Dariusz Zuba

25C-NBOMe – New potent hallucinogenic substance identified on the drug market

This publication reports analytical properties of a new hallucinogenic substance identified in blotter papers seized from the drug market, namely 25C-NBOMe [2-(4-chloro-2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine]. The identification was based on results of comprehensive study including several analytical methods, i.e., GC-EI-MS (without derivatization and after derivatization with TFAA), LC-ESI-QTOF-MS, FTIR and NMR. The GC-MS spectrum of 25C-NBOMe was similar to those obtained for other representatives of the 25-NBOMe series, with dominant ions observed at m/z=150, 121 and 91. Fragment ions analogic to those in 2C-C (4-chloro-2,5-dimethoxy-β-phenylethanamine) were also observed, but their intensities were low. Derivatization allowed the determination of molecular mass of the investigated substance. The exact molecular mass and chemical formula were confirmed by LC-QTOF-MS experiments and fragmentation pattern under electrospray ionization was determined. The MS/MS experiments confirmed that the investigated substance was N-(2-methoxy)benzyl derivative of 2C-C. The substance was also characterized by FTIR spectroscopy to corroborate its identity. Final elucidation of the structure was performed by NMR spectroscopy.

Analysis of UR-144 and its pyrolysis product in blood and their metabolites in urine.

UR-144 [(1-pentyl-1H-indol-3-yl)(2,2,3,3-tetramethylcyclopropyl)methanone] is a synthetic cannabinoid, which has been detected in many herbal blends, resinous samples and powders seized from the Polish drug market since the beginning of 2012. This paper presents the case of intoxication by this substance. A complete picture of the symptoms observed by a witness, paramedics and medical doctors are given. In the analysis of powder residues from the plastic bag seized from the intoxicated person by gas chromatography-mass spectrometry (GC-MS), UR-144 and its major pyrolysis product [1-(1-pentyl-1H-indol-3-yl)-3-methyl-2-(propan-2-yl)but-3-en-1-one] were detected. Both substances were also identified in a blood sample collected on admission of the patient to hospital using liquid chromatography-triple quadrupole tandem mass spectrometry (LC-QqQ-MS). Blood concentration of UR-144 was 6.1 ng/mL. A urine sample collected at the same time was analyzed by liquid chromatography-quadruple time-of-flight tandem mass spectrometry (LC-QTOF-MS). The parent substance and its pyrolysis products were not detected in urine, while their five metabolites were found. The experiments allowed the location of derivative groups to be established, and thus elucidate rough structures of the metabolites; a dihydroxylated metabolite of UR-144 and mono-, dihydroxylated and carboxylated metabolites of its pyrolysis product were identified.

Fatal intoxication with 3-methyl-N-methylcathinone (3-MMC) and 5-(2-aminopropyl)benzofuran (5-APB)

The emergence of a large number of new psychoactive substances (NPSs) in recent years poses a serious problem to clinical and forensic toxicologists. Here we report a patient who administrated ca. 500mg of 3-MMC (3-methyl-N-methylcathinone) and 400mg of 5-APB (5-(2-aminopropyl)benzofuran) in combination with 80g of ethyl alcohol. The clinical manifestations included agitation, seizures, hypertension, tachycardia, hyperthermia and bradycardia. The patient did not recover and died around 4h after the use of drugs. The cause of death was acute cardiovascular collapse that occurred following mixed intoxication with NPSs and alcohol. Toxicological analysis of post-mortem blood revealed 3-MMC and 5-APB in concentrations of 1.6μg/mL and 5.6μg/mL, respectively. Moreover, the serum alcohol concentration was 1.4g/L in ante-mortem sample collected 1h after admission to the hospital. This is the first report on blood concentration of 3-MMC and 5-APB in fatal intoxication.

Detection of buphedrone in biological and non-biological material – Two case reports

Buphedrone (2-(methylamino)-1-phenylbutan-1-one, α-methylamino-butyrophenone, MABP) is a positional isomer of mephedrone. In Poland, it was marketed in the second half of 2010 after the banning of mephedrone. Buphedrone is a stimulant that is snorted, smoked or taken orally. This substance was identified in 15 products seized by law enforcement after August 2010 and analysed in the Institute of Forensic Research (IFR). Buphedrone was the sole psychoactive substance in only 5 products. It was mixed mainly with 4-MEC and MDPV. This paper presents two cases in which both biological and non-biological materials were delivered to the IFR for toxicological analysis. In the first case, a passenger car crashed into a truck. The car driver suffered severe injuries resulting in his death. During external inspection of the deceased the police discovered several packages containing a white powder. In the second case, a man was arrested for possession of illicit drugs. Analysis of powders was carried out using gas chromatography-mass spectrometry (GC-MS) and high-pressure liquid chromatography with diode array detection (HPLC-DAD). The purity of buphedrone found in powder samples was in the range of 58-68%. Analyses of blood were carried out using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Buphedrone was found in the blood of the deceased at a concentration of 127 ng/mL and of the drug user/seller at 3 ng/mL.

Fatal Intoxication with Methoxetamine

Methoxetamine (MXE) is a new synthetic drug of abuse structurally related to ketamine and phencyclidine. A case of a 29‐year‐old male with acute toxicity related to the analytically confirmed use of MXE is reported. The man was found dead at his residence. Biological material was analyzed using liquid chromatography–tandem mass spectrometry. The concentration of MXE in urine of the deceased was 85 μg/mL. Despite the vial containing the blood sample being destroyed during transportation and the blood leaking out into the cardboard packaging, the blood level of MXE was estimated. After determination of the cardboard grammage (approx. 400 g/m3) and the mean mass of the blood obtained after drying (0.1785 ± 0.0173 g per 1 mL), the estimated blood concentration of MXE was found to be 5.8 μg/mL. The high concentration of MXE in blood and urine and the circumstances of the case indicate an unintentional, fatal intoxication with this substance.

Identification and characterization of 2,5-dimethoxy-3,4-dimethyl-β-phenethylamine (2C-G)--a new designer drug.

This study presents and discusses the mass spectrometric, infrared spectroscopic and nuclear magnetic resonance spectroscopic data of 2,5-dimethoxy-3,4-dimethyl-β-phenethylamine (2C-G), a new designer drug. A powder sample containing 2C-G was seized in Poland in 2011. The paper focuses on a comparison of the analytical features of 2C-G and other members of the 2C-series, in order to assess the possibility of unequivocal identification. The occurrence of intense peak at m/z = 178 and different intensities of the ions at m/z = 165 and 180 in the gas chromatography-electron impact-mass spectrometry (GC-EI/MS) spectrum of 2C-G made it possible to distinguish it from 2C-E. Differences in relative intensities of the ions at m/z = 192, 179 and 177 were observed for GC-EI/MS spectra of TFAA derivatives of 2C-G and 2C-E. An identical set of ions was recorded for these substances using the liquid chromatography-electrospray ionization/quadrupole time of flight mass spectrometry (LC-ESI/QTOFMS) method in both MS and tandem mass spectrometry (MS/MS) mode, but the distinction was possible based on differences in the ion intensities at m/z = 193.1223 and 178.0988. The Fourier transform infrared (FTIR) spectrum of 2C-G was significantly different from other members of the 2C-series, with a characteristic doublets at 993-1014 cm(-1) and 1099-1124 cm(-1) , and the ratio of bands at higher wavenumbers. Final elucidation of the structure of 2C-G was carried out by (1) H and (13) C NMR spectroscopy. The study indicated that the marketing of analogues of controlled substances poses a real analytical challenge for forensic laboratories, and the application of sophisticated methods is often required for unequivocal identification of a new substance.

Structural elucidation and identification of a new derivative of phenethylamine using quadrupole time-of-flight mass spectrometry.

RATIONALE In recent years, the phenomenon of uncontrolled distribution of new psychoactive substances that were marketed without prior toxicological studies has been observed. Because many designer drugs are related in chemical structure, the potential for misidentifying them is an important problem. It is therefore essential to develop an analytical procedure for unequivocal elucidation of the structures of these compounds. The issue has been discussed in the context of 25I-NBMD [2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2,3-methylenedioxyphenyl)methyl]ethanamine], a psychoactive substance first discovered on the drug market in 2012. METHODS The substance was extracted from blotter papers with methanol. Separation was achieved via liquid chromatography. Analysis was conducted by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-QTOFMS). Identification of the psychoactive component was supported by electron impact gas chromatography/mass spectrometry (GC/EI-MS). RESULTS The high accuracy of the LC/ESI-QTOFMS method allowed the molecular mass of the investigated substance (M(exp) = 441.0438 Da; mass error, ∆m = 0.2 ppm) and the formulae of ions formed during fragmentation to be determined. The main ions were recorded at m/z = 135.0440, 290.9876 and 305.9981. Structures of the obtained ions were elucidated in the tandem mass spectrometry (MS/MS) experiments by comparing them to mass spectra of previously detected derivatives of phenethylamine. CONCLUSIONS The performed study indicated the potential for using LC/QTOFMS method to identify new designer drugs. This technique can be used supplementary to standard GC/MS. Prior knowledge of the fragmentation mechanisms of phenethylamines allowed to predict the mass spectra of the novel substance--25I-NBMD.

[1-(Tetrahydropyran-4-ylmethyl)-1H-indol-3-yl]-(2,2,3,3-tetramethylcyclopropyl)methanone: a new synthetic cannabinoid identified on the drug market

A new synthetic cannabinoid, [1-(tetrahydropyran-4-ylmethyl)-1H-indol-3-yl]-(2,2,3,3-tetramethylcyclopropyl)methanone, was identified in several resinous samples seized by law enforcement officers in Poland. Its identification was based on liquid chromatography–electrospray ionization–quadrupole time-of-flight–mass spectrometry, gas chromatography–electron ionization–mass spectrometry, one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy, and Fourier-transform infrared spectroscopy. The reported substance was first developed by Abbott Laboratories and patented under the name “A-834,735”. It is a potent agonist of both CB1 and CB2 receptors. Although A-834,735 shows moderate selectivity to CB2 receptor, it exhibits a CB1 affinity similar to that of ∆9-tetrahydrocannabinol. The drug has recently become available in online shops. To our knowledge, this is the first report to disclose a synthetic cannabinoid containing a (tetrahydropyran-4-yl)methyl structure in products seized from the drug market.

Alpha- and beta-asarone in herbal medicinal products. A case study

The composition of pellets and tablets, which were sold as the preparations of Chinese natural medicine, was studied. The drugs were seized by the police due to the intoxication of a young woman. Analyses were performed by GC-MS (HP-5 ms column) and by HPLC (RP-18e Chromolith(®) monolithic column). Diverse content of asarone isomers was found in the tested material. The dose of alpha-asarone ranged from 0.49 to 42.5 μg per pellet, while its average concentration in the tablets was 51.4 μg. Beta-asarone was not detected in the pellets with low content of alpha isomer (below 1 μg), while its content in the remaining pellets ranged from 142 to 645 μg. The tablets contained higher doses of beta-asarone (1526 μg in average). The total content of asarones in most of the examined pellets and tablets exceeded 115 μg, which is the maximum acceptable daily intake. Taking into account the dosage written on packages that ranged from several to more than ten pellets/tablets a day, one can assume that the intake of such doses of asarone might be health-threatening.

Distinction of constitutional isomers of mephedrone by chromatographic and spectrometric methods

Many new psychoactive substances (NPS) have similar chemical structures. Separation and unequivocal identification of structural isomers is an important issue due to the fact that they can differ in pharmacology, toxicology, effects and action on the human body, as well as legal status. This paper is aimed at comparing the abilities of different analytical methods (GC-MS, UHPLC-DAD, LC-MS/MS), to distinguish mephedrone and its isomers: 3-MMC, 2-MMC, buphedrone, metamfepramone and ethcathinone. These techniques allowed for distinction between the isomers of mephedrone. In the GC-MS method, combining retention times with the mass spectra made the differentiation efficient. In the UHPLC-DAD conditions, it was impossible to distinguish the isomers based on their retention times, but the UV/VIS spectra were useful in identification. The separation conditions were optimised for the LC-MS/MS method. The identification of isomers was supported by the different intensity of product ions. The LODs for the GC-MS method (1000–2500 ng/mL) in scan mode were definitely the worst. The values for the UHPLC-DAD method were much better (1.4–2.7 ng/mL) making this assay suitable for the analysis of biological material. The LC-MS/MS method was sensitive enough (LODs = 0.03–0.39 ng/mL) for the analysis of biological material even a long time after the initial drug use.