Cannabis use and neurocognitive performance at 13-14 Years-Old: Optimizing assessment with hair toxicology in the Adolescent brain cognitive development (ABCD) study.

OBJECTIVE: Cannabis is widely used, including in early adolescence, with prevalence rates varying by measurement method (e.g., toxicology vs. self-report). Critical neurocognitive development occurs throughout adolescence. Given conflicting prior brain-behavior results in cannabis research, improved measurement of cannabis use in younger adolescents is needed. METHODS: Data from the Adolescent Brain Cognitive Development (ABCD) Study Year 4 follow-up (participant age: 13-14 years-old) included hair samples assessed by LC-MS/MS and GC-MS/MS, quantifying THCCOOH (THC metabolite), THC, and cannabidiol concentrations, and the NIH Toolbox Cognitive Battery. Youth whose hair was positive for cannabinoids or reported past-year cannabis use were included in a Cannabis Use (CU) group (n = 123) and matched with non-using Controls on sociodemographics (n = 123). Standard and nested ANCOVAs assessed group status predicting cognitive performance, controlling for family relationships. Follow-up correlations assessed cannabinoid hair concentration, self-reported cannabis use, and neurocognition. RESULTS: CU scored lower on Picture Memory (p = .03) than Controls. Within the CU group, THCCOOH negatively correlated with Picture Vocabulary (r = -0.20, p = .03) and Flanker Inhibitory Control and Attention (r = -0.19, p = .04), and past-year cannabis use was negatively associated with List Sorting Working Memory (r = -0.33, p = .0002) and Picture Sequence Memory (r = -0.19, p = .04) performances. CONCLUSIONS: Youth who had used cannabis showed lower scores on an episodic memory task, and more cannabis use was linked to poorer performances on verbal, inhibitory, working memory, and episodic memory tasks. Combining hair toxicology with self-report revealed more brain-behavior relationships than self-report data alone. These youth will be followed to determine long-term substance use and neurocognition trajectories.

An assessment of qualitative and quantitative cannabinoids analysis in selected commercially available cannabis oils in Argentina.

In recent years, the therapeutic use of cannabis products, especially cannabis oils, has increased significantly, due to the pharmacological potential of their cannabinoids, for the treatment of conditions, such as pain management, cancer, and epilepsy. In Argentina, patients with medical prescriptions can access to cannabis oil, through self-cultivation, a third-person (grower or importer), or a civil organization authorized for that purpose. However, these products remain largely unregulated in Argentina, and information available regarding labeling accuracy, especially cannabidiol (CBD)/ Δ9-tetrahydrocannabinol (Δ9-THC) concentrations are inconsistent or nonexistent, nor long-term product stability, and lot to lot variability. Understanding these properties is fundamental if these products are to be used in patients with a determinate pathology. Therefore, we analyzed commercially available cannabis oils (n: 500) in Argentina for qualitative and quantitative cannabinoids content. In order to provide a detailed overview of their cannabinoids profiles, and determine Δ9-THC, CBD, and cannabinol (CBN) concentrations, samples were diluted and analyzed by gas chromatography- mass spectrometry (GC/MS). Most of the samples tested positive for cannabinoids (n: 469) with Δ9-THC and CBD as the predominant cannabinoids. Among products tested, only 29.8% (n: 149) gave specific CBD label claims, and testing indicated a CBD tested positive of 70.5% (n: 105). For products (n: 17) with a THC-free label claim, testing indicated 76.5% (n: 13) of Δ9-THC positive, and cannabinoids were not detected in four products. Δ9-THC concentrations ranged from 0.1 to 143.0 mg/mL, CBD concentrations from 0.1 to 125.3 mg/mL, and CBN concentrations from 0.04 to 60.10 mg/mL; CBN/ Δ9-THC ratios ranged from 0.0012 to 2.31, and CBD/ Δ9-THC ratios from 0.0008 to 178.87. Furthermore, the (Δ9-THC + CBN)/CBD ratio of most samples was greater than one. In summary, our results indicate that cannabis oil products show wide variability in cannabinoids content, purity, and labeling.

Removal of residues of psychoactive substances during wastewater treatment, their occurrence in receiving river waters and environmental risk assessment.

Continuous consumption combined with incomplete removal during wastewater treatment means residues of psychoactive substances (licit drugs, medications of abuse and illicit drugs) are constantly introduced into the aquatic environment, where they have the potential to affect non-target organisms. In this study, 17 drug residues of psychoactive substances were determined in wastewater influent, effluent and in receiving rivers of six Slovene municipal wastewater treatment plants employing different treatment technologies. Variations in removal efficiencies (REs) during spring, summer and winter were explored, and ecotoxic effects were evaluated using in silico (Ecological Structure-Activity Relationships software-ECOSAR) and in vivo (algal growth inhibition test) methods. Drug residues were detected in influent and effluent in the ng/L to µg/L range. In receiving rivers, biomarkers were in the ng/L range, and there was good agreement between measured and predicted concentrations. On average, REs were highest for nicotine, 11-nor-9-carboxy-∆9-tetrahydrocannabinol (THC-COOH), cocaine residues, and amphetamine (>90 %) and lowest for methadone residues (<30 %). REs were comparable between treatments involving activated sludge and membrane bioreactors, while the moving biofilm bed reactor (MBBR) removed cotinine, cocaine, and benzoylecgonine to a lesser extent. Accordingly, higher levels of nicotine and cocaine residues were detected in river water receiving MBBR discharge. Although there were seasonal variations in REs and levels of drug residues in receiving rivers, no general pattern could be observed. No significant inhibition of algal growth (Chlamydomonas reinhardtii) was observed for the tested compounds (1 mg/L) during 72 h and 240 h of exposure, although effects on aquatic plants were predicted in silico. In addition, environmental risk assessment revealed that levels of nicotine, methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), morphine, and 3,4-methylenedioxymethamphetamine (MDMA) pose a risk to aquatic organisms. Since nicotine and EDDP can have acute and chronic effects, the authors support regular monitoring of receiving surface waters, followed up by regulatory actions.

Delta9-THC determination by the EU official method: evaluation of measurement uncertainty and compliance assessment of hemp samples.

Hemp cultivation is living a period of renewed interest worldwide after long years of opposition and abandonment. The European Union (EU) allows and subsidizes the growing of fiber and oilseed cultivars of Cannabis sativa L. with respect to the THC content limit of 0.2%. The EU method for the quantitative determination of Δ9-tetrahydrocannabinol (THC) content in hemp varieties provides to apply a tolerance of 0.03 g of THC per 100 g of sample concerning compliance assessment to that limit. However, the method does not report any precision data, especially useful as a function of THC content to evaluate measurement uncertainty and therefore to establish the conformity of hemp at different THC legal limits. Measurement uncertainty of the method by both bottom-up and top-down approach, besides repeatability and reproducibility, was investigated and estimated in the THC concentration range 0.2-1.0%, which includes the different legal limits set out for hemp around the world. We proposed Decision Rules for conformity of hemp showing that a non-compliant declaration beyond reasonable doubt should be stated when the THC content, as a mean result on a duplicate analysis, exceeds the limit by at least 11-15%, depending on THC limit. We highlighted other issues concerning practical aspects of hemp analysis, from sampling to evaluation of results, as well as the need to carry out collaborative studies on the EU method.

Low-Cost Active Thermography using Cellphone Infrared Cameras: from Early Detection of Dental Caries to Quantification of THC in Oral Fluid.

Active thermography (AT) is a widely studied non-destructive testing method for the characterization and evaluation of biological and industrial materials. Despite its broad range of potential applications, commercialization and wide-spread adaption of AT has long been impeded by the cost and size of infrared (IR) cameras. In this paper, we demonstrate that this cost and size limitation can be overcome using cell-phone attachment IR cameras. A software development kit (SDK) is developed that controls camera attributes through a simple USB interface and acquires camera frames at a constant frame rate up to 33 fps. To demonstrate the performance of our low-cost AT system, we report and discuss our experimental results on two high impact potential applications. The first set of experiments is conducted on a dental sample to investigate the clinical potential of the developed low-cost technology for detecting early dental caries, while the second set of experiments is conducted on the oral-fluid based lateral flow immunoassay to determine the viability of our technology for detecting and quantifying cannabis consumption at the point-of-care. Our results suggest achievement of reliable performance in the low-cost platform, comparable to those of costly and bulky research-grade systems, paving the way for translation of AT techniques to market.

[Plant-derived contaminants in food : Occurrence, effects and risk assessment]. / Pflanzliche Kontaminanten in Lebensmitteln : Vorkommen, Wirkung und Risikobewertung.

Among the various contaminants, the group of natural plant-derived substances in the modern food chain has been generating increasing concern in recent years. The adverse effects encountered may be diverse and pose risks of acute, subchronic or chronic toxicity. The underlying mechanisms of toxicity may be thresholded or be based on interactions with DNA, as for genotoxic carcinogens, for which the existence of a threshold cannot be assumed. This article gives an overview of the major plant-derived contaminants of present concern in the modern food chain and describes their mode of action and adverse effects.

Integrating environmental and self-report data to refine cannabis prevalence estimates in a major urban area of Switzerland.

BACKGROUND: Cannabis consumption is a topical subject because of discussions about reviewing current regulations. In this context, having a more comprehensive approach to assess and monitor prevalence and consumption is highly relevant. The objective of this work was to refine current estimates about prevalence of cannabis use by combining self-report data and results derived from wastewater analysis. METHODS: Self-report data was retrieved from surveys conducted in Switzerland and Europe. Wastewater samples were collected at the wastewater treatment plant of Lausanne, western Switzerland, over a 15 months period. The occurrence of 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH), a specific metabolite of delta-9-tetrahydrocannabinol (THC), was monitored. Bayesian hierarchical models were used to estimate consumption, prevalence and number of cannabis users in the investigated area. RESULTS: According to survey data, 12-months prevalence in western Switzerland was estimated to 6.2% of the population aged 15 or older, with an estimated daily cannabis consumption of 8.1gday(-1)·1000inhab(-1) (at 11.2% purity). The integrative model comprising self-report and wastewater data substantially reduced the uncertainty in the estimates and suggested a last-year prevalence of 9.4%, with a daily cannabis consumption of 14.0gday(-1)·1000inhab(-1). CONCLUSION: Although in the same order of magnitude, consumption and prevalence estimates obtained with the integrative model were 78% and 52% higher compared to self-report figures, respectively. Interestingly, these figures are similar to discrepancies observed when comparing self-reported alcohol consumption and sales or tax data. The suggested integrative model allowed to account for known sources of uncertainty and provided refined estimates of cannabis prevalence in a major urban area of Switzerland.

Limitations to the Dutch cannabis toleration policy: Assumptions underlying the reclassification of cannabis above 15% THC.

The Netherlands has seen an increase in Δ9-tetrahydrocannabinol (THC) concentrations from approximately 8% in the 1990s up to 20% in 2004. Increased cannabis potency may lead to higher THC-exposure and cannabis related harm. The Dutch government officially condones the sale of cannabis from so called 'coffee shops', and the Opium Act distinguishes cannabis as a Schedule II drug with 'acceptable risk' from other drugs with 'unacceptable risk' (Schedule I). Even in 1976, however, cannabis potency was taken into account by distinguishing hemp oil as a Schedule I drug. In 2011, an advisory committee recommended tightening up legislation, leading to a 2013 bill proposing the reclassification of high potency cannabis products with a THC content of 15% or more as a Schedule I drug. The purpose of this measure was twofold: to reduce public health risks and to reduce illegal cultivation and export of cannabis by increasing punishment. This paper focuses on the public health aspects and describes the (explicit and implicit) assumptions underlying this '15% THC measure', as well as to what extent these are supported by scientific research. Based on scientific literature and other sources of information, we conclude that the 15% measure can provide in theory a slight health benefit for specific groups of cannabis users (i.e., frequent users preferring strong cannabis, purchasing from coffee shops, using 'steady quantities' and not changing their smoking behaviour), but certainly not for all cannabis users. These gains should be weighed against the investment in enforcement and the risk of unintended (adverse) effects. Given the many assumptions and uncertainty about the nature and extent of the expected buying and smoking behaviour changes, the measure is a political choice and based on thin evidence.

Cannabinoid findings in children hair - what do they really tell us? An assessment in the light of three different analytical methods with focus on interpretation of Δ9-tetrahydrocannabinolic acid A concentrations.

Hair analysis for drugs and drugs of abuse is increasingly applied in child protection cases. To determine the potential risk to a child living in a household where drugs are consumed, not only can the hair of the parents be analyzed but also the hair of the child. In the case of hair analysis for cannabinoids, the differentiation between external contamination and systemic uptake is particularly difficult, since the drug is quite often handled extensively prior to consumption (e.g. when preparing a joint) and smoke causes a further risk for an external contamination. Δ9-tetrahydrocannabinolic acid A (THCA-A), the non-psychoactive biogenetic precursor of Δ9-tetrahydrocannabinol (THC), is a suitable marker for external contamination since it is not incorporated into the hair matrix through the bloodstream in relevant amounts. In the presented study, hair samples from 41 children, 4 teenagers, and 34 drug-consuming parents were analyzed for THCA-A, THC and cannabinol (CBN) applying methanolic extraction and a fully validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method (Method 1). For comparison, a part of the samples was also analyzed applying alkaline hydrolysis followed by liquid/liquid extraction and gas chromatography-mass spectrometry (GC-M)S (Method 2), or by headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) (Method 3). Furthermore, 458 seized marihuana samples and 180 seized hashish samples were analyzed for the same cannabinoids by gas-chromatography-flame ionization detector (GC-FID). In all but one of the hair samples, the concentration of THCA-A was higher than the concentration of THC and in 14 cases no THC could be detected despite the presence of THCA-A, suggesting that in almost all cases a significant external contamination had occurred. Within-family comparison showed a higher THCA-A/THC ratio in hair of children than of their consuming caregivers. Mean and median of this ratio of all hair samples (6.7 and 4.2) were between those of marihuana (11.0 and 8.3) and hashish (2.8 and 2.1) with a large variation in all samples. Comparison of the Methods 1 to 3 showed clearly that the choice of the analytical procedure has a strong influence on the quantitative results, mainly because of decarboxylation of THCA-A during hair hydrolysis by NaOH and other analytical steps, which lead to artifactually elevated THC concentrations. In conclusion, these findings suggest that the major part of the cannabinoids detected in the hair samples from children arose from an external contamination through 'passive' transfer by e.g. contaminated hands or surfaces and not from inhalation or deposition of side stream smoke.

The effects of medical marijuana laws on potency.

BACKGROUND: Marijuana potency has risen dramatically over the past two decades. In the United States, it is unclear whether state medical marijuana policies have contributed to this increase. METHODS: Employing a differences-in-differences model within a mediation framework, we analyzed data on n=39,157 marijuana samples seized by law enforcement in 51 U.S. jurisdictions between 1990 and 2010, producing estimates of the direct and indirect effects of state medical marijuana laws on potency, as measured by Δ(9)-tetrahydrocannabinol content. RESULTS: We found evidence that potency increased by a half percentage point on average after legalization of medical marijuana, although this result was not significant. When we examined specific medical marijuana supply provisions, results suggest that legal allowances for retail dispensaries had the strongest influence, significantly increasing potency by about one percentage point on average. Our mediation analyses examining the mechanisms through which medical marijuana laws influence potency found no evidence of direct regulatory impact. Rather, the results suggest that the impact of these laws occurs predominantly through a compositional shift in the share of the market captured by high-potency sinsemilla. CONCLUSION: Our findings have important implications for policymakers and those in the scientific community trying to understand the extent to which greater availability of higher potency marijuana increases the risk of negative public health outcomes, such as drugged driving and drug-induced psychoses. Future work should reconsider the impact of medical marijuana laws on health outcomes in light of dramatic and ongoing shifts in both marijuana potency and the medical marijuana policy environment.

Assessment of different mouthwashes on cannabis oral fluid concentrations.

Since the implementation of mandatory drug testing in drivers' oral fluid, several solutions to avoid an onsite positive result can be found on drug users' forums, especially for marijuana, including the use of different mouthwashes. Recently, a product for personal hygiene, Kleaner, has been sold for this purpose. The aims of this study were to assess the effect of water, whole milk, and Kleaner mouthwashes on tetrahydrocannabinol (THC) oral fluid concentrations, and those observed in passive smokers subjected to extreme contamination conditions. The study was performed on four days. On day 0, study information was given to the participants. On days 1, 2, and 3, 11 chronic cannabis users smoked their usual daily dose, and oral fluid specimens were collected before smoking (t=-0.5h) and at t=0.25, 0.5, 1, 3, 6, 12, and 24 h post-smoking. On day 1, participants rinsed their mouth with water before each specimen collection. On day 2, 5 participants rinsed their mouth with Kleaner and 6 with whole milk. On day 3, a specimen was collected before and after rinsing the mouth with water. Statistically significant lower concentrations were observed comparing concentrations in oral fluid specimens collected before and after a water rinse. However, maximum THC concentrations at t=0.25 h were >3-fold higher than the cut-off employed by the Spanish police (25 ng/mL) regardless of the use of any mouthwash. THC was also detected in the oral fluid of passive smokers subjected to extreme contamination conditions; however, concentrations were <25 ng/mL in all cases.

Valoración de los efectos fisiológicos y neuroquímicos de altas concentraciones de tetrahidrocannabinol en modelos biológicos / Assessment physiological and neurochemical effects of high concentrations of tetrahydrocannabinol in biological models

El objetivo de esta investigación fue evaluar los efectos fisiológicos y neuroquímicos en 60 ratones machos cepas Naval Medical Research Institute (NMRI) en edad adulto-joven con pesos promedios de 25,45 ± 3,05 g, sometidos durante seis semanas a dosis del principio psicoactivo de la marihuana el Δ-9-tetrahidrocannabinol en concentraciones entre 4 - 20%. Se realizaron tomas de sangre retroorbital para evaluar parámetros hematológicos y bioquímicos antes, durante y post experiencia. Se monitorearon medidas tales como: peso, ingesta de agua, alimentos, actividad locomotora horizontal y vertical, entre otros. Al final de la experiencia se realizo autopsia y toma de muestras de regiones cerebrales, para medir niveles de neurotransmisores aminoacidicos y dopamina. Estos resultados permiten concluir que altas concentraciones del principio psicoactivo de la marihuana hacen más dependiente al consumidor con los consecuentes daños fisiológicos y neurológicos. Esto lleva a que cada vez se necesite más droga para producir el mismo efecto.

The objective of this research was to evaluate the physiological and neurochemical effects in 60 (Naval Medical Research Institute) NMRI male mice strains in young adult - age average weight 25.45 ± 3.05 g, underwent six weeks at doses of the psychoactive ingredient in marijuana the Δ -9-tetrahydrocannabinol in concentrations between 4-20 %. Retroorbital blood shots were conducted to evaluate hematological and biochemical parameters before, during and post experience. Weight, water intake, food, horizontal and vertical locomotors activity include: measures such as monitored. At the end of the experience autopsy was conducted and sampling of brain regions to measure levels of amino acid neurotransmitters and dopamine. These results suggest that high concentrations of the psychoactive ingredient in marijuana consumers become more dependent with consequent physiological and neurological damage. This leads to more and more drugs is needed to produce the same effect.

Development of an electrospray LC-MS/MS method for quantification of Δ(9) -tetrahydrocannabinol and its main metabolite in oral fluid.

A fast and sensitive reference method for quantification of Δ(9) -tetrahydrocannabinol (THC) and its main metabolite 11-nor-9-carboxy-Δ(9) -tetrahydrocannabinol (THCCOOH) in oral fluid is described in this study. Samples were collected using an oral specimen collection device, followed by solid-phase extraction and liquid chromatography-tandem mass spectrometry analysis. Chromatographic separation of the analytes was achieved by gradient elution on a reversed-phase column with subsequent detection by electrospray triple quadrupole mass spectrometry in positive ionization multiple reaction monitoring mode. Quantification was performed by means of deuterated analogues of the analytes as internal standards. Total run time of the assay was 12 min. The method allowed sensitive quantification of both analytes at a limit of quantification of 0.2 ng/ml. This sensitivity is essential for analysis of samples collected with the Intercept Oral Fluid Collection device (OraSure) and an assay for simultaneous quantification of THC and THCCOOH in saliva has not yet been described. The calibration curves for THC and THCCOOH were linear in the range between 0.25 and 8 ng/ml (r(2) > 0.99). Ion suppression effects from endogenous or exogenous interferences were investigated using selected model substances (albumin, ascorbic acid, bilirubin, hemoglobin, breath spray, cigarette, chewing gum, chewing tobacco, candy, tooth whitening, and Tums antacid). These substances were chosen because of the high probability of their presence in the collected samples. None of the 11 endogenous model interferences altered the accuracy of analysis, demonstrating good robustness of the method with respect to interferences in common hygiene products, medicine, tobacco and naturally occurring endogenous substances.

Can the prevalence of high blood drug concentrations in a population be estimated by analysing oral fluid? A study of tetrahydrocannabinol and amphetamine.

AIM: To study several methods for estimating the prevalence of high blood concentrations of tetrahydrocannabinol and amphetamine in a population of drug users by analysing oral fluid (saliva). METHODS: Five methods were compared, including simple calculation procedures dividing the drug concentrations in oral fluid by average or median oral fluid/blood (OF/B) drug concentration ratios or linear regression coefficients, and more complex Monte Carlo simulations. Populations of 311 cannabis users and 197 amphetamine users from the Rosita-2 Project were studied. RESULTS: The results of a feasibility study suggested that the Monte Carlo simulations might give better accuracies than simple calculations if good data on OF/B ratios is available. If using only 20 randomly selected OF/B ratios, a Monte Carlo simulation gave the best accuracy but not the best precision. Dividing by the OF/B regression coefficient gave acceptable accuracy and precision, and was therefore the best method. None of the methods gave acceptable accuracy if the prevalence of high blood drug concentrations was less than 15%. CONCLUSION: Dividing the drug concentration in oral fluid by the OF/B regression coefficient gave an acceptable estimation of high blood drug concentrations in a population, and may therefore give valuable additional information on possible drug impairment, e.g. in roadside surveys of drugs and driving. If good data on the distribution of OF/B ratios are available, a Monte Carlo simulation may give better accuracy.

Assessment of the genetic stability of micropropagated plants of Cannabis sativa by ISSR markers.

Inter-simple sequence repeat (ISSR) markers were used to evaluate the genetic stability of the micropropagated plants of Cannabis sativa over 30 passages in culture and hardening in soil for 8 months. A total of 15 ISSR primers resulted in 115 distinct and reproducible bands. All the ISSR profiles from micropropagated plants were monomorphic and comparable to mother plants, confirming the genetic stability among clones and mother plants. Chemical analysis of cannabinoids, using gas chromatography/flame ionization detection (GC/FID), was done to further confirm whether the qualitative and quantitative differences in the major secondary metabolites exist between the mother plant and micropropagated plants. Six major cannabinoids - Delta(9)-THC, THCV, CBD, CBC, CBG, and CBN - were identified and compared with the mother plant. Our results clearly showed a similar cannabinoid profile and insignificant differences in THC content between the two types of plants. These results suggest that the micropropagation protocol developed by us for rapid IN VITRO multiplication is appropriate and applicable for clonal mass propagation of C. SATIVA.