Fatal intoxication with new synthetic cannabinoids 5F-MDMB-PICA and 4F-MDMB-BINACA-parent compounds and metabolite identification in blood, urine and cerebrospinal fluid.

Synthetic cannabinoids (SCs) remain one of the largest groups of new psychoactive substances. Recently, new synthetic cannabinoids 5F-MDMB-PICA and 4F-MDMB-BINACA are increasing in popularity. A 33-year-old man lost consciousness after smoking an unknown substance. A glass pipe and two lumps of substance that turned out to contain 5F-MDMB-PICA and 4F-MDMB-BINACA were found at the scene. Blood, urine and cerebrospinal fluid were collected during the examination of the body. The synthetic cannabinoids were isolated from autopsy materials by precipitation with acetonitrile and extraction with ethyl acetate. The screening and quantitative analyses were performed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The liquid chromatography-quadrupole/time of flight mass spectrometry (LC-Q/TOF) technique was used for metabolite identification. 5F-MDMB-PICA was detected and quantified in all analysed materials, whereas 4F-MDMB-BINACA was found only in cerebrospinal fluid. The determined concentrations of 5F-MDMB-PICA were 0.9 (blood), 0.1 (urine) and 3.2 ng/mL (cerebrospinal fluid). The concentration of 4F-MDMB-BINACA in cerebrospinal fluid was 0.1 ng/mL. The main metabolites of both compounds (hydrolysis and oxidative defluorination) were found in all analysed body fluids. Cerebrospinal fluid may be important alternative material in autopsy cases. Rapid elimination of 5F-MDMB-PICA and 4F-MDMB-BINACA compounds also means that the metabolite analysis can be crucial for the investigation. Laboratories must be made aware of their presence and incorporate these SCs and their metabolites into workflows for detection and confirmation. Ester hydrolysis and oxidative defluorination products can be found in blood, urine and cerebrospinal fluid making them useful biomarkers of intake.

NBOMes-Highly Potent and Toxic Alternatives of LSD.

Recently, a new class of psychedelic compounds named NBOMe (or 25X-NBOMe) has appeared on the illegal drug market. NBOMes are analogs of the 2C family of phenethylamine drugs, originally synthesized by Alexander Shulgin, that contain a N-(2-methoxy)benzyl substituent. The most frequently reported drugs from this group are 25I-NBOMe, 25B-NBOMe, and 25C-NBOMe. NBOMe compounds are ultrapotent and highly efficacious agonists of serotonin 5-HT2A and 5-HT2C receptors (Ki values in low nanomolar range) with more than 1000-fold selectivity for 5-HT2A compared with 5-HT1A. They display higher affinity for 5-HT2A receptors than their 2C counterparts and have markedly lower affinity, potency, and efficacy at the 5-HT2B receptor compared to 5-HT2A or 5-HT2C. The drugs are sold as blotter papers, or in powder, liquid, or tablet form, and they are administered sublingually/buccally, intravenously, via nasal insufflations, or by smoking. Since their introduction in the early 2010s, numerous reports have been published on clinical intoxications and fatalities resulting from the consumption of NBOMe compounds. Commonly observed adverse effects include visual and auditory hallucinations, confusion, anxiety, panic and fear, agitation, uncontrollable violent behavior, seizures, excited delirium, and sympathomimetic signs such mydriasis, tachycardia, hypertension, hyperthermia, and diaphoresis. Rhabdomyolysis, disseminated intravascular coagulation, hypoglycemia, metabolic acidosis, and multiorgan failure were also reported. This survey provides an updated overview of the pharmacological properties, pattern of use, metabolism, and desired effects associated with NBOMe use. Special emphasis is given to cases of non-fatal and lethal intoxication involving these compounds. As the analysis of NBOMes in biological materials can be challenging even for laboratories applying modern sensitive techniques, this paper also presents the analytical methods most commonly used for detection and identification of NBOMes and their metabolites.

Screening procedure for 38 fentanyl analogues and five other new opioids in whole blood by liquid chromatography-tandem mass spectrometry.

In recent years, many new opioids, particularly fentanyl analogues, have appeared on the drug market. The extreme potency of even low doses of these compounds leads to numerous fatal poisonings. This also results in the fact that only sophisticated techniques are capable of detecting fentanyl analogues at concentrations that can be expected in blood. In this context, the purpose of this study was to develop a fast liquid chromatography-tandem mass spectrometry screening method for the detection of fentanyl analogues, and other new synthetic opioid receptor agonists in whole blood. Blood samples were extracted with ethyl acetate under basic conditions. The separation was achieved with the gradient of the mobile phase composition and the gradient of the flow rate in 13 minutes. The detection of all compounds was based on dynamic multiple reaction monitoring. Most of the compounds were well differentiated by their retention times and/or transitions; however, separation of some isomers has not been achieved. The validation was performed for 21 compounds. The limits of detection were in the range 0.01-0.20 ng/mL. The developed procedure enables simultaneous qualitative screening, detection and identification of 38 fentanyl analogues and five other new opioids. The method was implemented to analyze authentic samples (positive; n = 3) demonstrating its suitability for this application. The procedure can be easily expanded to include new emerging opioids, which is an indispensable advantage in the dynamically developing drug market. The developed protocol can be adopted for routine work in both forensic and clinical analytical laboratories worldwide.

Simple screening procedure for 72 synthetic cannabinoids in whole blood by liquid chromatography-tandem mass spectrometry.

PURPOSE: In recent years, many synthetic cannabinoids (SCs) have appeared on the drug market. Despite the increasing number of SCs, there are few comprehensive screening methods for their detection in biological specimens. In this context, the purpose of this study was to develop a fast and simple liquid chromatography-tandem mass spectrometry screening procedure for detection and identification of SCs in whole blood. METHODS: The elaborated qualitative screening method allows the simultaneous detection and identification of 72 compounds from different chemical groups: naphthoylindoles, naphthoylindazoles, benzoylindoles, phenylacetylindoles, tetramethylcyclopropylindoles, indole-3-carboxylic acid esters, indole-3-carboxylic acid amides, indazole-3-carboxylic acid amides, and others. Whole-blood samples (0.2 mL) were precipitated with acetonitrile (0.6 mL). The separation was achieved with the gradient of the mobile phase composition (0.1% formic acid in acetonitrile and 0.1% formic acid in water) and the gradient of the flow rate (0.5-0.8 mL/min) in 16 min. Detection of all compounds was based on dynamic multiple reaction monitoring. RESULTS: Mass spectrometer parameters for all compounds were presented. All of the compounds were well-separated by their retention times and/or transitions. The limits of detection (LODs) for 50 compounds were in the range 0.01-0.48 ng/mL. CONCLUSIONS: Estimated LODs make this assay suitable for the analysis of biological material. The procedure can be easily expanded for more substances, which is an indispensable advantage in the dynamically developing drug market. It can have wide application in various analytical forensic and clinical laboratories.

Acute intoxication of four individuals following use of the synthetic cannabinoid MAB-CHMINACA.

CONTEXT: The largest group of new psychoactive substances (NPS) are synthetic cannabinoids (SC). Those that become controlled are immediately replaced by new uncontrolled substances. The recent resurgence of the NPS market in Poland resulted in a further amendment to the Drug Addiction Counteraction Act. This resulted in significant changes in the composition of "legal high" preparations, and consequently a large outbreak of intoxications with SC was reported in Poland at the beginning of July 2015. CASE DETAILS: This paper describes the circumstances of intoxication and toxicological findings in an acute intoxication of four individuals with MAB-CHMINACA. They each smoked tobacco mixed with powder from the package with the description "AM-2201". The adverse effects observed in the individuals included vomiting, seizures, limb twisting, muscle tremors, aggression, agitation, slurred speech, blood pressure spikes, wheezing, respiratory failure and losses of consciousness. Blood samples were analysed using liquid chromatography with mass spectrometry. Results from analysis performed on the blood samples showed the presence of MAB-CHMINACA, while AM-2201 was not found (LOD 0.09 ng/mL). The determined concentrations were 5.2, 1.3, 1.7 and 14.6 ng/mL, respectively. The analyses of the blood did not reveal any other substances (excluding medicines given in hospital). CONCLUSION: The presented cases show the health risks associated with MAB-CHMINACA use and confirm that "legal high" preparations do not always contain a substance represented on the package.

Fatal intoxication with synthetic cannabinoid MDMB-CHMICA.

MDMB-CHMICA is a synthetic cannabinoid that appeared on the European drug market in September 2014. This substance was found in Poland in the herbal mixture "Mocarz" ("Strongman"), which caused a large outbreak of intoxications at the beginning of July 2015. This paper describes the circumstances of death and toxicological findings in a fatal intoxication with MDMB-CHMICA (in combination with alcohol). Loss of consciousness and asystole occurred a few minutes after smoking the 'legal high'. The man died after 4 days of hospitalisation. The cause of death accepted by the medical examiner was multiple organ failure. MDMB-CHMICA was detected and quantified in blood (ante- and postmortem) and internal organs tissues. The samples were analysed using liquid chromatography with mass spectrometry (LC-MS/MS). The concentration of MDMB-CHMICA in antemortem blood was 5.6 ng/mL. Although the death occurred after 4 days from administration a relatively high concentration (2.6 ng/g) was estimated in the brain. Traces of this compound were also found in other postmortem materials (blood, stomach, liver, bile, and kidney). The presented case shows the health risks associated with MDMB-CHMICA use. The administration of this substance can lead to the number of organ failures, cardiac arrest and consequently death.

Simultaneous screening for and determination of 128 date-rape drugs in urine by gas chromatography-electron ionization-mass spectrometry.

Date-rape drugs (DRDs) are used for the purpose of "drugging" unsuspected victims and raping or robbing them while under the influence of the drug. The wide variety of substances used for criminal purposes, their low concentrations in body fluids and, often, a long time delay between the event and clinical examination make comprehensive screening analysis of biological materials collected from crime victims for the presence of these drugs very difficult. Detection of a drug used to facilitate sexual assault in biological fluids can be very important evidence of a committed crime. The purpose of this study was to develop a simple GC-EI-MS screening procedure for date-rape drugs in urine. Target analytes were isolated by solid-phase extraction. 2-mL urine samples were extracted and then derivatized by using BSTFA+1%TMCS reagent. Detection of all compounds was based on full-scan mass spectra and for each compound one ion was chosen for further quantification. The method allowed the simultaneous screening, detection and quantification of 128 compounds from different groups (number of compounds): opioids (20), amphetamines (11), GHB and related products (3), hallucinogens (9), benzodiazepines (18), antihistamines (9), antidepressants (14), selective serotonin-reuptake inhibitors (4), antipsychotics (7), barbiturates (7), other sedatives (5), muscle relaxants (2) and other drugs (19). The procedure can easily be expanded to encompass more substances. The developed method appeared to be suitable for screening for the target DRDs. The procedure was successfully applied to the analysis of authentic urine samples collected from victims of rapes and other crimes in routine casework.

Date-rape drugs scene in Poland.

Since the beginnings of twenty-first century in Poland increasing number of reports about the drug-facilitated sexual assaults have been observed. Many drugs have been identified as so-called "date-rape drugs", because of their pharmacological properties, especially inducing amnesia. These drugs are used for the purpose of "drugging" unsuspected victims and than raping them. In a typical scenario, the perpetrator surreptitiously adds "date-rape drug" to the alcoholic or non-alcoholic beverage of an unsuspecting person, who is subsequently sexually assaulted while under the influence of this substance. Many victims do not report the incident until several days after the event or even do not report it at all. They report the incident so late after the events because they often have problems with remember the course of incident. It causes that victim is not reliable witness for justice. Detection of "date-rape drugs" in biological fluids is unequivocal evidence of perpetration. Analysis of biological fluids collected from victims of rapes for presence of drugs was rare in Poland up to now. The aim of this study is to show the use of "date-rape drugs" in Poland. Materials for this study were from the routine casework elaborated at the Institute of Forensic Research in Kraków. APCI-LC-MS methods were applied for screening of biological fluids (blood and/or urine) for amphetamine and its 6 analogues, for 12 substances from benzodiazepine group and for quantification of the detected drugs. HPLC-DAD was used as a screening method for wide range of medicinal drugs, and NCI-GC-MS methods for determination of ketamine and tetrahydrocannabinols (delta9-tetrahydrocannabinol, 9THC) and its metabolite (11-nor-carboxy-delta9-tetrahydrocannabinol, THCCOOH). In 2000-2004, the biological fluids taken from 33 persons, both sexually assaulted or perpetrators were analysed. In 2000 and 2002 not any case of this type was registered, in 2001 only two cases were recorded. After 2003 significant increase in the number of these cases was observed. Eleven and twenty cases involving "date-rape drugs" were submitted to the Institute in 2003 and 2004, respectively. The most common substances detected in analysed materials were amphetamine (in concentrations ranged from 10 to 85 ng/ml) and 9THC (0.36-1.4 ng/ml). Alcohol (0.27-2.3% per hundred), MDMA (8-201 ng/ml), benzodiazepines (oxazepam, nordazepam, estazolam), propranolol and lidocaine were also found in blood and urine specimens.

Urinary excretion rates of ketamine and norketamine following therapeutic ketamine administration: method and detection window considerations.

Ketamine is widely used in veterinary medicine. Its medical application in humans is limited to children because in adults it induces severe psychedelic episodes. In recent years, teenagers have abused ketamine as a recreational and "club drug" because of its hallucinogenic and stimulant effects. Ketamine is also misused as a "date-rape" drug (to induce amnesia in unsuspecting victims). Sensitive gas chromatography-mass spectrometry-negative chemical ionization (GC-MS-NCI) and liquid chromatography-mass spectrometry-atmospheric pressure chemical ionization (LC-MS-APCI) methods were applied for the simultaneous quantification of ketamine and its major metabolite, norketamine, in urine. Urine samples were collected from hospitalized children who had received ketamine as an anesthetic. Individual urine samples were collected up to 16 days after drug administration. Using the GC-MS-NCI method, ketamine was detected in the urine of the children from only the day of drug administration up to 2 days after drug administration. Its concentrations ranged from 29 to 1410 ng/mL. Norketamine (measured in concentrations of 0.1-1442 ng/mL) was detected up to 14 days. Using the LC-MS-APCI method, norketamine was detected up to 6 days after drug administration, ranging in concentrations of 2-1559 ng/mL, while ketamine was detected up to 11 days (2-1204 ng/mL). In the urine taken from one child, ketamine was not detected through the entire 16-day period using both methods. The detection window for the analytes is highly dependent on the method used for determination and varies between individuals.

Detection of ketamine and norketamine in urine of nonhuman primates after a single dose of ketamine using microplate enzyme-linked immunosorbent assay (ELISA) and NCI-GC-MS.

The general anesthetic ketamine (Ketalar, Ketaject, Vetalar) (KET) is used in human and veterinary medicine for induction of anesthesia for short surgical procedures and routine veterinary examination. Its illicit use by teenagers in rave parties has been reported, and it has recently been identified as a substance associated with sexual assault. One aim of this paper was to study the elimination of KET and its major metabolite norketamine (NKET) in urine collected from five nonhuman primates that received a single dose (5 mg/kg, I.M.) of KET and to study elimination patterns to determine how long after drug administration KET and NKET can be detected. Another aim of this study was to develop and validate a highly sensitive negative ion chemical ionization-gas chromatography-mass spectrometry (NCI-GC-MS) method for the simultaneous quantitation of KET and its major metabolite NKET in urine and to analyze urine samples collected from the animals. The last aim of this study was to apply and evaluate a newly developed ELISA screening methodology for detection of KET and its metabolites in the same urine samples collected from primates which received a single dose of KET. In two monkeys, KET was detected in urine up to 3 days after drug administration (32-7070 ng/mL); in one monkey, it was detected up to 4 days (65-13,500 ng/mL); in one monkey, it was detected only on days 1 and 2 (4000 and 70 ng/mL, respectively); and in one monkey, it was detected 10 days after KET injection (22-35,000 ng/mL). NKET concentrations ranged from 63 pg/mL to 1.75 microg/mL, and it remained in the urine throughout the entire 35-day study period in 4 out of 5 animals. In one monkey, NKET was detected up to 31 days after KET administration. Urine analysis using ELISA revealed that KET and NKET can be easily detectable at 25 ng/mL. In one monkey, KET and its metabolites were detected in urine up to 4 days after drug administration, up to 7 days in two monkeys, up to 11 days in one monkey, and 16 days after KET injection in one monkey. Urine extraction followed by screening using ELISA methodology allowed for significant extension of the detection period in all animals from the study. It is believed that the KET elimination in urine of nonhuman primates is slightly faster than in humans. We propose that NCI-GC-MS be employed to detect NKET as a target compound in urine in toxicological investigations of drug-facilitated sexual assault when KET use by the perpetrator is suspected.