A multi-centre, tolerability study of a cannabidiol-enriched Cannabis Herbal Extract for chronic headaches in adolescents: The CAN-CHA protocol.

INTRODUCTION: Cannabis products have been used in the management of headaches in adults and may play a role in pediatric chronic pain. Canadian pediatricians report increasing use of cannabis for the management of chronic headaches, despite no well-controlled studies to inform its dosing, safety, and effectiveness. The aim of our clinical trial is to determine the dosing and safety of a Cannabidiol (CBD)-enriched Cannabis Herbal Extract (CHE) for the treatment of chronic headaches in adolescents. METHODS AND ANALYSIS: Youth, parents, and an expert steering committee co-designed this tolerability study. Twenty adolescents (aged 14 to 17 years), with a chronic migraine diagnosis for more than 6 months that has not responded to other therapies will be enrolled into an open label, dose escalation study across three Canadian sites. Study participants will receive escalating doses of a CBD-enriched CHE (MPL-001 with a THC:CBD of 1:25), starting at 0.2-0.4 mg/kg of CBD per day and escalating monthly up to 0.8-1.0 mg/kg of CBD per day. The primary objective of this study is to determine the safety and tolerability of CBD-enriched CHE in adolescents with chronic migraine. Secondary objectives of this study will inform the development of subsequent randomized controlled trials and include investigating the relationship between the dose escalation and change in the frequency of headache, impact and intensity of pain, changes in sleep, mood, function, and quality of life. Exploratory outcomes include investigating steady-state trough plasma levels of bioactive cannabinoids and investigating how pharmacogenetic profiles affect cannabinoid metabolism among adolescents receiving CBD-enriched CHE. DISCUSSION: This protocol was co-designed with youth and describes a tolerability clinical trial of CBD-enriched CHE in adolescents with chronic headaches that have not responded to conventional therapies. This study is the first clinical trial on cannabis products in adolescents with chronic headaches and will inform the development of future comparative effectiveness clinical trials. TRIAL REGISTRATION: CAN-CHA trial is registered with ClinicalTrials.gov with a number of register NCT05337033.

Tea Prepared from Dried Cannabis: What Do We Drink?

Besides many other uses, dried Cannabis may be used for "tea" preparation. This study focused on a comprehensive characterization of an aqueous infusion prepared according to a common practice from three fairly different Cannabis cultivars. The transfer of 42 phytocannabinoids and 12 major bioactive compounds (flavonoids) into the infusion was investigated using UHPLC-HRMS/MS. Phytocannabinoid acids were transferred generally in a higher extent compared to their counterparts; in the case of Δ9-THC, it was only in the range of 0.4-1.9% of content in the Cannabis used. A dramatic increase of phytocannabinoids, mainly of the neutral species, occurred when cream was added during steeping, and the transfer of Δ9-THC into "tea" achieved a range of 53-64%. Under such conditions, drinking a 250 mL cup of such tea by a 70 kg person might lead to multiple exceedance of the Acute Reference Dose (ARfD), 1 µg/kg b.w., even in the case when using hemp with a Δ9-THC content below 1% in dry weight for preparation.

The effects of Cannabis sativa and cannabinoids on the inhibition of pancreatic lipase - An enzyme involved in obesity.

INTRODUCTION: Obesity is a chronic noncommunicable disease characterized by excessive body fat that can have negative health consequences. Obesity is a complex disease caused by a combination of genetic, environmental, and lifestyle factors. It is characterized by a discrepancy between caloric intake and expenditure. Obesity increases the risk of acquiring major chronic diseases, including heart disease, stroke, cancer, and Type 2 diabetes mellitus (T2DM). Currently, the inhibition of pancreatic lipases (PL) is a promising pharmacological therapy for obesity and weight management. In this study, the inhibition of pancreatic lipase by Cannabis sativa (C. sativa) plant extract and cannabinoids was investigated. METHODS: The inhibitory effect was assessed using p-nitrophenyl butyrate (pNPB), and the results were obtained by calculating the percentage relative activity and assessed using one-way analysis of variance (ANOVA). Kinetic studies and spectroscopy techniques were used to evaluate the mode of inhibition. Diet-induced; and diabetic rat models were studied to evaluate the direct effects of C. sativa extract on PL activity. RESULTS: Kinetic analyses showed that the plant extracts inhibited pancreatic lipase, with tetrahydrocannabinol (THC) and cannabinol (CBN) being the potential cause of the inhibition noted for the C. sativa plant extract. CBN and THC inhibited the pancreatic lipase activity in a competitive manner, with the lowest residual enzyme activity of 52 % observed at a 10 µg/mL concentration of CBN and 39 % inhibition at a 25 µg/mL concentration of THC. Circular dichroism (CD) spectroscopy revealed that the inhibitors caused a change in the enzyme's secondary structure. At low concentrations, THC showed potential for synergistic inhibition with orlistat. C.sativa treatment in an in vivo rat model confirmed its inhibitory effects on pancreatic lipase activity. CONCLUSION: The findings in this study provided insight into the use of cannabinoids as pancreatic lipase inhibitors and the possibility of using these compounds to develop new pharmacological treatments for obesity.

Cannabis-infused foods: Phytonutrients, health, and safe product innovations.

Cannabis-infused foods are currently on the rise in markets all around the world. Meanwhile, there are concerns over the health implications for consumers. Studies have explored the therapeutic potential and nutritional and economic benefits of cannabis usage. Yet, the phytonutrients, processing methods, and health implications of cannabis-infused foods have not been well explored. This review evaluates existing evidence on the nutritional, processing, safety, and phytonutrient composition of cannabis-infused food products and their medicinal and functional prospects. Cannabis seeds contain the highest amount of dietary nutrients, while flowers contain the highest amount of bioactive constituents. Oils, butter, seeds, flowers, and leaf extracts are the plant forms currently incorporated into food products such as beverages, baked products, cooking ingredients, functional foods, nutraceuticals, and nootropics. Cannabis-infused foods have been found to offer therapeutic benefits for pain management, brain function, gut health, and certain cancers. Findings also show significant constraints associated with cannabis-infused foods regarding dosage guidelines, limited research, efficacy, and long-term health effects on consumers. This is further worsened by the lack of policies that regulate the industry. To realize the full potential of cannabis use in the food and health industries and in research, regulatory guidelines are needed to control dosages and improve its efficient use in these industries. This will go a long way to ensure the safety of cannabis users and enhance responsible production, marketing, and distribution.

Cannabis sativa in veterinary medicine: Foundations and therapeutic applications.

An increase in products containing phytocannabinoids, particularly cannabidiol, is often observed in human and veterinary markets following the legalization of hemp (cannabis) for industrial purposes. In veterinary medicine, derivatives of Cannabis sativa are used for managing pain (osteoarticular, oncological, and neuropathic), epilepsy, and behavioral disorders, as well as oncological, immune-mediated, cardiovascular, and respiratory diseases. In addition, there is growing interest in incorporating C. sativa into livestock feed. To elucidate the mechanisms of action of phytocannabinoids, a thorough understanding of the endocannabinoid system and its role in maintaining homeostasis is essential. Short-term use of phytocannabinoid products appears generally safe, but further research is required to understand the routes of administration, pharmacokinetics, and pharmacodynamics across various species. Although literature on phytocannabinoids in veterinary patients is limited, the available data suggest significant therapeutic potential.

Cannabis sativa en médecine vétérinaire : fondements et applications thérapeutiquesUne augmentation des produits contenant des phytocannabinoïdes, notamment du cannabidiol, est souvent observée sur les marchés humains et vétérinaires à la suite de la légalisation du chanvre (cannabis) à des fins industrielles. En médecine vétérinaire, les dérivés du Cannabis sativa sont utilisés pour gérer la douleur (ostéoarticulaire, oncologique et neuropathique), l'épilepsie et les troubles du comportement, ainsi que les maladies oncologiques, immunitaires, cardiovasculaires et respiratoires. En outre, l'incorporation de C. sativa dans l'alimentation du bétail suscite un intérêt croissant. Pour élucider les mécanismes d'action des phytocannabinoïdes, une compréhension approfondie du système endocannabinoïde et de son rôle dans le maintien de l'homéostasie est essentielle. L'utilisation à court terme de produits phytocannabinoïdes semble généralement sécuritaire, mais des recherches supplémentaires sont nécessaires pour comprendre les voies d'administration, la pharmacocinétique et la pharmacodynamique chez diverses espèces. Bien que la littérature sur les phytocannabinoïdes chez les patients vétérinaires soit limitée, les données disponibles suggèrent un potentiel thérapeutique important.(Traduit par Dr Serge Messier).

Profile of Polyphenols, Fatty Acids, and Terpenes in Henola Hemp Seeds Depending on the Method of Fertilization.

Botanical varieties of hemp differ in chemical composition, plant morphology, agronomy, and industrial suitability. Hemp is popular for cultivation for the production of cannabinoid oil, fiber production, biomass, etc. The fertilization process is one of the most important factors affecting the plant, both its condition and chemical composition. So far, research has been carried out proving that hemp is a valuable source of, among others: fatty acids, amino acids, acids, vitamins, numerous micro- and macroelements, and antioxidant compounds. In this experiment, it was decided to check the possibility of harvesting hemp panicles twice in one year. The purpose of this treatment is to use one plant to produce cannabidiol oil and grain. The main aim of the research was to determine bioactive compounds in hemp seeds and to determine whether the cultivation method affects their content and quantity. Based on the research conducted, it was observed that hemp can be grown in two directions at the same time and harvested twice because its health-promoting properties do not lose their value. It was found that regardless of whether hemp is grown solely for seeds or to obtain essential oils and then seeds, the type of fertilization does not affect the content of phenolic acids (e.g., syringic acid: 69.69-75.14 µg/100 g, vanillic acid: 1.47-1.63 µg/100 g). Based on the conducted research, it was found that essential oils can be obtained from one plant in the summer and seeds from Henola hemp cultivation in the autumn, because such a treatment does not affect the content of the discussed compounds.

Encapsulation of cannabidiol in hemp seed oleosomes.

Oleosomes are natural lipid droplets that can be extracted intact from oil seeds, forming oil/water emulsions. Their lipid cores, surrounded by a monolayer of phospholipids and proteins, make oleosomes suitable as carriers of hydrophobic bioactive compounds like cannabidiol (CBD). As CBD is crystalline at room temperature, it first has to be liquified to allow better encapsulation. This was done by heating (80 °C for 4 h) or by pre-solubilizing CBD in ethanol and then the liquified CBD was mixed with oleosome dispersions for the encapsulation. Both methods exhibit good encapsulation efficiency, but the results were significantly influenced by the ratio of CBD to lipid contents, regardless of the encapsulation method applied. At higher concentrations of CBD relative to that of the lipid in the oleosomes, the encapsulation efficiency decreased as saturation was attained. Moreover, the in vitro digestion analysis was conducted to investigate the potential of oleosomes as carriers to transport CBD. The relatively slow and steady release of CBD from oleosomes indicates that oleosomes are a slow-release carrier for hydrophobic functional ingredients. An important finding is that the encapsulation and in vitro digestive properties of the oleosomes remain unaffected by the presence of CBD, heating treatment or ethanol, which could bring more opportunities for the applications of oleosomes as carriers in various fields.

Inter-strain variability in responses to a single administration of the cannabidiol-rich cannabis extract in mice.

Cannabidiol (CBD) has gained widespread popularity; however, its pharmacological and toxicological profiles in the context of human genetic diversity remain largely unexplored. Here, we investigated the variability in metabolism and toxicity of CBD-rich cannabis extract (CRCE) in genetically diverse mouse models: C57BL/6J, B6C3F1/J, and NZO/HlLtJ strains. Mice received a single dose of CRCE containing 57.9% CBD at dosages of 0, 246, 738, and 2460 mg/kg of CBD. At 24 h after treatment, no appreciable histomorphological changes were detected in the liver. Plasma bilirubin levels increased markedly in all strains at the highest CBD dose. Mice in all treatment groups displayed significant but distinct increases in ALT and AST levels. While B6C3F1/J and NZO/HlLtJ mice had negligible plasma CBD levels at 738 mg/kg, C57BL/6J mice exhibited levels exceeding 7000 ng/mL. At 2460 mg/kg, high CBD concentrations were found in B6C3F1/J and C57BL/6J mice, but markedly lower levels were seen in NZO/HlLtJ mice. Gene expression profiling showed significant increases in Cyp2b10 across all strains but varying responses in Cyp1a1 expression, indicating strain-specific CYP dysregulation. Genetically diverse mice exhibited differential pharmacological and toxicological responses to CRCE, suggesting a high potential for inter-individual variability in the pharmacology and toxicology of CBD in humans.

Semen Cannabis and Oleum Hyperici: Antileishmanial activity against Leishmania tropica promastigotes and intracellular amastigotes.

The exploration of alternative agents and novel drug candidates for the effective treatment of cutaneous leishmaniasis has garnered significant attention, driven by the high cost, toxic effects, and the emergence of drug resistance associated with current therapeutic options. Plant extracts derived from Semen Cannabis, the seeds of the Cannabis sativa L. (hemp) plant, and Oleum Hyperici, the oily macerate of Hypericum perforatum L. (St. John's Wort) plant, were prepared by using solvents of varying polarity (n-hexane, chloroform, ethanol, and 60% aqueous ethanol). The primary objective of this study was to research in vitro and ex vivo antileishmanial efficacy of Semen Cannabis and Oleum Hyperici plant extracts against Leishmania tropica promastigotes and intracellular amastigotes. The efficacy of plant extracts against promastigotes were assessed using the cell counting by hemocytometer and the CellTiter-Glo assay. Additionally, their impact on infected THP-1 macrophages and the quantity of intracelluler amastigotes were investigated. Cytotoxicity was evaluated in THP-1 macrophages. Among the tested plant extracts, chloroform extract of Oleum Hyperici demonstrated significant antileishmanial activity against promastigotes (SI: 12.6) and intracellular amastigotes (SI: 16.8) of L. tropica without inducing cytotoxic effects and hold promise for further investigation as potential antileishmanial agents.

An initial exploration of machine learning for establishing associations between genetic markers and THC levels in Cannabis sativa samples.

Cannabis sativa, a globally commercialized plant used for medicinal, food, fiber production, and recreation, necessitates effective identification to distinguish legal and illegal varieties in forensic contexts. This research utilizes multivariate statistical models and Machine Learning approaches to establish correlations between specific genotypes and tetrahydrocannabinol (Δ9-THC) content (%) in C. sativa samples. 132 cannabis leaves samples were obtained from legal growers in Piedmont, Italy, and illegal drug seizures in Turin. Samples were genetically profiled using a 13-loci STR multiplex and their Δ9-THC content was detected through quantitative GC-MS analysis. This study aims to assess the use of supervised classification modelling on genetic data to distinguish cannabis samples into legal and illegal categories, revealing distinct clusters characterized by unique allele profiles and THC content. t-distributed Stochastic Neighbor Embedding (t-SNE), Random Forest (RF) and Partial Least Squares Regression (PLS-R) were executed for the machine learning modelling. All the tested models resulted effective discriminating between legal samples and illegal. Although further validation is necessary, this study presents a novel forensic investigative approach, potentially aiding law enforcement in significant marijuana seizures or tracking illicit drug trafficking routes.

Greenness assessment of analytical methods for determination of cannabinoids in oils using NEMI, Analytical Eco-Scale, AGREE and GAPI.

The cannabis plant is being increasingly researched due to its numerous therapeutic properties leading to the need for analytical techniques to assess substances present in extracts of the cannabis plant in carrier oils, such as medium chain triglycerides (MCT) oil. Awareness of the environmental impact of activities related to analysis led to the development of greenness assessment metrics. This study aimed to assess the environmental impact of analytical techniques applied in the analysis of cannabinoids in oil using Green Analytical Chemistry metrics. The first phase of the study consisted of a systematic literature review to identify high performance liquid chromatography and ultra high performance liquid chromatographic methods of analysis for cannabinoids in oil. In the second phase, the identified methods were assessed using the National Environmental Method Index (NEMI), Analytical Eco-scale, Analytical Greenness Calculator (AGREE) and Green Analytical Procedure Index (GAPI). Out of 124 identified studies, 8 were considered for the comparative analysis. The identified analytical methods consisted of high performance liquid chromatography (HPLC) using high resolution MS (n = 1), DAD (n = 2), UV (n = 1), UV and MS (n = 2) and MS/MS (n = 2) as detectors. When the analytical methods were assessed using the Analytical Eco-Scale, 7 out of 8 methods achieved a score ranging between 50 and 73, categorising them as acceptable green methods of analysis. One method achieved a total score of 80, categorising the method as an excellent green analysis. The application of Green Analytical Chemistry and respective metrics during the development of analytical methods contributes towards a reduction in the environmental footprint which results from related activities.

Microwave-assisted extraction, separation, and chromogenic detection of laced marijuana for presumptive point-of-interdiction testing.

Presumptive drug screening enables timely procurement of search and arrest warrants and represents a crucial first step in crime scene analysis. Screening also reduces the burden on forensic laboratories which often face insurmountable backlogs. In most scenarios, on-site presumptive drug screening relies on chemical field tests for initial identification. However, even when used appropriately, these test kits remain limited to subjective colorimetric analysis, produce false positive or negative results with excessive sample quantities, and are known to cross-react with numerous innocuous substances. Previous efforts to develop microfluidic devices that incorporate these chromogenic indicator reagents address only a few of the many challenges associated with these kits. This is especially true for samples where the drug of interest is present as a lacing agent. This work describes the development of a centrifugal microfluidic device capable of integrating facile sample preparation, by way of a 3D printed snap-on cartridge amenable to microwave assisted extraction, followed by chromatographic separation and chromogenic detection on-disc. As cannabis is among the most widely used controlled substance worldwide, and displays strong interference with these indicator reagents, mock samples of laced marijuana are used for a proof-of-concept demonstration. Post extraction, the microdevice completes high throughput metering just prior to simultaneous reaction with four of the most commonly employed microchemical tests, followed by objective image analysis in CIELAB (a device-independent color model). Separation and recovery of a representative controlled substance with 93% efficiency is achieved. Correct identification, according to hierarchical cluster analysis, of three illicit drugs (e.g., heroin, phencyclidine, and cocaine) in artificially laced samples is also demonstrated on-disc. The cost effective microdevice is capable of complete automation post-extraction, with a total analysis time (including extraction) of <8 min. Finally, sample consumption is minimized, thereby preventing the complete destruction of forensic evidence.

Dietary supplementation of Cannabis sativa residues in broiler chickens affects performance, carcass characteristics, intestinal morphology, blood biochemistry profile and oxidative stability.

Public interest in the cannabis plant has increased after its legalization in many countries. Cannabis sativa residues (CR) are a part of the plant waste in the cannabis industry. The CR contain medicinal properties that could be used as a feed additive in poultry production. The trial was conducted to investigate the effects of CR on growth performance, carcass characteristics, intestinal morphology, and blood biochemistry profile of broiler chickens. In a completely randomized design, 256 one-day-old male Ross 308 broilers were randomly allocated to 4 treatments with 8 replicates and 8 birds per replicate. These 4 dietary treatments included a basal diet with 0, 0.5, 1 and 2% CR for 40 d. The results showed that 2% CR supplementation reduced feed intake (FI) in the starter phase (d 3-23, P < 0.05). The birds in the CR groups had lower FI in the finishing phase (d 24-40, P < 0.01) and the whole raising period (d 3-40, P < 0.01) than the control. However, the body weight and carcass yield were not different (P > 0.05). In addition, the CR diet had no adverse effects on the blood biochemistry profile, including total cholesterol, triglycerides, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total protein, globulin, albumin, and direct bilirubin (P > 0.05). In addition, total bilirubin and malondialdehyde were better in the plasma of CR-supplemented birds than in the control groups (P < 0.05). The observations on intestinal morphology showed that CR supplementation improved the ratio between villus height and crypt depth in the ileum (P < 0.05). In conclusion, CR supplementation can improve intestinal morphology and oxidative stability of broiler chickens. This suggests that CR could potentially be used as an alternative feed additive in broiler production.

Anti-inflammatory effects of phytocannabinoids and terpenes on inflamed Tregs and Th17 cells in vitro.

AIMS: Phytocannabinoids and terpenes from Cannabis sativa have demonstrated limited anti-inflammatory and analgesic effects in several inflammatory conditions. In the current study, we test the hypothesis that phytocannabinoids exert immunomodulatory effects in vitro by decreasing inflammatory cytokine expression and activation. KEY METHODS: CD3/CD28 and lipopolysaccharide activated peripheral blood mononuclear cells (PBMCs) from healthy donors (n = 6) were treated with phytocannabinoid compounds and terpenes in vitro. Flow cytometry was used to determine regulatory T cell (Treg) and T helper 17 (Th17) cell responses to treatments. Cell pellets were harvested for qRT-PCR gene expression analysis of cytokines, cell activation markers, and inflammation-related receptors. Cell culture supernatants were analysed by ELISA to quantify IL-6, TNF-α and IL-10 secretion. MAIN FINDINGS: In an initial screen of 20 µM cannabinoids and terpenes which were coded to blind investigators, cannabigerol (GL4a), caryophyllene oxide (GL5a) and gamma-terpinene (GL6a) significantly reduced cytotoxicity and gene expression levels of IL6, IL10, TNF, TRPV1, CNR1, HTR1A, FOXP3, RORC and NFKΒ1. Tetrahydrocannabinol (GL7a) suppression of T cell activation was associated with downregulation of RORC and NFKΒ1 gene expression and reduced IL-6 (p < 0.0001) and IL10 (p < 0.01) secretion. Cannabidiol (GL1b) significantly suppressed activation of Tregs (p < 0.05) and Th17 cells (p < 0.05) in a follow-on in vitro dose-response study. IL-6 (p < 0.01) and IL-10 (p < 0.01) secretion was significantly reduced with 50 µM cannabidiol. SIGNIFICANCE: The study provides the first evidence that cannabidiol and tetrahydrocannabinol suppress extracellular expression of both anti- and pro-inflammatory cytokines in an in vitro PBMC model of inflammation.

[Genome-wide screening and bioinformatics analysis of Cannabis sativa LecRLK gene family].

Lectin receptor-like kinase(LecRLK) is a class of phytokinase with lectin conserved domain, which plays an important role in plant resistance to biological and abiotic stresses, as well as plant growth and development. Cannabis sativa is an important multi-purpose plant, widely used in food, textile, medicine, and other fields. Genome-wide screening and expression analysis of the LecRLK family of C. sativa were performed in this paper, so as to provide scientific reference for functional analysis of the LecRLK family of C. sativa. Based on BLAST and HMM methods, 93 LecRLKs were identified in the whole genome of C. sativa, including 69 G types, 23 L types, and one C types. Subsequently, a series of bioinformatics analyses were performed on the LecRLK family members, and the physicochemical properties of the protein of the LecRLK family members were initially revealed. The prediction of cis-acting elements of promoters in family members showed that family members were regulated by hormones and stress response. The expression analysis showed that some family members were highly expressed in the roots, which may participate in the process of stress resistance. Several members were highly expressed in female flowers and may be involved in female flower development. This study provides a theoretical basis for further study of LecRLK gene function. Meanwhile, the expression analysis screens candidate LecRLK members who may participate in the resistance of C. sativa, which provides a theoretical basis for the subsequent selection of C. sativa varieties against resistance.

Green plastics: Direct production from grocery wastes to bioplastics and structural characterization by using synchrotron FTIR.

Lignocellulosic bioplastics were produced using four different green wastes: hemp, parsley stem, pineapple leaves and walnut shell. Two different solutions were used to dissolve the green wastes: trifluoroacetic acid (TFA) and pure water. The changes in their natural structures and the solvent effect during the regeneration in biofilm formation were investigated by using Synchrotron FTIR Microspectroscopy (SR-µFTIR). The presence of cellulose, hemicellulose and lignin components in the water-based biofilms was confirmed. After dissolving in TFA, the spectra demonstrated some additional bands especially in the hemicellulose region. This is due to the hydrolysis of ester bonds and conversion to carboxylic acids. Principal component analysis showed grouping due to different solvents and polymer addition. Hemp-PVA (Polyvinyl Alcohol) composite biofilms were obtained by adding polyvinyl alcohol to the hemp solution to give extra strength to the hemp biofilms. It has been shown that water-based hemp-PVA biofilms do not cause any significant spectral changes, comparing with pure hemp and PVA spectra. However, after dissolving in TFA, unlike water-based biofilms, it appears that TFA molecules are retained by PVA through hydrogen bonds of TFA's carboxylic acid and hydroxyl groups and distinct spectral regions belong to TFA bands are clearly identified.

Insight into the mechanism of adsorption and release of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in sewage by modeling regional suspended particles for evaluation of the influence on monitoring of cannabis illicit abuse.

The adsorption of 11-nor-9-carboxy-∆9-tetrahydrocannabinol (THC-COOH) by the suspended particles in sewage makes it fail to accurately monitor cannabis abuse. In this work, the model sewage sample was prepared through equivalent mixing the sewage from 10 different sewage treatment plants in Guangdong province of China and used as a comprehensive representative for investigating the adsorption and release behavior of THC-COOH on the suspended particles under different temperature and pH. The solid-liquid distribution of THC-COOH in sewage depended strongly on the adsorption and release properties which were susceptible to the temperature and pH, specially adjusting pH to 11.0 could release more than 90 % of THC-COOH from the suspended particles. By means of the kinetics models, pseudo-second-order kinetic and Weber-Morris models revealed the mechanism of adsorption and release of THC-COOH in sewage that was a relatively reversible and controllable process with multiple interactions, and then it was further confirmed by the validation experiment in a variety of actual sewage samples. According to the suggested pH, the modification of the existing detection protocol prior to high performance liquid chromatography-tandem triple quadrupole mass spectrometry (HPLC-TQ-MS/MS), was successfully applied to determination of THC-COOH in the stimulated positive samples, and the recoveries and RSDs were respectively 95.48-99.79 % and 4.0-5.6 %. The finding could greatly help improving the accuracy of not only the detection of THC-COOH in sewage but also the estimation data of the consumption level of cannabis in the related regions.

Recent advances in the chromatographic analysis of endocannabinoids and phytocannabinoids in biological samples.

Endocannabinoid system, including endocannabinoid neurotransmitters (eCBs), has gained much attention over the last years due to its involvement with the pathophysiology of diseases and the potential use of Cannabis sativa (marijuana). The identification of eCBs and phytocannabinoids in biological samples for forensic, clinical, or therapeutic drug monitoring purposes constitutes a still significant challenge. In this scoping review, the recent advantages, and limitations of the eCBs and phytocannabinoids quantification in biological samples are described. Published studies from 2018-2023 were searched in 8 databases, and after screening and exclusions, the selected 38 articles had their data tabulated, summarized, and analyzed. The main characteristics of the eCBs and phytocannabinoids analyzed and the potential use of each biological sample were described, indicating gaps in the literature that still need to be explored. Well-established and innovative sample preparation protocols, and chromatographic separations, such as GC, HPLC, and UHPLC, are reviewed highlighting their respective advantages, drawbacks, and challenges. Lastly, future approaches, challenges, and tendencies in the quantification analysis of cannabinoids are discussed.

Comprehensive two-dimensional gas chromatography using a miniaturized multiloop splitter-based non-cryogenic artificial trapping (M-SNAT) modulation device for analysis of cannabis samples.

Multiloop splitter-based non-cryogenic artificial trapping (M-SNAT) modulation technique was developed, miniaturized and applied in comprehensive two-dimensional gas chromatography (GC×GC) for analysis of cannabis samples. The approach employed deactivated fuse silica (DFS) columns configured into multiple loop splitter system halving the perimeters of the progressively upstream loops. This splitter device was located between the first (1D) semi-nonpolar column outlet and a microfluidic Deans switch (DS). Each splitter loop splits a peak into two subpeaks having the same area with different void times. Three loops were then applied resulting in the number of the split subpeaks (nsplit) of 8 for each peak, and retention time differences between any two adjacent subpeaks (∆tR,split) were the same. By applying periodic heartcut event (H/C) within every artificial modulation period (PAM) of nsplit×∆tR,split, comprehensive split-and-trapped modulation profiles of analytes could be selectively transferred onto the second (2D) polar column (30 m) without cryogen consumption. This artificial modulation system was applied for analysis of cannabis samples with enhanced 2D peak capacity (2nc∼15). The established method was applied to analyse cannabis extracts using vegetable oils with or without frying process. This reveals 454 different peaks with 76, 92, 35 and 70 specific components specifically observed by using olive oil extraction (OE), fried OE, coconut oil extraction (CE) and fried CE, respectively.

Analysis of Flutianil and OC56635 Residues in Hemp Cannabis Matrices by LC-MS/MS.

In this paper, we present analytical methodologies for the determination of the thiazolidine fungicide flutianil (trade name GATTEN) and its primary metabolite OC56635 in hemp cannabis matrices. A total of nine crop matrices were tested: whole seed, fiber, flower buds, hemp hearts, hemp seed oil, hemp meal, hemp flour, ethanol extracted CBD resin (CBD-E), and supercritical CO2 extracted CBD resin (CBD-C). Processing of the CBD-E and CBD-C crop fractions was carried out in-house using methods detailed herein. Field sample analysis utilized sequential extractions, stacked solid phase extraction (SPE) column cleanups, and evaporation to prepare the samples for LC-MS/MS quantitation. Method validations for each fraction were carried out using untreated hemp matrices over a minimum of three levels, with lowest levels of method validation (LLMV) of 0.010 µg/g for all fractions except the CBD resins, for which LLMV was 0.020 µg/g. Flutianil-treated samples from nine field sites were collected from several crop production regions and analyzed to determine the distribution of incurred flutianil and OC56635 residues within the different hemp matrices. This data was generated in support of nationwide registration with the United States Environmental Protection Agency (USEPA).