A fatal case of MAM-2201 poisoning

A 59-year-old man was found dead in his house, where three sachets containing herbal blends were found on a table. The sachet contents were analyzed by gas chromatography–mass spectrometry and found to contain [1-(5-fluoropentyl)-1H-indol-3-yl](4-methyl-1-naphthalenyl)methanone (MAM-2201). The deceased was subjected to forensic autopsy. There were neither external injuries nor endogenous diseases judged by macroscopic and microscopic observations. Liquid chromatography–electrospray ionization–tandem mass spectrometry was used to quantitate the concentrations of MAM-2201 in postmortem samples using deuterated MAM-2201 as internal standard. The MAM-2201 concentrations were: 12.4 ng/ml in whole blood; 18.1 ng/g in the liver; 11.2 ng/g in the kidney; 4.3 ng/g in the brain; and 1,535 ng/g in the adipose tissue. We concluded that the man’s death was caused by acute intoxication with MAM-2201. In addition, we propose that the adipose tissue is the specimen of choice to detect MAM-2201 in the unchanged form. To our knowledge, this is the first report of a fatal MAM-2201 poisoning case. In addition, this report is also the first to describe the distribution of the drug in postmortem human tissues and blood.     

Identification and quantitation of JWH-213, a cannabimimetic indole, as a designer drug in a herbal product

In our survey of designer drugs in the Japanese market, a cannabimimetic indole was identified as a new active compound in a herbal product. The structure of this compound was elucidated by liquid chromatography–photodiode array–mass spectrometry (LC–PDA–MS), gas chromatography–mass spectrometry (GC–MS), high-resolution MS, and nuclear magnetic resonance (NMR) analyses. The compound was finally identified as (4-ethyl-1-naphthalenyl)(2-methyl-1-pentyl-1H-indol-3-yl)methanone (JWH-213), an indole-based cannabinoid receptor ligand. To our knowledge, this is the first finding of JWH-213 as a designer drug in a herbal product. The quantitative LC–PDA analysis showed that the JWH-213 content in the product was 252 mg/pack.     

A large amount of new designer drug diphenidine coexisting with a synthetic cannabinoid 5-fluoro-AB-PINACA found in a dubious herbal product

Traffic police brought five dubious herbal product packages to our laboratory for toxicological analysis. These products usually contain one or multiple kinds of synthetic cannabinoids. In one of the packages, we identified the coexistence of a new type of dubious drug (diphenidine) with the synthetic cannabinoid 5-fluoro-AB-PINACA. Conclusive identification was performed by comparison of the mass spectra of the test herb extracts with those of the reference standards of diphenidine and 5-fluoro-AB-PINACA by both gas chromatography–mass spectrometry (GC–MS) and electrospray ionization–tandem mass spectrometry. Both mass spectra of the test extracts coincided with those of the reference standards for each MS method. Diphenidine and 5-fluoro-AB-PINACA were quantitated in the herbal product by GC–MS using selected ion monitoring and the standard addition method. The content of diphenidine in the herbal product was as high as 289 ± 23.2 mg/g (n = 5); that of 5-fluoro-AB-PINACA was 55.5 ± 5.8 mg/g (n = 3). Diphenidine is known as an N-methyl-d-aspartate receptor channel blocker. Although its human toxicity has not been studied, it is likely to have severe psychotropic action in humans. The very high content of diphenidine in the present herbal package should prompt law enforcement agencies to be aware of the potential harmful effects of diphenidine itself and also when consumed in combination with other drugs of abuse.     

Characterization of the designer benzodiazepine pyrazolam and its detectability in human serum and urine

In 2012, online shops selling so-called research chemicals started offering pyrazolam, a new benzodiazepine that differs from phenazepam and etizolam, which have also recently appeared on the “gray market”, in that it is not marketed by pharmaceutical companies anywhere in the world. This article describes the characterization of pyrazolam (8-bromo-1-methyl-6-pyridin-2-yl-4H-[1,2,4]triazolo[4,3–a][1, 4]benzodiazepine) using gas chromatography-mass spectrometry, liquid chromatography-tandem mass spectrometry (LC–MS–MS), liquid chromatography quadrupole time-of-flight mass spectrometry (LC–Q–TOF–MS), and nuclear magnetic resonance spectroscopy. In addition, a study was carried out in which one of the authors ingested two 0.5-mg pyrazolam tablets. Serum and urine samples were then obtained to investigate the metabolism of pyrazolam and to obtain preliminary results for the elimination half-life and the detectability of a 1-mg dose in serum and urine using a highly sensitive LC–MS–MS method and immunoassays. The results showed an elimination half-life of about 17 h and no detectable metabolism. The parent compound was detected with the described LC–MS–MS method in serum for more than 50 h and in urine for approximately 6 days. Immunoassays showed cross-reactivity, but poor detection in the study samples demonstrated that consumption or administration of this presumably potent drug could go undetected unless instrumental analytical techniques are also used.     

Identification of (1-pentylindol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone and its 5-pentyl fluorinated analog in herbal incense seized for drug trafficking

Four herbal incense products were seized from suspected drug abusers in Korea. The major ingredients in the herbal incense samples were purified, and their structures were elucidated using gas chromatography–electron ionization–mass spectrometry (GC–EI–MS), liquid chromatography–time-of-flight–mass spectrometry (LC–TOF–MS), and 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. As a result, ingredients in the herbal incense were identified as (1-pentylindol-3-yl)-(2,2,3,3-tetramethylcyclopropyl)methanone and its 5-pentyl fluorinated analog [1-(5-fluoropentyl)indol-3-yl]-(2,2,3,3-tetramethylcyclopropyl)methanone. The former is being sold via the Internet as a "research chemical" named UR-144, and the latter is sold as 5F-UR-144. UR-144 is a selective full agonist of CB₂ cannabinoid receptor, and was first developed by Abbott Laboratories as an analgesic. It exhibits analgesic activity against both neuropathic and inflammatory pain associated mainly with the CB₂ receptor, but shows less psychotropic effects associated with the CB₁ receptor. Fluorination of the N-pentyl side chain of cannabimimetic compounds increases their cannabinoid receptor affinity such as with AM-2201, which shows both increased analgesic and psychotropic effects simultaneously. UR-144 and 5F-UR-144 can be classified as research chemicals based on their analgesic effects, but in practice are abused as psychotropic agents and can cause unexpected toxic effects. Thus, the trade and diversion of these chemicals should be monitored carefully for possible abuse. To our knowledge, this is the first report disclosing cyclopropylcarbonylindoles in herbal products.     

Simultaneous determination of tryptamine analogues in designer drugs using gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry

In recent years, a large number of tryptamine-based designer drugs have been encountered in forensic samples. We have developed simultaneous analytical methods for 14 tryptamine analogues using gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–tandem mass spectrometry (LC–MS–MS). Trimethylsilyl (TMS) derivatives of the analytes were separated on a DB-1ms column within 15 min. The structural isomers could be differentiated by electron ionization GC–MS. LC–MS–MS with a C₁₈ column could separate structural isomers of tryptamines except for a combination of 5-methoxy-N,N-diethyltryptamine and 5-methoxy-N-methyl-N-isopropyltryptamine. Higher collision energy gave different product ion spectra between the structural isomers. The results indicate that GC–MS is the first choice for identification of tryptamines, preferably after TMS derivatization, and LC–MS–MS can be used as a complementary approach for the unequivocal differentiation of tryptamine isomers.     

In vitro and in vivo metabolisms of 1-pentyl-3-(4-methyl-1-naphthoyl)indole (JWH-122)

1-Pentyl-3-(4-methyl-1-naphthoyl)indole (JWH-122) is an agonist of the cannabinoid receptors CB₁ and CB₂. In this study, the phase I and phase II metabolisms of JWH-122 were investigated using two models. In vitro studies using incubations of JWH-122 with human liver microsomes were performed to obtain metabolites of the drug at the initial step; 11 classes of metabolites were found and analyzed by liquid chromatography–mass spectrometry (LC–MS) and liquid chromatography–tandem mass spectrometry (LC–MS–MS). Hydroxylation(s) on the naphthalene moiety and/or the indole moiety of the molecule took place as such or in combination with dehydrogenation or cleavage of the N-pentyl side chain. Furthermore, dihydrodiol metabolites were formed probably via epoxide formation on the naphthalene moiety, irrespective of the combination with hydroxylation(s). A metabolite carrying a carboxyl group on the N-pentyl side chain was also detected. As the second step of the study, in vivo experiments using chimeric mice were performed; the mice were orally administered JWH-122, and their urine samples were collected, subjected to enzymatic hydrolysis, and analyzed by LC–MS and LC–MS–MS. The urine samples without hydrolysis were also analyzed for their molecular formulae in the conjugated forms by LC–high resolution MS. The in vivo model using chimeric mice confirmed most metabolite classes and clarified the phase II metabolism of JWH-122. It was concluded that all metabolites formed in vivo were excreted conjugated as glucuronide or sulfate, with conjugation rates above 50 %.     

Simultaneous and sensitive analysis of fourth-generation antidepressants in human plasma by ultra-performance liquid chromatography–tandem mass spectrometry

A sensitive method for simultaneous analysis of nine fourth-generation antidepressants in human plasma by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS–MS) is presented. The method was used with a Waters Acquity UPLC LC system, an Acquity TQD MS–MS system, and a Poroshell 120 EC-C18 separation column. For extraction of the target compounds, solid-phase extraction with Oasis HLB cartridges was used. All compounds were detected with retention times lower than 3 min. The calibration curves for the antidepressants spiked into human plasma showed similar good linearities in the range of subnanogram to nanogram per milliliter. The detection limits (signal-to-noise ratio = 3) were in the range of 0.2–100 pg/ml. The method gave satisfactory recovery rates, accuracy, and precision for quality control samples spiked with these drugs. To further validate the present method, 25 mg of milnacipran was orally administered to a healthy male volunteer, and the drug concentrations in plasma were measured in samples collected 0.5, 1, 2, 4, and 8 h after dosing. The concentrations were in the range of 31.2–56.8 ng/ml. To our knowledge, this is the most sensitive quantitative method for fourth-generation antidepressants so far reported, and should prove very useful in forensic and clinical toxicology. In addition, the quantitative analysis of tianeptine by LC–MS–MS is first described in this study.     

High-throughput analysis of ramelteon, agomelatine, and melatonin in human plasma by ultra-performance liquid chromatography–tandem mass spectrometry

A high-throughput method for analysis of ramelteon, agomelatine, and melatonin in human plasma by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS–MS) is presented. The LC system, MS–MS system, and separation column used were Waters Acquity UPLC, Acquity TQD, and Poroshell 120 EC-C18, respectively. For extraction of the target compounds, solid-phase extraction was performed with Oasis HLB cartridges. All compounds were detected with retention times of <3 min. The calibration curves for the compounds spiked into human plasma showed good linearities in the nanogram-per-milliliter range. The detection limit (signal-to-noise ratio = 3) was as low as 0.2–0.5 ng/ml. The method gave satisfactory recovery rates, accuracy, and precision for quality control samples spiked with these drugs. The present method should prove very useful in forensic and clinical toxicology and pharmacokinetic studies, because of its high sensitivity and rapidness. To our knowledge, this is the first trial to analyze ramelteon in a biological sample by LC–MS–MS.     

An LC–MS–MS method for quantitative analysis of six trimethoxyamphetamine designer drugs in rat plasma, and its application to a pharmacokinetic study

Trimethoxyamphetamines (TMAs) comprise a family of hallucinogenic drugs that includes various different positional isomers, which are important both for their hallucinogenic activity and their circulation in illicit drug markets. This report describes a method for identification and quantitation of six TMA isomers in rat plasma by solid-phase extraction and liquid chromatography–tandem mass spectrometry (LC–MS–MS) with electrospray ionization. Mescaline-d ₁ was used as internal standard. Multiple reaction monitoring on a triple quadrupole mass spectrometer operating in the positive ion mode was used for detection. The chromatographic system used a Varian Polaris C18-A column (2.0 × 100 mm i.d., 3 μm) and gradient elution with acetonitrile and 0.1 % formic acid in water. The calibration curves were linear over the concentration range from 10 to 200 ng/ml for all drugs with correlation coefficients that exceeded 0.998. The limits of detection and quantitation ranged from 1.1 to 2.3 ng/ml and from 6.9 to 10.2 ng/ml, respectively. The validation data, such as precision, accuracy, and recovery, showed good reproducibility and selectivity. This method was successfully applied to evaluating the pharmacokinetic profiles of TMAs in rats.     

Phosphorus-specific determination of glyphosate,glufosinate, and their hydrolysis products in biological samples by liquid chromatography–inductively coupled plasma–mass spectrometry

Anion exchange liquid chromatography (LC) with inductively coupled plasma–mass spectrometry (ICP–MS) was used for simultaneous determination of glyphosate and glufosinate (phosphonic and amino acid group-containing herbicides; PAAHs) and their hydrolysis products, aminomethylphosphonic acid (AMPA) and 3-methylphosphinicoacetic acid (MPPA), in biological samples. The target compounds were separated using an anion exchange resin column and gradient elution with sodium carbonate and sodium hydroxide system. The chromatographic eluates were passed through a membrane suppressor system and monitored by phosphorus-specific detection at m/z 31. PAAHs and their hydrolysis products were baseline separated from each other within 40 min of elution time, and phosphoric acid did not interfere with detection. The detection limits in serum samples were 0.1–0.7 µg/ml; those in urine samples, 0.2–1.6 µg/ml for the four compounds. The spiked recoveries for the four compounds were over 91 % in serum and urine samples. The detection of the compounds was not subject to interference from sample matrix components. The present method would be useful for toxicological analysis of PAAHs and their products in actual forensic practice. To our knowledge, this is the first trial to establish an LC–ICP–MS method for analysis of PAAHs and their products in biological samples .     

Validated liquid chromatography–mass spectrometry method for the quantitation of N-substituted derivatives of 3,4-methylenedioxyamphetamine in rat urine

3,4-Methylenedioxyamphetamines (MDAs) are central nervous system stimulants that are widely diffusing into the illegal market. Their identification is often difficult because of the high structural variety of new compounds. We describe a method for identifying and quantitating four MDAs in rat urine; three of them are new designer drugs: 3,4-methylenedioxy-N-isopropylamphetamine (MDIP), 3,4-methylenedioxy-N-cyclopropylmethylamphetamine (MDCPM), and 3,4-methylenedioxy-N-benzylamphetamine (MDBZ). The well-known and well-studied 3,4-methylenedioxy-N-ethylamphetamine (MDEA) was included as a reference. The quantitative analysis was performed by liquid chromatography–mass spectrometry (LC–MS) using 2,3-dimethoxyphenethylamine-d ₃ (2,3-DMPEA-d ₃) as internal standard. Samples were extracted by solid-phase extraction before injection. Chromatographic separation was achieved using a C₁₈ column (150 × 2.1 mm i.d., particle size 3 μm) by gradient elution with a mixture of 0.1 % formic acid in water and acetonitrile. The step gradient elution required a total run time of about 25 min. Regression equations were linear over the tested concentration range (10–500 ng/ml). The limits of detection and quantitation were in the ranges of 4.20–10.5 ng/ml and 12.4–23.4 ng/ml, respectively. The intraday and interday precision showed relative standard deviation values of 5.20–14.1 %. This LC–MS method proved to be robust and reliable, and was successfully applied to the quantitation of each drug in rat urine after oral administration of each drug (1.0 mg/kg). To our knowledge, this is the first report to identify and quantitate MDIP, MDCPM, and MDBZ in biological samples by MS.     

GHB-<Emphasis Type="Italic">O</Emphasis>-β-glucuronide in blood and urine is not a suitable tool for the extension of the detection window after GHB intake

Because of its small detection window, uncovering drug-facilitated crime after gamma-hydroxybutyric acid (GHB) intake remains a problem. The aim of this experiment was to determine endogenous concentrations of GHB and GHB-O-β-glucuronide (GHB-Gluc) in plasma and urine samples and to compare them with concentrations after GHB intake in humans. Plasma and urine samples of volunteers (n = 50) who had never taken GHB during their lifetime (control group) were collected, and endogenous concentrations of GHB and GHB-Gluc were determined. In addition, plasma and urine samples of patients (n = 3) therapeutically taking sodium oxybate (GHB-sodium salt) were collected prior to and at different time points after the intake. GHB was determined via a liquid chromatography (LC)–tandem mass spectrometry system operated in multiple reaction monitoring mode. GHB-Gluc was detected by LC–quadrupole time-of-flight mass spectrometry. In plasma and urine samples of the control group (n = 50), endogenous concentrations of GHB-Gluc ranged from 0.011 to 0.067 mg/L and from 0.16 to 7.1 mg/L, respectively, while unconjugated GHB concentrations were less than 2 mg/L in both matrices. In contrast, after sodium oxybate administration, GHB concentrations increased markedly, and fell to below the commonly used cutoff value (plasma 4 mg/L and urine 10 mg/L) after 6–8 h in all patients. GHB-Gluc concentrations showed no significant time-dependent increase in plasma samples. In urine, GHB-Gluc concentrations increased after GHB intake, but were generally not higher than the endogenous concentrations of the control group. Therefore, it can be concluded that GHB-Gluc concentrations are not a suitable marker for extending the detection window after GHB intake.     

Identification and quantitation of a new cathinone designer drug PV9 in an “aroma liquid” product,antemortem whole blood and urine specimens,and a postmortem whole blood specimen in a fatal poisoning case

A couple bought “aroma liquid” and “bath salt” type drugs at a dubious drug shop. Both of them orally took the liquid type drug; although the male subject showed no symptoms, the female subject suffered shivering, convulsions, and low levels of consciousness. The woman was taken to an emergency hospital to receive intensive medical treatment, but died about 20 h after admission. The aroma liquid solution, and the antemortem blood and urine collected during medical treatment at the hospital were brought to our laboratory by the police for analysis of the causative drug(s). In addition, a sample of postmortem femoral vein blood was collected from the cadaver. After some screening tests, we finally identified PV9 (α-POP) in all specimens by gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry (LC–MS–MS). The concentration of PV9 was 18.3 mg/ml in the aroma liquid solution, 45.7 ng/ml in the antemortem blood, 20.3 ng/ml in the antemortem urine, and 180 ng/ml in the postmortem femoral vein blood. The concentrations in antemortem blood and urine and in postmortem blood were greatly lowered by dilution during the intensive medical treatment, including intravenous drip infusion of a large volume of solution. The probable coexistence of a β-hydroxyl metabolite was also investigated by mass chromatography and analysis of fragment ions of the product ion spectrum obtained by LC–MS–MS. To our knowledge, this is the first reported identification and quantitation of PV9 in human specimens in a fatal PV9 poisoning case.     

SPME–GC–MS analysis of α-pyrrolidinovaleorophenone in blood in a fatal poisoning case

We provided toxicological analytical support for a fatal case of abuse of α-pyrrolidinovaleorophenone (α-PVP). Solid-phase microextraction (SPME) and capillary gas chromatography coupled to mass spectrometry (GC–MS) was employed to quantify the drug in whole blood. The whole blood concentration of the drug in the heart was 486 ng/ml. This is the first report of α-PVP intoxication as ascertained by mass spectrometric identification of α-PVP in whole blood.     

MALDI-TOF mass spectrometric determination of 11 phenothiazines with heavy side chains in urine

A rapid screening method was developed for the determination of phenothiazines by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF–MS). In this method, α-cyano-4-hydroxy cinnamic acid (CHCA) was used as the matrix to assist the ionization of phenothiazines. The identification of 11 phenothiazines with heavy side chains was achieved by observing their protonated molecular ions [M+H]⁺ at m/z 340–447. Quantification was achieved by using triflupromazine at m/z 353 as the internal standard (IS). The relative ionization efficiencies of 11 phenothiazines and IS on MALDI-TOF–MS were different from those achieved by ESI-MS, but the product ion spectra from MALDI-MS–MS were quite similar to those from ESI-MS–MS except in the case of flupentixol. The limit of detection was 0.3 ng/ml with a quantification range of 1–50 ng/ml urine in the best case; the limit of detection was 5 ng/ml with a quantification range of 10–100 ng/ml urine in the worst case for 10 phenothiazines except thiethylperazine. The present method provides a simple and high-throughput method for the screening of these phenothiazines using only 20 μl of urine. To our knowledge, this study is the first trial to analyze phenothiazines by MALDI-TOF–MS (–MS).     

Development and validation of a sensitive LC–MS/MS method to analyze NBOMes in dried blood spots: evaluation of long-term stability

Purpose

We evaluated the use of dried blood spots (DBS) to determine seven NBOMes by liquid chromatography–tandem mass spectrometry (LC–MS/MS), and also evaluated the stability of these compounds in this dried matrix.

Methods

An LC–MS/MS method was developed and fully validated to quantify seven NBOMes (25C-, 25H-, 25I-, 25B-, 25G-, 25D- and 25E-NBOMe) in DBS samples. The extraction procedure was optimized using mixture design experiment. Stability study was performed in two different concentrations over 180 days at three different storage temperatures.

Results

Good linearity, and limits of detection and quantitation of 0.05 and 0.1 ng/mL, respectively, were obtained. The interday imprecision (n = 15) and bias (n = 15) were not higher than 11.4 and 10.3%, respectively, and no carryover was observed. All analytes remained stable in DBS at ? 20, 4 °C and even at room temperatures for 180 days, except 25B-NBOMe and 25I-NBOMe which experienced degradation (22 and 21%, respectively) of the initial concentration at room temperature after 180 days of study. The method was applied to a DBS of an authentic postmortem blood from an NBOMe user, and it was found to be reliable with good selectivity and specificity.

Conclusions

DBS has been found to allow reliable, sensitive, accurate and robust detection and quantification of seven NBOMes via LC–MS/MS. Also, DBS provided great stability to most of the compounds at room temperature, and no degradation was observed for DBS kept at 4 and ? 20 °C. This is the first trial to analyze NBOMes in DBS samples to our knowledge.

     

Determination of ibotenic acid and muscimol, the Amanita mushroom toxins, in human serum by liquid chromatography–tandem mass spectrometry

In our previous article, we reported the analysis of ibotenic acid and muscimol in Amanita mushrooms by liquid chromatography–tandem mass spectrometry (LC–MS–MS). The levels of ibotenic acid and muscimol in the mushroom were as high as 210 and 107 μg/g, respectively. We have since tried to measure the same toxins in human serum obtained from a poisoned subject, who ingested the Amanita mushrooms, by the same method. However, the levels of the toxins in the human serum were about three orders of magnitude lower than those in Amanita mushrooms. In addition, the recovery rates for ibotenic acid and muscimol in human serum were found to be much lower than those in the previous study for the mushrooms. Therefore, we optimized the solid-phase extraction procedure again, and reevaluated the data for validation at much lower levels of ibotenic acid and muscimol in human serum. A 100-μl aliquot of human serum containing the target toxins was mixed with 100 ng of acivicin as internal standard (IS), 200 μl of distilled water, and 100 μl of 0.5 % ammonium hydroxide in distilled water, and vortexed well for 10 s. The mixture was loaded on an Oasis MAX 3cc (60 mg) extraction cartridge. The cartridge was washed with 0.5 ml of distilled water and 1.0 ml of methanol. The target compounds and IS were eluted with 4 ml of 0.05 % trifluoroacetic acid in methanol. The eluate was evaporated to dryness and reconstituted in methanol, and its small volume was subjected to LC–MS–MS analysis with the same TSK-GEL Amide-80 separation column. The LC elution was made in the gradient and isocratic modes. The selected reaction monitoring chromatograms showed clear peaks at 5.3, 3.5, and 3.6 min for ibotenic acid, muscimol, and IS, respectively; the blank serum sample without the target compounds or IS gave no peaks at the respective retention times except for an impurity peak at 6.5 min. There was good linearity from 10 to 1,000 ng/ml for both ibotenic acid and muscimol with correlation coefficients not <0.999. The detection limits (signal-to-noise ratio = 3) were 1.0 and 2.5 ng/ml for ibotenic acid and muscimol, respectively. The recovery rates of the target compounds in sera at five different concentrations were 87.9–103 %. The intraday and interday accuracy and precision data were also generally satisfactory. Using the modified method, the actual concentrations of ibotenic acid and muscimol were measured for a serum sample obtained from an ill patient thought to have ingested Amanita ibotengutake; they were 95.9 and 105 ng/ml, respectively. To our knowledge, this is the first report of analysis of ibotenic acid and muscimol in human serum by an MS technique, which we believe will be very useful in forensic and clinical toxicology.     

Two new synthetic cannabinoids, AM-2201 benzimidazole analog (FUBIMINA) and (4-methylpiperazin-1-yl)(1-pentyl-1H-indol-3-yl)methanone (MEPIRAPIM), and three phenethylamine derivatives, 25H-NBOMe 3,4,5-trimethoxybenzyl analog, 25B-NBOMe, and 2C-N-NBOMe, identified in illegal products

Two new types of synthetic cannabinoids, an AM-2201 benzimidazole analog (FUBIMINA, 1) and (4-methylpiperazin-1-yl)(1-pentyl-1H-indol-3-yl)methanone (MEPIRAPIM, 2), and three newly emerged phenethylamine derivatives, 25B-NBOMe (3), 2C-N-NBOMe (4), and a 25H-NBOMe 3,4,5-trimethoxybenzyl analog (5), were detected in illegal products distributed in Japan. The identification was based on liquid chromatography–mass spectrometry (LC–MS) and gas chromatography–mass spectrometry (GC–MS), high-resolution MS, and nuclear magnetic resonance analyses. Different from the representative synthetic cannabinoids, such as JWH-018, which have a naphthoylindole moiety, compounds 1 and 2 were completely new types of synthetic cannabinoids; compound 1 had a benzimidazole group in place of an indole group, and compound 2 had a 4-methylpiperazine group in place of the naphthyl group. Compounds 3 and 4 were N-o-methoxybenzyl derivatives of 2,5-dimethoxyphenethylamines (25-NBOMe series), which had been previously detected in European countries, but have newly emerged in Japan. Compound 5 had an N-trimethoxybenzyl group in place of an N-o-methoxybenzyl group. Data on the chemistry and pharmacology of compounds 1, 2, and 5 have never been reported to our knowledge.     

Another fatal case related to the recreational abuse of U-47700

The abuse of synthetic opioids has become a major threat in recent years. Several clinical reports and fatal case reports exist discussing life-threatening hypoventilation and fatal respiratory depression following the abuse of trans-3,4-dichloro-N-(2-(dimethylamino)cyclohexyl)-N-methylbenzamide (U-47700). The reported concentration of U-47700 in peripheral blood varies between 0.01 μg/mL and 1.46 μg/mL. These values depend on the mode of administration and whether the drug was used in combination with other drugs and/or pharmaceuticals. In the past, U-47700 was predominantly insufflated and not injected. The current study presents a non-targeted liquid chromatography/mass spectrometry (LC/MS)-based screening approach of urine and cerebrospinal fluid samples after intravenous injection of U-47700. Furthermore, quantitative values on U-47700 as obtained by liquid chromatography coupled to a linear ion trap (LC/ESI-QTRAPMS) are presented concerning femoral blood (0.29 μg/mL), urine (0.24 μg/mL), gastric contents (0.57 μg/mL), bile fluid (2.3 μg/mL), heart blood (1.25 μg/mL), liver (9.9 μg/g), cerebrospinal fluid (0.4 μg/mL), and hair (0.14 ng/mg). Thereof, concentrations in hair, gastric contents, bile fluid and cerebrospinal fluid have never been reported before. Drug paraphernalia were also analyzed by liquid chromatography coupled to a diode array detector (LC/DAD) and nuclear magnetic resonance spectrometer (NMR). The analyses show that the powder had a relatively high purity and was adulterated to a low degree. This is the first case report which lists concentration distributions of various specimens after intravenous injection. These findings as well as the U-47700 concentration are important to evaluate autopsy cases of U-47700 intoxication in the future.