Extraction Of Cannabinoids Research Articles

Effect of Organic Acid-Aided Extraction on Characteristics and Functional Properties of Pectin from Cannabis sativa L.

The extraction of cannabinoids from the inflorescence and leaves of Cannabis sativa L. is gaining interest from researchers, in addition to addressing the under-utilization of the by-products in the stems and roots of the trees. The present study investigated the recovery of pectin from the left-over parts of hemp tress using an eco-friendly method with the aid of organic acids. Different cannabis cultivars-Chalotte's Angels (CHA) and Hang-Krarog (HKR)-were used as plant materials. The stems of both cannabis cultivars contained more pectin than the roots, and tartaric acid-aided extraction provided higher yields than from citric acid. Extracting the acid solution affected some characteristics, thereby differentiating the functional properties of the derived pectin. Extraction using tartaric acid provided pectin with a higher galacturonic acid content, whereas pectin with a higher methylation degree could be prepared using citric acid. The pectin samples extracted from the stems of CHA (P-CHA) and HKR (P-HKR) had low methoxyl pectin. P-CHA had better free radical scavenging capability, whereas P-HKR showed more potent reducibility. Considering the functional properties, P-CHA showed greater emulsion formability and foaming activity, whereas P-HKR possessed a better thickening effect. The present work suggests the feasible utilization of P-CHA and P-HKR as food additives with bioactivity.

Optimization of microwave‐assisted extraction for quantification of cannabinoids in hemp tea by liquid chromatography‐mass spectrometry

AbstractMicrowave‐assisted extraction (MAE) of cannabinoids from hemp tea was optimized, for the first, using response surface methodology. The effect of temperature (50, 65, and 80°C), irradiation time (4, 7, and 10 min), and solvent‐to‐solid ratio (20, 30, and 40 mL of methanol/g of hemp tea) on cannabinoid extractability were investigated. The concentrations of five cannabinoids, namely Δ9‐tetrahydrocannabinol (Δ9‐THC), cannabidiol (CBD), cannabigerol (CBG), cannabichromene (CBC), and cannabinol (CBN), were selected as response variables. For the quantitative analysis, a liquid chromatography‐mass spectrometry method was developed and validated. The proposed analytical approach demonstrated satisfactory performance characteristics in terms of linearity (R2 ≥ 0.9998), precision (intra‐day: 1.99%–5.97% relative standard deviation [%RSD], inter‐day: 1.95%–6.08%RSD), sensitivity (limit of detection: 1.35–2.36 ng/g, limit of quantification: 4.05–7.08 ng/g) and carry‐over effect (signals ≤ 5.03%), with all cannabinoids eluting within 6 min. For comparison purposes, soxhlet extraction, ultrasound‐assisted extraction (UAE), and conventional‐stirring extraction were additionally performed. MAE proved to be a more effective technique for the extraction of CBD and CBN, while UAE managed to extract Δ9‐THC, CBG, and CBC at higher concentration levels.

Water Extracts from Industrial Hemp Waste Inhibit the Adhesion and Development of Candida Biofilm and Showed Antioxidant Activity on HT-29 Colon Cancer Cells.

The evolution of regulatory perspectives regarding the health and nutritional properties of industrial hemp-based products (Cannabis sativa L.) has pushed research to focus on the development of new methods for both the extraction and formulation of the bioactive compounds present in hemp extracts. While the psychoactive and medicinal properties of hemp-derived cannabinoid extracts are well known, much less has been investigated on the functional and antimicrobial properties of hemp extracts. Within the hemp value chain, various agricultural wastes and by-products are generated. These materials can be valorised through eco-innovations, ultimately promoting sustainable economic development. In this study, we explored the use of waste from industrial light cannabis production for the extraction of bioactive compounds without the addition of chemicals. The five extracts obtained were tested for their antimicrobial activity on both planktonic and sessile cells of pathogenic strains of the Candida albicans, Candida parapsilosis, and Candida tropicalis species and for their antioxidant activity on HT-29 colon cancer cells under oxidative stress. Our results demonstrated that these extracts display interesting properties both as antioxidants and in hindering the development of fungal biofilm, paving the way for further investigations into the sustainable valorisation of hemp waste for different biomedical applications.

Efficient Capture of Cannabis Terpenes in Olive Oil during Microwave-Assisted Cannabinoid Decarboxylation.

The development of selective extraction protocols for Cannabis-inflorescence constituents is still a significant challenge. The characteristic Cannabis fragrance can be mainly ascribed to monoterpenes, sesquiterpenes and oxygenated terpenoids. This work investigates the entrapment of Cannabis terpenes in olive oil from inflorescences via stripping under mild vacuum during the rapid microwave-assisted decarboxylation of cannabinoids (MW, 120 °C, 30 min) and after subsequent extraction of cannabinoids (60 and 100 °C). The profiles of the volatiles collected in the oil samples before and after the extraction step were evaluated using static headspace solid-phase microextraction (HS-SPME), followed by gas chromatography coupled to mass spectrometry (GC-MS). Between the three fractions obtained, the first shows the highest volatile content (~37,400 mg/kg oil), with α-pinene, β-pinene, β-myrcene, limonene and trans-β-caryophyllene as the main components. The MW-assisted extraction at 60 and 100 °C of inflorescences using the collected oil fractions allowed an increase of 70% and 86% of total terpene content, respectively. Considering the initial terpene amount of 91,324.7 ± 2774.4 mg/kg dry inflorescences, the percentage of recovery after decarboxylation was close to 58% (mainly monoterpenes), while it reached nearly 100% (including sesquiterpenes) after extraction. The selective and efficient extraction of volatile compounds, while avoiding direct contact between the matrix and extraction solvents, paves the way for specific applications in various aromatic plants. In this context, aromatized extracts can be employed to create innovative Cannabis-based products within the hemp processing industry, as well as in perfumery, cosmetics, dietary supplements, food, and the pharmaceutical industry.

Prevalence and Characteristics of Medicinal Cannabis Use among Chronic Pain Patients; A Post- Legalization Study in a Tertiary Care Setting in Thailand

Objective: Cannabinoid products have been applied for numerous medical conditions, including chronic pain. Thailand was the first country in South East Asia to legalize medical cannabinoids. This study aims to explore prevalence, characters, attitude, side effects of medical cannabinoid use, and pain-related outcome among the chronic cancer and non-cancer pain population at Siriraj Hospital. Materials and Methods: 200 chronic cancer pain and 670 chronic noncancer pain patients were collected by questionnaires and interviews. Data included demographic data, clinical diagnosis, pain treatment, knowledge, attitude, pattern of use, side effects and quality of life of cannabinoid extracts. Results: Prevalence of active cannabis user was 15% in chronic cancer pain and 3.1% in noncancer pain. Oil extract sublingual was the most common form. Pain control was the most common initial reason for usage. No serious side effects were reported. Common side effects were dry oral mucosa, drowsiness, and headache. The most common source was obtained from friends. 36% of the patients believed they had enough understanding of medical cannabis, while 68.5% agreed that it is appropriate to use in Thailand. In cancer patients, the Edmonton Symptom Assessment System (ESAS) subscale for lack of appetite, anxiety, and subscale for a brief pain inventory (BPI) for enjoyment of life were higher among active users. In patients with noncancer pain, only the mood subscale BPI was lower among active users. Conclusion: Medical cannabis usage is common compared with general population in Thai patients with chronic pain and may be associated with increased pain interference and cancer-related symptoms. Nonmedical license prescription and nonmedical license cannabis products were common in Thailand.

Solvent-assisted recrystallisation for the recovery of cannabinoids from Cannabis extraction by-products

A wax by-product removed from the initial stages of commercial Cannabis processing, which contains significant amounts (39–51% (w/w)) of cannabinoids, is currently underutilised due to a lack of information on wax composition and further processing options. Significant value might be generated from further recovery of the cannabinoids entrapped within the wax fraction, and from further processing of the wax into useable products. This study aimed to develop a method to recover the cannabinoids contained in the wax and to prepare the wax for further refinement into a valuable product. A method where the wax by-product was dissolved in ethyl acetate and then recrystalized was developed, which consisted of a heating and mixing stage followed by filtering and washing with additional solvent. The main variables for the experiments were the ratio of solvent to wax for dilution, and the ratio of solvent to wax for post-crystallisation washing. The recrystallisation method was successful in recovering the cannabinoids from the wax and the effectiveness of the recovery was significantly impacted by the solvent addition. The method was validated on two samples of industrial by-products sourced from the cannabinoid extraction industry, and the highest recovery over two consecutive dissolution-recrystallisation cycles was 68.3% and 96.3%, for wax A and B respectively. In both cases, maximum recoveries were achieved with the experiments corresponding with the highest solvent addition. It was therefore concluded that the use of more solvent during recrystallisation, especially for the washing step during filtering, was favourable as it resulted in a higher recovery of cannabinoids from the wax by-product. Furthermore, wax carry-over to the recovered filtrate was sufficiently low (< 3% (w/w)), and was easily removed by centrifugation. The majority of the carried-over lipophilic components were fatty acids, with a maximum concentration of 17.39 mg/g in the filtrate removed after the first stage of recrystallisation with wax A. The other components (n-alkanes, fatty alcohols) were in significantly lower abundance compared to the fatty acid fraction. The recovery when the wax was filtered without undergoing recrystallisation was 33.5%, significantly less than the recovery of the recrystallisation method. This work successfully demonstrated a methodology for simple cannabinoid recovery from Cannabis processing by-products, with the potential to create significant additional value from what is currently regarded as a waste in this industry.

Polymeric ionic liquid sorbent coatings in thin film microextraction: Insight into sorbent selectivity for pesticides and cannabinoids

Polymeric ionic liquid (PIL) sorbent coatings consisting of polymerizable cations and anions were employed as sorbent coatings in thin film microextraction (TFME) for the extraction of pesticides and cannabinoids. The blades consisted of a thin film of PIL sorbents chemically bonded to vinyltrimethoxysilane-functionalized nitinol sheets. The imidazolium- or ammonium-based PIL sorbents contained aromatic benzyl moieties as well as polar hydroxyl groups or aliphatic functional groups within the chemical structure of the IL monomer. The chemical structure of the IL crosslinkers of the PILs were kept constant across each sorbent, except for the anion, which consisted of either bis[(trifluoromethyl)sulfonyl]imide ([NTf2−]), p-styrenesulfonate ([SS−]), or 3-sulfopropyl acrylate ([SPA−]). Temperature, salt content, and methanol content were optimized as extraction conditions to maximize pesticide-cannabinoid selectivity using Doehlert design of experiments (DOE). Effects of these three factors on selectivity and extraction efficiency are discussed. The optimal extraction conditions consisting of sample temperature (31°C), sodium chloride (30% w/v), and methanol content (0.25% v/v) are compared to initial sorbent screening conditions at a sample temperature of 40°C, 15% (w/v) sodium chloride, and 2.5% (v/v) methanol content. PIL sorbent swelling behavior at different salt and methanol content conditions and its effect on extraction efficiency are hypothesized. Selectivity factors for the sorbents indicated that aromatic moieties within the IL monomer may enhance pesticide-cannabinoid selectivity under optimized conditions, but the extraction efficiency of pesticides that are known to coelute with cannabinoids in the chromatographic separation may be enhanced by employing sorbent coatings with [SPA−] anions.

Ultrasound-assisted dispersive solid-liquid microextraction with eutectic solvents for the determination of cannabinoids in different hemp products

The wide range of applications of hemp products, together with the environmental benefits that come from hemp cultivation are driving up the market demand for Cannabis sativa L. plant. One of the main restrictions for hemp cultivation and marketing concerns the content of delta-9-tetrahydrocannabidiol (Δ9-THC), which is known to have psychotomimetic effect. If the recent growing of hemp market is beneficial by an economic and environmental point of view, it is necessary to develop reliable analytical methods for the chemical characterization of hemp products, to guarantee the safety of use for the customers.This study aimed to develop a simple ultrasound-assisted dispersive solid-liquid microextraction (UA-DSLME) method for the extraction of cannabinoids in hemp products, using eutectic solvents (ESs) as extraction material. Two types of ESs were compared: one prepared with a [Ch+][Br-]-modified salts as hydrogen bond acceptor and one based on natural terpenoids. The ultrasound-assisted dispersive solid-liquid microextraction method was optimized to be applied for the analysis of aerial parts of hemp collected before flowering, hemp inflorescences and a commercial sample called CBD oil, and proved to be robust and versatile. Under optimal conditions, only 100 µL of ES and 2 mL of water as co-solvent were used in the US-assisted extraction, before the analysis in the UHPLC-PDA system. The developed approach allowed to obtain the same chemical profile of conventional methods, while improving the greenness of the method and the enrichment of the marker analytes. To overcome the strong matrix effect for cannabinoids, a matrix-matched calibration was used. Blank matrices of the samples under study were easily obtained by performing an exhaustive extraction of the marker analytes in the hemp samples. These matrices were successfully used for validation, achieving accuracy values between 82% and 118%.

The Development, Validation, and Application of a UHPLC-HESI-MS Method for the Determination of 17 Cannabinoids in Cannabis sativa L. var. sativa Plant Material.

Cannabinoids are an important group of secondary metabolites found in the plant Cannabis sativa L. The growing interest in the use of hemp in food production (e.g., hemp teas, hemp cookies) makes it necessary to develop a method for determining these compounds in the plant, both fresh and dried. The selection of a suitable extraction liquid for the extraction of cannabinoids and the development of a method for the determination of 17 cannabinoids is a prelude to the development of an effective method for the extraction of these compounds. In the present study, a novel, simple, and efficient method was developed and validated for the determination of up to 17 cannabinoids in fresh plant parts (inflorescences and leaves) of Cannabis sativa L. and in dried material, including hemp teas. Analyses were performed using ultra-high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry setup operating with a heated electrospray interface (UHPLC-HESI-MS). Based on the comparison, methanol was selected as the best for the extraction of cannabinoids from fresh and dried material. The efficiency and validity of the method were assessed using certified reference material (dried Cannabis) and confirmed by z-score from participation in an international proficiency test conducted by ASTM International for dried hemp.

Improvement of cannabidiolic acid synthetase activity through molecular docking and site-directed mutagenesis

Cannabidiol (CBD), a nonhallucinogenic but therapeutic compound, is widely used in fields such as medicine, food, health products, and cosmetics. Cannabidiolic acid synthase (CBDAS) can exclusively catalyze the conversion of cannabigerolic acid (CBGA), which is the common precursor of cannabinoids, to CBDA, which is then decarboxylated to form CBD. In order to determine which amino acids in the CBDAS protein undergo mutations that can significantly enhance the catalytic activity of the enzyme, molecular docking and experimental validation were performed in this study. The CBDAS gene was cloned from a high-CBD-content hemp germplasm; this 1635 base pair gene encodes 544 amino acids. Saturation and alanine scanning mutagenesis and combinatorial mutation analysis showed that a single amino acid mutation (C176W) and three dual mutations (H69G+H114L, H114L+C176W and G183V+N482W) in CBDAS reduced the binding energy of the mutant for the substrate CBGA. The molecular docking results indicated that for the mutants CBDASH114L+C176W and CBDASG183V+N482W, the interaction force with CBGA was increased, the bond distance was shortened, and some amino acids that interacted with the substrate were also altered compared with the WT. For the mutants CBDASC176W and CBDASH69G+H114L, compared with the WT, the interaction force with CBGA was decreased, or the interaction was eliminated. The results of enzyme catalytic activity measurement showed that compared with the WT, the catalytic activities of CBDASG183V+N482W and CBDASH114L+C176W were significantly increased, and that of CBDASH69G+H114L was significantly decreased (P < 0.05), which was generally consistent with the expected results from molecular docking. The crude leaf extract, when used as the substrate, produced more CBDA and CBD than an equal concentration of the CBGA standard. Obviously, it is possible that the recombinant CBDAS produced with yeast mutants can be used for secondary production of CBD in the process of cannabinoid extraction with the remaining CBGA in the leaves as the substrate. The research results provide a theoretical basis for the molecular modification of other cannabinoid synthetases.

Medicinal Cannabis: Extended Stability of Cannabis Extracts Produced Using a New Ethanol-based Extraction Method.

Cannabis as a therapeutic agent is increasing in popularity all around the globe, particularly in Western countries, and its potential is now well assessed. On the other hand, each country has its own regulation for the preparation of cannabis macerated oils; in Italy, there are only a few preparation methods allowed. With this work, we aim to perform a stability study of cannabis oils produced with a novel method for the extraction of cannabinoids from cannabis inflorescence. Three different varieties of cannabis were used, with and without the adding of tocopherol acetate as an antioxidant. Cannabinoids were extracted using ethanol at room temperature; then, the solvent was evaporated under reduced pressure and the preparations reconstituted with olive oil. In this work, we assessed the stability of both cannabinoids and terpenes in these formulas over 8 months. Cannabinoid stability was assessed by monitoring the concentrations of THC and CBD, while terpene stability was assessed by monitoring β-Caryophyllene and α-Humulene concentrations. Stability of the extracts was not influenced by the presence of tocopherol acetate, though refrigeration seems to be detrimental for a long storage of products, especially regarding THC concentrations. The improvements offered by this method reside in the flexibility in controlling the concentration of the extract and the ability to produce highly concentrated oils, alongside the possibility to produce standardized oils despite the variability of the starting plant material.

Fast and sensitive LC-MS/MS method for quantification of cannabinoids and their metabolites in plasma of cattle fed hemp.

Hemp-based materials have gained interest as alternative feed ingredients for livestock. However, safety concerns arise regarding the transfer of cannabinoids from the plant to the animals. Addressing these concerns requires the use of methods capable of detecting and quantifying cannabinoids in livestock. In this study, a fast and sensitive method was developed for quantification of cannabinoids and cannabinoid metabolites in cattle plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The extraction of cannabinoids from the plasma matrix was achieved by combining the Captiva Enhanced Matrix Removal-Lipid clean-up and salting-out assisted liquid-liquid extraction procedure. The developed method underwent validation using various analytical parameters, and the results demonstrated good accuracy, precision, specificity, and high sensitivity. The method was applied to real plasma samples obtained from cattle fed hemp for 2 weeks, and successfully detected various cannabinoids, including delta-9-tetrahydrocannabinol. Furthermore, the study revealed that 7-carboxy cannabidiol, a metabolite of cannabidiol, was the predominant cannabinoid present in the cattle plasma throughout the feeding period, which could remain detectable for weeks after the hemp feeding had ended.

A new prototype reactor for the fast microwave-assisted decarboxylation and extraction of cannabinoids in olive oil from Cannabis inflorescences

Several different Cannabis-based pharmaceutical preparations have been approved for medical treatments. Standardization to ensure specified phytocannabinoid contents and to maintain the integrity of the product is therefore required. Olive oil is clearly a safe and edible solvent suitable for the galenic preparation of ready-to-use Cannabis extracts. Pharmacists can use several non-standard procedures for this purpose, most of which involve the heat treatment of the plant matrix (i.e. the female inflorescences) for 2 h. In order to allow the “neutral” and more biologically active forms of the major phytocannabinoids to be extracted, prior thermal treatment of the plant matrix is desirable. This work investigates a new prototype microwave (MW) reactor (Ethos Lean) specifically designed for the decarboxylation of the acidic cannabinoids in Cannabis inflorescences and their subsequent extraction into oil. Both of the steps to produce the oily extract were carried out in the Ethos Lean's cavity with special accessories; a rotating drum for the decarboxylation process and a glass reactor with a stirrer for the extraction step. The variables considered in the process optimization were time and temperature, according to which the instrument automatically calibrated the power output. In collaboration with a hospital pharmacy, the efficiency of the device has been evaluated by comparing the results obtained with those of exhaustive decarboxylation in a conventional oven and ethanol extraction. A comparison was also made with conventional procedures in olive oil. Thanks to the rotating drum, which is sensitive to dielectric heating, the complete and homogeneous decarboxylation of phytocannabinoids was rapidly achieved (30 min, 120 °C), even with discrete quantities of inflorescences (150 g) and without releasing the characteristic intense odor into the laboratory. The exhaustiveness (100% yields of both CBD and THC) of MW-assisted extraction process in olive oil was achieved with 30 min irradiation at 90 °C. Furthermore, the possible decarboxylation occurring during MW-assisted extraction in oil and the effect of cooling time on the final yields have also been evaluated.

Fluorinated-Squaramide Covalent Organic Frameworks for High-Performance and Interference-Free Extraction of Synthetic Cannabinoids.

Synthetic cannabinoids (SCs), one of the largest groups of new psychoactive substances (NPSs), have emerged as a significant public health threat in different regions worldwide. Analyzing SCs in water samples is critical to estimate their consumption and control. However, due to their low background concentration and the coexistence of complex matrix, the selective and effective enrichment of SCs is still challenging. In this study, a series of fluorinated-squaramide-based covalent organic frameworks (COF: FSQ-2, FSQ-3, and FSQ-4) are synthesized, and the as-prepared FSQ-4 exhibits strong affinity to different SCs. The proper pore size (1.4nm) and pre-located functional groups (hydrogen-bond donors, hydrogen-bond acceptors, and fluorophilic segments) work synergistically for efficient SCs capture. Remarkably, when coupled FSQ-4 with solid-phase microextraction (SPME), trace-level (part per trillion, 10-9 ) determination of 13 SCs can be easily achieved, representing one of the best results among NPS analyses, and the excellent extraction performance can be maintained under various interfering conditions.

Solvent extraction of Cannabis sativa under cryogenic conditions

Extraction of natural products using safe, selective and green solvents is of major importance for their use as ingredients in foods, beverages, cosmetics and pharmaceutical products. In the particular case of cannabis, ethanol at ambient conditions has been found to be very effective in the extraction of cannabinoids, terpenoids, and other valuable metabolites; however, it also extracts chlorophyll, vegetable oils and waxes. Alternatively, the selectivity towards cannabinoids could be increased by operating at lower temperatures. In this regard, this work focused on the cryogenic extraction of leaves and flowers of Cannabis sativa L. with ethanol. Extractions were carried out in a rotary tumbler under batch operation at different temperatures (−80 °C to 20 °C) and solvent:solid ratios (4:1 to 25:1 mL/g). The content of cannabinoids in samples was measured via U-HPLC under isocratic elution using a mixture of a 0.1% wt. formic acid aqueous solution and pure acetonitrile, using a diode array detector at 280 nm, and with an external standard method. Terpenes were quantified by GC using headspace injection, FID detection, and using external standards for calibration. The yields with respect to dry biomass were in-between 0.43% and 4.22% wt. The corresponding cannabinoids content with respect to dry material ranged from 0.25% to 2.67% wt., and these mainly corresponded to cannabidiol and cannabinol. The content of terpenoids was in between 0.01 and 1% wt., and the major fraction corresponded to monoterpenes d-Limonene, β-Myrcene, α-Terpinene, Terpinolene, and p-Cymene. Cryogenic extracts were characterized by a translucent aspect with minor content of coloring compounds and other impurities. Preferred operating conditions corresponded to −20 °C and a solvent:solid ratio of 16:1 mL/g; under these conditions and with respect to dry biomass, the extraction yield was 3.47% wt., the cannabinoids content was 0.67% wt., and the CBD content was 0.59% wt.

Feeding spent hemp biomass to lactating dairy cows: Effects on performance, milk components and quality, blood parameters, and nitrogen metabolism

The legalization of industrial hemp by the 2018 Farm Bill in the United States has driven a sharp increase in its cultivation, including for cannabinoid extraction. Spent hemp biomass (SHB), produced from the extraction of cannabinoids, can potentially be used as feed for dairy cows; however, it is still illegal to do so in the United States, according to the US Food and Drug Administration Center for Veterinary Medicine, due to the presence of cannabinoids and the lack of data on the effect on animals. To assess the safety of this byproduct as feed for dairy cows, late-lactation Jersey cows (245 ± 37 d in milk; 483 ± 38 kg body weight; 10 multiparous and 8 primiparous) received a basal total mixed ration (TMR) diet plus 13% alfalfa pellet (CON) or 13% pelleted SHB for 4 wk (intervention period [IP]) followed by 4 wk of withdrawal period (WP), where all cows received only the basal TMR during WP. The dry matter intake (DMI), body weight, body condition score, milk yield, milk components, and fatty acid profile, blood parameters, N metabolism, methane emission, and activity were measured. Results indicated that feeding SHB decreased DMI mainly due to the low palatability of the SHB pellet, as the cows consumed only 7.4% of the total TMR with 13.0% SHB pellet offered in the ration. However, milk yield was not affected during the IP and was higher than CON during the WP, leading to higher milk yield/DMI. Milk components were not affected, except for a tendency in decreased fat percentage. Milk fat produced by cows fed SHB had a higher proportion of oleate and bacteria-derived fatty acids than CON. The activity of the cows was not affected, except for a shorter overall lying time in SHB versus CON cows during the IP. Blood parameters related to immune function were not affected. Compared with CON, cows fed SHB had a lower cholesterol concentration during the whole experiment and higher β-hydroxybutyric acid during the WP, while a likely low-grade inflammation during the IP was indicated by higher ceruloplasmin and reactive oxidative metabolites. Other parameters related to liver health and inflammatory response were unaffected, except for a tendency for higher activity of alkaline phosphatase during IP and a lower activity of gamma-glutamyl transferase during WP in the SHB group versus CON. The bilirubin concentration was increased in cows fed SHB, suggesting a possible decrease in the clearance ability of the liver. Digestibility of the dry matter and protein and methane emission were not affected by feeding SHB. The urea, purine derivatives, and creatinine concentration in urine was unaffected, but cows fed SHB had higher N use efficiency and lower urine volume. Altogether, our data revealed a relatively low palatability of SHB affecting DMI with minimal biological effects, except for a likely low-grade inflammation, a higher N use efficiency, and a possible decrease in liver clearance. Overall, the data support the use of SHB as a safe feed ingredient for lactating dairy cows.

Cannabidiol alleviates perfluorooctane sulfonate-induced macrophage extracellular trap mediate inflammation and fibrosis in mice liver

As a new type of persistent organic pollutant, perfluorooctane sulphonate (PFOS) has received extensive attention worldwide. Cannabidiol (CBD) is a non-psychoactive natural cannabinoid extract that has been proved to have antioxidation, regulation of inflammation and other functions. However, the effects of PFOS on liver injury and whether CBD can alleviate PFOS-induced liver injury are still unclear. Therefore, in this study, we used CBD (10 mg/kg) and/or PFOS (5 mg/kg) to intraperitoneally inject mice for 30 days. We found that PFOS exposure led to inflammatory infiltration in the liver of mice, increased the formation of macrophage extracellular trap (MET), and promoted fibrosis. In vitro, we established a coculture system of RAW264.7, AML12 and LX-2 cells, and treated them with CBD (10 μM) and/or PFOS (200 μM). The results showed that PFOS could also induce the expression of MET, inflammation and fibrosis marker genes in vitro. Coiled-coil domain containing protein 25 (CCD25), as a MET-DNA sensor, was used to investigate its ability to regulate inflammation and fibrosis, we knocked down CCDC25 and its downstream proteins (integrin-linked kinase, ILK) by siRNA technology, and used QNZ to inhibit NF-κB pathway. The results showed that the knockdown of CCDC25 and ILK and the inhibition of NF-κB pathway could inhibit MET-induced inflammation and fibrosis marker gene expression. In summary, we found that PFOS-induced MET can promote inflammation and fibrosis through the CCDC25-ILK-NF-κB signaling axis, while the treatment of CBD showed a protective effect, and it is proved by Macromolecular docking that this protective effect is achieved by combining CBD with peptidylarginine deiminase 4 (PAD4) to alleviate the release of MET. Therefore, regulating the formation of MET and the CCDC25-ILK-NF-κB signaling axis is an innovative treatment option that can effectively reduce hepatotoxicity. Our study reveals the mechanism of PFOS-induced hepatotoxicity and provides promising insights into the protective role of CBD in this process.

Quality Control of 11 Cannabinoids by Ultraperformance Liquid Chromatography Coupled with Mass Spectrometry (UPLC-MS/MS).

Cannabinoid extraction from Cannabis sativa L. (hemp) for nonmedical purposes has become popular in the United States. Concerns, however, have been raised regarding the accuracy of the labels for cannabinoid levels in the commercial products. In this study, we developed rapid, sensitive, selective, accurate, and validated liquid chromatography-tandem mass spectrometry for the quantification of cannabinoids. The methods are for determining 11 cannabinoids in cannabis (hemp) extracted in oil form, and we investigated the accuracy of the labeling and thermal stability regarding the cannabinoids on 17 oil cannabis samples. In the UPLC chromatogram, we see a good resolution and there is no matrix effect and the accuracy were 98.2% to 102.6%, and the precision was 0.52%-8.18%. The linearity of the calibration curves in methanol was with a regression r2 ≥ 0.99. The lowest of detection (LOD) was 5-25 ng/mL, and the limit of quantification (LOQ) was 10-50 ng/mL. The study showed that only 30% of the commercial samples were within the acceptable range of +/-10% compared to the labeled ingredient concentrations. The thermal stability test profile showed a change in the concentration of cannabinoids in each sample at 37°C for one week, with an average loss of cannabinoids up to 15%. The validated method proved to be selective, accurate, and precise, with acceptable linearity within the calibration range with no matrix effect. The stability profile data indicated that high temperatures could change the quality of commercial samples.

Oral Tetrahydrocannabinol (THC):Cannabinoid (CBD) Cannabis Extract Adjuvant for Reducing Chemotherapy-Induced Nausea and Vomiting (CINV): A Randomized, Double-Blinded, Placebo-Controlled, Crossover Trial.

To evaluate the effects of tetrahydrocannabinol (THC):cannabinoid (CBD) (1:1) oil in reducing chemotherapy-induced nausea and vomiting (CINV) in gynecologic cancer patients who received moderate-to-high emetogenic chemotherapy. This was a randomized, double-blinded, crossover and placebo-controlled trial. The study was conducted at the Gynecologic Oncology Units, Bhumibol Adulyadej Hospital (BAH), Royal Thai Air Force, Bangkok, Thailand, between August and November 2022. Participants had gynecologic cancer and received moderate-to-high emetogenic chemotherapy. Subjects were randomized and divided into two groups (A and B) based on the block of four randomization method. In the first cycle, groups A and B received THC:CBD extract oil 1:1 (TCEO) and placebo before chemotherapy administration. In the second cycle, groups A and B received placebo and TCEO before chemotherapy administration. Both groups received per protocol antiemetic medication during chemotherapy. Nausea score and side effects were recorded. A total of 60 cases were recruited. After exclusion, 54 cases were included in the study. The mean age of participants was 54.4 years. The mean body mass index (BMI) was 26.5 kg/m2. Fifty-nine (21/54) percent cases were the advanced stages of cancer. The nausea score of TCEO and placebo groups were 2.11 and 2.99, respectively (P < 0.05). More than half of the participants (36/54) reported dizziness and sedation side effects. Dry mouth, confusion, anxiety, and palpitation of both groups were comparable. The cannabinoid extract (THC:CBD) was an appropriate adjuvant agent to reduce CINV in patients with gynecologic cancer who received high-emetogenic chemotherapy. Dizziness and sedation were the major side effects.

Metal organic frameworks as advanced materials for the paper-based extraction of synthetic cannabinoids in oral fluids

In this manuscript, a paper-based device modified with metal-organic frameworks was applied for the extraction of synthetic cannabinoid receptor agonists (SCRAs) in oral fluids, taking profit of the several interaction mechanisms between MOF-analytes, followed by their quantification by LC-fluorescence detection. The resulting composite, named as NH2[email protected], was characterized by different techniques such as infrared, scanning electron microscopy, among others. The extraction procedure was optimized by using a full factorial design after an appropriate selection of the significant variables. Once the method was optimized, the analytical parameters were studied including liner range (LOQ-400 µg L−1), limit of detection (0.7 to 2.8 µg L−1) and intra- and inter-batch precision (<8 and <20%, expressed as RSD, respectively). Next, the accuracy of the method was assessed at three different concentration levels (25, 75 and 100 µg L−1), obtaining values comprised from 64 to 98%. Finally, the method was applied for the analysis of oral fluids (n=25) (15 positives and 10 negative samples) and sensitivity, specificity and accuracy (efficiency) of the methodology were calculated obtaining 80, 100 and 88%, respectively.