Detection of anabolic-androgenic steroids in e-cigarettes seized from prisons: A case study.

The administration of new psychoactive substances (NPS), in particular synthetic cannabinoid receptor agonists (SCRAs), via e-cigarettes, within prison settings has been well publicized. This study provides an overview of five e-cigarette case samples seized from Scottish prisons between May 2022 and July 2023 where the anabolic-androgenic steroids (AASs) mestanolone and oxandrolone were identified following gas chromatography-mass spectrometry (GC-MS) analysis. These e-cigarette samples represented 2.9% of all samples containing e-cigarette cartridges (n = 170) and 9.4% of all samples found to contain AASs (n = 53) seized during the same time period. The AASs were detected in combination with other drugs, including cocaine, Δ9-tetrahydrocannabinol (Δ9-THC), SCRAs and nicotine. This represents a new and novel route of administration for AASs.

Log D7.4 and plasma protein binding of synthetic cannabinoid receptor agonists and a comparison of experimental and predicted lipophilicity.

The emergence of new synthetic cannabinoid receptor agonists (SCRAs) onto the illicit drugs market continues to cause harm, and the overall availability of physicochemical and pharmacokinetic data for new psychoactive substances is lacking. The lipophilicity of 23 SCRAs and the plasma protein binding (PPB) of 11 SCRAs was determined. Lipophilicity was determined using a validated chromatographic hydrophobicity index (CHI) log D method; tested SCRAs showed moderate to high lipophilicity, with experimental log D7.4 ranging from 2.48 (AB-FUBINACA) to 4.95 (4F-ABUTINACA). These results were also compared to in silico predictions generated using seven commercially available software packages and online tools (Canvas; ChemDraw; Gastroplus; MoKa; PreADMET; SwissADME; and XlogP). Licenced, dedicated software packages provided more accurate lipophilicity predictions than those which were free or had prediction as a secondary function; however, the latter still provided competitive estimates in most cases. PPB of tested SCRAs, as determined by equilibrium dialysis, was in the upper range of the lipophilicity scale, ranging from 90.8% (ADB-BUTINACA) to 99.9% (BZO-HEXOXIZID). The high PPB of these drugs may contribute to reduced rate of clearance and extended durations of pharmacological effects compared to lesser-bound SCRAs. The presented data improve understanding of the behaviour of these drugs in the body. Ultimately, similar data and predictions may be used in the prediction of the structure and properties of drugs yet to emerge on the illicit market.

The metabolic profile of the synthetic cannabinoid receptor agonist ADB-HEXINACA using human hepatocytes, LC-QTOF-MS and synthesized reference standards.

Synthetic cannabinoid receptor agonists (SCRAs) remain a major public health concern, with their use implicated in intoxications and drug-related deaths worldwide. Increasing our systematic understanding of SCRA metabolism supports clinical and forensic toxicology casework, facilitating the timely identification of analytical targets for toxicological screening procedures and confirmatory analysis. This is particularly important as new SCRAs continue to emerge on the illicit drug market. In this work, the metabolism of ADB-HEXINACA (ADB-HINACA, N-[1-amino-3,3-dimethyl-1-oxobutan-2-yl]-1-hexyl-1H-indazole-3-carboxamide), which has increased in prevalence in the United Kingdom and other jurisdictions, was investigated using in vitro techniques. The (S)-enantiomer of ADB-HEXINACA was incubated with pooled human hepatocytes over 3 hours to identify unique and abundant metabolites using liquid chromatography-quadrupole time-of-flight mass spectrometry. In total, 16 metabolites were identified, resulting from mono-hydroxylation, di-hydroxylation, ketone formation (mono-hydroxylation then dehydrogenation), carboxylic acid formation, terminal amide hydrolysis, dihydrodiol formation, glucuronidation and combinations thereof. The majority of metabolism took place on the hexyl tail, forming ketone and mono-hydroxylated products. The major metabolite was the 5-oxo-hexyl product (M9), while the most significant mono-hydroxylation product was the 4-hydroxy-hexyl product (M8), both of which were confirmed by comparison to in-house synthesized reference standards. The 5-hydroxy-hexyl (M6) and 6-hydroxy-hexyl (M7) metabolites were not chromatographically resolved, and the 5-hydroxy-hexyl product was the second largest mono-hydroxylated metabolite. The structures of the terminal amide hydrolysis products without (M16, third largest metabolite) and with the 5-positioned ketone (M13) were also confirmed by comparison to synthesized reference standards, along with the 4-oxo-hexyl metabolite (M11). The 5-oxo-hexyl and 4-hydroxy-hexyl metabolites are suggested as biomarkers for ADB-HEXINACA consumption.

Extended Stability Evaluation of Selected Cathinones.

Understanding the stability of drugs in a forensic toxicology setting is critical for the evaluation of drug concentrations. Synthetic cathinones are new psychoactive substances structurally derived from cathinone, the psychoactive component of Catha edulis ("khat"), a shrub that is indigenous to the Middle East and East Africa. Previous research has evaluated the stability of synthetic cathinones in biological matrices, including blood preserved with the combination of NaF and K2C2O4 used in gray-top tubes. However, it does not assess their stability in blood preserved with Na2EDTA, used for some clinical samples. Further, stability in unpreserved urine samples was only studied for two weeks. This research evaluates the stabilities of four Schedule I synthetic cathinones: mephedrone, MDPV (3,4-methylenedioxypyrovalerone), naphyrone, and α-PVP (alpha-pyrrolidinopentiophenone) at 20°C (room temperature), 4°C (refrigerator), and -20°C (freezer). Stability was assessed in methanolic and acetonitrile solutions, as well as in Na2EDTA-preserved blood and unpreserved urine. Solutions (1 mg/L) of each drug in each matrix stored in aliquots (100 µL, solvents; 1.2 mL, biological samples; n = 12) at each of the three temperatures for triplicate analysis on days 3, 7, 14, and 30. On day 0 of each study, three additional aliquots of each solution were analyzed. Biological samples underwent solid-phase extraction before analysis. All samples were analyzed in full-scan by gas chromatography-mass spectrometry (GC-MS). The results of this study show that under room temperature and refrigerator storage conditions, mephedrone, naphyrone, and MDPV will degrade in methanol. This degradation starts are early as day 3. Additionally, all four drugs will degrade in Na2EDTA-preserved human whole blood samples in at least one evaluated storage environment. However, when in acetonitrile-based working solutions and unpreserved urine samples, they proved to be more stable. Methanolic working solutions and samples of Na2EDTA-preserved blood containing these cathinones should be stored in the freezer and used or tested with urgency to ensure that quantitative sample analysis is as accurate as possible in forensic casework.

Association between cannabis use and sexual behavior among adolescents aged 12-15 years in 21 low- and middle-income countries.

AIMS: To investigate the relationship between cannabis use and two sexual behaviors (ever had sex, multiple partners) in a large representative sample of adolescents aged 12-15 years from 21 low- and-middle income countries. METHODS: Data from 84,867 adolescents aged 12-15 years participating in the Global School-based Student Health Survey were analyzed. Participants reported lifetime frequency of cannabis use (analyzed as 0, 1-2, 3-19 or ≥ 20 times), whether they had ever had sexual intercourse (yes/no) and, if yes, their lifetime number of sexual partners. We used multivariable logistic regression to analyze associations, adjusting for a range of relevant covariates. RESULTS: 12.7% of the sample reported having had sexual intercourse, and of these adolescents, 53.1% had had multiple sexual partners. The prevalence of lifetime cannabis use of 1-2 times, 3-19 times, and ≥ 20 times were 1.1%, 1.2%, and 0.4%, respectively. Those who reported using cannabis 1-2 times, 3-9 times, and ≥ 20 times had 2.32 (95%CI = 1.47-3.65), 2.34 (95%CI = 1.34-4.07), and 5.45 (95%CI = 2.22-13.40) times higher odds of having had sexual intercourse than those who had never used cannabis. Among those who had ever had sexual intercourse, the respective odds ratios (95%CIs) for having multiple sexual partners were 1.56 (0.93-2.62), 1.70 (0.92-3.14), and 5.66 (2.97-10.82). There were no significant interactions by sex for these associations. CONCLUSIONS: Adolescents from LMIC who use cannabis are more likely to have ever had sexual intercourse than those who do not. Among those who have had sexual intercourse, those who use cannabis are more likely to have had multiple sexual partners.

Gas Chromatography-Mass Spectrometry Method for the Quantitative Identification of 23 New Psychoactive Substances in Blood and Urine.

New psychoactive substances (NPSs) have become an integral part of the recreational drug market with "new" compounds being reported by the European Monitoring Centre for Drugs and Drug Addiction weekly. Due to the changing nature of NPSs, it is impractical to carry out single analyte or even simple class quantitation. Although several gas chromatography-mass spectrometry (GC-MS) methods have been developed these are typically class specific. We present a validated GC-MS method for the quantitation of 2-DPMP, 3-MeO-PCE, 3-MeO-PCP, 5-APB, 6-APB, benzedrone, butylone, ethylone, flephedrone, methiopropamine, MDPV, mephedrone, methoxetamine, methylone, naphyrone, 25B-NBOME, 25C-NBOME, 25D-NBOMe, 25E-NBOME, 25H-NBOME, 25I-NBOME, Mescaline-NBOME and 25P-NBOME in blood and urine samples. Sample preparation was carried out using solid-phase extraction followed by derivatisation and analysis by GC-MS. Parameters investigated for validation included bias, precision, linear calibration model, carryover, interferences, limit of detection, limit of quantification, and autosampler and freeze/thaw stability. All drugs yielded successful results for each of these parameters as per SWGTOX guidelines. The GC-MS method was used for the reanalysis of 12 blood samples (eight cases) where 25I-NBOMe, 25C-NBOMe, methoxetamine and methylone had previously been detected by NMS laboratories. This GC-MS method was able to quantitatively detect these drugs in 75% of the blood samples, 42% of which contained either 25C-NBOMe or 25I-NBOMe. This method accurately allows for the simultaneous quantification of a wide variety of compounds via GC-MS, in particular NBOMe compounds which are typically analysed by liquid chromatography-tandem mass spectrometry which is not available in all laboratories.

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.