4 research outputs found
Activity-Based Detection of Consumption of Synthetic Cannabinoids in Authentic Urine Samples Using a Stable Cannabinoid Reporter System
Synthetic
cannabinoids (SCs) continue to be the largest group of
new psychoactive substances (NPS) monitored by the European Monitoring
Center of Drugs and Drugs of Abuse (EMCDDA). The identification and
subsequent prohibition of single SCs has driven clandestine chemists
to produce analogues of increasing structural diversity, intended
to evade legislation. That structural diversity, combined with the
mostly unknown metabolic profiles of these new SCs, poses a big challenge
for the conventional targeted analytical assays, as it is difficult
to screen for âunknownâ compounds. Therefore, an alternative
screening method, not directly based on the structure but on the activity
of the SC, may offer a solution for this problem. We generated stable
CB1 and CB2 receptor activation assays based on functional complementation
of a split NanoLuc luciferase and used these to test an expanded set
of recent SCs (UR-144, XLR-11, and their thermal degradation products;
AB-CHMINACA and ADB-CHMINACA) and their major phase I metabolites.
By doing so, we demonstrate that several major metabolites of these
SCs retain their activity at the cannabinoid receptors. These active
metabolites may prolong the parent compoundâs psychotropic
and physiological effects and may contribute to the toxicity profile.
Utility of the generated stable cell systems as a first-line screening
tool for SCs in urine was also demonstrated using a relatively large
set of authentic urine samples. Our data indicate that the stable
CB reporter assays detect CB receptor activation by extracts of urine
in which SCs (or their metabolites) are present at low- or subnanomolar
(ng/mL) level. Hence, the developed assays do not only allow activity
profiling of SCs and their metabolites, it may also serve as a screening
tool, complementing targeted and untargeted analytical assays and
preceding analytical (mass spectrometry based) confirmation
Activity-Based Detection of Consumption of Synthetic Cannabinoids in Authentic Urine Samples Using a Stable Cannabinoid Reporter System
Synthetic
cannabinoids (SCs) continue to be the largest group of
new psychoactive substances (NPS) monitored by the European Monitoring
Center of Drugs and Drugs of Abuse (EMCDDA). The identification and
subsequent prohibition of single SCs has driven clandestine chemists
to produce analogues of increasing structural diversity, intended
to evade legislation. That structural diversity, combined with the
mostly unknown metabolic profiles of these new SCs, poses a big challenge
for the conventional targeted analytical assays, as it is difficult
to screen for âunknownâ compounds. Therefore, an alternative
screening method, not directly based on the structure but on the activity
of the SC, may offer a solution for this problem. We generated stable
CB1 and CB2 receptor activation assays based on functional complementation
of a split NanoLuc luciferase and used these to test an expanded set
of recent SCs (UR-144, XLR-11, and their thermal degradation products;
AB-CHMINACA and ADB-CHMINACA) and their major phase I metabolites.
By doing so, we demonstrate that several major metabolites of these
SCs retain their activity at the cannabinoid receptors. These active
metabolites may prolong the parent compoundâs psychotropic
and physiological effects and may contribute to the toxicity profile.
Utility of the generated stable cell systems as a first-line screening
tool for SCs in urine was also demonstrated using a relatively large
set of authentic urine samples. Our data indicate that the stable
CB reporter assays detect CB receptor activation by extracts of urine
in which SCs (or their metabolites) are present at low- or subnanomolar
(ng/mL) level. Hence, the developed assays do not only allow activity
profiling of SCs and their metabolites, it may also serve as a screening
tool, complementing targeted and untargeted analytical assays and
preceding analytical (mass spectrometry based) confirmation
Detection of the recently emerged synthetic cannabinoid 4F-MDMB-BINACA in "legal high" products and human urine specimens
Synthetic cannabinoids (SCs) remain one of the largest groups of new psychoactive substances (NPS) on the European drug market. Although the number of new derivatives occurring on the market has dropped in the last two years, newly emerging NPS still represent a challenge for laboratories performing forensic drug analysis in biological matrices. The newly emerged SC 4F-MDMB-BINACA has been reported by several law enforcement agencies in Europe and the USA since November 2018. This work aimed at revealing urinary markers to prove uptake of 4F-MDMB-BINACA and differentiate from the use of structurally similar SCs. Phase-I metabolites detected in human urine specimens were confirmed by phase-I metabolites generated in vitro using a pooled human liver microsomes (pHLM) assay. Seized materials and test-purchased "legal high" products were analyzed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-quadrupole-time-of-flight-mass spectrometry (LC-qToF-MS). Human urine specimens and pHLM assay extracts were measured with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) and confirmed by LC-qToF-MS. In January 2019, the Institute of Legal Medicine in Erlangen (Germany) identified 4F-MDMB-BINACA in three herbal blends. During the same time period, the described SC was identified in a research chemical purchased online. Investigation of phase-I metabolism led to the metabolites M10 (ester hydrolysis) and M11 (ester hydrolysis and dehydrogenation) as reliable urinary markers. Widespread distribution on the German drug market was proven by analysis of urine samples from abstinence control programs and by frequent detection of 4F-MDMB-BINACA in "herbal blends" and "'research chemicals" purchased via the Internet
Synthetic cannabinoids in hairâPrevalence of use in abstinence control programs for driver's license regranting in Germany
Although the use, structural variety, and prevalence of synthetic cannabinoids (SCs) have steadily increased on the drug market, they are rarely analyzed in abstinence control programs for driver's license regranting. The aim of this study was to determine the SC prevalence in these programs by analyzing hair samples collected between March 2020 and March 2021 from various regions in Germany, mainly Bavaria (40%). Specimens were analyzed quantitatively for drugs of abuse and qualitatively for 107 SCs. Hair samples were screened by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and to search for unknown SC analogs, positive samples were additionally screened by liquid chromatography-high resolution time of flight mass spectrometry (LC-qTOF/MS). The analysis of 5097 hair samples resulted in 181 SC detections (3.6%), showing a wide range of 44 SCs, with up to 13 different compounds found in a single sample. The most prevalent compounds were 5F-MDMB-PICA and MDMB-4en-PINACA; furthermore, 10 new substances not initially covered by LC-MS/MS analysis were detected by LC-qTOF/MS. The SC positivity rate was comparable to cocaine (5.4%) and amphetamine (2.6%). Only in 35 cases (0.7%), SC analysis was requested by the clients, highlighting the insufficient coverage of SC consumption in the studied collective. In summary, hair sample analysis proved to be a valuable tool to monitor the use of SCs. In order to keep pace with newly emerging SC analogs, an up-to-date analytical method is essential. Prospectively, SCs should be more routinely screened in hair analysis for abstinence control to avoid cannabis substitution by SCs.Synthetic cannabinoids (SCs) have steadily increased on the drug market but are rarely analyzed in abstinence control programs for driver's license regranting. Between March 2020 and March 2021, hair samples from this collective were retrospectively screened for SCs by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, and those testing positive were additionally screened by liquid chromatography-high resolution time of flight mass spectrometry (LC-qTOF/MS).imag