Journal of analytical toxicology最新文献

Synthetic cannabinoid receptor agonists (SCRAs) 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, SCRAs 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 (DLLME) was employed to determine twelve SCRAs in OF. For 200 µL of sample (mixture of OF and Quantisal™ buffer), 200 µL of ice-cold acetonitrile were used as the dispersive solvent, and 100 µL of ethyl acetate were 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.

The continuous emergence of new psychoactive substances on the illicit drug market provides challenges for forensic and clinical analytics. Reliable detection of a previous ingestion of these drugs in human urine samples requires elucidation of target metabolites and, in the case of gas chromatography-mass spectrometry (GC-MS), the knowledge of their derivatized mass spectra. The study presented here focused on the two pyrrolidinophenones 3,4-methylenedioxy-α-pyrrolidinohexanophenone (MDPHP) and 3,4-methylenedioxy-α-pyrrolidinoheptanophenone (MDPHpP), which could be identified in 25 and 3 authentic cases, respectively. Using a standard analytical procedure by means of full-scan GC-MS after acid hydrolysis and acetylation, phase I metabolites of both substances were identified in authentic urine samples by elucidation of their mass spectral fragmentation patterns. The postulated phase I metabolic steps of MDPHP and MDPHpP comprised demethylenation followed by methylation of the methylenedioxy moiety, oxidation of the pyrrolidine ring, N,N-bisdealkylation of the pyrrolidine ring to its primary amine, hydroxylation of the aliphatic side chain. Various combinations were detected. Acetylated mass spectra of the metabolites were provided for both substances. The analogy in mass fragmentation of the proposed metabolites for the homologous parent compounds indicated a high plausibility. Based on the frequency of occurrence and abundances of the metabolites in the urine samples, target analytes for both substances and base peak fragment ions for specific mass search could be recommended for the mentioned procedure: m/z 140, 154 and 86 for MDPHP and m/z 154, 168 and 100 for MDPHpP. The study could support the detection of these new substances in forensic and clinical cases.

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 anti-doping 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 2 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 anti-doping violation. In a case of hidden administration, trazodone was identified at 9 and 24 pg/mg in 2 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 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.

Phosphatidylethanol (PEth) consists of phospholipids synthesized in erythrocyte cell membranes in the presence of ethanol and serves as a sensitive and specific indicator of alcohol consumption. Further research on PEth formation, degradation, and stability in postmortem (PM) samples would support its routine application in forensic toxicology. A supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) method was developed and validated to quantify PEth 16:0/18:1 in blood. PEth 16:0/18:1 was extracted from blood (0.25 g) using an 8:2 (v/v) heptane:2-propanol solvent mixture. Method validation results met ANSI/ASB 036 guidelines. Recovery was >48%, and matrix effects were <20%. The linear range was 10-2500 ng/g, and LLOQ was 10 ng/g. Bias was ±17.7%, and precision was <17.1% for all QC levels. Carryover, endogenous and exogenous interferences were negligible. Extracts were stable beyond 72 hours. In a proof-of-concept study reanalyzing 35 PM case samples, PEth concentrations ranged between 32.6-2476 ng/g. Short-term stability studies showed that fortified bovine blood (200 ng/g) preserved with 0.4% sodium fluoride (NaF) stored at room temperature had a 6.6% concentration drop after 48 hours, while blood stored at 4°C decreased by 13.5% over 14 days. Additionally, human PEth-positive blood preserved with 0.4% NaF showed a 6.7% decrease in in vivo PEth concentrations compared to a 17.5% decrease in heparin-preserved blood after 14 days at 4°C, supporting the use of 0.4% NaF in reducing PEth degradation over time. An in vitro model was also developed to simulate early PM PEth changes. Results found that PEth formation occurred in an ethanol concentration-dependent manner with minimal degradation, and considerations should be taken when interpreting PEth concentrations in cases with long PM interval, and if the decedent had a high BAC level and was left at elevated temperatures. This is the first SFC-MS/MS method successfully developed and validated for analysis of PEth in PM samples.

The legalization of hemp and the commercialization of hemp-based extracts has resulted in numerous cannabinoids appearing in consumer products. Although human pharmacological data are lacking for many of these isomers, analogs, and homologs of delta-9-tetrahydrocannabinol (∆9-THC), these cannabinoids may be capable of inducing cannabimimetic effects. Structural similarities also pose unique analytical challenges due to overlapping retention times and ion transitions used to distinguish between various parent drugs and metabolites. Therefore, traditional cannabinoid assays containing Δ9-THC, 11-hydroxy-Δ9-THC (11-OH-Δ9-THC), and 11-nor-9-carboxy-Δ9-THC (Δ9-THCCOOH) are no longer sufficient to confront this new threat to public safety. A new method has been developed and validated to quantitatively confirm Δ8- and Δ9-THC, their hydroxylated and carboxylated metabolites, and 9(R)- and 9(S)-hexahydrocannabinol (HHC) stereoisomers in blood and qualitatively identify these analytes in urine. This method is also capable of qualitatively confirming ∆9,11-THC (exo-THC), HHC and Δ6a,1°a-THC /Δ1°-THC carboxylated metabolites, and Δ8 and Δ9 THC homologs including tetrahydrocannabivarin (THCV), it's metabolite 11-nor-carboxy-THCV, tetrahydrocannabutol (THCB), tetrahydrocannabihexol (THCH; Δ8-THCH in urine only), tetrahydrocannabiphorol (THCP), THC-C8, as well as 9(R) and 9(S)-hexahydrocannabiphorol (HHCP) in blood and urine. Limits of detection were 1 ng/mL for non-carboxylated analytes and 5 ng/mL for carboxylated analytes. Calibration curves for parent and hydroxylated THC isomers and HHC stereoisomers were established from 1-50 ng/mL, whereas the calibration curve for the carboxylated THC isomers was 5-250 ng/mL. This method separates all analytes of interest from potential synthesis byproducts such as ∆8-iso-THC, ∆4(8)-iso-THC, and exo-THC. Unambiguous identification of these cannabinoids will increase forensic toxicology reporting accuracy while navigating the changing landscape of cannabis regulation and product formulation.

This study aimed to develop and validate a green method for the detection of ketamine and its metabolites in human urine samples and subsequently determine which metabolite is more suitable for judgment of ketamine abuse. Ketamine and its metabolites were extracted from urine samples with 1-undecanol using liquid-liquid microextraction based on the solidification of floating organic droplet extraction. The extracts were directly analyzed using gas chromatography-mass spectrometry (GC-MS) without derivatization. The sample pretreatment procedure prior to GC-MS analysis was simple, fast, and required little solvent consumption. Parameters that affect the extraction efficiency, including pH of the urine sample and centrifugation speed, were optimized. Under the optimized conditions, the limits of detection for norketamine, dehydronorketamine, and ketamine were 1.5, 1.7 and 1.0 ng/mL, respectively. The method was used to analyze eight real urine samples from drug abusers and exhibited acceptable accuracy and precision. By analyzing real samples, this study revealed that detection of dehydronorketamine in urine samples for the judgment of ketamine abuse may be more suitable than detection of norketamine or ketamine. The method used in this study addresses the need for green analytical techniques in toxicology.

Alcohol is the most abused substance in Western society, resulting in major economic losses and negative health consequences. Therefore, there is a need for a selective and robust detection method for alcohol consumption in various clinical and forensic settings. This study aimed to validate a mass spectrometry method for quantifying phosphatidylethanol (PEth) and perform retrospective data analysis from the patient population of a national reference laboratory. Quantification of PEth in whole blood was accomplished using an LC-MS-MS assay. Isotopically labeled internal standard for the two PEth homologues was added to the whole-blood specimen, followed by protein precipitation with a mixture of acetonitrile and isopropyl alcohol. After centrifugation, an aliquot of the supernatant was buffered with ammonium acetate before LC-MS-MS analysis on an Agilent 6470 triple quadrupole mass spectrometer coupled to an Agilent 1260 Infinity II LC system. This LC-MS-MS assay was validated for clinical use in accordance with Clinical & Laboratory Standards Institute guidelines. The analytical measurement range, 10-2000 ng/mL, was linear with R2 of 0.999. The within-run and total imprecision was < 5% CV for the low (20 ng/mL), medium (200 ng/mL), and high QC (1000 ng/mL). Results from accuracy and method comparison experiments met the bias criteria of ±15%. Retrospective data analysis showed ∼27% of patients had PEth concentrations <20 ng/mL. Males and females had similar positivity rates for PEth and the positivity rate of women of reproductive age (15-44 years old) was 35% in comparison to 25% in women 45-89 years old. This study's LC-MS-MS method showed acceptable analytical performance in quantifying PEth as a sensitive and specific biomarker for evaluating alcohol consumption. Results from this study may provide an opportunity to educate women of reproductive age on drinking during pregnancy and the long-term effects of alcohol use.

Carbon monoxide (CO) is a common gaseous toxin that causes severe poisoning symptoms. Accurate detection of the formation of carboxyhemoglobin (COHb) in the blood is very important for the identification of CO poisoning. In this review, the effects of exogenous toxins, including dichloromethane (DCM), nitrite, and hydrogen sulfide, on the determination of COHb by spectrophotometry are summarized by comparing epidemiological data, case studies, and analytical methods. The mechanism of the effects of these exogenous poisons on COHb detection is described, and the extent of their influence on the clinical diagnosis and forensic identification of CO poisoning is discussed. We suggest that emergency medicine and forensic science practices need to improve the understanding of these toxins and optimize clinical diagnosis and evaluation strategies to address the effects of toxins on the determination of COHb.

Novel psychoactive substances (NPSs) have historically been difficult compounds to analyze in forensic toxicology. The identification, detection, and quantitation of these analytes and their metabolites have been difficult due to their rapid emergence, short lifespan, and various potencies. Advancements in analytical instrumentation are fundamental to mitigating these NPS challenges by providing reliable identification and sensitivity. This review discusses the pros and cons of various analytical instruments that have played a pivotal role in NPS analysis. As analytical technology advanced, the ability to analyze for NPS became easier with high-resolution mass spectrometry (MS); however, traditional immunoassays are still beneficial for some NPS classes such as benzodiazepines. Over 200 articles from 2010-23 were reviewed, and 180 were utilized for this review. Journal articles were categorized according to the technology used during analysis: immunoassay, gas chromatography-MS, liquid chromatography-MS-low resolution, and liquid chromatography-MS-high resolution to allow for quick references based on a laboratory's technologies. Journal articles were organized in table format to outline the authors, NPS drug classes, and instrumentation used, among other important information.