Journal of analytical toxicology最新文献

Transdermal drug delivery has been of particular interest to pharmaceutical research for decades, but is also becoming increasingly relevant in the field of sports drug testing. As shown in the past, the (unintentional) oral ingestion of trace amounts of prohibited selective androgen receptor modulators (SARMs), e.g. due to product contamination, can lead to an adverse analytical finding (AAF) in doping controls. Another site of exposure is presented by the skin, as it provides a large surface for drug penetration. However, the extent of diffusion through various layers of the skin and into the blood vessels depends, among other things, on the physicochemical and biological properties of a substance. The objective of this project was to simulate a transdermal contamination scenario and investigate the skin penetration and subsequent metabolism of microdoses of three commonly used SARMs: LGD-4033, RAD140, and S-23. For this purpose, an administration study was conducted, in which either 10 or 50 µg of the substances were applied to the lower forearm of 5 volunteers each. The collected urine samples were analyzed via LC-MS/MS following enzymatic hydrolysis and solid-phase extraction. This methodical approach is distinguished by its high sensitivity, enabling the detection of at least 5 pg/mL for LGD-4033 and S-23. After 10 µg administration, LGD-4033 and S-23 as well as associated metabolites were detected, while RAD140 was only detected in urine samples of one subject (n = 5). Following the application of 50 µg, RAD140 was detected in all subjects (n = 5) for up to 9 days, and additional metabolites of LGD-4033 and S-23 were identified. The long-term metabolite of LGD-4033 (M5b) was detected up to 12 days after the dermal administration of 10 µg, and up to 25 days after application of 50 µg, while S-23 was traceable for up to 16 respectively 24 days. It was demonstrated for all three SARMs that they penetrate the skin and may-even in trace amounts-produce AAFs when administered transdermally. Information on urinary concentrations and metabolism following transdermal administration of SARMs may assist in the interpretation of AAFs, particularly when dermal contamination or intentional doping via the skin is discussed.

Tobacco cigarette smoking is the leading cause of preventable diseases and death in the US. Exposure to secondhand smoke (SHS) can also cause heart disease, lung cancer, and respiratory illness. Cotinine (COT) and trans-3'-hydroxycotinine (HCT) are the primary metabolites of nicotine, the main addictive alkaloid in tobacco products. For many years, we have measured serum levels of COT and HCT in National Health and Nutritional Examination Survey (NHANES) participants to monitor exposure of the U.S. population to active smoking and SHS. As exposure to SHS is decreasing, a more sensitive analytical method is needed to detect the lower levels of these biomarkers for SHS assessment. We developed and validated a new automated method for the detection of COT and HCT in human serum. We implemented a new liquid handling automation system to aliquot and prepare samples using supported liquid extraction. Samples were analyzed by liquid chromatography-tandem mass spectrometry. The new automated sample preparation method increases sample throughput by reducing sample cleanup time to 2 hours for preparing a 96-well plate. The method has excellent sensitivity, specificity, precision (<10%), and accuracy (±15%). We were able to lower the estimated limit of detection (LOD) for COT by 33% and HCT by 73% from our previous LOD. The new LODs for COT and HCT are 0.010 ng/mL and 0.004 ng/mL, respectively. These lower LODs would enable better detection of SHS in future NHANES surveys.

Trazodone, a medicine registered for human, is a serotonin agonist-antagonist. At low dose, the drug is sedative due to its antagonist properties. At high dose, it is an agonist with anxiolytic and antidepressant actions. Trazodone can be administered to the horse to reduce anxiety. However, according to the antidoping rules for horses, the presence of trazodone in blood or urine is considered as a violation, which will produce a suspension of both the athlete and the horse as the drug is listed banned on the International Federation of Horseracing Authorities prohibited substances list. As a hair test can provide more evidence or supplementary information to an adverse analytical finding or to document drug exposure, our forensic laboratory received two specimens with a request for trazodone identification. After mane collection, trazodone was analysed by a new LC-MS/MS method involving pH 9.5 borate buffer overnight incubation of 20 mg of specimen in presence of clozapine-d4 used as internal standard, followed by solvents extraction. Linearity was verified from 1 to 100 pg/mg (R2 = 0.9967). Limit of detection of the method was 0.1 pg/mg. Trazodone was measured at 0.4 pg/mg in the mane of a horse suspended after an antidoping violation. In a case of hidden administration, trazodone was identified at 9 and 24 pg/mg in two consecutive mane hair segments. Although no controlled study allows interpretation, particularly about the frequency of exposure and the dose that entered in the body, this is the first evidence that trazodone can be incorporated in the mane of horses.

Synthetic cathinones (SCs) were confirmed as the second most prevalent class of New Psychoactive Substances (NPS) in 2024, underscoring their widespread availability and use. Notably, the SCs seizure and related intoxication cases increased within the European Union, involving mostly 3-chloromethcathinone, 3-methylmethcathinone, 2-methylmethcathinone, and N-ethylnorpentedrone. Italy has observed a distinct trend, with methylendioxy pyrrolidinohexanophenone (MDPHP) emerging as a significant concern. This technical note aims to provide a comparative analysis of the most recent data concerning the emergence and the spread of MDPHP in Italy. Data from the National Early Warning System on Drugs indicate that the total amount of seized MDPHP increased from 2021 to 2024. In 2023, MDPHP-related intoxication cases peaked at 47, considering either alone or in combination with other drugs, followed by a slight decrease in 2024. In Italy, MDPHP seems to dominate the Italian black market of SCs and with two fatalities reported. This increased demand for MDPHP raises health concerns, indicating the necessity to enhance drug-related harm reduction services and improve a multidisciplinary network that provides data for understanding the complex phenomenon of NPS.

4-Phosporyloxy-N, N-dimethyltryptamine (psilocybin) is a psychedelic tryptamine found in certain mushroom species that has shown efficacy in the treatment of various psychiatric disorders. In conjunction with the renewed interest in therapeutic effects of psychedelics, there has been an increase in psilocybin-like designer tryptamines appearing in non-medical drug markets. The present study aimed to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detecting and quantifying 4-position ring-substituted tryptamines and their 4-hydroxy metabolites in plasma. Specifically, we investigated 4-phosphoryloxy-N, N-dimethyltryptamine (psilocybin), 4-acetoxy-N, N-dimethyltryptamine (psilacetin), 4-propionoxy-N, N-dimethyltryptamine (4-Pro-DMT) and their shared metabolite 4-hydroxy-N, N-dimethyltryptamine (psilocin), along with 4-methyl carbonato-N, N-di-n-propyltryptamine (4-MeCO3-DPT) and its metabolite 4-hydroxy-N, N-di-n-propyltryptamine (4-HO-DPT). Mass spectrometry analysis employed electrospray ionization (ESI) in positive mode, with two multiple reaction monitoring (MRM) transitions per analyte. Plasma samples were acidified with ascorbic acid, followed by protein precipitation with acetonitrile. Linearity was achieved across a concentration range of 0.5-100 ng/mL for all analytes, except psilocybin, which displayed linearity from 5 to 100 ng/mL. Validation results demonstrated acceptable bias (±20%) and imprecision (<20%) for all analytes. Matrix effects, evaluated in 10 samples (CV <18.3%), indicated minimal interference, although ion enhancement was observed for psilocin (31.9%) and psilocybin (45.7%). Extraction efficiency across all tryptamines was approximately 50%. The assay method was used to quantitate plasma samples from male rats treated with 1.0 mg/kg s.c. of the prodrug psilacetin, and collected before and 5, 30, 60, 120 and 240 min after injection. No psilacetin was detected, and psilocin concentrations ranged from non-detected up to 32.7 ng/mL. Overall, we successfully developed a sensitive and specific method for the detection and quantification of six tryptamines in plasma, providing a robust tool for future research and clinical applications.

Postmortem alcohol production by microorganisms has been known to increase blood alcohol concentration, potentially leading to erroneous interpretation. Here, we present a peculiar case where postmortem toxicology detected a high ethanol concentration only in the stomach content. An 87-year-old bedridden woman was taken to senior day care facility, where the nurse attempted to take her vitals but noticed she was not breathing. The facility called for an ambulance, but she was pronounced dead soon after arriving at the hospital. She was found to be severely dehydrated, and her blood tests indicated poor health. Autopsy was performed 2 days after death, and postmortem alcohol analysis detected a high ethanol concentration only in the stomach content. Our aim was to investigate the causative agent of ethanol production; microbiological analysis identified the yeast species Candida glabrata as the responsible microorganism.

A drug-related fatality involving 3,4-methylenedioxy-α-pyrrolidinohexanophenone (MDPHP) is here reported. Belonging to the class of synthetic cathinones (SCs), MDPHP is a 3,4-methylenedioxy-derived designer (MDDs) drug with a pyrrolidine moiety and an alkyl portion with six carbon atoms. Other MDD pyrrolidine derivatives belong to the alkyl homologous series (C3-C5) and are known as 3,4-methylenedioxy-α-pyrrolidinopropiophenone (MDPPP), 3,4-methylenedioxy-α-pyrrolidinobutyrophenone (MDPBP) and 3,4-methylenedioxypyrovalerone (MDPV). MDDs are psychostimulant drugs of abuse that primarily act on monoamine transporters; little is known about their off-target liability. Recently, MDPHP has gained attention due to increasing seizures and involvement in human intoxications, but currently there is a lack of data about its pharmaco-toxicological effects. In the case reported here, a 58-year-old man with a history of MDPV addiction was found dead in a waterway. While no evidence of natural disease or trauma was found to account for the death, toxicological analysis revealed the presence of MDPHP in addition to MDPPP, MDPV, MDPBP, clonazepam, and citalopram. Since no standards of MDPPP and MDPBP were available at the time of the analysis, LC-QTOF analysis of the drugs and their metabolites were performed. The following concentrations of MDPHP were reported: 350 ng/mL in femoral blood (FB), 110 ng/mL in cardiac blood (CB), 1900 ng/mL in urine, 3000 ng/mL in bile, 490 ng/g in kidney, 80 ng/g in liver, 480 ng/g in lung, 98 ng/g in brain, 700 ng/mL in gastric content and 8 ng/mg in pubic hair. Other MDDs concentrations in biological fluids and tissue were significantly lower than MDPHP suggesting their presence as synthetic impurities. Finally, to better understand the binding properties of the abovementioned MDDs to several documented transporters and receptors, an in silico evaluation was performed. The medical examiner reported that the cause of death was an acute multidrug intoxication by MDPHP and clonazepam in presence of MDPPP, MDPV, MDPBP and citalopram.

A streamlined liquid chromatography quadrupole time-of-flight mass spectrometry method utilizing protein precipitation and filtration extraction was developed to achieve rapid and reliable screening and confirmation for blood and urine matrices. This method targets 946 drugs and metabolites across 35 drug classes via sequential window acquisition of all theoretical mass spectra with variable customized windows to enhance spectral clarity, and was validated per established guidelines to ensure high accuracy and reproducibility. Combined with complementary in-house methods, this approach meets and exceeds the testing requirements outlined in ANSI/ASB standards and recommendations for postmortem, drug-facilitated crime, and Tier I and II driving under the influence of drug analyses. The method demonstrated efficient and sensitive performance, achieving limits of detection as low as 0.1 ng/mL. It accurately identified expected detections across 67 proficiency test samples and 224 authentic case samples, with high accuracy and reliability in the detection of both traditional drugs and novel psychoactive substances. The method employs an in-house built library and incorporates in-batch standards analyzed alongside case samples to ensure contemporaneous identification criteria, making it suitable for confirmation and reporting purposes. By expanding the analytical capabilities to include a vast range of analytes, this method improves the likelihood of identifying substances that may otherwise go undetected and reduces the need for multiple separate tests, thereby enhancing the overall effectiveness of toxicological investigations.