Journal of analytical toxicology最新文献

Tobacco cigarette smoking is the leading cause of preventable diseases and death in the USA. 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 US 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 and 0.004 ng/mL, respectively. These lower LODs would enable better detection of SHS in future NHANES surveys.

Oral fluid is considered a favorable matrix for the identification of drug intake mainly because of its simple, observed, non-invasive collection. Fentanyl and fentanyl analog use, misuse, overdose, and deaths are currently occurring at an alarming rate in the USA. The law enforcement community, the Food and Drug Administration (FDA) and the Centers for Disease Control (CDC) are all keenly aware of the urgency in addressing an unmet public health need to identify opioid overdose in individuals as rapidly as possible. As part of a National Institute of Justice grant, the present study was intended to develop and validate an environmentally friendly, rapid, sensitive quantitative method using liquid chromatography coupled to tandem mass spectral detection (LC-MS/MS) for fentanyl and fentanyl analogs in oral fluid collected using the nform rapid test device. Oral fluid samples were subjected to liquid-liquid extraction incorporating bio-renewable solvents where possible, reducing the environmental footprint of the assay. A buffer/salt free mobile phase was employed consisting of 0.1% formic acid in water (95%): 0.1% formic acid in methanol (5%) at a flow rate of 0.8 mL/min; the run time was 4.5 minutes, again reducing environmental impact in terms of salt and solvent usage. The method included fentanyl, 4-anilino-N-phenethylpiperidine; (4-ANPP; desproprionyl fentanyl), acetyl fentanyl, carfentanil, p-fluorofentanyl, valeryl fentanyl, p-fluorobutyrylfentanyl, furanyl fentanyl and benzoyl fentanyl as well as xylazine, which is often detected with fentanyl. The method was validated according to ANSI/ASB 036 (2019) Standard Practices for Method Validation in Forensic Toxicology.

Traditional immunoassay (IA)-based drug screens are limited in their scope of analysis and specificity. Their reliance on the cross-reactivity of antibody reagents is a limiting factor, particularly in light of the emergence of new therapeutics, emerging drugs, and new psychoactive substances (NPS). High resolution mass spectrometry (HRMS)-based techniques can offer improved versatility and specificity and increase the scope of analytical testing. In this study, a validated HRMS screening procedure was used to reanalyze adjudicated blood samples previously tested by immunoassay. IA methods employed enzyme-linked immunosorbent assay (ELISA) and enzyme multiplied immunotechnique (EMIT®). The comprehensive HRMS screen utilized supported liquid extraction (SLE) and liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Specimens previously tested by IA were reanalyzed using the HRMS screen following long term storage. The LC-QTOF-MS toxicology screen produced an additional 709 positive drug findings (67 compounds) among a population of 919 previously analyzed blood specimens. This study highlights the analytical benefits of MS-based toxicological screening and the advantages of data acquisition modalities that permit retrospective data interrogation.

This study evaluated the performance of the Immunalysis Tapentadol 343 Urine Enzyme Immunoassay (EIA) screening kit, focusing on the prevalence of false-positive results due to cross-reactivity with tramadol. Tapentadol is a dual-action analgesic, modulating μ-opioid receptors and inhibiting norepinephrine reuptake, while tramadol, a structurally related compound, is a weak μ-opioid receptor agonist and norepinephrine/serotonin reuptake inhibitor. Cross-reactivity between these compounds can complicate urine drug screening results for adherence monitoring in chronic pain management. A total of 28 samples initially produced false-positive results for tapentadol BNl using the Immunalysis Tapentadol 343 Urine EIA screening kit. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to confirm the absence of tapentadol. Of the false-positive samples, 61% contained tramadol at concentrations below the manufacturer-reported cross-reactivity threshold of 60 000 ng/mL, indicating assay limitations in specificity. To address this issue, a newly reformulated Immunalysis Tapentadol 343UR Urine HEIA kit was evaluated for tramadol cross-reactivity. Upon retesting the 28 false-positive samples with the reformulated kit, no false positives were detected, with results consistent with LC-MS/MS confirmation. The rate of false-positive tapentadol screens in urine has substantially reduced since the implementation of the new tapentadol kit in routine testing. These findings demonstrate the importance of assay verification to assess cross-reactivity, particularly for structurally related compounds. The reformulated Immunalysis Tapentadol 343UR kit shows improved specificity, reducing false-positive rates and enhancing the accuracy of tapentadol detection in clinical and forensic toxicology applications.

Hair analysis is a valuable tool in forensic toxicology, providing extended detection windows and critical insights into drug testing, usage trends, and drug-facilitated crimes. This systematic review was conducted using Scopus, Web of Science, and PubMed databases from March 2017 to September 2024, and evaluated 19 studies (16 research articles and 3 case reports) on the detection of γ-hydroxybutyrate (GHB) in hair. This review examines recent studies on GHB concentrations in hair, focusing on both endogenous and exogenous concentrations resulting from illicit and prescribed use, as well as the analytical methods employed. This review includes decontamination parameters, extraction techniques, and sample sizes used during the analytical method. New studies report that endogenous GHB levels range from 0.2 to 5.5 ng/mg, while exogenous levels vary widely from 0.3 to 239.6 ng/mg. Additionally, published results indicate that the frequency of use may be more significant than the dosage for exogenous GHB to be incorporated into the hair. A novel adjacent segmentation method has been proposed to differentiate endogenous from exogenous GHB, identifying local peaks within adjacent hair segments. Research into GHB-glucuronide as a biomarker has found it unreliable due to inconsistent correlations with exogenous use. Further research is needed to refine the interpretation of GHB levels in forensic applications.

N, N-Dimethyltryptamine (DMT) is a hallucinogen found in the South American Psychotria viridis plant and is the major psychoactive ingredient in the brew ayahuasca. In this report, we performed a review of the surrounding literature and detail two deaths which recently occurred in the UK following DMT use. A literature search of both academic (PubMed, GoogleScholar) and media (using Google search engine) publications was performed to identify previously reported fatalities following DMT use. The National Programme on Substance Use Mortality (NPSUM) was also searched for deaths which have occurred in the UK following DMT use. Literature search-There have been three previous reports of fatalities following DMT use, all deemed accidental in nature, with DMT consumption taking place as part of an ayahuasca ceremony in two of these cases. NPSUM cases-Two cases were identified (Case Report 1 [CR1] & Case Report 2 [CR2]), neither of which occurred in the context of an ayahuasca ceremony. DMT was detected and quantified in femoral blood in both cases (CR1 0.23 mg/l; CR2 0.24 mg/l). There was evidence of polydrug use in both cases (CR1 n = 6; CR2 n = 9), which in each case included additional compounds which can increase serotonergic drive (CR1 cocaine, amphetamine; CR2 venlafaxine, mirtazapine). There have been two recent deaths following DMT use in the UK, both in the context of polydrug use which may have caused death due to excessive serotonergic innervation leading to serotonin syndrome. Polydrug use is increasingly common in the UK, and users of unregulated drugs should caution their use in combination with other unregulated drugs and also any prescribed medications.

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.

Postmortem vitreous humor may be used for toxicological analysis if blood and urine are unavailable or where postmortem blood is thought to be affected by postmortem changes. Use of vitreous humor has been restricted by the available sample volume and instrument sensitivity. However, the advent of combined screening and quantitative methodologies using liquid chromatography-high-resolution mass spectrometry makes analysis of vitreous humor possible. This study examines an existing combined screening and quantitative methodology to determine if it is suitable for use with vitreous humor. Analysis of standard solutions containing 48 compounds showed % difference between expected and measured values in the range -15.59 to 20.81, -15.73 to 18.34, -14.32 to 19.77, and -19.90 to 19.78 for very low, low, mid and high range standard solutions respectively. Intraassay %CV was in the range 0.93 to 10.10, 1.35 to 15.19, 3.07 to 11.56, and 2.04 to 8.29 and interassay %CV was 0.96 to 17.40, 3.68 to 17.03, 3.94 to 17.12, and 4.87 to 16.55. Limits of quantitation range from 0.002 to 0.5 and limits of detection from 0.0008 to 0.06 mg/L. There was no significant interference from ion suppression or isobaric compounds and very little carryover. Dilution 1:2, 1:5 and 1:10 with vitreous humor gave acceptable results. Comparison of screening results from 129 postmortem cases showed that most compounds detected in blood and/or urine were also detected in vitreous humor. Compounds more readily detected in vitreous humor included 6-monoacetylmorphine, cocaine, codeine, dihydrocodeine, olanzapine, desmethylzopiclone, diazepam, cocaethylene, and desmethylmirtazapine. Compounds more readily identified in blood and/or urine included desmethylsertraline, EDDP, nordiazepam, papaverine, paracetamol, and morphine. The assay is suitable for screening and quantitation of drugs and their metabolites in vitreous humor and can be used where blood and urine are unavailable, or where the analysis of vitreous humor may provide useful information.

Drug overdoses are among the most common causes of death in the United States, with synthetic opioids such as fentanyl implicated in the majority of overdose fatalities in the last 10 years. As such, effective rapid assays capable of screening against large drugs of abuse panels that include synthetic opioids are critical tools for detecting drug abuse. The iScreen™ Urine Test FUO Drug Screen Cup (Abbott Laboratories) is a multiplexed lateral flow device designed for the preliminary qualitative screening drugs of abuse in urine for forensic applications, and can be used to simultaneously screen urine specimens for up to 17 different drugs of abuse, with multiple potential configurations of assays and cutoffs to support 22 different assay/cutoff combinations. This investigation focused on evaluating the performance of the iScreen™ Urine Test FUO Drug Screen Cup for analytical sensitivity, cross-reactivity characteristics for structurally-related compounds, and method comparison versus the gold standard method (GC-MS or LC-MS/MS). Analytical sensitivity testing demonstrated ≥95% accuracy for all 22 assays during evaluation against both negative and 3x cutoff positive control specimens, and all 22 assays achieved ≥89% concordance with the established reference methodologies in testing with positive and negative human urine specimens.