Journal of analytical toxicology最新文献

Hydroxyzine (Vistaril©) is an unscheduled, first-generation antihistamine prescribed for nausea/vomiting, atopic dermatitis or eczema, and anxiety. It can produce adverse central nervous system (CNS) depressant effects such as fatigue, sedation, and impaired memory and concentration. Several clinical studies have shown hydroxyzine is able to impair driving and psychomotor function; however, no case series have been published highlighting driving performance and roadside observations in driving under the influence of drugs investigations (DUID). Between January 2017 and October 2024, 319 blood specimens submitted for DUID testing confirmed positive for hydroxyzine. Mean and median blood concentrations were 70 ± 79 and 48 ng/mL, respectively, with a range of 8.0-600 ng/mL. More than half of hydroxyzine positive drivers were female (57%) with a mean age of 42 years. Common drug combinations in this study involved antidepressants (74%), opioids (44%), and anticonvulsants (38%). Only seven cases have been reported involving a single substance, and four of these investigations have been presented. Behavioral observations included incoordination, slow and slurred speech, and difficulty following instructions on standardized field sobriety tests. Driving observations included erratic driving, crashes, driving in opposite lanes of travel, and running stop signs. This is concerning since hydroxyzine became the most identified antihistamine in this population in 2023 outpacing diphenhydramine.

Suvorexant, lemborexant, and daridorexant are dual orexin receptor antagonists (DORAs) used to treat insomnia, offering a lower abuse potential due to their lack of gamma-aminobutyric acid activity compared to traditional sleep medications. Still, DORAs remain drugs of forensic interest due to their accessibility, long half-lives, and potential risk for next-day residual drowsiness, impaired motor coordination, and decreased alertness, which may feature prominently in cases of driving impairment or drug-facilitated sexual assault. Thus, developing analytical methods to detect these compounds, particularly the more novel lemborexant and daridorexant, is crucial for forensic toxicological testing. This study aimed to develop and validate an LC-MS/MS method for quantifying daridorexant, lemborexant, and suvorexant in blood. An acidic/neutral liquid-liquid extraction (LLE) using N-butyl chloride was optimized to isolate the three DORAs and the internal standard, suvorexant-d6, from bovine blood. All accuracy, precision, and key validation parameters met acceptability requirements per ANSI/ASB 036. LLE recovery was >94%, with the calibration range between 0.25 and 500 ng/mL for all analytes. The LLOQ was 0.25 ng/mL. Matrix effects were -54.2 to 75.7%. Bias ranged between -10.9% and 8.8%, while %CV was <17.7%. Two-fold dilution integrity studies yielded a bias <-7.6%, with %CV <7.6%. Exogenous/endogenous interferences were negligible. Re-injection of the blank following the highest calibrator was free from carryover. Extracts were stable beyond 48 h when stored at 4°C. A proof-of-concept study using authentic blood samples containing suvorexant, compared to previously reported concentrations, showed no consistent decrease over time, highlighting the need for further studies to determine optimal storage conditions for long-term stability. Although further studies are needed with authentic samples containing lemborexant or daridorexant, this validated method supports broader adoption in forensic toxicology, enhancing the detection and monitoring of these emerging sedative-hypnotics in forensic investigations.

Synthetic cannabinoid receptor agonists (SCRA) comprise a class of new psychoactive substances (NPS) that rank second in terms of notified substances to the United Nations Office on Drugs and Crime. Moreover, SCRA are the most prevalent NPS in Brazilian territory. Given the risks they pose to public health, there is a pressing need to develop simple and rapid sample preparation methods in alternative biological matrices that are easy to handle and collect, such as oral fluid (OF). In this study, dispersive liquid-liquid microextraction was employed to determine 12 SCRAs in OF. For 200 µl of the sample (mixture of OF and Quantisal™ buffer), 200 µl of ice-cold acetonitrile was used as the dispersive solvent, and 100 µl of ethyl acetate was used as the extraction solvent. The limits of detection ranged from 0.5 to 2 ng/mL, while the limits of quantification were 2 ng/mL for ADB-FUBIATA and 1 ng/mL for the other analytes. The working range was 1-100 ng/mL, except for ADB-FUBIATA, which had a range of 2-100 ng/mL. The coefficients of variation for quantified analytes were <11.3% for within-run precision, <12.6% for between-run precision, and <15.8% for accuracy across all controls. The developed method was applied to six suspected samples, and one sample yielded a positive result with 39.9 ng/mL of MDMB-4en-PINACA, the most prevalent SCRA in São Paulo State, Brazil.

Amatoxin-containing mushrooms, which contribute to many intoxications each year, are of particular interest for clinicians and toxicologists, as patients require special treatment in hospitals. To confirm the presence of amatoxins, approaches for their fast, sensitive, and reliable identification must be available. Solid-phase extraction followed by liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS) is widely applied for analysis of amatoxins as this combination provides suitable sensitivity, specificity, and mass accuracy. Nevertheless, time-consuming preparatory steps and expensive equipment are required. Therefore, a lateral flow immunoassay (LFIA) for the trace detection of α-, β-, and γ-amanitin was established and evaluated using dog urine. In this study, we answered the questions whether this LFIA can be transferred to human urine samples, and whether this LFIA can be used as a supporting tool prior to LC-HRMS/MS confirmation. Result interpretation by eye and using digitally acquired pixel intensity ratios was investigated with respect to analytical sensitivity. The LFIA detects amatoxins in human urine after visual evaluation to as little as 5 ng/mL (α-amanitin-10 ng/mL, β-amanitin-50 ng/mL, γ-amanitin-5 ng/mL). After digital analysis, pixel intensity ratios were determined to evaluate the LFIA as positive, negative, or trace result. Detection limits were redefined ranging from 1 ng/mL (α- and γ-amanitin) to 3 ng/mL (β-amanitin). For the proof-of-concept, 73 human urine samples submitted to the authors´ laboratory for toxicological analysis were analyzed using the LFIA and LC-HRMS/MS. Only three out of 73 urine samples were tested false positive with the LFIA as LC-HRMS/MS confirmation revealed no detection of amatoxins. Sixteen urine samples were evaluated as trace results and confirmed negative using LC-HRMS/MS except for one case which was positive for α-amanitin but negative for β-amanitin. Although particularly positive and trace results of the LFIA still need to be confirmed, the negative LFIA results correlated well with LC-HRMS/MS.

E-vapour products generate aerosols typically containing nicotine, flavour ingredients, and aerosol formers (propylene glycol and vegetable glycerine). While many flavour ingredients are 'generally recognized as safe (GRAS)' for oral use in the food industry, there exist knowledge gaps on their effects when delivered by the inhalation route. Due to the large number of available ingredients and potential combinations used to create e-liquids, toxicological and analytical evaluation of each flavour ingredient is impractical. Moreover, chemical characterization requires analytical methods to be developed and validated to measure key ingredients, as well as stability assessments to demonstrate that these test materials were stable during the testing period, which is equally challenging. In this study, we present a pragmatic approach of preparing 'preblends' prior to making a test formulation, containing 38 flavour ingredients as an example case, ahead of preclinical toxicity testing. We used the preblends to simplify the preparation and the characterization of test formulations, establishing the stability criteria for the subsequent toxicity testing. We prepared preblends by dividing the 38 flavour ingredients into five preblend groups based on structural moiety, solubility, and chemical reactivity. These preblends were mixed to make two different 'final' test formulations (containing all 38 flavour ingredients with and without nicotine). We evaluated the stability of the preblends and the two test formulations prior to the subsequent in vivo inhalation studies. Based on the analytical assessment, all the preblends were stable up to 4 weeks at 0°C-4°C. When all preblends were mixed, the test formulation was stable up to 3 days in the presence of nicotine and 10 days without nicotine when stored at 0°C-4°C. These stability results were used to set the frequency of preparing the test formulations for the in vivo inhalation studies, ensuring stability of test materials prior to the biological testing.

Stimulant use while driving is a high-risk factor for collisions and fatalities. In recent years, several strategies to curtail impaired driving were employed on highways, including on-site oral fluid testing. This study evaluated four roadside oral fluid testing devices (AquilaScan®, Dräger DrugTest®, Druglizer®, and DrugWipe®) for the detection of amphetamine, methamphetamine, and cocaine in oral fluid from Brazilian drivers. Overall, 8985 screening tests were conducted, and LC-MS-MS analysis was performed on 46% of the oral fluid samples. Screening and confirmatory test results were compared considering the manufacturers' and Substance Abuse and Mental Health Services Administration's recommended cutoff concentrations. Performance reliability data are available for well-known oral fluid screening devices such as the Dräger DrugTest® or Securetec DrugWipe®, but most evaluations were based on highly prevalent cannabinoid results. In many cases, there were insufficient data to evaluate performance of other drug classes, including reliability data for amphetamines and cocaine that are presented here. Approximately, 3.0% of samples screened positive for amphetamine, 0.9% for methamphetamine, and 2.6% for cocaine. Efficiency was higher than 93.9% for all devices, but other parameters varied considerably, with sensitivity 56.4-100% and positive predictive value 4.2-87.1%. When considering the recommended minimum of 80% performance criteria suggested by the Driving Under the Influence of Drugs, Alcohol, and Medicines study, the Dräger DrugTest® was the only device to achieve satisfactory sensitivity, specificity, and efficiency for these stimulants at multiple evaluated cutoffs. Given the observed variability between devices, a detailed evaluation of the analytical performance of roadside oral fluid testing devices is advised before implementation in traffic enforcement actions.

In this study, six single hairs, each measuring 9-11 cm in length, from a victim of a single criminal thallium (Tl) poisoning incident were analyzed in 0.3 cm segments applying a validated inductively coupled plasma mass spectrometry (ICP-MS) method. The results hold significant forensic value, particularly in cases involving limited sample materials. Despite the very low weight of 0.3-cm single hair (12-16 µg), we found that the sensitivity of ICP-MS is sufficient to carry out the section-by-section analysis even in one single hair for determining Tl in poisoning cases. The measured Tl concentrations in this case were determined to be in the range of 0.6-6.5 µg/g hair, which are 100-fold beyond what is normally found in the German population. The consistent decrease in concentration from proximal to distal segments could be interpreted by predominant Tl incorporation via sweat. This finding is discussed in comparison with previous studies on Tl hair concentrations and in relation to the toxic effects of thallium on hair growth and sweat gland function. Due to the low limit of detection of 0.008 µg Tl/g hair, we conclude that Tl poisoning can be detected by the analysis of single hairs in 0.3 cm segments. However, proving repeated or continuous exposure to the toxin may be challenging due to its incorporation from sweat.

Protonitazene is a highly potent synthetic opioid that has recently emerged in the illicit drug supply. Data on its metabolism and the detection of its metabolites in postmortem specimens are limited. This study retrospectively analyzed 14 postmortem cases to investigate metabolite profiles in blood and urine using high-resolution mass spectrometry. Detected metabolites included 5-aminoprotonitazene, N-desethylprotonitazene, and 4-hydroxynitazene. Additionally, a novel metabolite, hydroxyprotonitazene, previously only observed in liver microsome models, was identified. Significant variability in the metabolites detected across cases was observed, likely influenced by differences in metabolism, postmortem changes, sampling methods, and metabolite stability. This study underscores the challenges of detecting protonitazene exposure and highlights the necessity of targeting multiple metabolites, as no single marker reliably indicated protonitazene use. Further research is needed to confirm metabolite identities, evaluate their stability, and understand their role in protonitazene toxicity.