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).

Hemp hull fiber and two constituent compounds, N-trans-caffeoyltyramine and N-trans-feruloyltyramine, shape the human gut microbiome in vitro.

Mounting evidence supports the potential of dietary bioactives to reduce chronic disease risk. N-trans-caffeoyltyramine (NCT) and N-trans-feruloyltyramine (NFT) have been hypothesized to drive regulation of gut permeability, but these components have not yet been studied in the context of the human gut microbiome. This work examined whether purified NCT and NFT, or a hemp hull product containing NCT and NFT (Brightseed® Bio Gut Fiber™), can impact the gut microbiome using an in vitro fermentation assay. Representative human gut microbiomes were treated with Bio Gut Fiber™ or NCT and NFT and compared to starch and methylcellulose, as controls, in vitro. Stronger changes were exerted by Bio Gut Fiber™, NCT, and NFT. Communities treated with Bio Gut Fiber™ saw increased productivity and diversity. We found a dose-dependent effect of NCT and NFT on microbial communities. Here, we describe novel potential for hemp-derived bioactives to shape the gut microbiome.

In vivo comparative study of hemp straw exposure and cannabidiol oil administration in horse urine.

The non-psychoactive cannabinoids cannabidiol (CBD) and cannabidiolic acid (CBDA) are available on the market in different forms, mostly for their anti-inflammatory and potential analgesic properties. These substances are prohibited during equine competitions. CBD and CBDA are naturally present in hemp straw, commonly used as a bedding substitute for wheat straw. Unfortunately, horses can eat it, which therefore could lead to a possible risk of positive findings for CBD/CBDA in biological samples after doping control tests. The goals of this study were, first, to provide recommendations on the use of hemp straw before competition and, second, to assess if discrimination between hemp bedding exposure and CBD oil administration is possible. Several CBD equine in vivo studies have been conducted, including one on hemp straw used as bedding and one after administration of CBD oil by topical and sublingual routes. In hemp straw, CBDA was detected in higher quantities than CBD, and other cannabinoids have been observed. After hemp straw exposure, CBDA was also detected in higher quantities than CBD in all urine samples. It appeared that hemp straw should not be used as bedding for equine competition except if a delay of at least 48 h is respected. Regarding the CBD oil product analysis, CBD was the main compound detected. After administration, 7-hydroxy CBD was identified in the urine. In conclusion, based on these data, we highlighted that it could be possible to discriminate the exposure of a horse to hemp straw from an administration of a CBD oil product thanks to the main presence of CBDA.

Diet supplementation with hemp (Cannabis sativa L.) inflorescences: effects on quanti-qualitative milk yield and fatty acid profile on grazing dairy goats.

Hemp (Cannabis sativa L.) is an annual plant belonging to the family of Cannabaceae with several varieties characterized by different fatty acid profile, content in flavonoids, polyphenols, and cannabinoid compounds. Hemp is mostly used in livestock nutrition as oil or as protein cake, but not as inflorescences. The aim of this study was to evaluate the effect of dietary hemp inflorescences on milk yield and composition in grazing dairy goats. Twenty Camosciata delle Alpi goats at their 3rd parity and with a mean body weight of 45.2 ± 2.0 kg, immediately after kidding, were equally allocated into two groups (G: Grazing and GH: grazing and hemp). For three months, all goats were fed on a permanent pasture and received 700/head/day of concentrate; diet of group GH was supplemented with 20 g/head/day of hemp inflorescences. Goats' body weight did not change during the trial. Individual milk yield was daily recorded and samples collected every 20 days for chemical composition and fatty acid profile analysis. No significant differences were found for milk yield and chemical composition. Caproic (C6:0) (1.80 vs. 1.74%; p < 0.01) and lauric acids (C12:0) were significantly higher in milk of group GH (4.83 vs. 4.32%; p < 0.01) as well as linoleic (C18:2) (2.04 vs. 1.93%; p < 0.05), adrenic acid (C22:4) (0.046 vs. 0.031%, p < 0.05), omega-6/omega-3 ratio (3.17 vs. 2.93, p < 0.05) and total conjugated linoleic acids (CLAs) (0.435 vs. 0.417%; p < 0.01). The results of this study suggest that the supplementation of grazing goats' diet with hemp inflorescences may enhance the milk nutritional characteristics by increasing its content of CLAs and other beneficial fatty acids.

Development of Delivery Systems with Prebiotic and Neuroprotective Potential of Industrial-Grade Cannabis sativa L.

This study delves into the transformative effects of supercritical carbon dioxide (scCO2) cannabis extracts and prebiotic substances (dextran, inulin, trehalose) on gut bacteria, coupled with a focus on neuroprotection. Extracts derived from the Bialobrzeska variety of Cannabis sativa, utilising supercritical fluid extraction (SFE), resulted in notable cannabinoid concentrations (cannabidiol (CBD): 6.675 ± 0.166; tetrahydrocannabinol (THC): 0.180 ± 0.006; cannabigerol (CBG): 0.434 ± 0.014; cannabichromene (CBC): 0.490 ± 0.017; cannabinol (CBN): 1.696 ± 0.047 mg/gD). The assessment encompassed antioxidant activity via four in vitro assays and neuroprotective effects against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The extract boasting the highest cannabinoid content exhibited remarkable antioxidant potential and significant inhibitory activity against both enzymes. Further investigation into prebiotic deliveries revealed their proficiency in fostering the growth of beneficial gut bacteria while maintaining antioxidant and neuroprotective functionalities. This study sheds light on the active compounds present in the Bialobrzeska variety, showcasing their therapeutic potential within prebiotic systems. Notably, the antioxidant, neuroprotective, and prebiotic properties observed underscore the promising therapeutic applications of these extracts. The results offer valuable insights for potential interventions in antioxidant, neuroprotective, and prebiotic domains. In addition, subsequent analyses of cannabinoid concentrations post-cultivation revealed nuanced changes, emphasising the need for further exploration into the dynamic interactions between cannabinoids and the gut microbiota.

Soluble hemp protein-xylose conjugates fabricated by high-pressure homogenization and pH-shifting treatments.

BACKGROUND: The process of Maillard conjugation occurs with plant proteins and sugars and can be influenced by several factors, such as processing time, pH, and shear force. By utilizing cavitation processes such as high-pressure homogenization (HPH) and pH-shifting, it is possible to regulate the degree of grafting, functional characteristics, and structural changes in the formation of conjugates. The present study aimed to improve the hemp protein concentrate (HPC) through two different conjugation techniques: HPH and pH-shifting-assisted processes. RESULTS: The best conjugation conditions for the conventional method were identified as a 1:2 HPC to xylose ratio, a pH of 10, and 3 h of treatment at 70 °C. The use of HPH and pH 12-shifting methods resulted in a remarkable 2.5-fold increase in grafting degree, requiring less processing time. Fourier transform infrared spectra confirmed the formation of conjugates. Conjugates produced through HPH with pH 12-shifting (MPHX) transformed into soluble glycoproteins with a particle size of 74 nm. MPHX solubility increased by 5.7-fold than HPC, reaching 85.7%, with a more negatively charged surface at -32.4 mV. Microimages showed cracked and sharp forms for conjugated proteins compared to untreated HPC. Additionally, MPHX conjugates demonstrated superior properties in emulsion stability, foaming capacity, and antioxidant activity compared to HPC and classical conjugates. CONCLUSION: The use of HPH and pH-shifting-assisted Maillard conjugation was highly effective in enhancing the functional attributes of hemp protein conjugates. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

CBD oil by-product (Hemp flakes): Evaluation for nutritional composition, heavy metals and functionality as a food ingredient.

Background: The recent interest among consumers in industrial hemp due to health and wellness benefits has led to several products from industrial hemp, including cannabidiol (CBD) oil. CBD oil extraction from hemp buds and flowers generates by-product biomass (hemp flakes), often posing disposal challenges and with little or no applications. We hypothesized that hemp flakes possess residual compounds with nutritional and health value that could be used to improve utilization. Methods: Locally sourced hemp flakes were compared to three commercial hemp protein products. The nutritional composition (proximate analysis), heavy metals (Al, Cu, As, Pb, Co, Cd), and functional composition (phenolic and antioxidant properties-total phenolic compounds (TPC), total flavonoid compounds (TFC), ferric reducing antioxidant potential (FRAP), 1,1-diphenyl-1-picrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC)), (CBD, cannabiodiolic acid-CBDA, cannabichromene-CBC, cannabigerol-CBG, and cannabinol-CBN) contents were determined and compared. Findings: Hemp flakes had a similar nutritional composition to commercial hemp protein products, with heavy metal levels within FDA allowed limits. The by-product had significantly higher CBDA levels than commercial products. Overall, hemp flakes had comparable nutrient composition and antioxidant capabilities. Based on the protein composition of hemp flakes (31.62 %) versus the highest commercial product (43 %), hemp flakes are an acceptable functional food ingredient.

An investigation into the sensory properties of luffa (Luffa cylindrica (L.)) seeds and a comparison to other seeds (flax, sunflower, chia, and hemp).

Consumers are interested in new sustainable ingredients but are unwilling to accept undesirable sensory properties in their food products. Luffa (Luffa cylindrica) is mainly harvested and processed for its fibrous network, which is used as an exfoliator, while its seeds are usually discarded. However, the seeds have been found to have various nutritional benefits. As such, this study investigated the sensory properties of luffa seed powder added to yogurt and compared it to other seed powder (flax, sunflower, chia, and hemp). Consumers (n = 107) evaluated their liking of the different seeds added to yogurt using hedonic scales and the sensory properties using check-all-that-apply (CATA). The luffa seeds when mixed with yogurt were associated with off-colour, off-flavour, metallic, strong flavour, bitter, salty, earthy and decreased consumer liking. The flax and sunflower seeds were found to be sweet, nutty, cooked, mild flavour, and to have a smooth texture. The overall liking scores for the flax and sunflower seed samples were significantly higher than the luffa and hemp samples. Future studies should investigate different drying and roasting treatments to improve the sensory properties of the luffa seeds.

Efficacy and tolerability of full spectrum hemp oil in dogs living with pain in common household settings.

Lameness and restricted mobility are a significant concern in companion animals experiencing chronic pain, inflammation, or age-related pathologies. The growing awareness of health risks and side effects associated with the long-term use of prescription analgesics requires different management strategies to address these issues. In this study, we conducted a crossover evaluation of the effect of full spectrum hemp oil dosed orally at 2 mg/kg BID phytocannabinoids for 8 weeks in dogs (n = 37) living with pain in common household settings. Owner-reported canine pain, home activity, accelerometer-based activity, walkway-based gait, and tolerability were assessed at each phase of the study. Secondary endpoints included changes in blood biochemistry, liver enzymes, inflammatory biomarkers, and plasma metabolites. The intervention was positively associated with a decrease in pain scores (-46.2%, p = 0.0016), increased ability to walk up and down the stairs (10.6-14.7%, p < 0.05), and improved daily activity (25.9%, p = 0.0038). Decreases in plasma levels of proinflammatory cytokines TNF-α, IL-6, and IL-8 were also observed. Taken together, these findings suggest that the benefits of nutritional supplementation with hemp oil could include control of pain, greater mobility, and an overall improvement in the animal wellbeing.

Comparison on the mechanism and potency of hepatotoxicity among hemp extract and its four major constituent cannabinoids.

Cannabidiol (CBD) has been reported to induce hepatotoxicity in clinical trials and research studies; however, little is known about the safety of other nonintoxicating cannabinoids. New approach methodologies (NAMs) based on bioinformatic analysis of high-throughput transcriptomic data are gaining increasing importance in risk assessment and regulatory decision-making of data-poor chemicals. In the current study, we conducted a concentration response transcriptomic analysis of hemp extract and its four major constituent cannabinoids [CBD, cannabichromene (CBC), cannabigerol (CBG), and cannabinol (CBN)] in hepatocytes derived from human induced pluripotent stem cells (iPSCs). Each compound impacted a distinctive combination of biological functions and pathways. However, all the cannabinoids impaired liver metabolism and caused oxidative stress in the cells. Benchmark concentration (BMC) analysis showed potencies in transcriptional activity of the cannabinoids were in the order of CBN > CBD > CBC > CBG, consistent with the order of their cytotoxicity IC50 values. Patterns of transcriptomic changes induced by hemp extract and its median overall BMC were very similar to CBD but differed significantly from other cannabinoids, suggesting that potential adverse effects of hemp extract were largely due to its major constituent CBD. Lastly, transcriptomic point-of-departure (tPoD) values were determined for each of the compounds, with the value for CBD (0.106 µM) being concordant with a previously reported one derived from apical endpoints of clinical and animal studies. Taken together, the current study demonstrates the potential utility of transcriptomic BMC analysis as a NAM for hazard assessment of data-poor chemicals, improves our understanding of the possible health effects of hemp extract and its constituent cannabinoids, and provides important tPoD data that could contribute to inform human safety assessment of these cannabinoid compounds.

Palearctic flea beetle and pest of hops and Cannabis, Psylliodesattenuata (Coleoptera, Chrysomelidae, Galerucinae), new to North America.

Background: The univoltine leaf beetle Psylliodesattenuata (Koch, 1803) is a pest of Cannabis and Humulus (Cannabaceae) and native to the Palaearctic Region, known from eastern Asia to western Europe. New information: First North American records are presented for P.attenuata from Canada: Ontario and Québec. Adult beetle feeding damage to hops Humuluslupulus L. (Cannabacaea) plants is recorded from Québec. Diagnostic information is presented to distinguish P.attenuata from other North American Chrysomelidae and a preliminary assessment of its potential to spread in North America is presented. While our climate analysis is limited by a lack of data, it appears P.attenuata is physiologically capable of persisting throughout the range of Humulus in North America.The United States of America and Canada are now known to be home to 71 or more species of adventive Chrysomelidae.

Valorisation of industrial hemp (Cannabis sativa L.) residues and cheese whey into volatile fatty acids for single cell protein production.

The production of single cell protein (SCP) using lignocellulosic materials stands out as a promising route in the circular bioeconomy transition. However, multiple steps are necessary for lignocellulosics-to-SCP processes, involving chemical pretreatments and specific aerobic cultures. Whereas there are no studies that investigated the SCP production from lignocellulosics by using only biological processes and microbial biomass able to work both anaerobically and aerobically. In this view, the valorisation of industrial hemp (Cannabis sativa L.) biomass residues (HBRs), specifically hurds and a mix of leaves and inflorescences, combined with cheese whey (CW) was investigated through a semi-continuous acidogenic co-fermentation process (co-AF). The aim of this study was to maximise HBRs conversion into VFAs to be further used as carbon-rich substrates for SCP production. Different process conditions were tested by either removing CW or increasing the amount of HBRs in terms of VS (i.e., two and four times) to evaluate the performance of the co-AF process. Increasing HBRs resulted in a proportional increase in VFA production up to 3115 mg HAc L-1, with experimental production nearly 40% higher than theoretical predictions. The synergy between HBRs and CW was demonstrated, proving the latter as essential to improve the biodegradability of the former. The produced VFAs were subsequently tested as substrates for SCP synthesis in batch aerobic tests. A biomass concentration of 2.43 g TSS L-1 was achieved with a C/N ratio of 5.0 and a pH of 9.0 after two days of aerobic fermentation, reaching a protein content of 42% (g protein per g TSS). These results demonstrate the overall feasibility of the VFA-mediated HBR-to-SCP valorisation process.

Larvicidal Activity of Hemp Extracts and Cannabidiol against the Yellow Fever Mosquito Aedes aegypti.

To mitigate pyrethroid resistance in mosquito vectors of emerging and re-emerging human pathogens, there is an urgent need to discover insecticides with novel modes of action. Natural alternatives, such as extracts derived from plants, may serve as substitutes for traditional synthetic insecticides if they prove to be sustainable, cost-effective, and safe for non-target organisms. Hemp (Cannabis sativa) is a sustainable plant known to produce various secondary metabolites with insecticidal properties, including terpenoids and flavonoids. The goal of this study was to assess the larvicidal activity of hemp leaf extract on mosquito larvae from both pyrethroid-susceptible (PS) and pyrethroid-resistant (PR) strains of Aedes aegypti. Another goal was to identify which components of the extract were responsible for any observed larvicidal activity. We found that a methanol extract of hemp leaves induced similar concentration-dependent larvicidal activity against PS (LC50: 4.4 ppm) and PR (LC50: 4.3 ppm) strains within 48 h. Partitioning of the leaf extract between methanol and hexane fractions revealed that full larvicidal activity was restricted to the methanol fraction. Analysis of this fraction by gas chromatography-mass spectrometry and nuclear magnetic resonance showed it to be dominated by cannabidiol (CBD). Larvicidal assays using authentic CBD confirmed this compound was primarily responsible for the toxicity of the hemp leaf extract against both strains. We conclude that hemp leaf extracts and CBD have the potential to serve as viable sources for the development of novel mosquito larvicides.

Hemp-based psychoactive use and risk among young adults: Trend theory, policy, and regulation beyond legality.

BACKGROUND: Hemp-based psychoactives comprise a burgeoning legal substance market with rising trends in use across the U.S. and Canada as well as parts of Europe. Currently, scant empirical research on the use of these Novel Psychoactive Substances (NPS) exists. By examining policy, market conditions, consumer uptake, and related risks, this research extends theory on drug trends by describing the regulatory environments that shape the emergence and popularity of psychoactive hemp-based products among young adults. METHODS: Relying on ethnographic fieldwork of hemp-based markets across 3 U.S. state hemp and cannabis policy contexts as well as in-depth interviews with 40 young adult cannabis consumers recruited across 10 U.S. states, we examined how regulatory conditions shape hemp-based psychoactive markets, trends in their use, and associated risks. RESULTS: Young adults are motivated to consume hemp-based psychoactives due to the regulatory and market environments that facilitate the production of highly potent products that are inexpensive and easily accessible. States that regulate hemp-based psychoactives as cannabis, do not provide hemp markets with a competitive advantage and as such see minimal uptake. In the absence of hemp specific policies, substantial variations in product potencies, insufficient dosing information, and unscrupulous product packaging practices may increase related risks. CONCLUSION: Trend theory provides insight into the complex relationships that exist between drug policy, markets, and the proliferation of legal highs. Understanding the contextual significance that both market and regulatory conditions have on legal drug production, distribution, and consumption may better inform approaches to reduce the risks commonly associated with novel psychoactive substances like hemp-based psychoactives.

Posaconazole-hemp seed oil loaded nanomicelles for invasive fungal disease.

Invasive fungal infections (IFI) pose a significant health burden, leading to high morbidity, mortality, and treatment costs. This study aims to develop and characterize nanomicelles for the codelivery of posaconazole and hemp seed oil for IFI via the oral route. The nanomicelles were prepared using a nanoprecipitation method and optimized through the Box Behnken design. The optimized nanomicelles resulted in satisfactory results for zeta potential, size, PDI, entrapment efficiency, TEM, and stability studies. FTIR and DSC results confirm the compatibility and amorphous state of the prepared nanomicelles. Confocal laser scanning microscopy showed that the optimized nanomicelles penetrated the tissue more deeply (44.9µm) than the suspension (25µm). The drug-loaded nanomicelles exhibited sustained cumulative drug release of 95.48 ± 3.27% for 24 h. The nanomicelles showed significant inhibition against Aspergillus niger and Candida albicans (22.4 ± 0.21 and 32.2 ± 0.46 mm, respectively). The pharmacokinetic study on Wistar rats exhibited a 1.8-fold increase in relative bioavailability for the nanomicelles compared to the suspension. These results confirm their therapeutic efficacy and lay the groundwork for future research and clinical applications, providing a promising synergistic antifungal nanomicelles approach for treating IFIs.

Eco-Friendly 3D-Printed Concrete Made with Waste and Organic Artificial Aggregates.

In this research, the results of an experimental study on the use of three alternative components for creating artificial aggregates (AAs) (granules) and their usage in 3D-printed concrete (3DPC) are examined. This study combines AAs made from organic components like hemp shives (HSs), pyrolyzed coal (charcoal), waste/municipal solid waste incinerator bottom slag (BS), and a mix of a reference 3DPC with the aforementioned AAs. Particularly, to enhance these properties to make low-carbon 3DPC, in this research, the potential of using AAs as lightweight aggregates was increased to 14% in terms of the mass of the concrete. Each mix was tested in terms of its printability via a preliminary test in a 3D printing laboratory. For an additional comparison with the aforementioned cases, 3DPC was mixed with unprocessed hemp shives, charcoal, and BS. Furthermore, their strength was measured at 28 days, and lastly, their durability parameters and shrinkage were experimentally investigated. Cross-sections of the fragments were studied under a scanning electron microscope. In this study, we achieved improvements in the mechanical properties of AAs for their development and implementation as an innovative way to reduce carbon in 3DPC.

Performance Evaluation of Thermal Insulation Materials from Sheep's Wool and Hemp Fibres.

In the current work, the performance properties of natural-fibre-based thermal insulation materials were examined. For this purpose, three different compositions of natural fibres were prepared: pure sheep wool (SW), wool and industrial hemp (SW/HF) fibres, and pure industrial hemp (HF) fibres. Low-melt bicomponent polylactide (PLA) fibres were used as a binding material. For specimens prepared from natural fibres, the dependence of the thermal conductivity, the tensile strength along and across the direction of product formation, and the short-term water absorption on the density of the specimens and the flammability parameters were determined. In addition, to reduce the water absorption and flammability, the specimens were coated with hydrophobic agents and flame retardants. The obtained research results were also statistically processed. The analysis of the results showed that the thermal conductivity of natural-fibre-based thermal insulation materials varied within the range of 0.0333 ÷ 0.0438 W/(m·K), the tensile strength varied from 2.5 to 130 kPa, the short-term water absorption varied from 0.5 to 8.5 kg/m2, and the water vapour diffusion resistance factor varied from 2.537 to 2.667. It was additionally determined that all the studied products were flammable. The water absorption and flammability values were significantly reduced by the use of hydrophobic agents and flame retardants.

Hemp Fiber-Reinforced Polymers Composite Jacketing Technique for Sustainable and Environment-Friendly Concrete.

This research suggested natural hemp fiber-reinforced ropes (FRR) polymer usage to reinforce recycled aggregate square concrete columns that contain fired-clay solid brick aggregates in order to reduce the high costs associated with synthetic fiber-reinforced polymers (FRPs). A total of 24 square columns of concrete were fabricated to conduct this study. The samples were tested under a monotonic axial compression load. The variables of interest were the strength of unconfined concrete and the number of FRR layers. According to the results, the strengthened specimens demonstrated an increased compressive strength and ductility. Notably, the specimens with the smallest unconfined strength demonstrated the largest improvement in compressive strength and ductility. Particularly, the compressive strength and strain were enhanced by up to 181% and 564%, respectively. In order to predict the ultimate confined compressive stress and strain, this study investigated a number of analytical stress-strain models. A comparison of experimental and theoretical findings deduced that only a limited number of strength models resulted in close predictions, whereas an even larger scatter was observed for strain prediction. Machine learning was employed by using neural networks to predict the compressive strength. A dataset comprising 142 specimens strengthened with hemp FRP was extracted from the literature. The neural network was trained on the extracted dataset, and its performance was evaluated for the experimental results of this study, which demonstrated a close agreement.

Integrated metabolomics and proteomics analyses to reveal anticancer mechanism of hemp oil extract in colorectal cancer.

Cannabis sativa L., with a rich history in Chinese folk medicine, includes hemp strains that offer substantial economic and medical benefits due to their non-addictive properties. Hemp has demonstrated various pharmaceutical activities, including anti-inflammatory, antioxidant, and anti-tumor effects. This study explores the potential of hemp oil extract (HOE) in treating colorectal cancer (CRC). Despite its promise, the specific anticancer mechanisms of HOE have not been well understood. To elucidate these mechanisms, we employed mass spectrometry-based metabolomics and proteomics to investigate the global effects of HOE on CRC cells. Additionally, bioinformatics approaches, including bulk RNA-seq and single-cell RNA-seq, were used to identify gene expression differences and cellular heterogeneity. The results were validated using flow cytometry, western blotting, and immunohistochemistry. Our findings reveal that HOE induces significant alterations in purine metabolism pathways, down-regulates c-MYC, and inhibits the expression of cell cycle-related proteins such as CCND1, CDK4, and CDK6, leading to cell cycle arrest in the G1 phase. This comprehensive analysis demonstrates that HOE effectively blocks the cell cycle in the G1 phase, thereby inhibiting colorectal cancer cell proliferation. These findings provide experimental evidence supporting the potential therapeutic use of hemp in medicine.

Cannabinoid residuals in tissues of lambs fed spent hemp biomass and consumer's exposure assessment.

Spent hemp biomass (SHB) contains trace amounts of cannabinoids, including Δ9-tetrahydrocannabinol (Δ9-THC), that may accumulate in the tissues of animals consuming SHB. We measured cannabinoid residues in the liver, adipose tissue, and muscle of finishing lambs fed either 10% or 20% SHB for 8 weeks, or 4 weeks followed by 4 weeks SHB withdrawal. We detected multiple cannabinoids in the liver at a similar proportion to the SHB. However, CBD and Δ9-THC were enriched >20-fold in the adipose and muscle, compared to their proportion in SHB. The highest concentration of Δ9-THC was detected in adipose tissue and was 7.4-times higher than in muscle. Most cannabinoids were undetectable in tissues after 4 weeks of clearance. The consumers' exposure assessment on Δ9-THC revealed tissue levels of total THC (THCA+Δ9-THC) that exceed the acute reference dose of 1 µg/kg BW across population groups. When consuming meat from the lambs fed 10% and 20% SHB, the maximum total THC exposure was 2.03 and 7.32 µg/kg BW, respectively, equal to or below the Lowest Observed Adverse Effect Level of 36 µg/kg BW, the No Observed Adverse Effect Level of 12 µg/kg BW or a tolerable dose intake of 7 µg/kg BW.