The use of multiple keratinous matrices (head hair, axillary hair, and toenail clippings) can help narrowing a period of drug exposure: experience with a criminal case involving 25I-NBOMe and 4-MMC.

The objective of this publication is to present the interest of collecting several keratinous specimens in order to document possible drug impairment at the time of the assault, when knowledge solely occurred 7 months after. A subject committed a murder and within minutes after the crime self-inflicted serious wounds. He was charged to the hospital where he slowly recovered. After several weeks, he was sent to prison. During this period, intelligence indicated possible drug impairment at the time of the assault after using 25I-NBOMe and 4-MMC. Head hair (4 cm), axillary hair, and toenails were collected 7 months after the crime. New psychoactive substances were tested in each specimen using LC-MS/MS, which revealed the presence of 25I-NBOMe and 4-MMC in axillary hair (2 and 6 pg/mg) and toenails (1 and 5 pg/mg). However, the perpetrator claimed that the positive findings were due to contamination in prison. Therefore, the head hair was also tested and results returned negative (LOQ at 1 pg/mg), demonstrating absence of contamination during the last 4 months before collection. Combining the window of drug detection in axillary hair (about 4 to 8 months) and the one of toenail clippings (up to 8 months), and excluding drug exposure during the previous 4 months as well as external contamination as the head hair results were negative, allowed us to conclude that the positive findings in axillary hair and toenails are more likely than not consistent with consumption of both 25I-NBOMe and 4-MMC at the time of the crime.

Nanosensor for Sensitive Detection of the New Psychedelic Drug 25I-NBOMe.

This work reports the synthesis, characterization, and sensing behavior of a hybrid nanodevice for the detection of the potent abuse drug 25I-NBOMe. The system is based on mesoporous silica nanoparticles, loaded with a fluorescent dye, functionalized with a serotonin derivative and capped with the 5-HT2A receptor antibody. In the presence of 25I-NBOMe the capping antibody is displaced, leading to pore opening and rhodamine B release. This delivery was ascribed to 5-HT2A receptor antibody detachment from the surface due to its stronger coordination with 25I-NBOMe present in the solution. The prepared nanodevice allowed the sensitive (limit of detection of 0.6 µm) and selective recognition of the 25I-NBOMe drug (cocaine, heroin, mescaline, lysergic acid diethylamide, MDMA, and morphine were unable to induce pore opening and rhodamine B release). This nanodevice acts as a highly sensitive and selective fluorometric probe for the 25I-NBOMe illicit drug in artificial saliva and in sweets.

Multimodal imaging of hallucinogens 25C- and 25I-NBOMe on blotter papers.

Due to the much lower production costs but similar effects to lysergic acid diethylamide (LSD), phenethylamine derivatives are sold as a cheaper replacement or deceptively as LSD itself. These potent hallucinogenic substances can lead to severe intoxication, thus a more profound understanding of their use is required. This includes the elucidation of the manufacturing processes for the commonly used blotter papers and the assessment of the risk of overdosing because of a heterogeneous distribution on the blotter papers. Besides the rapid detection of the analytes, the manufacturing process was elucidated by three different imaging techniques and liquid chromatography-mass spectrometry (LC-MS). A blotter paper sample, containing the two hallucinogenic phenethylamine derivatives 25I-NBOMe and 25C-NBOMe, was analyzed by complementary techniques such as micro x-ray fluorescence (µXRF), laser ablation (LA)-inductively coupled plasma-optical emission spectroscopy (ICP-OES), matrix assisted laser desorption ionization (MALDI)-MS, and with LC-MS after extraction. Using the signal from chlorine and iodine within the compounds, µXRF proved to be the fastest, cheapest and easiest method for identification, requiring no sample preparation at all. LA-ICP-OES provided three-dimensional information of the elements in the blotter paper. These results helped to confirm the assumption that manufacturers spray the compounds onto the paper. Whereas µXRF and LA-ICP-OES detected signals for chlorine and iodine, MALDI-MS-imaging showed the molecular distribution of both analytes. LC-MS analyses as a complementary method support the imaging results. Quantitative results for different drug hotspots revealed a heterogeneous distribution of the drugs on the blotter paper implying an inherent risk of overdosing for consumers.

Retrospective Demonstration of 25I-NBOMe Acute Poisoning Using Hair Analysis.

BACKGROUND: The abuse of new psychoactive substances or NPS has been dramatically increasing all around the world since the last half of the year 2000 and has become a serious public health problem. NPS are a challenge for the worldwide forensic community due to the difficulties to accurately document the cases. The N-benzylmethoxy (NBOMe) group is a new class of hallucinogenic designer drugs and has gained importance in recent years. 25I-NBOMe (2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)- methyl]ethanamine) is an analog of the 2C series of psychedelic phenethylamine drugs that contain an N-methoxybenzyl substituent, which significantly affects their pharmacological activities. It is a potent agonist of 5-HTA receptors and a severe hallucinogenic drug, with numerous irreversible psychedelic effects which can last from 5 to 10 hours. It is consumed most often in the form of drops or blotters by the transmucosal, sublingual or intranasal routes. The active dosage is very low, supposed to be less than 100 µg. The literature is poor in reporting cases where 25I-NBOMe was identified. Only very few clinical cases of over dosages were published, suggesting a low prevalence of this compound. METHODS: We present a retrospective demonstration of 25I-NBOMe acute poisoning with dramatic outcome, using hair analysis. Two hair strands, measuring 9.5 cm, were collected 6.5 months after drug consumption during a forensic clinical evaluation of brain dysfunctions after cardiorespiratory arrest and were analyzed by ultra-high performance liquid chromatography system coupled to a tandem mass spectrometry (UPLC-MS/MS) and using two specific transitions: m/z 428.1 > 121.2 (quantification) and 428.1 > 90.6 (confirmation). Hair strands were segmented to determine the historic pattern of drug use and differentiate a single exposure from a chronic exposure. The hair test result for 25I-NBOMe was the following: not detected (0-2 cm), not detected (2-4 cm), 1.0 pg/mg (4-6 cm), 4.9 pg/mg (6-8 cm) and not detected (8-9.5 cm). RESULT: The result of the segment 6-8 cm coincides with the date of consumption (calculated with a hair growth rate at 1 cm/month) and the low concentration detected in the segment 4-6 cm probably corresponds to the contribution of dormant hair. The toxicological significance of the measured concentrations is difficult to determine because this is the first case dealing with hair analysis for 25I-NBOMe. CONCLUSION: The use of hair analysis for NPS is still at the initial stages. In particular, little is known about the incorporation into the keratin matrix after intake and the correlation between dosage frequency of use, and hair concentrations. Under these circumstances, NPS hair analysis should be cautiously interpreted by experienced forensic toxicologist.

Application of a Urine and Hair Validated LC-MS-MS Method to Determine the Effect of Hair Color on the Incorporation of 25B-NBOMe, 25C-NBOMe and 25I-NBOMe into Hair in the Rat.

NBOMes are a group of new psychoactive substances derived from phenethylamines. Recreational abuse is thought to have begun in 2010 and they are commonly associated with the "club drug" scene. They are administered in liquid form or as blotters due to their high potency. An LC-MS-MS method was validated using Scientific Working Group for Forensic Toxicology parameters for the detection of 25B-, 25C- and 4-iodo-2,5-dimethoxy-N-[(2-methoxyphenyl)methyl]-benzeneethanamine (25I-NBOMe) using 4-bromo-2,5-dimethoxy-N-[(2-methoxyphenyl)methyl]-benzeneethanamine (25B-NBOMe)-D3 as internal standard for urine and hair. Calibration graphs with R2 values >0.99 were observed for urine and hair for concentrations ranging from 0.1 to 100 ng/mL and 0.025 to 2.5 ng/mg, respectively. Urine LODs ranged from 5 to 25 pg/mL and had an LOQ of 50 pg/mL. Hair LOD and LOQs ranged from 3 to 5 pg/mg and 6.25 to 12.5 pg/mg, respectively. Intra- and inter-day precision was <20% and accuracy was within ±20% for both matrices. The method was shown to be selective for both exogenous and endogenous compounds. No matrix effects were observed for either matrix. LLE recovery ranged from 90 to 103% for urine samples and solid phase extraction recovery ranged from 80 to 107% for hair samples. Long-Evans rats (n = 55) were administered 25B-, 25C- or 25I-NBOMe at doses ranging from 30 to 300 µg/kg over a period of 10 days. Rats were shaved prior to their first dose and re-shaved after the 10-day period. Hair was separated by color (black: n = 55 and white: n = 55) and analyzed using the validated LC-MS-MS method to assess the impact hair color has on the incorporation of these drugs. All drugs were successfully detected in black hair. 25B-NBOMe from rats receiving the highest dose and 25C-NBOMe from rats receiving the medium and high doses were quantified in white hair. 25I-NBOMe was detected but fell below the limit of quantification. A dose-dependent concentration increase was observed in the black hair. All pooled urine samples tested positive for their expected NBOMes.

Rapid Screening Method for New Psychoactive Substances of Forensic Interest: Electrochemistry and Analytical Determination of Phenethylamines Derivatives (NBOMe) via Cyclic and Differential Pulse Voltammetry.

The NBOMe derivatives are phenethylamines derived from the 2C class of hallucinogens. Only a few human pharmacologic studies have been conducted on these drugs, and several cases of intoxication and deaths have been reported. Presently, NBOMe are not a part of the routine drugs-of-abuse screening procedure for many police forces, and there are no rapid immunoassay screening tests that can detect the presence of those compounds. In this Article, the voltammetric behavior of 25B NBOMe and 25I NBOMe were investigated and their electroanalytical characteristics determined for the first time. A novel, fast, and sensitive screening method for the identification of the two most common NBOMes (25B-NBOMe and 25I-NBOMe) in real samples is reported. The method uses the electrochemical oxidation of these molecules to produce an analytical signal that can be related to the NBOMe concentration with an average lower limit of quantitation of 0.01 mg/mL for both of them. The method is selective enough to identify the two compounds individually, even given the great similarity in their structure.

Characterization of hallucinogenic phenethylamines using high-resolution mass spectrometry for non-targeted screening purposes.

Hallucinogenic phenethylamines such as 2,5-dimethoxyphenethylamines (2C-X) and their N-(2-methoxybenzyl) derivatives (25X-NBOMe) have seen an increase in novel analogues in recent years. These rapidly changing analogues make it difficult for laboratories to rely on traditional targeted screening methods to detect unknown new psychoactive substances (NPS). In this study, twelve 2C-X, six 2,5-dimethoxyamphetamines (DOX), and fourteen 25X-NBOMe derivatives, including two deuterated derivatives (2C-B-d6 and 25I-NBOMe-d9 ), were analyzed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Collision-induced dissociation (CID) experiments were performed using collision energies set at 10, 20, and 40 eV. For 2C-X and DOX derivatives, common losses were observed including neutral and radical losses such as NH3 (17.0265 Da), •CH6 N (32.0500 Da), C2 H7 N (45.0578 Da) and C2 H9 N (47.0735 Da). 2C-X derivatives displayed common product ions at m/z 164.0837 ([C10 H12 O2 ]+• ), 149.0603 ([C9 H9 O2 ]+ ), and 134.0732 ([C9 H10 O]+• ) while DOX derivatives had common product ions at m/z 178.0994 ([C11 H14 O2 ]+• ), 163.0754 ([C10 H11 O2 ]+ ), 147.0804 ([C10 H11 O]+ ), and 135.0810 ([C9 H11 O]+ ). 25X-NBOMe had characteristic product ions at m/z 121.0654 ([C8 H9 O]+ ) and 91.0548 ([C7 H7 ]+ ) with minor common losses corresponding to 2-methylanisole (C8 H10 O, 122.0732 Da), 2-methoxybenzylamine (C8 H11 NO, 137.0847 Da), and •C9 H14 NO (152.1074 Da). Novel analogues of the selected classes can be detected by applying neutral loss filters (NLFs) and extracting the common product ions. Copyright © 2017 John Wiley & Sons, Ltd.

Analysis of 25I-NBOMe, 25B-NBOMe, 25C-NBOMe and Other Dimethoxyphenyl-N-[(2-Methoxyphenyl) Methyl]Ethanamine Derivatives on Blotter Paper.

In recent years, N-methoxybenzyl-methoxyphenylethylamine (NBOMe) derivatives, a class of designer hallucinogenic drugs, have become popular drugs of abuse. These drugs have been the cause of severe intoxications and even deaths. They act as 5-HT2A receptors agonists and have been reported to produce serotonin-like syndrome with bizarre behavior, severe agitation and seizures persisting for as long as 3 days. The most commonly reported derivatives are 25I-NBOMe, 25B-NBOMe and 25C-NBOMe, respectively 2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl) methyl]ethanamine, N-(2-methoxybenzyl)-2,5-dimethoxy-4-bromophenethylamine and N-(2-methoxybenzyl)-2,5-dimethoxy-4-chlorophenethylamine. Like many low dose hallucinogenic drugs these compounds are often sold on blotter paper. Three different types of commercially available blotter papers reported to contain NBOMe derivatives were obtained. These blotter papers were screened using Direct Analysis in Real Time AccuTOF(TM) mass spectrometry followed by confirmation and quantification by high-performance liquid chromatography triple quadrapole mass spectrometry. The major drug present on each of the three blotter products was different, 25I-NBOMe, 25C-NBOMe or 25B-NBOMe. The blotter papers were also found to have minute amounts of two or three NBOMe derivative impurities of 25H-NBOMe, 25I-NBOMe, 25C-NBOMe, 25B-NBOMe and/or 25D-NBOMe.

A Case Review of the First Analytically Confirmed 25I-NBOMe-Related Death in Washington State.

This case was submitted to the Washington State Patrol Toxicology Laboratory in September 2014. A 15-year-old male went to a party where he ingested 25I-NBOMe and mushrooms. A short time later, he started to vomit and began seizing until he eventually passed out. Resuscitation efforts were made, but were unsuccessful. He was transported to a local hospital, where he died three days later of multi-system organ failure following cardiopulmonary arrest. The hospital admission samples were negative for ethanol and basic drugs and their metabolites. The hospital serum confirmed positive for delta-9-tetrahydrocannabinol (THC) and carboxy-THC at 4.1 and 83 ng/mL, respectively. On the basis of the case history, the hospital blood and urine were sent to NMS Labs for NBOMe and psilocin confirmation. The blood was positive for 25I-NBOMe, and the urine was positive for 25C-, 25H- and 25I-NBOMe, as well as, psilocin. Antemortem and postmortem blood were also sent to AIT Laboratories for NBOMe confirmation. The antemortem blood confirmed positive for 25I-NBOMe with a concentration of 0.76 ng/mL. The manner of death was ruled an accident as a result of combined 25I-NBOMe and psilocin intoxication.

Test purchase, identification and synthesis of 2-amino-1-(4-bromo-2, 5-dimethoxyphenyl)ethan-1-one (bk-2C-B).

2-Amino-1-(4-bromo-2,5-dimethoxyphenyl)ethan-1-one (bk-2C-B) has been recently offered for purchase by a variety of Internet retailers. This substance may be considered a cathinone analogue of the phenethylamine 2-(4-bromo-2,5-dimethoxyphenyl)ethan-1-amine (2C-B) which suggests that it may have psychoactive effects in humans. A test purchase of bk-2C-B was carried out and its identity was confirmed by a range of analytical techniques including nuclear magnetic resonance spectroscopy, gas and liquid chromatography, and high-resolution mass spectrometry. Confirmation was also obtained from the synthesis of bk-2C-B based on the implementation of the Delépine reaction in which the α-brominated intermediate was reacted with hexamethylenetetramine to afford the primary amine. Analysis of underivatized bk-2C-B by gas chromatography-mass spectrometry (GC-MS) showed that there was potential for artificial formation of 1-(4-bromo-2,5-dimethoxyphenyl)ethanone and a pyrazine dimer, these substances were not detected when employing liquid chromatographic analysis. Ion chromatography and X-ray crystallography analysis confirmed that the purchased bk-2C-B consisted of a hydrochloride and hydrobromide salt mixture, which indicated that it might have been prepared by the hexamethylenetetramine route followed by hydrochloric acid hydrolysis of the quaternary ammonium salt. X-ray crystallography also revealed that the purchased (mixed HCl/HBr salt) and synthesized bk-2C-B (HCl salt) exists as polymorphs.

Fatal mephedrone intoxication--a case report.

A death caused by a new designer drug, 4-methylmethcathinone (mephedrone), is reported. Eight small plastic bags containing white powder were found in the jacket of a young dead male. Spot tests conducted by the police officer indicated the presence of 4-bromo-2,5-dimethoxyphenethylamine (2C-B) in the powders. Laboratory routine screening analyses of blood and vitreous humor did not reveal any positive results; therefore, 2C-B was excluded. Analysis of powders was conducted using gas chromatography-mass spectrometry and high-pressure liquid chromatography with diode array detection. The purity of mephedrone found in all powder samples was in the range of 80.4-87.3%. In connection with these findings, blood and vitreous humor samples were analyzed for mephedrone. Analyses were conducted using liquid chromatography-tandem mass spectrometry. Mephedrone was found in blood and vitreous humor at the concentrations of 5.5 and 7.1 µg/mL, respectively, revealing that this was a fatal mephedrone intoxication.

4-Bromo-2,5-dimethoxyphenethylamine (2C-B): presence in the recreational drug market in Spain, pattern of use and subjective effects.

4-Bromo-2,5-dimethoxyphenethylamine (2C-B) is a psychoactive analogue of mescaline that is becoming increasingly popular as a rave and club drug. We investigated its presence in the illicit drug market in Spain, its pattern of use and profile of subjective effects. Drug material was analysed for 2C-B and information on pattern of use and subjective effects was obtained from recreational users. Scores were statistically compared with previously collected data on psychostimulants (d-amphetamine), entactogens (MDMA) and psychedelics (ayahuasca and Salvia divinorum). The percentage of samples containing 2C-B doubled between 2006 and 2009, evolved from powder to tablet form and showed low falsification rates. Respondents reported taking 2C-B orally in doses of about 20 mg. Subjective effects involved perceptual modifications analogous to those observed after ayahuasca and salvia but absent after amphetamine and MDMA. Pleasure and sociability effects did not differ from those after MDMA and incapacitation was lower than for the psychedelics used as comparators. In conclusion, we found 2C-B is consistently present in the illicit drug market in Spain. While it elicits perceptual modifications that are analogous to other psychedelics, the lower impairment and higher pleasurable effects make it comparable with entactogens.

Morbidity involving the hallucinogenic designer amines MDA and 2C-I.

A case is presented of a 39-year-old woman who suffered severe debilitation because of a hemorrhagic stroke in the context of substance abuse. The patient presented to the emergency room with rapidly diminishing mental status, hypertension, and vasoconstriction; her friends provided a history of ingestion of cocaine, 3,4-methylenedioxymethamphetamine (MDMA), and 2C-I, a novel designer amine. A multi-targeted LC-MS/MS method for sympathomimetic amines and related drugs in urine detected and quantified 2C-I and MDA, while ruling out MDMA. The cause of the stroke was determined to be an underlying cerebrovascular abnormality called Moyamoya, secondary to substance abuse. In clinical laboratories, gas chromatography-mass spectrometry or liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmation of a positive amphetamine immunoassay is usually directed only towards amphetamine, methamphetamine, MDMA and MDA. This report demonstrates the utility of testing for a wider menu of compounds using LC-MS/MS in order to better characterize the prevalence and toxicities of novel amines such as 2C-I.

2C-B: a new psychoactive phenylethylamine recently discovered in Ecstasy tablets sold on the Swiss black market.

This study sought to identify, by means of several analytical methods (GC-MS, HPLC-DAD, CE-DAD, FTIR, and NMR), 4-bromo-2,5-dimethoxyphenethylamine (2C-B), which was found in two sets of tablets obtained from the Swiss black market. Unequivocal identification of 2C-B was only achieved by a combination of mass spectrometric and NMR analysis. Quantitation of 2C-B was performed by HPLC-DAD and CE-DAD. The amounts of 2C-B found in the tablets (3-8 mg) were in the range of the minimum quantity required to induce the effects characteristic of this drug.

Problems in the development of radioimmunoassay of catecholamines.

This review describes experiments to produce antibodies towards the catecholamines and some catecholamine metabolites with the intention of developing radioimmunological methods. First, attempts to produce antibodies to the catecholamines themselves are described. In the course of synthesis of immunogens of catecholamines, it was necessary to exercise special care - e.g. the introduction of protecting groups - owing to the great susceptibility of the catechol structure to oxidation. Despite many efforts, only one working group (Miwa et al.) has reported in a series of papers the successful production of antibodies to catecholamines, but they did not develop a radioimmunoassay. In contrast, antibodies to some metabolites of the catecholamines - such as 3,4-dimethoxyphenylethylamine, the 3-O-methylated catecholamines (normetanephrine, metaneprine, and 3-methoxytyramine) as well as 3-methoxy-4-hydroxyphenylethyleneglycol - have been produced, whose avidity and specificity were high enough to permit the development of sensitive radioimmunoassays.