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An increasing number of countries have started to decriminalize or legalize the consumption of cannabis for recreational and medical purposes. The active ingredients in cannabis, termed cannabinoids, affect multiple functions in the human body, including coordination, motor skills, memory, response time to external stimuli, and even judgment. Cannabinoids are a unique class of terpeno-phenolic compounds, with 120 molecules discovered so far. There are certain situations when people under the influence of cannabis may be a risk to themselves or the public safety. Over the past two decades, there has been a growing research interest in detecting cannabinoids from various biological matrices. There is a need to develop a rapid, accurate, and reliable method of detecting cannabinoids in oral fluid as it can reveal the recent intake in comparison with urine specimens, which only show a history of consumption. Significant improvements are continuously made in the analytical formats of various technologies, mainly concerning improving their sensitivity, miniaturization, and making them more user-friendly. Additionally, sample collection and pretreatment have been extensively studied, and specific devices for collecting oral fluid specimens have been perfected to allow rapid and effective sample collection. This review presents the recent findings regarding the use of oral fluid specimens as the preferred biological matrix for cannabinoid detection in a point-of-care biosensor diagnostic device. A critical review is presented, discussing the findings from a collection of review and research articles, as well as publicly available data from companies that manufacture oral fluid screening devices. Firstly, the various conventional methods used to detect cannabinoids in biological matrices are presented. Secondly, the detection of cannabinoids using point-of-care biosensors is discussed, emphasizing oral fluid specimens. This review presents the current pressing technological challenges and highlights the gaps where new technological solutions can be implemented. [ABSTRACT FROM AUTHOR]

This study aimed to measure the prevalence of drugs of abuse (DOA) among the goods carriage drivers associated with the southern State of India, Kerala. Point-of-collection testing (POCT) of oral fluid collected from the participants (n = 249) was done using the Evidence MultiSTAT DOA Oral Fluid II Assay kits and the Evidence MultiSTAT analyzer. Out of the total samples, 53 (21.29%) were positive for one or more DOA. A high prevalence of tetrahydrocannabinol (THC) (10.04%) and synthetic cannabinoids were detected in the samples. The use of ketamine, alpha-PVP, LSD, methamphetamine, opiate, 6-MAM, benzodiazepines I, methadone, PCP, tramadol, and amphetamine was also detected and their frequency of use ranged between 4.02 and 0.80%. An association between drug abuse and distance of travel was found in drivers in this study, χ2 (5, N = 249) = 123.5, p < 0.001. Confirmatory analysis using ultra-high performance liquid chromatography-tandem mass spectrometry showed excellent agreement with the results of the screening test. This was the first study conducted among drivers in India for the detection of DOA. Tetrahydrocannabinol (THC) was used more by the goods carriage drivers associated with Kerala State, India. The use of psychoactive substances significantly increased with the distance of travel. Point-of-collection testing (POCT) by the biochip array technology is an efficient method for the detection of these substances. [ABSTRACT FROM AUTHOR]

Fentanyl has emerged as the most prolific drug in the ongoing opioid epidemic and has greatly impacted traffic safety in recent years. This study aimed to evaluate fentanyl prevalence and concentrations in blood and oral fluid in driving under the influence of drugs (DUID) cases in three different regions (i.e., Alabama, Orange County, CA, and Houston, TX) from 2017 to 2022. Furthermore, traffic fatalities were evaluated for Alabama and Orange County, CA. Fentanyl positivity in DUID and traffic fatalities increased for most years in this study. In Alabama, the prevalence of fentanyl DUID cases increased 4‐fold in 2022 compared to 2017. Orange County's increase from 2017 to 2022 was 14‐fold. In Houston, the increase was approximately 2‐fold from 2019 to 2022. The greatest increase for all laboratories coincided with the start of the COVID‐19 pandemic. In 2022, the median fentanyl DUID blood concentrations were 4.7, 11, and 4.7 ng/mL in Alabama, Orange County, and Houston, respectively. Most fentanyl cases were polydrug cases (≥90%). Methamphetamine, THC, and alprazolam were the most frequently detected drugs in combination with fentanyl. Alabama has collected oral fluid and blood in DUID cases since 2018. The detection of fentanyl in oral fluid was comparable to blood. However, 59% and 8.7% of fentanyl‐positive cases had concentrations of >20 ng/mL in oral fluid and blood, respectively. Therefore, oral fluid as an alternative or supplemental specimen to blood is an attractive approach for fentanyl in DUID cases. This study contributes to understanding recent fentanyl trends and their impact on traffic safety. [ABSTRACT FROM AUTHOR]

Cannabis is used globally for both recreational and medical purposes. The use of cannabis can impair driving ability and increase the risk of fatal accidents. Therefore, operating motor vehicles and machinery under the influence of cannabis is illegal in most jurisdictions. With many governments easing legal restrictions on the use of cannabis, the demand for cannabis intoxication testing is increasing globally. Identification of prior cannabis use is currently accomplished using a variety of biological matrices, including urine, blood, sweat, hair, and oral fluid. Despite the ability to identify prior exposure to or intake of cannabis, each of these matrices has limitations in assessing the current intoxication state of an individual, an essential aspect of many current legislations. Exhaled breath represents a potentially useful matrix for measuring recent use, however, current uncertainty in breath levels indicative of impairment and the technical limitations associated with rapid, on‐site analysis of breath matrices have restricted the widespread deployment of cannabis breath analyzers. These challenges are compounded by the relatively short intoxication window of cannabis relative to its lifetime in the body, reinforcing the need for rapid toxicological screening. This review summarizes the state of cannabis intoxication testing, highlighting the challenges associated with the current forensic testing techniques, and the potential for breath analysis, which has gained interest recently from both academic researchers and commercial entities. This article is categorized under:Toxicology > Drug AnalysisToxicology > Cannabis [ABSTRACT FROM AUTHOR]

Cannabis sativa L. is a substance widely used around the world for recreational and medicinal purposes. Oral fluid has been investigated as an alternative biological matrix for demonstrating the illegal use of cannabis, particularly in situations where its recent use needs to be identified. In the last two decades, many methods have been developed to detect and quantify cannabinoids in oral fluid, especially for Δ9‐tetrahydrocannabinol, the primary psychoactive substance of cannabis. However, some aspects must be considered in the use of these techniques, such as cannabinoids recoveries or extraction efficiency from different oral fluid collection devices/containers. Pharmacokinetic studies have shown that the presence of minor cannabinoids and metabolites in the analysis of oral fluid may be valuable in interpreting tests, which indicates the need to improve the sensitivity of detecting low concentrations. The aim of this review is to summarize and to describe the methodologies for the quantitative analysis of cannabinoids in oral fluid that have previously been investigated. A systematic search for articles was performed of four different databases, using the descriptor "cannabinoids and oral fluid". Forty‐seven studies that examined quantitative methods were identified. The analytical data described in these articles, including oral fluid collection, sample preparation, cannabinoids recovery and extraction efficiency, detection instruments, and quantification limits, were analyzed. The discussion of these particular features of cannabinoid analysis in oral fluid could help to improve or to develop methods for use in Forensic Toxicology. [ABSTRACT FROM AUTHOR]

Increased prevalence of cannabis consumption and impaired driving are a growing public safety concern. Some states adopted per se driving laws, making it illegal to drive with more than a specified blood concentration of ∆9-tetrahydrocannabinol (THC) in a biological fluid (typically blood). Blood THC concentrations decrease significantly (∼90%) with delays in specimen collection, suggesting the use of alternative matrices, such as oral fluid (OF). We characterized 10 cannabinoids' concentrations, including THC metabolites, in blood and OF from 191 frequent and occasional users by liquid chromatography with tandem mass spectrometry for up to 6 h after ad libitum smoking. Subjects self-titrated when smoking placebo, 5.9 or 13.4% THC cannabis. Higher maximum blood THC concentrations (Cmax) were observed in individuals who received the 5.9% THC versus the 13.4% THC plant material. In blood, the Cmax of multiple analytes, including THC and its metabolites, were increased in frequent compared to occasional users, whereas there were no significant differences in OF Cmax. Blood THC remained detectable (≥5 ng/mL) at the final sample collection for 14% of individuals who smoked either the 5.9 or 13.4% THC cigarette, whereas 54% had detectable THC in OF when applying the same cutoff. Occasional and frequent cannabis users' profiles were compared, THC was detectable for significantly longer duration in blood and OF from frequent users. Detection rates between frequent and occasional users at multiple per se cutoffs showed larger differences in blood versus OF. Understanding cannabinoid profiles of frequent and occasional users and the subsequent impact on detectability with current drug per se driving limits is important to support forensic interpretations and the development of scientifically supported driving under the influence of cannabis laws. [ABSTRACT FROM AUTHOR]

A liquid chromatography–mass spectrometry method using dried oral fluid spots was developed and validated for the simultaneous quantification of cocaine, benzoylecgonine, cocaethylene, amphetamine, and 3,4‐methylenedioxymethamphetamine. The oral fluid was applied to a Whatman 903 grade paper and submitted to a drying time of 2.5 h. The extraction procedure was optimized by chemometric approach using simplex centroid design. Spots were extracted with a mixture of acetonitrile, buffer, and methanol. Calibration curves covered a linear concentration range of 40–500 ng/mL. Validation parameters of linearity, precision, accuracy, selectivity, carryover, matrix effects, and stability were evaluated and showed satisfactory results. Spot homogeneity was also satisfactory, with less than 15% of deviation from nominal concentration. Spot volume did not influence accuracy when less than 100 μL of the sample was applied to the spot. The validation of the proposed method suggests a potential application in different scenarios in toxicology. [ABSTRACT FROM AUTHOR]

With advances in analytical technology and new research informing result interpretation, oral fluid (OF) testing has gained acceptance over the past decades as an alternative biological matrix for detecting drugs in forensic and clinical settings. OF testing offers simple, rapid, non-invasive, observed specimen collection. This article offers a review of the scientific literature covering analytical methods and interpretation published over the past two decades for amphetamines, cannabis, cocaine, opioids, and benzodiazepines. Several analytical methods have been published for individual drug classes and, increasingly, for multiple drug classes. The method of OF collection can have a significant impact on the resultant drug concentration. Drug concentrations for amphetamines, cannabis, cocaine, opioids, and benzodiazepines are reviewed in the context of the dosing condition and the collection method. Time of last detection is evaluated against several agencies' cutoffs, including the proposed Substance Abuse and Mental Health Services Administration, European Workplace Drug Testing Society and Driving Under the Influence of Drugs, Alcohol and Medicines cutoffs. A significant correlation was frequently observed between matrices (i.e. between OF and plasma or blood concentrations); however, high intra-subject and inter-subject variability precludes prediction of blood concentrations from OF concentrations. This article will assist individuals in understanding the relative merits and limitations of various methods of OF collection, analysis and interpretation. [ABSTRACT FROM AUTHOR]

Long-term testosterone replacement therapy is mainly monitored by trough levels of serum testosterone (S-T), while urinary testosterone (U-T) is used by forensic toxicology to evaluate testosterone doping. Testosterone in saliva (Sal-T) may provide additional information and simplify the sample collection. We aimed to investigate the relationships between testosterone measured in saliva, serum and urine during standard treatment with 1,000 mg testosterone undecanoate ( TU) every 12th week during 1 year. This was an observational study. Males with primary and secondary hypogonadism ( HG; n = 23), subjects with gender dysphoria ( GD FtM; n = 15) and a healthy control group of men ( n = 32) were investigated. Sal-T, S-T and U-T were measured before and after TU injections. Sal-T was determined with Salimetrics® enzyme immunoassay, S-T with Roche Elecsys® testosterone II assay and U-T by gas chromatography-mass spectrometry. Sal-T correlated significantly with S-T and calculated free testosterone in both controls and patients ( HG men and GD FtM), while Sal-T to U-T showed weaker correlations. Trough values of Sal-T after 12 months were significantly higher in the GD FtM group (0.77 ± 0.35 nmol/L) compared to HG men (0.53 ± 0.22 nmol/L) and controls (0.46 ± 0.15 nmol/L), while no differences between S-T and U-T trough values were found. Markedly elevated concentrations of salivary testosterone, 7-14 days after injection, were observed, especially in the GD FtM group. This study demonstrates that Sal-T might be a useful clinical tool to monitor long-term testosterone replacement therapy and might give additional information in forensic cases. [ABSTRACT FROM AUTHOR]

Oral fluid is an alternative matrix to blood for preliminary screening of drugs of abuse. Dried oral fluid spot may allow simple sampling, storage, transportation, and exclusion of enzymatic and hydrolytic degradation of analytes. The purpose of present study was to optimize drug extraction from oral fluid and dried samples to evaluate the determination of illicit drugs. The efficiency of extraction was assessed using pooled oral fluid spiked with amphetamine, methamphetamine, 3,4-methylenedioxy-methamphetamine, Δ9-tetrahydrocannabinol, cocaine, morphine, methadone, and clonazepam. The results indicate that simultaneous extraction of the drugs may be performed by ethyl acetate in the presence of sodium hydroxide. This procedure was used for dried oral fluid with pretreatment using methanol and hydrochloric acid to provide acceptable recovery values exceeding 70%. The applicability of the protocol was confirmed using oral fluid provided by subjects with recent use of illicit drugs. The results showed that drugs of abuse in oral fluid may be determined as the dried material and is a promising alternative to liquid sampling. [ABSTRACT FROM AUTHOR]

Metabolomic analysis of easily accessible biofluids has provided numerous biomarkers in urine and blood for biodosimetric purposes. In this pilot study we assessed saliva for its utility in biodosimetry using a mouse model. Mice were exposed to 0.5, 3 and 8 Gy total-body gamma irradiation and saliva was collected on day 1 and 7 postirradiation. Global metabolomic profiling was conducted through liquid chromatography mass spectrometry and metabolites were positively identified using tandem mass spectrometry. Multivariate data analysis revealed distinct metabolic profiles for all groups at day 1, whereas at day 7 the two lower dose profiles appeared to have minimal differences. Metabolites that were identified include amino acids and fatty acids, and intermediates of the nicotinate and nicotinamide metabolism. The specificity and sensitivity of the radiation signature, as expected, was higher for the 8 Gy dose at both time points, as determined through generation of receiver operating characteristic curves. To the best of our knowledge, this is the first metabolomics study in saliva of irradiated mice to demonstrate the utility of this biofluid as a potential matrix for identification of radiation and dose-specific biomarkers. [ABSTRACT FROM AUTHOR]

A fast and sensitive methodology has been developed for the evaluation of the 3,4-methylenedioxypyrovalerone (MDPV) consumed. Based on ion mobility spectrometry (IMS), MDPV was directly determined in nasal fluids with a limit of detection (LOD) in the order of 22 ng mL, which corresponds to an absolute amount of 33 ng of MDPV per swab. MDPV was also determined after liquid-liquid microextraction (LLME) in oral fluids to avoid matrix effects, obtaining a LOD value of 4.4 ng mL in oral fluid samples. The IMS spectrum for MDPV exhibited a peak with K = 1.210 ± 0.005 cmV s at a drift time of 14.62 ms, the total analysis time being 4.5 min per oral fluid and 1.5 min per nasal fluid sample. Samples must be analyzed within 24 h following collection and dissolution in 2-propanol, based on the complementary stability studies. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Oral fluid (OF) is increasingly used for clinical, forensic and work- place drug testing as an alternative to urine. Uncertainties surround- ing OF collection device performance, drug stability and testing reproducibility may be partially responsible for delays in the imple- mentation of OF testing in regulated drug testing programs. Stability of D9-tetrahydrocannabinol (THC) fortified and authentic specimens was examined after routine collection, transport and laboratory test- ing. Acceptable recovery and stability were observed when THC-for- tified OF (1.5 and 4.5 ng/mL) was applied to Oral-Eze devices. Neat OF samples collected with Oral-Eze, processed per the package in- sert, and fortified with THC (3 and 6 ng/mL) were stable ( +20%) at room temperature (21 - 258C), refrigerated (2 - 88C) and frozen (225 to 2158C) conditions up to 1 month, while samples collected with Intercept devices showed decreases at refrigerated and room temperatures. After long-term refrigerated or frozen storage, maxi- mum reductions in THC concentrations were 42% for Oral-Eze and 69% for Intercept. After ≥1 year frozen storage, 80.7% of laboratory specimens positive for THC (3 ng/mL cut-off ) by GC - MS were recon- firmed positive (within 25%), with an average THC decrease of 4.2%. Specimens (n 5 47) processed with Oral-Eze (diluted) and tested via enzyme immunoassay were concordant with LC - MS-MS results and showed 100% sensitivity and 95% specificity. Paired specimens col- lected with Oral-Eze and Intercept exhibited 98% overall agreement between the immunoassay test systems. Collectively, these data demonstrate consistent and reproducible recovery and stability of THC in OF after collection, transport and laboratory testing using the Oral-Eze OF Collection System. [ABSTRACT FROM AUTHOR]