Journal of analytical toxicology最新文献

We report a case of systemic organophosphate (OP) poisoning in a young child following the use of a household shampoo containing diazinon. In previous reported cases, diagnosis is typically made rapidly and indirectly through decreased cholinesterase activity and identification of OPs in commercial containers. In our case, however, the diagnosis was significantly delayed due to nonspecific clinical signs and a language barrier between the family and medical staff, resulting in the development of an intermediate syndrome. Initial symptoms included gastrointestinal disturbances, followed several hours later by neurological and respiratory complications. Plasma, urinary and hair arsenic levels were first investigated due to the use of a product which may contain organometallic copper acetoarsenite complex. All arsenic concentrations were within physiological concentrations. Clinical presentation and biological cholinesterase activity (313 U/L vs. reference 1,900-3,800 U/L) later supported the diagnosis of OP poisoning. This hypothesis was reinforced by the analysis of the household shampoo indicating the presence of diazinon. Furthermore, despite the delayed diagnosis, non-targeted high-resolution mass spectrometry (LC-HR-MS/MS) identified diazinon in plasma and multiple phase I metabolites, including the specific marker IMPY, in both plasma and urine. Four previously undescribed metabolites and one phase II metabolite, hydroxy-IMPY-glucuronide, were also detected. Pralidoxime methylsulfate (Contrathion™) was administered on day 4, following toxicological confirmation and lack of clinical improvement. Although late, this treatment led to gradual neurological recovery, suggesting possible slow "aging" of diazinon-inhibited acetylcholinesterase. Nonetheless, the child developed persistent sequelae including hypotonia, peripheral neuropathies, and respiratory dysfunction consistent with intermediate syndrome. This case highlights the importance of considering OP poisoning even in atypical presentations and illustrates the utility of non-targeted HRMS screening in providing direct evidence of exposure when conventional approaches are inconclusive.

Semi-synthetic cannabinoids are a class of new psychoactive substances (NPSs) with structural similarities to the main psychoactive phytocannabinoid Δ9-tetrahydrocannabinol (Δ9-THC) found in Cannabis sativa L. The first semi-synthetic cannabinoids, which were used as legal substitutes for marijuana were Δ8-tetrahydrocannabinol (Δ8-THC) and hexahydrocannabinol (HHC). Δ8-THC emerged around 2019 on the recreational drug market in the United States after it became legal due to an ambiguity in the Agricultural Improvement Act 2018 (Farm Bill 2018). It was never legal outside the United States as the isomers of THC are regulated in the United Single Convention on Narcotic Drugs from 1971. HHC, a hydrogenated derivative of THC, followed as a legal substitute on the European recreational drug market. Many countries already placed HHC in their narcotic substance law, which lead to the emergence of other structurally related derivatives of THC. An existing rapid screening method for the qualitative analysis of various new psychoactive substances was expanded for semi-synthetic cannabinoids in whole blood using a LC-QTOF-MS system. This method was validated for 24 different phytocannabinoids and semi-synthetic cannabinoids in blood. Recovery rates of the analytes from a liquid-liquid-extraction ranged from 87-118%, matrix effects ranged from 24-93%, and limits of detection (LOD) ranged from 0.8-16 ng/mL.

In case of standard biological postmortem matrices such as blood or urine being unavailable, vitreous humor has been shown to be a versatile matrix used as alternative for postmortem systematic toxicological analysis for the detection of drugs and their metabolites. This study focused on the qualitative detection of drugs in vitreous humor using an untargeted metabolite based LC-M/MS screening approach. Vitreous humor samples were retrospectively analyzed by LC-MS/MS after being worked-up by protein precipitation. In n = 96 samples 418 detections were recorded. Those corresponded to 121 different xenobiotics. Cardiovascular drugs being the most frequently detected drug class followed by (local) anesthetics, neuroleptics, and antidepressants. Drug exposure was indicated 227 times (∼ 54 %) solely by the presence of the parent compound. One hundred thirty seven times (∼ 32 %) a parent compound and corresponding metabolite(s) were detected. Fifty four times (∼ 13 %) a drug exposure was indicated solely by the detection of metabolites. Out of those 54 detections, 32 times corresponding metabolites were not common commercially available. Therefore, the authors suggest that metabolite information should be considered for a vitreous humor based screening approach. Screening for metabolites will lead to ∼13 % more detections, if the corresponding metabolites are covered by the applied screening approach. The obtained results pointed out, that some compounds are exclusively found in form of their corresponding phase II metabolites. No linkage between protein binding or molecular weight was found for the detection of a compound in vitreous humor either by parent compound or metabolite(s).

In hair drug testing for cocaine, the presence of hydroxycocaine metabolites has been suggested as a means of distinguishing between externally-contaminated hair and positive hair due to ingestion of drug. However, hydroxy compounds can also be produced by the action of certain hair products containing peroxides or other reactive chemicals. In this study, hair samples contaminated with cocaine, benzoylecgonine, methamphetamine, and amphetamine were treated with three hair products: a relaxer containing calcium hydroxide, bleaching colorant containing hydrogen and ammonium hydroxide, and a dye containing hydrogen peroxide to determine the effect of the chemical agents on the drug compounds. Authentic positive hair from people who used drugs were treated with the same hair products. Analysis of the contaminated hair showed that the relaxer removed most of the contaminating drugs with trace production of hydroxy compounds. The peroxide-containing hair products resulted in less reduction in parent drug concentrations and higher concentrations of hydroxy compounds. Analysis of hydroxy to parent compound ratios, specifically for para- and meta- hydroxycocaine, showed that no samples produced ratios at 0.05% or higher-a ratio that has been suggested as indicating drug ingestion. With the authentic drug-positive hair, treatment with the products reduced parent and metabolite concentrations while not affecting hydroxy metabolite to parent ratios.

C6-keto-opioids, such as hydrocodone, hydromorphone, oxycodone, and oxymorphone, are a group of semi-synthetic morphine-like analgesics with extensive applications in clinical settings and high potential for abuse and misuse. Therefore, they have become targets of workplace forensic urine drug testing (UDT) for years. Due to the undesired C6-C7 keto-enol tautomerization, the C6 ketone often needs to be deactivated prior to further derivatization for GC-MS analysis. Although it has been over two decades since the method of converting the C6 ketone to its methoxime-derivative was initially reported, little information has been published regarding the resulting Z/E-methoxime-derivative isomers' formation mechanism, stereo-configurations, or relative kinetic or thermodynamic features. Mixed Z/E-methoxime-derivative isomers create a potential peak resolution issue for GC-MS-based C6-keto-opioids identification and quantification, since the two isomers are often difficult to be completely separated by GC and they share common fragmentation pathways. We here provided the first detailed report and qualitative conformational analyses of the Z/E-methoxime-derivative isomers of C6-keto-opioids and their isomerization under the non-aqueous Brønsted-Lowry acidic conditions. By in-depth studying the C6-keto-opioids Z/E-methoxime-derivative isomers, we were able to gain important insights into potential reaction condition optimization with an attempt to reduce the formation of the minor methoxime-derivative isomer, thus, to minimize the potential interferences caused by co-existing of the two isomers and further improve the method's limit of detection (LOD) and/or limit of quantification (LOQ). Our report offered valuable information that could facilitate other laboratories using the similar derivatization procedures for GS-MS-based C6-keto-oipoids testing to improve their testing method sensitivity and enhance their analysis product quality.

In some cases, the determination of carboxyhemoglobin (COHb) concentration in postmortem blood using standard spectrophotometric methods, may be either impossible or yield misleading results. In forensic laboratories, there is an increasing need to analyze COHb saturation in autopsy blood samples, the quality of which is often compromised by various factors. Therefore, a gas chromatographic method has been developed and validated to confirm the results of an established spectrophotometric method and to determine the concentration of COHb in postmortem samples affected by processes such as putrefaction or thermocoagulation. This method is based on automated headspace analysis of a blood sample treated with a solution of potassium hexacyanoferrate and saponin. It employs a combination of two capillary columns for chromatographic separation, followed by a highly sensitive detection system that incorporates a methanizer to convert carbon monoxide (CO) to methane couple to a flame ionization detector (FID). The percentage of COHb concentration is calculated using the ratio of the analyzed blood to that of fully CO-saturated blood. Careful optimization of the method has reduced the sample requirement to 100 mg while maintaining high sensitivity. Additionally, the sample pretreatment process has been simplified to ensure that the method can be easily integrated into the routine operation of a conventional forensic laboratory. The method demonstrates linearity across a concentration range of 0.1% to 100% COHb. The determined values of LOD (< 0.01% COHb) and LOQ (0.1% COHb) reflect excellent sensitivity. Overall, the validation study confirmed the method for its intended purpose. The method is particularly effective in reliably determining COHb content in putrefied and similarly degraded samples. Its robustness has been further validated through the analysis of dozens of real samples over more than four years.

Marijuana (cannabis) is generally considered the most frequently misused substance during pregnancy. The prevalence in the use of either medical or non-medical marijuana for relief of pregnancy-related symptoms is increasing, as well as the use of cannabis-related products containing cannabidiol (CBD) and semi-synthetic cannabinoids (SSCs). Δ9-tetrahydrocannabinol (THC) and CBD are highly lipophilic substances and will readily pass into breastmilk upon ingestion. The solubility of THC and CBD in lipids poses significant analytical challenges in extracting and identifying these substances in breastmilk. The aim of this study was to develop a new and sensitive assay utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the detection of cannabinoids in breastmilk. The method was optimized to quantitate Δ8-THC, Δ9-THC, cannabigerol (CBG), CBD, and cannabidiolic acid (CBDA) and validated with the guidance of the American Academy of Forensic Sciences Standards Board (ASB) Standard 036. The assay was then used to analyze breastmilk samples (N = 57) collected postpartum from female patients enrolled in a study assessing use behaviors of medical marijuana, non-medical marijuana, and CBD. All analytes passed validation criteria. Calibration curves for all analytes ranged 0.5-400 ng/mL, with the LOD and LLOQ of the method set at the lowest calibrator concentration. Δ9-THC was quantitated in 19 samples (33.3%) with a concentration range of 0.5-291 ng/mL. Δ8-THC was detected in one sample (1.8%) at 0.8 ng/mL, while CBD was observed in 3 samples at a concentration

Advancing knowledge of endocannabinoid receptor agonists and the federal legalization of hemp has created a cannabinoid market of naturally abundant phytocannabinoids to a wide array of semi-synthetic and synthetic cannabinoid analogs. Public safety and toxicological concerns exist from lack of regulation, limited pharmacological and metabolomic data, and minimal knowledge of detection ability. Structural similarities of the cannabinoid analogs may allow detection on immunoassays including enzyme-linked immunosorbent assays (ELISA) and homogenous enzyme immunoassays (HEIA), screening platforms in forensic toxicology laboratories for rapid presumptive testing. The cross-reactivity of 27 cannabinoid analogs and 26 commercially available metabolites was evaluated using the Medica EasyRA Enzymatic Immunoassay Analyzer with the Immunalysis Cannabinoids (THC) and Synthetic Cannabinoids 1-3 kits. These analogs were also evaluated using the Dynex DSX Automated ELISA System with the OraSure Technologies Cannabinoids Intercept Microplate EIA. The cannabinoid kits target 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (Δ9-THCCOOH) at a 50 ng/mL cutoff, and the synthetic cannabinoid kits target the N-pentanoic acid metabolite of JWH-018, UR-144, and AB-PINACA at a 10 ng/mL cutoff. Cross-reactivity was evaluated at concentrations of 20, 50, 100, 500, and 1000 ng/mL in urine in triplicate. Absence of cross-reactivity at 1000 ng/mL was considered undetectable. No cross-reactivity was detected on the synthetic cannabinoid kits. Cross-reactivity to Δ9-THCCOOH kits was variable with Δ8-THCCOOH and R-HHCCOOH cross-reacting at the cutoff on the ELISA, with several additional phase I metabolites cross-reacting at 100 ng/mL on both platforms. Analogs lacking the Δ9-THC tricyclic structure and pyran ring cyclization including cannabidiol were undetectable. Alicyclic bond location and alkyl chain length variably affected cross-reactivity, with alkyl lengths 2-4 having increased cross-reactivity comparatively. Compound chirality was also observed to effect instrumental response, with the ELISA having increased cross-reactivity and instrumental response to R-isomers. As knowledge and prevalence of analogs increases, it is crucial to understand the impact on utilized testing platforms.

In 2021, the U.S. Bureau of Justice Statistics (BJS) published results for the 2018 Census of Medical Examiner and Coroner Offices (CMEC) that provided an update on the medicolegal death investigation system in the USA. The 2018 Census collected data regarding toxicology service provisions, staffing, infrastructure, and practices, some of which were not included in the 2021 published BJS report from more than 1600 responding medical examiner/coroner offices (MECs). The 2018 CMEC was conducted from June 2019 through March 2020 by mail, online, and email. Toxicology-related CMEC data were obtained from BJS's publicly accessible dataset and evaluated in this study. Results from this study include information on toxicology service capability across MECs, including the number and salary of forensic toxicologists, toxicology retention time schedules, laboratory accreditation, professional certification, drug screening practices at the death scene, and whether they request confirmation testing. Overall, internal capabilities for toxicology testing were rare in 2018, with only 78 MECs (5.9%) reporting this function. Large MECs, serving a population of 250 000 or more, comprised about 15% of MECs that responded to the toxicology testing questions, with the rest being evenly divided between MECs that serve small (<25 000) and medium-sized (25 000-249 999) populations. Overall, 57.4% (n = 761) of MECs indicated that their forensic toxicology testing strategy has changed because of the increase in drug-related deaths, 53.9% of MECs (n = 715) perform drug screening tests, and 95.1% (n = 674) confirmed these tests with laboratory toxicology testing. Less than half of MECs reported that they had a toxicology specimen retention schedule (45.3%) or a computerized case management system (44.8%). These data are key to understanding (i) postmortem toxicology policies and practices, (ii) how these practices have evolved, (iii) MEC infrastructure, and (iv) the national importance of these data considering the ongoing drug overdose crisis.

Toxicology testing is an integral component of postmortem, drug-facilitated crime, and driving under the influence investigations. Recommendations pertaining to traditional matrices, sample amounts, and collection container types are well documented in the literature and guidance documents. However, not all cases have traditional toxicological specimens available (e.g. blood with a fluoride additive), and thus require nontraditional toxicology test options. In these cases, a forensic laboratory is contacted to determine if nontraditional objects, such as clothing, bedding, automotive, personal hygiene, or household items, stained with biological material, are suitable for analysis. Comprehensive method validation, as required for routine toxicology tests, is not practical to complete for these items, but this should not deter the toxicology laboratory from taking on this work. Herein, we describe a developed and implemented process for qualitative analysis of biological fluids on/in objects, which ensures the robustness and reliability of reported results. The specific procedures used, which include sample preparation, the incorporation of specialized quality control samples made from the items themselves, analytical acceptance criteria, and reporting considerations are thoroughly detailed. Positive findings from cases were obtained for a variety of drugs, encompassing illicit, prescription, novel psychoactive substances, and over-the-counter medications. Some examples include identification of zolpidem from vomit on clothing; cocaine, cocaine metabolites, levamisole, codeine, acetaminophen, and caffeine in stains on bedding; and diphenhydramine, doxylamine, and dextromethorphan in stains on a mattress pad cover. This methodology is fit-for-purpose and suitable for the toxicological investigation of these unique specimens without any significant limitations. This testing process may be used to identify past drug exposure, associate drug exposure to a particular location or scene, and/or provide insight into an event when a missing person has not been found.