Chemical Characterization and Evaluation of the Antibacterial Activity of Essential Oils from Fibre-Type Cannabis sativa L. (Hemp)

Leghissa A., Hildenbrand Z.L., Schug K.A.

The study of Cannabis sativa (L.) is growing in importance due to the many medicinal and pharmacological benefits of the plant. These benefits are exhibited by its most abundant constituents, the cannabinoid and terpene molecules. Cannabis-based products can be found in many different shapes and forms, such as raw plant material and extracts, which can be consumed by themselves or prepared in different ways to make them more attractive for human consumption. Among the breadth of these products, the most challenging to analyze are edibles due to the complexity of the matrices, which vary among recipes and can potentially require a myriad of sample preparation and cleanup techniques before analysis. The consumption of edibles has rapidly increased due to increased availability, ease of use, and reduced adverse health effects relative to smoking. Even so, some authorities are moving to ban edibles because of the difficulty associated with their quality control. Different extraction techniques may be required based on the type of matrix and the possible related interferences present. Here, the most common techniques for food sample preparation are reviewed, with an eye towards their use for analysis of cannabis edibles.

Marchetti L., Brighenti V., Rossi M., Sperlea J., Pellati F., Bertelli D.

Cannabis sativa L. is a dioecious plant belonging to the Cannabaceae family. The discovery of the presence of many biologically-active metabolites (cannabinoids) in fibre-type Cannabis (hemp) has recently given rise to the valorisation of this variety. In this context, the present study was aimed at the multi-component analysis and determination of the main non-psychoactive cannabinoids (cannabidiol, cannabidiolic acid, cannabigerol and cannabigerolic acid) in female inflorescences of different hemp varieties by means of 13C quantitative nuclear magnetic resonance spectroscopy (qNMR). The method proposed here for the first time for the determination of cannabinoids provided reliable results in a competitive time with respect to the more consolidated HPLC technique. In fact, it gave sufficiently precise and sensitive results, with LOQ values lower than 750 μg/mL, which is easily achievable with concentrated extracts, without affecting the quality of 13C-qNMR spectra. In conclusion, this method can be considered as a promising and appropriate tool for the comprehensive chemical analysis of bioactive cannabinoids in hemp and other derived products in order to ensure their quality, efficacy and safety.

Borgonetti V., Governa P., Montopoli M., Biagi M.

The interest in Cannabis sativa L. phytocomplex as a medicinal tool is a recently-emerging topic. Neurodegenerative diseases represent a promising field of application for cannabis and its preparations, as most of this pathologic conditions relies on an inflammatory etiology. Several cannabis constituents display anti-inflammatory effects targeting multiple pathways. In this review, a comprehensive overview of the available literature on C. sativa constituents activities in neuroinflammation is given. On the basis that the anti-inflammatory activity of cannabis is not attributable to only a single constituent, we discuss the possible advantages of administering the whole phytocomplex in order to fully exploit the “entourage effect” in neuroinflammatory-related conditions.

Nagy D.U., Cianfaglione K., Maggi F., Sut S., Dall'Acqua S.

Spontaneous forms of hemp (Cannabis sativa L., often reported as Cannabis sativa var. spontanea Vavilov) with a low content of psychoactive cannabinoids can be considered as a valuable source of other phytoconstituents to be used in nutraceuticals or for their health promoting properties. Chemical data on this hemp variety are rather scarce. In this article, we report a comprehensive phytochemical characterization of leaves, male and female inflorescences of C. sativa growing wild in Hungary. For the purpose, the essential oil along with polar extracts were analyzed using GC/MS, NMR and LC-DAD-MS techniques, respectively. The results indicated that female inflorescence essential oil contains high amounts of the CB2 agonists, (E)-caryophyllene (28.3 %) and cannabidiol (CBD; 24.9 %), whereas leaves and male inflorescence essential oils contained lower amounts of both compounds. HPLC/MS allowed to quantify cannabidiol (CBD) and cannabidiolic acid (CBD-A) in the ethyl acetate extracts from leaves, male and female inflorescences; they were 0.3, 0.8 and 0.9 %, and 0.2, 0.3 and 0.4 %, respectively. Flavonoids of this spontaneous form of hemp were formed by C-glycosides and glucuronic acids of kaempferol and apigenin with a total content of 3.8, 6.1 and 7.8 mg/g in methanolic extracts from leaves, male and female inflorescences, respectively. Based on these results, spontaneous C. sativa may represent an important source of CB2 agonists and bioflavonoids to be used in nutraceuticals, cosmetics and pharmaceuticals.

Wang C., Wiedinmyer C., Ashworth K., Harley P.C., Ortega J., Vizuete W.

The legal commercialization of Cannabis for recreational and medical use in certain US states has effectively created a new and nearly unregulated cultivation industry. Within the city limits of Denver, Colorado, there are now more than 600 registered Cannabis spp. cultivation facilities (CCFs) for recreational and medical uses, each containing thousands of plants. Ambient measurements collected inside growing operations pre-legalization have found concentrations as high as 50–100 ppbv of terpenes; a group of highly reactive biogenic volatile organic compounds (BVOCs) and known precursors for the formation of ozone and particulate matter (PM). Due to its illicit nature there has been insufficient experimental data produced to determine Cannabis spp. emission rates. This study used, for the first time, an enclosure chamber and live Cannabis spp. plants during a 90-day growing period consisting of four different strains of Cannabis spp.: Critical Mass, Lemon Wheel, Elephant Purple, and Rockstar Kush. These measurements enabled characterization of terpenes and estimates of emission capacity (EC, μgC g−1 hr−1) at standard conditions. During peak growth, the percentages of individual BVOC emissions were dominated by β-myrcene (18–60%), eucalyptol (17–38%), and d-limonene (3–10%) for all strains. Our results showed large variability in the rate and composition of terpene emissions across different strains. For the Critical Mass and Lemon Wheel, the dominant terpenoid was eucalyptol (32% and 38%), and it was β-myrcene (60% and 45%) for the Elephant Purple and Rockstar Kush. Critical Mass produced the highest terpene emission capacity (8.7 μgC g−1 hr−1) and Rockstar Kush the lowest (4.9 μgC g−1 hr−1). With 600 CCFs in Denver, and assuming 10,000 plants per CCF, an emission capacity of 8.7 μgC g−1 hr−1 would more than double the existing rate of BVOC emissions to 520 metric ton year−1. Using Maximum Incremental Reactivity (MIR) values the total ozone formation potential from all these emitted species could produce 2100 metric tons year−1 of ozone, and based on published secondary organic aerosols yields 131 metric tons year−1 of PM. It is likely that the ECs calculated here are lower than those achieved in CCFs where growing conditions are optimized for rapid growth and higher biomass yields. Further studies including a greater number of the 620 available Cannabis spp. strains and a wider range of treatments are needed to generate a representative dataset. Such a dataset could then better enable assessments of the potential impacts of this new industry on indoor and regional air quality.

Fiorini D., Molle A., Nabissi M., Santini G., Benelli G., Maggi F.

Byproducts of industrial hemp (Cannabis sativa L.), including inflorescences, represent an exploitable material to produce niche products for the pharmaceutical, nutraceutical, cosmetic and pesticide industry. One of them is the essential oil, whose composition can be properly modulated on an industrial level by optimizing the extractive conditions and sample pretreatment. This allows to achieve high concentrations of bioactive compounds, such as cannabidiol (CBD) and sesquiterpenes [e.g. (E)-caryophyllene]. In the present work, we evaluated the effects of type of distillation apparatus, status of conservation of the plant material, grinding and sample-pretreatment with microwave and heat, on the hemp essential oil chemical profile obtained from the monoecious cultivar Felina 32. Seven marker compounds, including the monoterpenes α-pinene, myrcene and terpinolene, the sesquiterpenes (E)-caryophyllene, α-humulene and caryophyllene oxide, and the cannabinoid CBD were quantified in the different hemp essential oil samples by gas chromatography-flame ionization detection (GC-FID) analysis, whereas the overall chemical profiles were achieved by gas chromatography-mass spectrometry (GC–MS) analysis. Results showed that hydrodistillation (HD) in comparison with steam distillation (SD) gave a higher content of cannabinoids. Drying was fundamental to induce decarboxylation of cannabinoid acids to the relative alcoholic forms, coupled with an increase of the sesquiterpene fraction. The optimization of sample pretreatments pointed out that the exposure of dry inflorescences to microwave heating at 900 W power for 1 min was the best method to increase the abundance of bioactive compounds in the essential oil, with special reference to CBD, (E)-caryophyllene and caryophyllene oxide. Overall, these results give new insights into the exploitation of hemp byproducts in different fields such as pharmaceuticals, nutraceuticals and eco-friendly insecticides.

Protti M., Brighenti V., Battaglia M.R., Anceschi L., Pellati F., Mercolini L.

Cannabis sativa L. represents one of the most widely used source of drugs and drugs of abuse worldwide. Its biologically active compounds are mainly cannabinoids, including Δ9-tetrahydrocannabinol (THC), which is responsible for the psychoactive effects, tetrahydrocannabinolic acid (THCA), cannabinol (CBN), cannabidiol (CBD), and cannabidiolic acid (CBDA). Together with recreational and drug-type (or medicinal) Cannabis, some new products have been recently released into the market as fiber-type Cannabis variants (also known as hemp or industrial hemp) with low THC content and high content of nonpsychoactive CBD. In this research work, the aim was to characterize Cannabis recreational and drug-type samples by quantifying their active principles, after the development and validation of a suitable analytical method. In addition to the Cannabis samples described above, fiber-type plant varieties were also analyzed to monitor their content of nonpsychoactive compounds for both pharmaceutical and nutraceutical purposes. To do this, a highly efficient HPLC-DAD-MS/MS method, with an electrospray ionization (ESI) source and a triple-quadrupole mass analyzer acquiring in the multiple reaction monitoring (MRM) mode also coupled to a diode array detector (DAD), was developed and applied. Satisfactory validation results were obtained in terms of precision (RSD < 6.0% for all the analytes) and accuracy (>92.1% for all the compounds). The proposed methodology represents a versatile and reliable tool to assess both psychoactive and nonpsychoactive cannabinoid levels in Cannabis samples for a more rational use in both medicinal chemistry and nutraceutics.

Ibrahim E., Wang M., Radwan M., Wanas A., Majumdar C., Avula B., Wang Y., Khan I., Chandra S., Lata H., Hadad G., Abdel Salam R., Ibrahim A., Ahmed S., ElSohly M.

AbstractTerpenes are the major components of the essential oils present in various Cannabis sativa L. varieties. These compounds are responsible for the distinctive aromas and flavors. Besides the quantification of the cannabinoids, determination of the terpenes in C. sativa strains could be of importance for the plant selection process. At the University of Mississippi, a GC-MS method has been developed and validated for the quantification of terpenes in cannabis plant material, viz., α-pinene, β-pinene, β-myrcene, limonene, terpinolene, linalool, α-terpineol, β-caryophyllene, α-humulene, and caryophyllene oxide. The method was optimized and fully validated according to AOAC (Association of Official Analytical Chemists) guidelines against reference standards of selected terpenes. Samples were prepared by extraction of the plant material with ethyl acetate containing n-tridecane solution (100 µg/mL) as the internal standard. The concentration-response relationship for all analyzed terpenes using the developed method was linear with r2 values > 0.99. The average recoveries for all terpenes in spiked indoor cultivated samples were between 95.0 – 105.7%, with the exception of terpinolene (67 – 70%). The measured repeatability and intermediate precisions (% relative standard deviation) in all varieties ranged from 0.32 to 8.47%. The limit of detection and limit of quantitation for all targeted terpenes were determined to be 0.25 and 0.75 µg/mL, respectively. The proposed method is highly selective, reliable, and accurate and has been applied to the simultaneous determination of these major terpenes in the C. sativa biomass produced by our facility at the University of Mississippi as well as in confiscated marijuana samples.

Zengin G., Menghini L., Di Sotto A., Mancinelli R., Sisto F., Carradori S., Cesa S., Fraschetti C., Filippi A., Angiolella L., Locatelli M., Mannina L., Ingallina C., Puca V., D’Antonio M., et. al.

Due to renewed interest in the cultivation and production of Italian Cannabis sativa L., we proposed a multi-methodological approach to explore chemically and biologically both the essential oil and the aromatic water of this plant. We reported the chemical composition in terms of cannabinoid content, volatile component, phenolic and flavonoid pattern, and color characteristics. Then, we demonstrated the ethnopharmacological relevance of this plant cultivated in Italy as a source of antioxidant compounds toward a large panel of enzymes (pancreatic lipase, α-amylase, α-glucosidase, and cholinesterases) and selected clinically relevant, multidrug-sensible, and multidrug-resistant microbial strains (Staphylococcus aureus, Helicobacter pylori, Candida, and Malassezia spp.), evaluating the cytotoxic effects against normal and malignant cell lines. Preliminary in vivo cytotoxicity was also performed on Galleria mellonella larvae. The results corroborate the use of this natural product as a rich source of important biologically active molecules with particular emphasis on the role exerted by naringenin, one of the most important secondary metabolites.

Pellati F., Borgonetti V., Brighenti V., Biagi M., Benvenuti S., Corsi L.

In the last decades, a lot of attention has been paid to the compounds present in medicinal Cannabis sativa L., such as Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD), and their effects on inflammation and cancer-related pain. The National Cancer Institute (NCI) currently recognizes medicinal C. sativa as an effective treatment for providing relief in a number of symptoms associated with cancer, including pain, loss of appetite, nausea and vomiting, and anxiety. Several studies have described CBD as a multitarget molecule, acting as an adaptogen, and as a modulator, in different ways, depending on the type and location of disequilibrium both in the brain and in the body, mainly interacting with specific receptor proteins CB1 and CB2. CBD is present in both medicinal and fibre-type C. sativa plants, but, unlike Δ9-THC, it is completely nonpsychoactive. Fibre-type C. sativa (hemp) differs from medicinal C. sativa, since it contains only few levels of Δ9-THC and high levels of CBD and related nonpsychoactive compounds. In recent years, a number of preclinical researches have been focused on the role of CBD as an anticancer molecule, suggesting CBD (and CBD-like molecules present in the hemp extract) as a possible candidate for future clinical trials. CBD has been found to possess antioxidant activity in many studies, thus suggesting a possible role in the prevention of both neurodegenerative and cardiovascular diseases. In animal models, CBD has been shown to inhibit the progression of several cancer types. Moreover, it has been found that coadministration of CBD and Δ9-THC, followed by radiation therapy, causes an increase of autophagy and apoptosis in cancer cells. In addition, CBD is able to inhibit cell proliferation and to increase apoptosis in different types of cancer models. These activities seem to involve also alternative pathways, such as the interactions with TRPV and GRP55 receptor complexes. Moreover, the finding that the acidic precursor of CBD (cannabidiolic acid, CBDA) is able to inhibit the migration of breast cancer cells and to downregulate the proto-oncogene c-fos and the cyclooxygenase-2 (COX-2) highlights the possibility that CBDA might act on a common pathway of inflammation and cancer mechanisms, which might be responsible for its anticancer activity. In the light of all these findings, in this review we explore the effects and the molecular mechanisms of CBD on inflammation and cancer processes, highlighting also the role of minor cannabinoids and noncannabinoids constituents of Δ9-THC deprived hemp.

Pellati F., Brighenti V., Sperlea J., Marchetti L., Bertelli D., Benvenuti S.

Cannabis sativa L. is a dioecious plant belonging to the Cannabaceae family. The main phytochemicals that are found in this plant are represented by cannabinoids, flavones, and terpenes. Some biological activities of cannabinoids are known to be enhanced by the presence of terpenes and flavonoids in the extracts, due to a synergistic action. In the light of all the above, the present study was aimed at the multi-component analysis of the bioactive compounds present in fibre-type C. sativa (hemp) inflorescences of different varieties by means of innovative HPLC and GC methods. In particular, the profiling of non-psychoactive cannabinoids was carried out by means of HPLC-UV/DAD, ESI-MS, and MS2. The content of prenylated flavones in hemp extracts, including cannflavins A and B, was also evaluated by HPLC. The study on Cannabis volatile compounds was performed by developing a new method based on headspace solid-phase microextraction (HS-SPME) coupled with GC-MS and GC-FID. Cannabidiolic acid (CBDA) and cannabidiol (CBD) were found to be the most abundant cannabinoids in the hemp samples analysed, while β-myrcene and β-caryophyllene were the major terpenes. As regards flavonoids, cannflavin A was observed to be the main compound in almost all the samples. The methods developed in this work are suitable for the comprehensive chemical analysis of both hemp plant material and related pharmaceutical or nutraceutical products in order to ensure their quality, efficacy, and safety.

Corsi L., Pellati F., Brighenti V., Plessi N., Benvenuti S.

Background: Fibre-type Cannabis sativa L. (hemp) usually contains cannabidiolic acid and cannabidiol as the main non-psychoactive cannabinoids. Even though there is evidence of the neuroprotective activity of pure cannabidiol, no in vitro studies have reported so far the role of hemp extracts on neuroprotection. The objective of this study was to evaluate the neuroprotective effect of hemp extracts in in vitro cellular models of neurotoxicity. Methods: One extract was obtained from raw hemp inflorescences, while the other was prepared from the same plant material submitted to a decarboxylation process. The composition of both these extracts was evaluated by HPLC-UV/DAD. Human neuroblastoma SH-SY5Y and microglial BV-2 cell lines treated with rotenone were selected as the model of neurodegeneration. The neuroprotection of hemp extracts was assessed also in serum-free conditions both in the presence and in the absence of rotenone as the toxic agent by using the same cell lines. The neuroprotective potential of cannabidiol was tested in parallel. Results: The decarboxylated hemp extract possesses a mild neuroprotective activity on BV-2 cells treated with rotenone, higher than that of pure cannabidiol. As regards serum-free experiments, the nondecarboxylated hemp extract was the most effective neuroprotective agent toward SH-SY5Y cells, while BV-2 cells were better protected from the toxic insult by the decarboxylated extract and cannabidiol. Conclusion: Both hemp extracts and pure cannabidiol displayed a moderate neuroprotective activity in the neurotoxicity models considered in this study; in addition, they showed a trophic effect on SHSY5Y cells.

Marini E., Magi G., Ferretti G., Bacchetti T., Giuliani A., Pugnaloni A., Rippo M.R., Facinelli B.

Anti-virulence strategies are being explored as a novel approach to combat pathogens. Such strategies include inhibition of surface adhesion, tissue invasion, toxin production, and/or interference with the gene regulation of other virulence traits. Listeria monocytogenes, the causative agent of listeriosis, is a facultative intracellular food pathogen characterized by a wide distribution in the environment. Its ability to persist within biofilms and to develop resistance to sanitizers is the cause of significant problems in food processing plants and of steep costs for the food industry. In humans, the treatment of listeriosis is hampered by the intracellular location of listeriae and the poor intracellular penetration of some antibiotics. Eleven L. monocytogenes isolates from patients who were diagnosed with invasive listeriosis in Italy in 2014-2016 were studied. This in vitro and in vivo study explored the antibacterial and anti-virulence properties of a steam-distilled essential oil of C. sativa L., which is being intensively investigated for its high content in powerful bioactive phytochemicals. Susceptibility experiments demonstrated a moderate bactericidal activity of the essential oil (Minimum Bactericidal Concentration >2048 μg/mL). Assessment of the effects of sublethal concentrations of the essential oil on L. monocytogenes virulence traits demonstrated a significant action on motility. Listeriae were non-motile after exposure to the essential oil. Light and scanning electron microscopy documented aggregates of listeriae with the flagella trapped inside the cluster. Real-time RT-PCR experiments showed downregulation of flagellar motility genes and of the regulatory gene prfA. The ability to form biofilm and to invade Caco-2 cells was also significantly reduced. Galleria mellonella larvae infected with L. monocytogenes grown in presence of sublethal concentrations of the essential oil showed much higher survival rates compared with controls, suggesting that the extract inhibited tissue invasion. Food contamination with L. monocytogenes is a major concern for the food industry, particularly for plants making ready-to-eat and processed food. The present work provides a baseline in the study of the anti-virulence properties of the C. sativa EO against L. monocytogenes. Further studies are needed to understand if it could be used as an alternative agent for the control of L. monocytogenes in food processing plants.

Calvi L., Pentimalli D., Panseri S., Giupponi L., Gelmini F., Beretta G., Vitali D., Bruno M., Zilio E., Pavlovic R., Giorgi A.

There are at least 554 identified compounds in C. sativa L., among them 113 phytocannabinoids and 120 terpenes. Phytocomplex composition differences between the pharmaceutical properties of different medical cannabis chemotype have been attributed to strict interactions, defined as 'entourage effect', between cannabinoids and terpenes as a result of synergic action. The chemical complexity of its bioactive constituents highlight the need for standardised and well-defined analytical approaches able to characterise the plant chemotype, the herbal drug quality as well as to monitor the quality of pharmaceutical cannabis extracts and preparations. Hence, in the first part of this study an analytical procedures involving the combination of headspace-solid-phase microextraction (HS-SPME) coupled to GC-MS and High Resolution Mass-Spectrometry LC-HRMS (Orbitrap®) were set up, validated and applied for the in-depth profiling and fingerprinting of cannabinoids and terpenes in two authorised medical grade varieties of Cannabis sativa L. inflorescences (Bedrocan® and Bediol®) and in obtained macerated oils. To better understand the trend of all volatile compounds and cannabinoids during oil storage a new procedure for cannabis macerated oil preparation without any thermal step was tested and compared with the existing conventional methods to assess the potentially detrimental effect of heating on overall product quality.

Citti C., Braghiroli D., Vandelli M.A., Cannazza G.

Cannabis products have recently regained much attention due to the high pharmacological potential of their cannabinoid content. In this review, the most widely used sample preparation strategies for the extraction of cannabinoids are described for the specific application to either plant materials or biological matrices. Several analytical techniques are described pointing out their respective advantages and drawbacks. In particular, chromatographic methods, such as TLC, GC and HPLC, are discussed and compared in terms of selectivity and sensitivity. Various detection methods are also presented based on the specific aim of the cannabinoids analysis. Lastly, critical considerations are mentioned with the aim to deliver useful suggestions for the selection of the optimal and most suitable method of analysis of cannabinoids in either biomedical or cannabis derived samples.

Kačániová M., Vukic M.D., Vukovic N.L., Terentjeva M., Ban Z., Li L., Bianchi A., Ben Sad R., Ben Hsouna A., Elizondo-Luévano J.H., Čmiková N., Garzoli S.

Bai Y., You N., Tian H., Zhao X.

Hemp seed oil is nutritious. Besides cannabinoids, it has a higher Omega-3 fatty acid content, making it an ideal and healthy edible oil. Roasting is a key factor affecting the flavor of hemp seed oil. In this study, solvent-assisted flavor evaporation (SAFE) combined with gas chromatography–mass spectrometry (GC-O-MS) was used to analyze the volatile compounds of hemp seed oil after the hemp seeds were roasted at different temperatures (120 °C, 140 °C, 160 °C, 180 °C). α-pinene, (1S)-(1)-β-pinene, myrcene, and (E)-β-ocimene, which are typical odorants for herbs and fresh flavors, had an active odor value (OAV) ≥ 1. The sweet, nutty, and toasty flavors became more pronounced as the temperature increased. 2-pentylfuran provided the caramel flavor, and 2,5-dimethylpyrazine, 2-ethyl-6-methylpyrazine, and 2-ethyl-5-methylpyrazine provided the toasty flavor. The combination of sensory evaluation and data analysis was effective in differentiating the odors of hemp seed oil extracted at different temperatures. Reconstitution and deletion experiments showed that these odorants could mimic the overall aroma profile. This article provides a theoretical basis for the effect of over-roasting on the flavor of hemp seed oil.

Dudley A., Kassama L., Jackson‐Davis A., Kuang X., Cebert E.

AbstractMedicinal plants are an excellent source of bioactive components and are gaining significant attention for food protection due to their bioactive properties. In this context, hemp (Cannabis sp.) is being explored for such applications because of its well‐known antibacterial and antioxidant activities. However, the bioactive efficacy of cultivars currently grown in Northern Alabama has not been widely studied. Therefore, the purpose of this study was to evaluate the physicochemical properties of two hemp cultivars (Hemp 5 and Hemp 17). Hemp inflorescences grown at the Winfred Thomas Agricultural Research Station were used in this study. The antioxidant activity and content of the extracts were determined by ferric reducing antioxidant power, radical scavenging activity, total phenolic content, and total flavonoid content. Antibacterial activity against cocktails of Listeria monocytogenes (LM) and Salmonella enterica (SE) was evaluated by optical density and disc diffusion. All treatments were analyzed in triplicate, and analysis of variance was conducted with statistical significance based on p ≤ 0.05. Results indicated that cultivars and defatting significantly affected (p ≤ 0.05) the antioxidant properties, with Hemp 17 DF (defatted) having the highest RSA (70.51 ± 4.24%) compared to the positive control of ascorbic acid (83.81 ± 5.85%). Antibacterial results based on optical density indicated that hemp extracts had a significantly (p ≤ 0.05) lower optical density compared to the negative controls (LM and SE). These findings suggest that Northern Alabama hemp cultivars can potentially be utilized to enhance food safety and quality.Practical ApplicationNorthern Alabama cultivars of hemp extract can be utilized for the enhancement of the safety and quality application due to inherent antioxidant and antimicrobial properties.

Rezghiyan A., Esmaeili H., Farzaneh M.

Various practical strategies have been employed to mitigate the detrimental effects of water deficit stress on plants such as application of nano-stimulants. Nanosilicon plays a crucial role in alleviating the deleterious impacts of both abiotic and biotic stresses in plants by modulating various phyto-morphological and physiological processes. This study aimed to examine the combined effects of drought stress and nanosilicon application on the morphological traits and essential oil content and compositions of hemp (Cannabis sativa L.), in which four-week-old seedlings were subjected to irrigation treatments at four levels, including 100% (control), 80% (mild stress), 60% (moderate stress) and 40% (severe stress) field capacity and nanosilicon at three concentrations (0, 0.5 and 1.5 mM) in a completely randomized factorial design experiment with three replications for 40 days. The results showed that the maximum plant height (109.07 cm), number of nodes (33.3), and number of flowering branches (29.4) were recorded under the treatment of 1.5 mM nanosilicon and 100% FC. The lowest fresh and dry weights of aerial parts were associated to the severe drought stress (40% FC) without nanosilicon application. The mild water stress (80% FC) combined with foliar application of 1.5 mM nanosilicon led to highest EO content (0.17%) compared with the other treatments. However, the highest content of cannabidiol in the essential oil was achieved in the severe water stress (40% FC) and treatment of 0.5 mM nanosilicon. The results showed that the application of nanosilicon improved the morphological characteristics and also changed the content and compositions of the hemp plants under drought stress conditions.

Dey M., Bera S., Tyagi P., Pal L.

AbstractThis review focuses on antimicrobial packaging for food safety, critically examining the activity and efficacy of cannabinoids against commonly found microorganisms and exploring their antimicrobial mechanisms. Specifically, the review considers cannabinoids derived from industrial hemp plants, which are characterized by low levels of psychoactive components. It also outlines viable strategies to control the sustained release of cannabinoids from the packaging, enabling extended storage and enhanced safety of food products. Research demonstrates that cannabinoids are effective against both foodborne bacteria and fungi, with their antimicrobial action primarily attributed to microbial membrane instability. Cannabinoids can be utilized to prepare effective antimicrobial films and edible coatings; however, the number of studies in this area remains limited. The potential of cannabinoids to contribute to intelligent packaging systems is also discussed, with an emphasis on the regulatory aspects and challenges associated with incorporating cannabinoids into food packaging. Finally, the review identifies future research directions to address current limitations and advance hemp‐based antimicrobial food packaging solutions.

Fulvio F., Pieracci Y., Ascrizzi R., Bassolino L., Flamini G., Paris R.

Terpenes, volatile compounds known for their aromatic and therapeutic properties, play a pivotal role in shaping the overall chemical profile of Cannabis sativa L. Their biosynthesis in planta occurs in trichomes and involves the 2-C-methyl-D-erythritol 4-phosphate (MEP) and the mevalonic acid (MVA) pathways, responsible for producing the substrates utilized by a family of enzymes, the terpene synthases (TPS), for terpene production. In this work, a comprehensive approach combining chemical analyses of the volatile compounds characterizing the aroma of the inflorescences three C. sativa genotypes collected at three stages of maturity and the transcriptional analyses of key genes involved in the terpene biosynthesis was adopted to study this pathway. The results revealed different terpene profiles among genotypes, which were characterized by peculiar compounds belonging to the sesqui- (CINBOL and Fibrante) or monoterpene (Ermo) categories. Both structural and putative regulatory genes were analysed by RT-qPCR, revealing distinct transcriptional profiles of Terpene Synthases, contributing to the diversity of mono and sesquiterpenes synthesized. Furthermore, the research delved into potential regulatory genes associated with trichome formation, a crucial factor influencing terpene accumulation. This integrated approach highlighted complex mechanisms governing terpene accumulation in cannabis, while also offering potential regulators putatively involved in this pathway.

Gugleva V., Mihaylova R., Kamenova K., Zheleva-Dimitrova D., Stefanova D., Tzankova V., Zaharieva M.M., Najdenski H., Forys A., Trzebicka B., Petrov P.D., Momekova D.

The study investigates the development and characterization of dual-loaded niosomes incorporated into ion-sensitive in situ gel as a potential drug delivery platform for ophthalmic application. Cannabidiol (CBD) and epigallocatechin-3-gallate (EGCG) simultaneously loaded niosomes were prepared via the thin film hydration (TFH) method followed by pulsatile sonication and were subjected to comprehensive physicochemical evaluation. The optimal composition was included in a gellan gum-based in situ gel, and the antimicrobial activity, in vitro toxicity in a suitable corneal epithelial model (HaCaT cell line), and antioxidant potential of the hybrid system were further assessed. Dual-loaded niosomes based on Span 60, Tween 60, and cholesterol (3.5:3.5:3 mol/mol) were characterized by appropriate size (250 nm), high entrapment efficiency values for both compounds (85% for CBD and 50% for EGCG) and sustained release profiles. The developed hybrid in situ gel exhibited suitable rheological characteristics to enhance the residence time on the ocular surface. The conducted microbiological studies reveal superior inhibition of methicillin-resistant Staphylococcus aureus (MRSA) adhesion by means of the niosomal in situ gel compared to the blank gel and untreated control. Regarding the antioxidant potential, the dual loading of CBD and EGCG in niosomes enhances their protective properties, and the inclusion of niosomes in gel form preserves these effects. The obtained outcomes indicate the developed niosomal in situ gel as a promising drug delivery platform in ophthalmology.

Nethengwe M., Maphosa Y., Ahiante B., Oyenihi A.

Zhou J., Kamei K., Marin E., Pezzotti G., Ma Y., Li Y., Zhu W.

Niyangoda D., Aung M.L., Qader M., Tesfaye W., Bushell M., Chiong F., Tsai D., Ahmad D., Samarawickrema I., Sinnollareddy M., Thomas J.

Background: Two major bacterial pathogens, Staphylococcus aureus and Streptococcus pyogenes, are becoming increasingly antibiotic-resistant. Despite the urgency, only a few new antibiotics have been approved to address these infections. Although cannabinoids have been noted for their antibacterial properties, a comprehensive review of their effects on these bacteria has been lacking. Objective: This systematic review examines the antibacterial activity of cannabinoids against S. aureus, including methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) strains, and S. pyogenes. Methods: Databases, including CINAHL, Cochrane, Medline, Scopus, Web of Science, and LILACS, were searched. Of 3510 records, 24 studies met the inclusion criteria, reporting on the minimum inhibitory concentration (MIC) and minimum bactericidal concentration of cannabinoids. Results: Cannabidiol (CBD) emerged as the most effective cannabinoid, with MICs ranging from 0.65 to 32 mg/L against S. aureus, 0.5 to 4 mg/L for MRSA, and 1 to 2 mg/L for VRSA. Other cannabinoids, such as cannabichromene, cannabigerol (CBG), and delta-9-tetrahydrocannabinol (Δ9-THC), also exhibited significant antistaphylococcal activity. CBD, CBG, and Δ9-THC also showed efficacy against S. pyogenes, with MICs between 0.6 and 50 mg/L. Synergistic effects were observed when CBD and essential oils from Cannabis sativa when combined with other antibacterial agents. Conclusion: Cannabinoids’ antibacterial potency is closely linked to their structure–activity relationships, with features like the monoterpene region, aromatic alkyl side chain, and aromatic carboxylic groups enhancing efficacy, particularly in CBD and its cyclic forms. These results highlight the potential of cannabinoids in developing therapies for resistant strains, though further research is needed to confirm their clinical effectiveness.

Saleem U., Asrar M., Jabeen F., Makhdoom Hussain S., Hussain D.

Ribeiro A., Alsayyed R., Oliveira D., Loureiro R., Cabral-Marques H.

Cannabis sativa L. has garnered attention as a potential source for new antimicrobial agents, particularly due to the increased prevalence of microbial resistance to conventional antimicrobials and the emergence of multidrug-resistant pathogens. This review, conducted according to the PRISMA 2020 statement, systematically analyzed the antimicrobial properties of C. sativa extracts and cannabinoids against various bacteria, fungi, viruses, and parasites. Data were collected from the scientific literature (102 papers) and clinical trials (5 studies) from 2014 to June 2024. Findings revealed that cannabinoids, especially CBD, demonstrate significant antimicrobial activity against Gram-positive bacteria like MRSA, Gram-negative bacteria such as Pseudomonas aeruginosa, various Candida species, SARS-CoV-2, and HIV. Additionally, CBD showed efficacy against parasitic infections like Echinococcus granulosus and Leishmania species. These results suggest that cannabinoids may represent a new class of antimicrobial agents with unique and diverse mechanisms of action, potentially effective in broad-spectrum therapies. This study highlights the urgent need for further research and standardized clinical trials to validate these findings and to develop cannabinoid-based treatments. The antimicrobial properties of C. sativa align with WHO priorities and support global health initiatives, offering promising avenues for addressing antimicrobial resistance and improving public health outcomes.

Pieracci Y., Bozzini M.F., Campanini C., Muscatello B., De Martino L., Nocera F.P., Fulvio F., Montanari M., Flamini G., Fratini F.

Nwanade C.F., Wang M., Pei T., Meng J., Yu Z., Liu J.

Plant essential oils (EOs)-based acaricides have been recognized as environmentally-friendly alternatives to synthetic acaricides because of their low toxicity against non-target species. Despite this, there are knowledge gaps regarding the toxicity mechanisms of plant EOs against non-target species. Here, the toxicology and enzymatic mechanism of Citrus reticulata and Citrus lemon EOs were evaluated against the vector pest, Haemaphysalis longicornis, and non-target ladybird beetle, Harmonia axyridis. Both EOs were mainly composed of d-Limonene, followed by β-Myrcene and γ-Terpinene in C. reticulata, and (-)-β-Pinene and γ-Terpinene in C. lemon. Citrus reticulata and C. lemon EOs were toxic to Hae. longicornis, with 50 % lethal concentration (LC

Niyangoda D., Muayad M., Tesfaye W., Bushell M., Ahmad D., Samarawickrema I., Sinclair J., Kebriti S., Maida V., Thomas J.

This systematic review critically evaluates preclinical and clinical data on the antibacterial and wound healing properties of cannabinoids in integument wounds. Comprehensive searches were conducted across multiple databases, including CINAHL, Cochrane library, Medline, Embase, PubMed, Web of Science, and LILACS, encompassing records up to May 22, 2024. Eighteen studies met the inclusion criteria. Eleven were animal studies, predominantly utilizing murine models (n = 10) and one equine model, involving 437 animals. The seven human studies ranged from case reports to randomized controlled trials, encompassing 92 participants aged six months to ninety years, with sample sizes varying from 1 to 69 patients. The studies examined the effects of various cannabinoid formulations, including combinations with other plant extracts, crude extracts, and purified and synthetic cannabis-based medications administered topically, intraperitoneally, orally, or sublingually. Four animal and three human studies reported complete wound closure. Hemp fruit oil extract, cannabidiol (CBD), and GP1a resulted in complete wound closure in twenty-three (range: 5–84) days with a healing rate of 66–86% within ten days in animal studies. One human study documented a wound healing rate of 3.3 cm2 over 30 days, while three studies on chronic, non-healing wounds reported an average healing time of 54 (21–150) days for 17 patients by oral oils with tetrahydrocannabinol (THC) and CBD and topical gels with THC, CBD, and terpenes. CBD and tetrahydrocannabidiol demonstrated significant potential in reducing bacterial loads in murine models. However, further high-quality research is imperative to fully elucidate the therapeutic potential of cannabinoids in the treatment of bacterial skin infections and wounds. Additionally, it is crucial to delineate the impact of medicinal cannabis on the various phases of wound healing. This study was registered in PROSPERO (CRD42021255413).