Drug Testing and Analysis最新文献

New psychoactive substances (NPS), especially synthetic cannabinoid receptor agonists (SCRA), are increasingly smuggled into prisons via infused mail. Consumption of those substances by inmates in prisons is associated with increased aggression, violence, and organized crime. Onsite detection of infused mail often is challenging. Because the infused papers do not show any visible stains or olfactory changes, physical inspection is often insufficient. The applicability of further conventional on-site detection methods like immunoassays and sniffer dogs is severely limited. Because of the rapidly changing supply of already circulating and newly emerging NPS, it is impractical to impossible to keep up with the development of immunoassays or the training of sniffer dogs. Hence, confiscated mail samples are routinely tested by either liquid chromatography or gas chromatography coupled to mass spectrometry (MS), which is costly and time-consuming. In this study, recent advancements in the hyphenation of laser ablation (LA) and molecular MS were investigated regarding the possible application for the rapid and easy detection of NPS in prison mail. Utilizing an in-house developed LA-MS hyphenation based on atmospheric pressure chemical ionization (APCI), 31 mail samples confiscated in German prisons were analyzed. It was possible to correctly identify 27 samples containing SCRAs. For these positive samples, it was also possible to detect the specific compounds each paper was infused with. The use of LA-APCI-MS has simplified sample preparation and reduced analysis time per sample to 1 min.

Hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs) represent a novel class of therapeutic substances that increase erythropoiesis. Due to their performance-enhancing effects and potential risk of abuse, these agents were added to the World Anti-Doping Agency (WADA) Prohibited List in 2011. Enarodustat is a novel HIF-PHI and has been approved for clinical use in China in 2023. This study primarily aimed to characterize its major urinary metabolites for antidoping purposes. A single oral dose of 40-mg enarodustat was administered to a volunteer. Urine samples were collected over 28 days and processed using solid-phase extraction (SPE). Analytical methods included liquid chromatography-high resolution mass spectrometry (LC-HRMS) under both positive and negative electrospray ionization conditions, complemented by in vitro metabolism studies using human liver microsomes (HLMs) for the characterization and identification of metabolites. A total of eight metabolites were detected, including Phase I products such as parent compound (PC) isomer (M1), monohydroxylation (M2), dihydroxylation (M3), and dehydrogenation (M4) metabolites, as well as Phase II conjugates involving methylation (M5), glycosylation (M6), glucuronidation (M7), and monohydroxylation-sulfation (M8) in vivo. Among these, M3-M5 are novel metabolites. In addition, compared with other metabolites, PC and M2 exhibited longer detection windows, suggesting they are valuable biomarkers for doping control purposes. The study elucidates enarodustat's metabolic pathways and provides a foundation for developing sensitive detection methods. Future work should focus on synthesizing reference materials to identify metabolite structures.

Phenibut is a new psychoactive substance (NPS) first synthesized in Russia in 1963 as a derivative of gamma-aminobutyric acid. Originally developed for therapeutic use, it has gained popularity for nonmedical purposes, including recreational and cognitive enhancement. In Brazil, phenibut is uncontrolled and easily purchased online. This study used wastewater-based epidemiology (WBE) to investigate phenibut use patterns in two northeastern Brazilian cities. Composite daily wastewater samples were collected from two treatment plants (WWTPs), Recife (WWTP_A) and Olinda (WWTP_B), during two periods in 2023: Carnival and a reference week. Samples underwent solid-phase extraction (SPE) and analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Phenibut concentrations were converted to population-normalized mass loads (PNMLs, mg/day/1000 inhabitants). The highest phenibut levels and PNMLs (up to 4.06 mg/day/1000 inhabitants) occurred during Carnival at WWTP_A, located in a major tourist area, suggesting recreational use. During the reference week, PNMLs ranged from detection limits to 2.29 mg/day/1000 inhabitants on weekdays, indicating possible functional or cognitive enhancement use. These findings reveal two distinct use patterns: recreational peaks during Carnival weekends and possible functional use on weekdays outside festive periods. This is the first evidence of phenibut detection in Brazilian wastewater and its temporal use patterns. The results highlight WBE's value in monitoring NPS trends and suggest recreational use predominates during large events. This underscores the need for public health attention and regulatory monitoring of uncontrolled substances with abuse potential.

Gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) is the method of choice to detect the abuse of synthetic forms of naturally occurring steroids for doping control purposes. GC-C-IRMS relies on a multistep sample cleanup to ensure each target analyte peak is chromatographically pure before combustion. To achieve that, liquid-liquid or solid phase extraction (SPE) is commonly used in combination with preparative liquid chromatography (LC). In this work, a procedure for isolation, purification, and analysis of endogenous steroids by GC-C-IRMS was developed and validated. The key elements were successive application of strong cation and strong anion exchange SPE with enzymatic hydrolysis in between to strip the ionic species from urine and decrease matrix complexity prior to LC cleanup; preparative two-dimensional LC, where only the testosterone fraction required secondary purification (40 min total run time per sample); and derivatization of selected fractions with formic acid to yield formate esters, followed by GC-C-IRMS analysis. Formylation afforded excellent separation between 5α- and 5β-androstanediols and simplified the detection of the endogenous reference compound pregnanetriol by converting it to a volatile artifact, tentatively identified as 3α,20-diformoxy-17-methyl-18-nor-5β,17α-pregn-13-ene. The overall method performance benefited from the customization of the GC-C-IRMS combustion interface, which improved robustness and facilitated the transfer of sample components into the hot zone of the oxidation reactor, minimizing peak tailing. The former was achieved by keeping the oxygen flow through the reactor at all times, obviating the need for periodic oxidation, and the latter-by implementing a direct capillary-in-capillary connection of the chromatographic column to the oxidation reactor.

Cannabis sativa is one of the oldest and most versatile plants with many facets ranging from intoxicant to medicine. Legalisation of medicinal cannabis leads to an increasing complexity of specific forensic questions to distinguish between recreational and medicinal use, for example, in context with participation in road traffic. Hence, there is a recent interest in finding objective markers that enable the differentiability of cannabis flowers. Terpenes, volatile hydrocarbons with a modular construction principle of isoprene subunits, are currently suggested as a second substance class alongside phytocannabinoids for the classification of cannabis material. A headspace full evaporation technique gas chromatography mass spectrometry (HS-FET-GC/MS) methodology was successfully validated according to forensic guidelines for the analysis of 45 terpenes in cannabis flowers including 16 monoterpenes, 16 monoterpenoids, 7 sesquiterpenes and 6 sesquiterpenoids. FET-sampling was developed in detail experimentally, revealing evidence of thermal instability of higher-boiling terpenes. Validation included selectivity, linearity of calibration (ranges 10-2000 μg/g), analytical limits (at least 6 μg/g), accuracy (bias) as well as intraday and interday precision. The use of a retention time index mixture as an internal standard and measurement in SIM-scan mode also allows for the qualitative identification of further terpenes present in cannabis. Application to a set of cannabis strains with similar Δ⁹-THC content demonstrated differences and similarities in their terpene profiles.

Using a targeted metabolic investigation approach, a new, previously undescribed metabolite, which is a pyrrole derivative of LGD-4033, has been detected and coded as M8. This metabolite can be detected in postadministration human urine samples up to 6 days after administration. It has also been detected in post-administration samples, mimicking supplement contamination, after repeated 10 μg doses detectable for ≥ 120 h after administration. Given M8's structural similarity to LGD-4033, its androgen receptor (AR) agonist/antagonist properties were studied using in silico molecular docking and functional in vitro AR transactivation assays in the PC3(AR)₂ cell model, alongside other selected LGD-4033 metabolites. The results indicate that M8 can act as a potent AR antagonist, whereas M2c was reconfirmed as a potent AR agonist. Therefore, we propose the inclusion of M2c in ITP doping control methods, as it could be used as an LGD-4033 alternative and may be introduced into the black market. Additionally, the detection of M8, which is an early-stage excreted metabolite, is valuable for estimating sample collection time relative to LGD-4033 intake. When combined with the evaluation of other long-term metabolites like M5b, M5a, M2c, and M2d, M8 detection can aid in distinguishing adverse analytical findings, associated abuse through regular dosing, from unintentional doping caused by certain contamination scenarios or abuse through microdosing.

This paper describes the detection of nandrolone decanoate and its metabolites in mane hair collected from horses that have been treated with nandrolone decanoate (Deca-Durabolin). The intramuscular administration study of nandrolone decanoate in three Thoroughbred castrated horses (each received 800 mg weekly for 3 consecutive weeks) was previously conducted to investigate its metabolism and detection time in plasma and urine for doping control purposes. In this work, segmental analysis of the post-administration hair has revealed that (i) nandrolone decanoate and its metabolites, nandrolone and 4-estrene-3,17-dione, could be detected in horse mane after intramuscular administration, and (ii) the spreading of these substances along the entire hair strand was observed, which suggested the involvement of sweat or sebum incorporation.

The racehorse industry has strict regulations regarding the detection of prohibited substances in horses. Metformin, a diabetes medication, is a prohibited substance that has been reported in post-race blood and urine samples collected from racehorses. For further characterization of the disposition of metformin, 12 Thoroughbred horses were administered metformin orally and intravenously in a randomized, balanced, two-way crossover design. Serum and urine samples were collected, and drug concentrations determined via liquid chromatography-tandem mass spectrometry. The serum data were analyzed using both noncompartmental analysis and a population pharmacokinetic model. Metformin concentrations were below the LOQ (0.25 ng/mL) in six of 12 horses by Day 11 postadministration, and below the LOQ for all horses on Day 25. The maximum serum concentration of metformin (mean ± SD) was 941.0 ± 467.8 ng/mL, and the mean terminal t_1/2 was 85.8 ± 15.1 h. Based on Monte Carlo simulations the time that serum metformin concentrations fell below the proposed HISA Anti-Doping and Medication Control (ADMC) minimum reporting level (MRL; 0.5 ng/mL) in a simulated population of 1000 Thoroughbred horses was 475 hrs (~20 days). Metformin concentrations in urine fluctuated significantly between and within individual horses, and there was not a consistent relationship between blood and urine samples across time points. Results of the present study demonstrate a prolonged detection time; thus, a prolonged withdrawal time is needed to prevent a positive finding following exposure to metformin. Additionally, these results suggest that blood is the preferred matrix for regulatory purposes due to the inconsistent elimination in urine.

This study aimed to explore how PEth and other commonly used alcohol biomarkers (CDT, AST, ALT, and GGT) respond to regular consumption of what has been generally considered to correspond to low to moderate amounts of alcohol over a 2-week period. A total of 21 voluntary participants (aged 31-69 years) took part in a 2-week drinking study. Group 1 (n = 11) consumed one glass of wine daily (16 g of alcohol), close to the present Swedish limit for hazardous alcohol consumption, while Group 2 (n = 10) consumed two glasses daily (32 g of alcohol). Alcohol biomarkers were measured at baseline and at three further occasions during the study. After 1 week of alcohol consumption, all participants had measurable concentrations (> 0.005 μmol/L, ≈3.5 ng/mL) of both PEth-homologues (PEth 16:0/18:1 and PEth 16:0/18:2). After 1- and 2-week periods, significant differences in PEth levels were observed between Group 1 and Group 2. The correlation between the two PEth-homologues was strong and increased as the study progressed. In contrast, other biomarkers showed little to no change during the study period. Both PEth-homologues appear capable of identifying hazardous alcohol consumption. The current Swedish reporting threshold for PEth 16:0/18:1 (0.05 μmol/L, ≈35 ng/mL) demonstrates high specificity but low sensitivity in identifying hazardous alcohol consumption involving regular/daily intake. The sensitivity of the other biomarkers is insufficient for detecting alcohol consumption at this level.

Although incineration is currently the primary method for the disposal of seized illicit drugs, alternative methods for the disposal of illicit drugs may be necessary to provide safer and more accessible alternatives. Chemical oxidation processes have been identified as a promising alternative method to degrade illicit drugs. Using commercially available reagents and established industry processes, chemical degradation holds potential as an alternative drug disposal technique. This study investigated the oxidants ozone, sodium hypochlorite, trichloroisocyanuric acid, hydrogen peroxide, OXONE, sodium percarbonate, and peracetic acid for their potential to degrade illicit amphetamine-type stimulants using β-phenethylamine (PEA) as an exploratory analog. Oxidants and conditions that showed the highest degradation efficiency with PEA were applied to methamphetamine, amphetamine, 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyamphetamine (MDA). Transformation products were identified using gas chromatography-mass spectrometry, and degradation was quantified using a fit-for-purpose method via liquid-chromatography quadrupole time-of-flight mass spectrometry. Of the oxidants explored, ozone performed the best, leading to high degradation efficiencies of methamphetamine (95%), amphetamine (86%), MDMA (100%), and MDA (100%) after 72 h of exposure. Sodium hypochlorite was also highly effective for the degradation of methamphetamine and amphetamine, while trichloroisocyanuric acid was particularly effective for MDMA and MDA. All the major transformation products of degradation were tentatively identified, with only one of 10 listed as a controlled, scheduled, or restricted substance. This research demonstrates how chemical degradation can provide a novel alternative to incineration for the destruction of amphetamine-type stimulants, providing a sustainable, long-term, and accessible method of illicit drug disposal.