Validation and application of an automated multitarget LC-MS/MS method for drugs of abuse testing using exhaled breath as specimen.

Aerosol microparticles in exhaled breath carry non-volatile compounds from the deeper parts of the lung. When captured and analyzed, these aerosol microparticles constitute a non-invasive and readily available specimen for drugs of abuse testing. The present study aimed to evaluate a simple breath collection device in a clinical setting. The device divides a breath sample into three parallel "collectors" that can be individually analyzed. Urine was used as the reference specimen, and parallel specimens were collected from 99 patients undergoing methadone maintenance treatment. Methadone was used as the primary validation parameter. A sensitive multi-analyte method using tandem liquid chromatography - mass spectrometry was developed and validated as part of the project. The method was successfully validated for 36 analytes with a limit of detection of 1 pg/collector for most compounds. Based on the validation results tetrahydrocannabinol THC), cannabidiol (CBD), and lysergic acid diethylamide (LSD) are suitable for qualitative analysis, but all other analytes can be quantitively assessed by the method. Methadone was positive in urine in 97 cases and detected in exhaled breath in 98 cases. Median methadone concentration was 64 pg/collector. The methadone metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) was detected in 90 % of the cases but below 10 pg/collector in most. Amphetamine was also present in the urine in 17 cases and in exhaled breath in 16 cases. Several other substances were detected in the exhaled breath and urine samples, but at a lower frequency. This study concluded that the device provides a specimen from exhaled breath, that is useful for drugs of abuse testing. The results show that high analytical sensitivity is needed to achieve good detectability and detection time after intake.

Appearance of hexahydrocannabinols as recreational drugs and implications for cannabis drug testing - focus on HHC, HHC-P, HHC-O and HHC-H.

This study investigated the effects of hexahydrocannabinol (HHC) and other unclassified cannabinoids, which were recently introduced to the recreational drug market, on cannabis drug testing in urine and oral fluid samples. After the appearance of HHC in Sweden in 2022, the number of posts about HHC on an online drug discussion forum increased significantly in the spring of 2023, indicating increased interest and use. In parallel, the frequency of false positive screening tests for tetrahydrocannabinol (THC) in oral fluid, and for its carboxy metabolite (THC-COOH) in urine, rose from <2% to >10%. This suggested that HHC cross-reacted with the antibodies in the immunoassay screening, which was confirmed in spiking experiments with HHC, HHC-COOH, HHC acetate (HHC-O), hexahydrocannabihexol (HHC-H), hexahydrocannabiphorol (HHC-P), and THC-P. When HHC and HHC-P were classified as narcotics in Sweden on 11 July 2023, they disappeared from the online and street shops market and were replaced by other unregulated variants (e.g. HHC-O and THC-P). In urine samples submitted for routine cannabis drug testing, HHC-COOH concentrations up to 205 (mean 60, median 27) µg/L were observed. To conclude, cannabis drug testing cannot rely on results from immunoassay screening, as it cannot distinguish between different tetra- and hexahydrocannabinols, some being classified but others unregulated. The current trend for increased use of unregulated cannabinols will likely increase the proportion of positive cannabis screening results that need to be confirmed with mass spectrometric methods. However, the observed cross-reactivity also means a way to pick up use of new cannabinoids that otherwise risk going undetected.

Automated extraction and LC-MS-MS analysis of 11-nor-9-carboxy-tetrahydrocannabinol isomers and prevalence in authentic urine specimens.

11-Nor-9-carboxy-Δ9-tetrahydrocannabinol (Δ9-THCCOOH) is the most frequently detected illicit drug metabolite in the military drug testing program. An increasing number of specimens containing unresolved Δ8-THCCOOH prompted the addition of this analyte to the Department of Defense drug testing panel. A method was developed and validated for the quantitative confirmation of the carboxylated metabolites of Δ8- and Δ9-THC in urine samples utilizing automated pipette tip dispersive solid-phase extraction and analysis by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Analytes were separated isocratically over an 8.5-min runtime and detected on an MS-MS equipped with an electrospray ionization source operated in negative mode. A single point calibrator (15 ng/mL) forced through zero demonstrated linearity from 3 to 1,000 ng/mL. Intra- and inter-day precision were ≤9.1%, and bias was within ±14.1% for Δ8-THCCOOH and Δ9-THCCOOH. No interferences were found after challenging the method with different over-the-counter drugs, prescription pharmaceuticals, drugs of abuse and several cannabinoids and cannabinoid metabolites, including Δ10-THCCOOH. Urine specimens presumptively positive by immunoassay (n = 2,939; 50 ng/mL Δ9-THCCOOH cutoff) were confirmed with this analytical method. Δ8-THCCOOH and Δ9-THCCOOH were present together above the 15 ng/mL cutoff in 33% of specimens. However, nearly one-third of the specimens analyzed were positive for Δ8-THCCOOH only. This manuscript describes the first validated automated extraction and confirmation method for Δ8- and Δ9-THCCOOH in urine that provides adequate analyte separation in urine specimens with extreme isomer abundance ratios.

Rapid Screening of New Psychoactive Substances Using pDART-QqQ-MS.

Drug abuse is a severe social problem worldwide. Particularly, the issue of new psychoactive substances (NPSs) have increasingly emerged. NPSs are structural or functional analogs of traditional illicit drugs, such as cocaine, cannabis, and amphetamine; these molecules provide the same or more severe neurological effects. Usually, immunoassays are utilized in the preliminary screening method. However, NPSs have poor detectability in commercially available immunoassay kits. Meanwhile, various chromatography combined with the mass spectrometry platform have been developed to quantify NPSs. Still, a significant amount of time and resources are required during these procedures. Therefore, we established a rapid analytical platform for NPSs employing paper-loaded direct analysis in real time triple quadrupole mass spectrometry (pDART-QqQ-MS). We implemented this platform for the semiquantitative analysis of forensic drug tests in urine. This platform significantly shrinks the analytical time of a single sample within 30 s and requires a low volume of the specimen. The platform can detect 21 NPSs in urine mixtures at a lower limit of qualification of concentration ranging from 20 to 75 nanograms per milliliter (ng mL-1) and is lower than the cutoff value of currently available immune-based devices for detecting multiple drugs (1000 ng mL-1). Urine samples from drug addicts have been collected to verify the platform's effectiveness. By combining efficiency and accuracy, our platform offers a promising solution for addressing the challenges posed by NPSs in drug abuse detection.

Extended Detection Window for Gamma-Hydroxybutyrate in the Urine of an Elderly Woman.

Gamma-hydroxybutyrate (GHB) is a central nervous system depressant that has gained popularity as an illicit recreational drug. We describe a case of an elderly woman who was found unconscious in her home. The paramedics initially suspected an intracranial incident. A head computed tomography was negative, as was the initial urinary drug screening. The diagnosis of GHB intoxication was made with the detection of GHB in a urine sample obtained 28-29 hours after the assumed time of intake. Our case underscores the importance of considering drug testing in a broad range of patients and shows that elderly patients may have an extended detection window of GHB.

Identification and Quantitative Analysis of 2-Fluoromethamphetamine and Its Metabolites in Human Urine.

Various synthetic drugs have appeared over the past years across the world, and phenethylamine derivatives are among them; indeed, aromatic fluoro analogs of methamphetamine and amphetamine have been in the illicit drug market since the early 2000s. Although they are currently widely abused across the world, little information is available on their metabolism and toxicology. Recently, we came across an alleged 2-fluoromethamphetamine (2-FMA) drug abuse case. The urine obtained from the alleged abuser was analyzed as part of a criminal investigation. 2-FMA, 2-fluoroamphetamine (2-FAP) and some related compounds were detected by liquid chromatography-tandem mass spectrometry. In forensic science, both an "unchanged" drug and its metabolite(s) need to be detected in urine to verify the illicit drug use. Notably, the detection of 2-FAP, which is a plausible 2-FMA metabolite, is insufficient as evidence of 2-FMA use because 2-FAP is widely available and may be present as such in taken liquids. In this study, we synthesized analytical standards for N-hydroxy 2-FMA (N-OH-2-FMA) and two diastereomers of 2-fluoroephedrine, which are plausible metabolites of 2-FMA. Using these standards, the urine specimen was found to contain N-OH-2FMA and one diastereomer of 2-fluoroephedrine; moreover, the concentrations of these compounds were successfully determined. The results of our study suggest that N-hydroxylation and aliphatic hydroxylation are the characteristic metabolic pathways of 2-FMA compared with that of methamphetamine. This evidence indicates that both N-OH-2-FMA and 2-fluoroephedrine are plausible candidates as analytical targets for drug-use certification in forensic science.

An ultra-sensitive UHPLC-QqQ-MS/MS method for determination of 54 benzodiazepines (pharmaceutical drugs, NPS and metabolites) and z-drugs in biological samples.

Benzodiazepines exhibit central nervous system depressive activity as well as sedative, hypnotic, and anticonvulsant properties, which enable to use them as medical treatment in anxiety, epilepsy, insomnia and alcohol withdrawal syndrome. However, from 2000s illegal benzodiazepine derivatives have started to emerge on illicit drug market as new psychoactive substances (NPSs) monitored in many countries. Analysis of both pharmaceutical drugs and NPSs from benzodiazepines group could be challenging, as usually very low concentrations need to be determined. Thus, an ultra-sensitive UHPLC-QqQ-MS/MS method was developed for simultaneous determination of 54 benzodiazepines (pharmaceutical drugs, NPS and their metabolites) and 3 z-drugs with one metabolite in biological fluid samples. The lower limit of quantification for most substances was 50 pg/mL, whereas for 17 substances as low as 10 pg/mL was achieved. Together with reduced sample volume to 100 µL it makes the developed method suitable for a sensitive multidrug toxicological analysis. Presented method was applied in routine toxicological practice as well as for the determination of benzodiazepines, z-drugs and their metabolites in 25 authentic biological fluids (blood, urine, vitreous humor and bile), both antemortem and postmortem. 19 different compounds, including benzodiazepines, their metabolites and z-drugs were determined. Antemortem blood concentrations were within 0.2-114.5 ng/mL, whereas concentrations in antemortem urine samples were 0.03-102.6 ng/mL. In postmortem specimens, concentrations ranged within 0.2-473.2 ng/mL, 0.5-94.1 ng/mL, 1.3-208.8 ng/mL and 41.5-42.0 ng/mL in blood, vitreous humor, urine and bile, respectively. The developed method is suitable for a forensic toxicology analysis, as well as clinical toxicology which is evidenced by the positive results of international proficiency tests.

Update: Correlation of Fentanyl Positive Drug Screens with other Medications in Patients from Pain, Rehabilitation and Behavioral Programs.

OBJECTIVE: To monitor fentanyl polydrug use over past six years. METHOD: Calculate percent of fentanyl and other drugs positive in urine drug tests. RESULTS: Percent of fentanyl positive drug tests remained unchanged, but increases in fentanyl/methamphetamine and fentanyl/marijuana were observed. CONCLUSIONS: Fentanyl laced illicit drugs remain a major substance abuse problem.

Rates and types of urine drug screen false negative results compared with confirmatory toxicology testing in major trauma patients.

BACKGROUND: Trauma centers are required to screen patients for alcohol use, and if necessary, intervene and refer to treatment (SBIRT). Similar screening for illicit drug use is recommended but not required. Urine drug screening (UDS) underestimates problematic substance use. This study aimed to estimate the types and rates of UDS false negatives (FN) compared to comprehensive testing by liquid chromatography-mass spectrometry (LC-MS) in trauma patients. METHODS: We performed a prospective cohort study of deidentified urine samples from adult trauma and burn activation patients. Both UDS and LC-MS comprehensive testing of >200 analytes were performed by a reference laboratory on all samples. Iatrogenic medications were excluded from the FN count. Crosstab analyses were conducted for UDS versus LC-MS outcomes to establish FN types and rates. We dichotomized the results by creating an "intentionality" variable (intentional injuries by self/others versus accidental injuries). A series of crosstabs with odds ratios considered intentionality by substance class and demographics. Statistically significant variables by Chi-Square were assessed by logistic regression. RESULTS: Psychoactive FN were detected in 56/100 urine samples analyzed; the most frequent included anticonvulsants (primarily gabapentin, N = 13), opioid agonists (N = 12), antihistamines (primarily diphenhydramine, N = 10), and phenethylamines (primarily bupropion, N = 5). Nonpsychoactive FN were detected in 70/100 samples; the most common were nicotine (N = 33), caffeine (N = 23), acetaminophen (N = 22), and antidepressants (N = 12). Of substance classes included in the UDS and also tested by LC-MS, FN occurred for opiates (3%), amphetamines (5%) and opioids (25%). Polypharmacy was associated with fall injuries in elderly patients. Cocaine (p = 0.015) and cannabinoids (p = 0.002) were significantly associated with intentionality. CONCLUSIONS: Our results indicate that FN for potentially important psychoactive and nonpsychoactive substances are common when toxicologic testing is limited to routine UDS in trauma patients. We recommend expanding SBIRT in this patient population to include misuse of tobacco products, prescription analgesics, and over-the-counter antihistamines.

NPS detection in prison: A systematic literature review of use, drug form, and analytical approaches.

This paper presents a systematic literature review on the detection of new psychoactive substances (NPS) in prison settings. It includes the most frequently reported NPS classes, the routes and forms used for smuggling, and the methods employed to analyse biological and non-biological samples. The search was carried out using MEDLINE (EBSCO), Scopus (ELSEVIER), PubMed (NCBI), and Web of Science (Clarivate) databases, along with reports from the grey literature in line with the PRISMA-S guidelines. A total of 2708 records were identified, of which 50 met the inclusion criteria. Findings showed the most prevalent NPS class reported in prison was synthetic cannabinoids (SCs). The most frequently reported SCs in non-biological samples were 4F-MDMB-BINACA, MDMB-4en-PINACA, and 5F-ADB. These were smuggled mainly through the postal services deposited on paper or herbal matrices. Concentrations of SCs detected on seized paper ranged between 0.05 and 1.17 mg/cm2 . The SCs most frequently reported in biological specimens (i.e., urine, blood, saliva, and wastewater) were 5F-MDMB-PICA, 4F-MDMB-BINACA, and MDMB-4en-PINACA. Concentrations of SCs reported in femoral blood and serum were 0.12-0.48 ng/ml and 34-17 ng/ml, respectively. Hyphenated techniques were predominantly employed and generally successful for the detection of NPS in biological (i.e., LC-HRMS/MS) and non-biological samples (i.e., LC-HRMS/MS and GC-MS). The onsite technique IMS showed promise for detecting SCs in various forms; however, immunoassays were not recommended. Future work should focus on accurate in-field detection of SCs deposited on paper and in urine and saliva to improve real-time decision-making, as well as wastewater and air monitoring for overall drug use trends.

Emergency department visits due to new psychoactive substances and other illicit drugs in Taiwan: preliminary results of the Taiwan Emergency Department Drug Abuse Surveillance (TEDAS) project.

INTRODUCTION: The use of new psychoactive substances (NPSs) has markedly increased worldwide; thus, it is important to monitor NPS-related effects. The Taiwan Emergency Department Drug Abuse Surveillance (TEDAS) project aims to assess the patterns of recreational drug use in patients presenting to emergency departments (EDs) across the country. Here, we report the preliminary results of this project. METHODS: This observational study included the collection and analysis of urine samples and assessment of the clinical presentation of patients from 79 EDs across Taiwan. Clinical features were recorded through a questionnaire filled by attending doctors or nurses who collected urine samples for clinical diagnosis. Urine samples were analyzed for 110 drugs and metabolites using electrospray ionization liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: Between February and November 2019, a total of 2649 patients were enrolled for urine drug analysis. A total of 675 cases older than 12 years (males, n = 480) had NPS or other illicit drugs detected in their urine samples. Overall, 1271 counts of drugs, among which 717 (56.4%) were NPS. At least one NPS was detected in 340 patients (50.4%), and 292 cases were positive for multiple drugs. The most frequently detected drug was methamphetamine/amphetamine, followed by synthetic cathinones, ketamine and its two analogs, and opioids. The most common drug combination was cathinones plus ketamine and/or its analogs (n = 56). Younger patients (OR = 3.3, p≤.0001) and women (OR = 1.5, p = .01) were more likely to have NPS detected in their urine samples. NPS-positive cases frequently experienced chest pain (OR = 2.6, p = .03), tachycardia (OR = 2.6, p = .0002), and suicide attempt/non-suicidal self-harm (OR = 1.8, p = .004), whereas depressed consciousness (OR = 0.5, p = .001) was less frequent among NPS-positive cases than among other illicit drug-positive cases. CONCLUSIONS: The TEDAS project provides a nationwide epidemiological profile of recreational drug use in Taiwan. More than half of the recreational drugs were NPSs, which were comprehensively detected using LC-MS/MS.

Illicit substances identified in the urine of 11.170 suspected drug users in North Tunisia.

INTRODUCTION: according to the latest World Drug Report, 271 million people worldwide (5.5% of the global population) aged 15-64 years are drug users. Drug addiction and trafficking became an urgent public health problem that affects human health and social life. A cross-sectional study was conducted from January 2016 to December 2018, to identify the socio-demographic profile of drug users captured by the anti-narcotics squad in North of Tunisia (North African country) and to type, through toxicological analysis, the nature of the substances consumed. METHODS: data were collected from expertise files of 11170 suspected drug users. Fresh urine samples were collected from suspected drugs users and submitted in the toxicology laboratory of the center for Urgent Medical Assistance (Tunis) for forensic urinalysis. Drugs screening was carried out by immunochemical methods. Positive samples were analyzed with gas chromatography coupled to mass selective detector (GC-MS) for confirmation. RESULTS: the investigation revealed that drug users were mainly males 97.4% (sex ratio 37), the median age was 29 ± 7.91 years, 91.3% were singles and 79% were daily workers. On a total of 11170 urine samples screened, 5 409 (48.4%) were positives for illicit drugs. The prevalence of positive samples was 55.4% in 2016; 50.45% in 2017 and 40.8% in 2018. Cannabis was the most widely consumed drug (95%), followed by Benzodiazepines (1.2%), Buprenorphine (1%), cocaine (0.95%), MDMA (0.24%) and opiates (0.13%). Polydrug abuse was observed in 79 specimens (1.5%). CONCLUSION: this study provides an overview of the illicit drug consumption in the north of Tunisia (knowing that nowadays epidemiological data are almost same since 2016) in order to set up an effective policy to fight against drugs and addictive behaviors and to provide health professionals with the epidemiological elements necessary for better medical care of drug users.

Simultaneous LC-MS/MS screening for multiple phenethylamine-type conventional drugs and new psychoactive substances in urine.

New psychoactive substances are being launched in the drug market at a rapidly growing pace. More than 950 new psychoactive substances have been reported to the United Nations Office on Drugs and Crime. The development of new psychoactive substance abuse has drawn risks on public health and safety. Phenethylamines, along with other stimulants, accounted for the majority of the new psychoactive substances being reported in the past decade. This study presents a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous screening of 74 conventional and artificial phenethylamines in urine samples. The chromatographic analysis was performed by a direct dilute-and-shoot procedure using a Phenomenex Kinetex® Phenyl-Hexyl column (10 cm × 2.1 mm i.d., 1.7 µm) and two mobile phases (A: 0.1% formic acid aqueous solution with 5 mM ammonium acetate, B: 0.1% formic acid methanolic solution). The mass fragments were collected under the multiple reaction monitoring mode. The linearity range located in 1.0-50.0 ng/mL for quantitative analysis. The limit of detection and lower limit of quantification for 74 phenethylamines were 0.5 ng/mL and 1.0 ng/mL, respectively. The method was validated and further applied to analyze authentic urine samples. Twenty samples were tested positive of seven phenethylamines from 67 samples, whereas the contents detected were 9.8 ng/mL to 147.1 µg/mL with dilution factors of 40 to 20,000 folds.

A comprehensive study on the performance of different retention mechanisms in sport drug testing by liquid chromatography tandem mass spectrometry.

Anti-doping substances listed by the World Anti-Doping Agency (WADA) include hundreds of compounds of very different physico-chemical properties. Anti-doping control laboratories need to screen all these substances in the so-called Initial Testing Procedures (ITPs) what is very challenging from an analytical point of view. ITPs are mostly based on reversed-phase (RP) liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) using C18 columns, which feature poor retention and peak tailing for polar and basic compounds, respectively. While studies on this field dealing with the comparison of stationary phases are focused on certain chemical classes, this research provides a wide multi-target approach. For this purpose, a representative group of 93 anti-doping agents (log P from -2.4 to 9.2) included in ten different classes of prohibited substances was selected. A comprehensive study on the performance of six columns and four eluents on different separation parameters (retention factors, asymmetry factors, co-elutions, total run times) and matrix effects (signal enhancement or suppression) was performed for LC-MS/MS-based ITPs. Columns working in both RP [C18, C8, phenyl hexyl (PH), pentafluorophenyl (PFP) and mixed-mode hydrophilic/RP (HILIC-RP)) and hydrophilic (HILIC)] modes were investigated. Eluents contained methanol or acetonitrile as organic modifiers, with or without the addition of ammonium acetate. The best column-mobile phase binomial for ITPs was PFP using water-methanol (0.1% formic acid) as eluent, while HILIC was the best option for highly polar non-aromatic anti-doping agents, which were poorly addressed by PFP. Excellent good peak shapes and relative acceptable matrix interferences were obtained for HILIC-RP, which was tested for the first time for the analysis of anti-doping agents, although the number of compounds eluting too fast was too high. On the whole, the alkyl phase C18 showed the worst performance and although C8 and PH were better, their performance did not surpass that of PFP. Possible retention mechanisms underlying separation in the different stationary phases were discussed. This research provides valuable information to anti-doping control labs for improving LC-MS/MS-based ITPs and it proposes PFP as a suitable alternative to the already established C18.

Urine drug screening: A guide to monitoring Tx with controlled substances.

Avoid error by ordering the appropriate test at a risk-based frequency. Be alert to sources of false-positives and adulteration. Be careful not to overreact to unexpected results.

Trend differences over 20 years between two methadone maintenance clinics, one with and one without cannabis legalization.

BACKGROUND: Although methadone maintenance treatment (MMT) guidelines are well established, patients' characteristics and outcome change over time may be affected by the legality of cannabis. OBJECTIVE: To study trend changes between two clinics over 20 years from Las Vegas (LV) and 27 years from Tel Aviv (TA). METHODS: Patients' characteristics at admission, including drugs in urine at first and 13th month were obtained from their medical charts. Changes by year of admission and cumulative retention were analyzed. RESULTS: The LV MMT clinic (1724 patients) had a lower one-year retention rate compared to the TA MMT clinic (1014 patients) (46.4% vs. 74.4%, respectively, p < 0.0005), and a higher rate of opioid stop after one year (75.9% vs. 68.8%, respectively, p = 0.003). The age at MMT admission and the retention rates decreased in LV and increased in TA. The prevalence of cannabis and benzodiazepine misuse on MMT admission increased in LV with no change recorded in TA. Cocaine on MMT admission decreased in LV and increased in TA, while amphetamine use increased in LV and decreased in TA. Cox models multivariate analyses found cannabis on admission to predict shorter retention in LV (as younger age male and amphetamines), and cannabis after one year in TA (as did cocaine and opiates after one year and BDZ on admission). CONCLUSION: Although cannabis prevalence increased only in LV where it was legalized, it was associated with poor outcomes in both clinics. Younger age, a known poor outcome predictor, may be related to decreased retention in LV.

Are point-of-care urine drug testing devices suitable for antenatal drug screening? A study verifying the Alere® Drug Screen Test Cup for the detection of six classes of drug in a pregnant population.

BACKGROUND: Currently, there are no national guidelines for antenatal drug testing. At Colchester Hospital, we use a strategy of screen-only using point-of-care testing to detect illicit drug use in pregnancy. To determine the suitability of this approach, we have compared the results of urine analysis by point-of-care testing with another NHS specialist clinical toxicology service that uses confirmation mass spectrometry. METHODS: A total of 482 anonymized random urine specimens from antenatal clinics were tested for six drug classes: amphetamine, benzodiazepines, buprenorphine, cocaine, methadone and opiates using the Alere™ Drug Screen Urine Test Cup. The manufacturer's claims for positive cut-off and result stability were verified using spiked blank urine. Confirmatory testing was performed using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for detection of 26 individual drugs. RESULTS: Of 473 urine samples with adequate volume for point-of-care screening, 4.4% tested positive: 19 opiate and 2 cocaine. Concordance between point-of-care screening and UPLC-MS/MS confirmation was 97.9% for all drugs and 78.9% for opiates. Using spiked urine, only positive results for opiates were stable when read up to the manufacturer's recommended time of 60 min. CONCLUSIONS: The key advantages of using point-of-care devices to detect drug use in pregnancy are that is convenient and cheap. However, the clinical utility of point-of-care testing is limited by its poor sensitivity. Best practice is to confirm results using a more specific and sensitive method. As a result of this study, we are now reviewing our own procedures to consider introducing routine confirmation by mass spectrometry.

In Vitro Phase I Metabolism of the Recently Emerged Synthetic MDMB-4en-PINACA and Its Detection in Human Urine Samples.

MDMB-4en-PINACA (methyl (S)-3,3-dimethyl-2-(1-(pent-4-en-1-yl)-1H-indazole-3-carboxamido)butanoate) is a recently emerged synthetic cannabinoid in Turkey. MDMB-4en-PINACA was detected in herbal material investigated by the Council of Forensic Medicine, Istanbul Narcotics Department in Turkey in April 2019. MDMB-4en-PINACA was added to the drug abuse list and quickly reported in biological samples after its first detection. In this study, the in vitro metabolism of MDMB-4en-PINACA was investigated by using a pooled human liver microsomes (HLMs) assay and liquid chromatography-high-resolution mass spectrometry (LC-HRMS). MDMB-4en-PINACA (5 µmol/L) was incubated with HLMs for up to 1 h, and the metabolites were identified using LC-HRMS and software-assisted data mining. The in vivo metabolism was investigated by the analysis of 22 authentic urine samples and compared to the data received from the in vitro metabolism study. Less than 7.5% of the MDMB-4en-PINACA parent compound remained after the 1 h incubation. We identified 14 metabolites, which were formed via double bond oxidation, ester hydrolysis, N-dealkylation, hydroxylation, dehydrogenation and further oxidation to N-pentanoic acid or a combination of these reactions in vitro. In 10 urine samples (total n = 22), MDMB-4en-PINACA was detected as the parent drug. Three of the identified main metabolites, double bond oxidation in combination with ester hydrolysis and hydroxylation metabolite (M3), MDMB-4en-PINACA butanoic acid (M14) and monohydroxypentyl-MDMB-4en-PINACA (M12), were suggested as suitable urinary markers. In vitro screening of 2,150 authentic urine samples for these identified MDMB-4en-PINACA metabolites resulted in 56 cases of confirmed MDMB-4en-PINACA consumption (2.6%).

Evaluation and applicability of Alere iCup DX 14 for rapid postmortem urine drug screening at autopsy.

Performing point-of-care urine drug screen testing at autopsy by a forensic pathologist may provide an early indication of the presence of analytes of interest during autopsy. An evaluation for the screening of 14 classes of common drugs of abuse in postmortem urine by the point-of-care screening device, Alere iCup DX 14, is presented. One hundred ninety postmortem urine samples were screened with the iCup occurring at autopsy by the forensic pathologist. Positive and negative results obtained from the screening kit were evaluated against confirmatory test results obtained using routine forensic toxicology analyses that employed LC-MS/MS and GC-MS to detect a combination of over 85 common drugs of abuse and medications. Sensitivity for each respective iCup drug class ranged from 66% (buprenorphine) to 100% (methadone, tricyclic antidepressants). Specificity for each respective iCup drug class ranged from 89% (benzodiazepines) to 100% (amphetamines, barbiturates, buprenorphine, 3,4-methylenedioxymethamphetamine, methadone). Positive predictive values ranged from 44% (benzodiazepines) to 100% (amphetamines, barbiturates, buprenorphine, methylenedioxymethamphetamine, methadone), while negative predictive values ranged from 96% (methamphetamine) to 100% (barbiturates, methadone, tricyclic antidepressants). A high false-positive rate was yielded by the benzodiazepine class. The lack of fentanyl screening in the point-of-care device is a significant limitation considering its prolific prevalence in forensic casework. The results obtained in the study should be acknowledged when considering the use of the Alere iCup DX 14 in the context of postmortem casework to help indicate potential drug use contemporaneously with autopsy and when requiring such preliminary results prior to the release of a final forensic toxicology report.

Urinary excretion profile of methiopropamine in mice following intraperitoneal administration: A liquid chromatography-tandem mass spectrometry investigation.

We have considered the urinary excretion profile of methiopropamine (MPA), a thiophene ring-based structural analog of methamphetamine with similar stimulant effects, with the aim of selecting the most appropriate marker(s) of intake that may be useful in forensic analysis. For this purpose, in vitro studies were preliminarily performed on human liver microsomes for tracing the phase I metabolic pathways of MPA, preselecting the best candidates as potential target analytes, and designing the optimal experimental strategy. In vivo studies were then conducted on mice, after the intraperitoneal administration of a 10-mg/kg dose. Urine samples were collected every 3 h in the first 9 h and, subsequently, from 24 to 36 h, and stored at -80°C until further analysis. The measurements were performed using a targeted procedure based on liquid/liquid extraction followed by liquid chromatography-tandem mass spectrometry analysis. Our results show that in the time interval 0-9 h after administration, MPA was extensively oxidized mainly to nor-MPA, oxo-MPA, and two hydroxylated metabolites (ie, hydroxy-aryl-methiopropamine and hydroxy-alkyl-methiopropamine). All phase I metabolites underwent phase II metabolism, with the formation of nor-hydroxy-methiopropamine only in phase II, confirmed by the results obtained after enzymatic hydrolysis with ß-glucuronidase and arylsulfatase. In the time interval 24-36 h after administration, only unchanged MPA and nor-MPA were detected, suggesting that these two markers are those endowed with the highest diagnostic value. The method was validated for these two principal markers, proving to be fit for anti-doping, toxicological, and forensic analyses.