5 mg/kg per day, respectively), and the scores of stereotypy were measured on the first, second, fourth, and sixth day. The increase of the stereotypy score was observed in the NADEP-treated group, but it was less than that after treatment with METH. NADEP administration (5 and 10 mg/kg) resulted in a significant increase of DA and 5-HT and a decrease of DOPAC and HVA in the striatum tissues, which were collected 1 h after administration, but the changes of the compounds were less than those after treatment with METH (5 mg/kg). Unlike the METH-treated group, the changed DA, 5-HT, DOPAC, and HVA levels of the NADEP- treated group (5 and 10 mg/kg) were soon recovered within 6 h after administration. Both NADEP and METH had no significant effect on the 5-HIAA concentrations of the brain tissues. These results suggest that NADEP has significant psychostimulatory effects, though the effects were less than those of METH. Thus, NADEP should be carefully monitored to avoid abuse as a psychoactive drug.

Urine samples for cases of drug abuse are regularly screened by gas chromatography-mass spectrometry (GC-MS) after enzyme hydrolysis, liquid-liquid extraction, and trimethylsilyl (TMS) derivatization. During such routine analysis, two compounds (labeled as X1 and X2) were detected in urine samples of opium and illicit heroin users as their TMS derivatives, which led us to tentatively characterize the compounds. It was found that the compounds were noscapine metabolites as determined by GC-MS in both electron-impact ionization (EI) and chemical ionization modes. Together with the analysis of fragment ions in the EI mass spectra, the X1 and X2 compounds were further assigned as 6,7-dihydroxynoscapine and its desmethyl product at either position 6′ or 7′, respectively. The two tentatively characterized compounds were found to be useful as markers for excluding medicinal morphine use and for refuting claims of codeine use or Golden-Seal preparation use often made by heroin addicts to evade conviction. By addition of the above two metabolites, the arsenal of markers for opium and illicit heroin use has now been strengthened.

We describe a case of poisoning by arsenic trioxide of a young man found dead at home. There were no obvious external signs of arsenic poisoning; but we observed marked endocardial hemorrhages, hepatomegaly, diffuse gastric mucosal hemorrhages, and slight brain edema at autopsy. The Reinsch test for the stomach contents and liver homogenate was positive for arsenic. Wavelength-dispersive X-ray fluorescence spectrometry combined with the Reinsch test showed that fatal levels of arsenic were present in blood and tissues. The cause of death was diagnosed as circulatory collapse caused by arsenic trioxide.

We encountered a curious case in which two male subjects self-administered mepirapim plus acetyl fentanyl by different routes, i.e., intravenously and by inhalation. We have thus established a detailed procedure for quantification of mepirapim and acetyl fentanyl in whole blood and urine specimens using gas chromatography (GC)–tandem mass spectrometry (MS/MS). The GC–MS/MS method was validated for linearity, extraction recovery, accuracy, and precision. Liquid chromatography–MS/MS was also used for identification of the target compounds. Good linearity and reproducibility were achieved in the range of 20–1000 ng/g for both target compounds in both matrices. The concentrations of mepirapim in heart whole blood, femoral vein whole blood, and urine of the deceased individual with administration by intravenous injection were 593, 567, and 527 ng/g, respectively; those of acetyl fentanyl were 155, 125, and 126 ng/g, respectively. The mepirapim and acetyl fentanyl concentrations in the urine specimen of the surviving individual who had administered them by inhalation were 4900 and 570 ng/g, respectively. To our knowledge, with the exception of a brief mention of a mepirapim concentration in a serum sample in emergency medicine, there are no reported data on the identification and quantification of mepirapim in biological samples. Mepirapim is a new synthetic cannabinoid. The concentration profiles of unchanged mepirapim in whole blood and urine were quite different and unique. A detailed clarification of such uniqueness is under way in our laboratory.

Two compounds newly found in the seizures by drug enforcement agencies were identified and characterized by various instrumental analytical methods. The obtained powder samples were analyzed by gas chromatography–mass spectrometry (GC–MS), liquid chromatography–mass spectrometryn (LC–MSn), nuclear magnetic resonance (NMR) spectroscopy, infrared and Raman spectroscopy and X-ray crystallography. The two compounds were tentatively identified as 4-chloro-α-PVP and 4-MDMC by GC–MS, and LC–MS/MS. The confirmation of the results was made by NMR spectroscopy. The X-ray crystallography gave information that 4-chloro-α-PVP and 4-MDMC were in salted forms with sulfate and hydrochloride, respectively; in addition, both compounds existed as racemic mixtures. We could identify 4-chloro-α-PVP and 4-MDMC in the seizure powder samples by various analytical methods. X-ray crystallography was especially useful for identifying the salted forms and enantiomeric forms.