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Metabolic fingerprinting of Cannabis sativa L., cannabinoids and terpenoids for chemotaxonomic and drug standardization purposes
... The same method and criterion were used for all the cannabis extracts. (27), and geraniol (30) and (+)-pulegone (31) in the second dimens Compound numbers are given according to Table S1. ...
... As expected, the monoterpenes, monoter noids, sesquiterpenes, and sesquiterpenoids were eluted in separate regions in the ch matograms of all six cannabis extracts using GC × GC, as explained above. Figure 2 sho (27), and geraniol (30) and (+)-pulegone (31) in the second dimension. Compound numbers are given according to Table S1. ...
... Even though each study design was different, we were able to detect around four times more terpenes and terpenoids using GC × GC as compared to the reported GC studies [4,[18][19][20]30,35,38,43,44,46,47,49,[56][57][58][59]. The number of terpenes and terpenoids detected in the reported GC studies ranged from 23 to 109. ...
Cannabis contains a wide range of terpenes and terpenoids that are mainly responsible for their distinctive aroma and flavor. These compounds have also demonstrated therapeutic effects either alone and/or as synergistic compounds with other terpenes, terpenoids, and/or cannabinoids. Several studies have attempted to fully characterize terpenes and terpenoids in cannabis; however, most of these studies used one-dimensional gas chromatography, which often results in the co-elution of the compounds. In the present study, we analyzed terpenes and terpenoids in the dried flowers of six cannabis strains using a two-dimensional gas chromatograph time-of-flight mass spectrometer (GC × GC-TOFMS). A total of 146 terpenes and terpenoids were detected across all six cannabis strains with an enhanced separation of 16 terpenes and terpenoids in the second dimension. Additionally, we achieved enhanced separation of four terpenes and terpenoids from a standard mixture in the second dimension. Chemical differences were observed in the number and relative abundance of monoterpenes, monoterpenoids, sesquiterpenes, and sesquiterpenoids in all six strains. We were also able to identify four new terpenoids in cannabis, which are reported here for the first time.
... The use of controlled lighting cycles, specialized soil, fine-tuned nutrients and pest control eliminate many of the environmental variables and will make geographical assigning of the plant difficult if not impossible. Fischedick et al. analyzed 11 cultivars of cannabis for 36 compounds and managed to discriminate the various cultivars with PCA [14]. Higher levels of cannabinoids correlated positively to higher levels of terpenoids (R 2 =0.7688). ...
... Calculation of THCmax, CBDmax and CBGmax were performed as described in our previous paper [18]. Terpene content was determined using GC-FID according to the same approach as Fischedick et al. using retention time comparison with authentic reference, mass spectra, and literature data [14]. A 5% diphenyl and 95% dimethyl polysiloxane column (SHRX5, 15 m, 0.25 mm ID, 0.25 µm film thickness, Shimadzu Scientific Instruments, Columbia, MD, USA) and helium carrier gas (Airgas, Radnor, PA, USA) was used for separation. ...
... The sum of terpenes was plotted against the sum of THCmax and CBDmax and a R 2 value of 0.4248 which is lower than the correlation of 0.77 found by Fischedick et al. [14]. The graph can be found in supplementary information (Figure 1). ...
... Cannabis sativa, Cannabis sativa L., is the oldest therapeutic plant (Hanif et al., 2017a), it's also referred to as a ceremonial drug (Fischedick et al., 2010). It is an ancient plant, as shown by its existence in 4500-year-old Egyptian tombs (Bonini et al., 2018). ...
... It has the right amount of both types of fats (Deferne and Pate, 1996). Cannabis sativa seed oil has a pleasant taste and a perfect and balanced ratio of omega 3 and omega 6 fatty acids, which are called polyunsaturated fatty acids (PUFAs) (Fischedick et al., 2010). This makes it different from other seed oils, which don't have this ratio. ...
... The experiment was conducted on harvested C. sativa plants that had been diluted to different concentrations. To measure the extract's total antioxidant capacity, the solvent was not eliminated throughout the liquid/ liquid extraction method (Fischedick et al., 2010). The researchers compared plant samples from Gujranwala, Narowal, and Lahore. ...
... The production of secondary metabolites widely varies in their concentrations and phytochemical profiles, even within the same genotype cultivated under controlled conditions, as well as even between different seasons and cycles. In a study developed by Fischedick and Hazekamp (2010) [51], under strictly controlled indoor growing conditions and in a standard lot, an average variation was observed of 7.6% and 5.5% for phytocannabinoids and 11% and 4% for terpenoids in Bedrocan and Bedica cannabis plant varieties, respectively. ...
... The production of secondary metabolites widely varies in their concentrations and phytochemical profiles, even within the same genotype cultivated under controlled conditions, as well as even between different seasons and cycles. In a study developed by Fischedick and Hazekamp (2010) [51], under strictly controlled indoor growing conditions and in a standard lot, an average variation was observed of 7.6% and 5.5% for phytocannabinoids and 11% and 4% for terpenoids in Bedrocan and Bedica cannabis plant varieties, respectively. ...
... One main difficulty in the production of plant secondary metabolites is the standardization of these compounds. Herbal products can never be perfectly standardized for active component content; however, variability in chemical composition may also be a cause of concern for medicinal users [51] because the maintenance of the concentration and profile of secondary metabolites is important to maintain the stability of the treatment. The knowledge of how abiotic factors and management techniques influence the production of phytocannabinoids can help in the development of management protocols that provide greater productivity, quality and stability in the production of these compounds. ...
The main characteristic of Cannabis sativa L. is the production of compounds of medicinal interest known as phytocannabinoids. Environmental factors and crop management practices are directly related to the yield of these compounds. Knowing how these factors influence the production of phytocannabinoids is essential to promote greater metabolite yield and stability. In this review, we aim to examine current cannabis agronomic research topics to identify the available information and the main gaps that need to be filled in future research. This paper introduces the importance of C. sativa L., approaching state-of-the-art research and evaluating the influence of crop management and environment conditions on yield and phytocannabinoid production, including (i) pruning; (ii) light and plant density; (iii) ontogeny; (iv) temperature, altitude, and CO2 concentration; (v) fertilization and substrate; and (vi) water availability, and presents concluding remarks to shed light on future directions.
... Identificación y cuantificación de terpenos por GC-FID: En este estudio se utilizó un Cromatógrafo de gases con detector de ionización de llama Agilent (GC-FID 7890B), usando las columnas HP-5 e Innowax para la optimización del método analítico para la identificación y cuantificación de terpenos presentes en aceites de Cannabis. Se ensayaron numerosas condiciones, de acuerdo a lo descrito en la bibliografía existente para elaborar dos métodos analíticos apropiados para las características de las columnas utilizadas (18)(19)(20)(21)(22)(23)(24)(25) . Para seleccionar estos métodos, también se consideraron los tiempos de retención de estándares disponibles comercialmente, tiempo que demanda el análisis completo de la muestra, resolución de los picos del cromatograma, linealidad de las curvas de calibración y reproducibilidad. ...
... Para las curvas de calibración, se prepararon soluciones de los estándares empleando metanol grado HPLC como diluyente, en un rango de 0,0005 a 2,0 mg/ml. Este rango fue seleccionado teniendo en cuenta la sensibilidad del equipo (límite de detección y lecturas por debajo de la saturación del detector), la concentración de los terpenos en las muestras de aceites de Cannabis que fueron analizadas para optimizar los métodos de análisis en nuestro laboratorio y las concentraciones reportadas en la bibliografía (16,18) . En este sentido, las muestras de aceites fueron diluidas 20 veces para quedar comprendidas en este rango de concentraciones. ...
Introducción: Cannabis sativa es una especie vegetal con gran número de principios activos, por lo que la lista de sus usos terapéuticos se está ampliando. En este sentido, hay numerosas evidencias del posible uso medicinal de los terpenos, así como del sinergismo de ellos con los cannabinoides (efecto séquito). Así, a medida que más países contemplan la legalización y autorización del cannabis medicinal, el número de laboratorios de extracción y análisis de cannabis aumenta para satisfacer la demanda, requiriéndose herramientas analíticas adecuadas. Metodología: En respuesta a numerosas consultas de médicos, laboratorios de análisis y usuarios, el laboratorio de cromatografía del PROBIEN ha seleccionado dos métodos para el análisis de terpenos en aceite de Cannabis por la técnica de cromatografía gaseosa (GC-FID). Se describen los métodos usando las columnas HP-5 e Innowax. Se empleó el método del estándar externo para la determinación cuantitativa de β-Pineno, Myrceno, p-Cymeno, Limoneno, Linalool, α-Terpineol, Nerol y Geraniol. Resultados: se observó una buena separación de picos y reproducibilidad, apropiadas para la identificación y cuantificación de los principales terpenos en extractos de Cannabis. La relación área/concentración fue lineal en el rango de 0,0005 a 2,0 mg/ml. Conclusión principal: los métodos descritos permiten la identificación y cuantificación de los terpenos mayoritarios en aceite de Cannabis para un control de calidad adecuado.
... From these, THC, responsible for the psychotropic effects [16], and cannabidiol (CBD) are the predominant forms of cannabinoids in drug-type and fiber-type varieties of cannabis, respectively. Other phenolic components of cannabis include flavonoids, spiroindanes, dihydrostilbenes, phenanthrenes and dihydrophenanthrenes [15,[17][18][19][20][21]. The concentration of these compounds depends on many factors such as plant variety, growth conditions and harvest time. ...
... However, C. sativa is a single species, and hundreds of cultivars have been developed for increasing or decreasing THC content [24]. There have been some attempts to classify cultivars based on chemical composition [19,[25][26][27], and different effects of cannabis have been attributed to different values of the ratio of THC/CBD [28]. ...
Currently, much effort is being placed into obtaining extracts and/or essential oils from Cannabis sativa L. for specific therapeutic purposes or pharmacological compositions. These potential applications depend mainly on the phytochemical composition of the oils, which in turn are determined by the type of C. sativa and the extraction method used to obtain the oils. In this work, we have evaluated the contents of secondary metabolites, delta-9-tetrahydrocannabinol (THC), and cannabidiol (CBD), in addition to the total phenolic, flavonoids, and anthraquinone content in oils obtained using solid–liquid extraction (SLE) and supercritical fluid extraction (SCF). Different varieties of C. sativa were chosen by using the ratio of THC to CBD concentrations. Additionally, antioxidant, antifungal and anticancer activities on different cancer cell lines were evaluated in vitro. The results indicate that oils extracted by SLE, with high contents of CBD, flavonoids, and phenolic compounds, exhibit a high antioxidant capacity and induce a high decrease in the cell viability of the tested breast cancer cell line (MCF-7). The observed biological activities are attributed to the entourage effect, in which CBD, phenols and flavonoids play a key role. Therefore, it is concluded that the right selection of C. sativa variety and the solvent for SLE extraction method could be used to obtain the optimal oil composition to develop a natural anticancer agent.
... According to Nagy, et al. [86], these essential oils are mainly composed of (E)-caryophyllene, α-humulene, β-selinene and α-selinene, with values of 29, 10, 4 and 3%, respectively. Additionally, Fischedick, et al. [110] reported the presence of mono-and sesquiterpenes in Cannabis flower essential oils. This study featured the quantification of monoterpenes and proved that they dominate the chemical composition, with a concentration of 28.3 mg/g of dry weight. ...
... Furthermore, sesquiterpenes are represented by β-caryophyllene and αhumulene. These sesquiterpenes represent a concentration ranging between 0.5 and 10.1 mg/g of dry weight [110]. Polyphenols and cannabinoids have also been identified in the methanolic extract of flowers. ...
For more than a century, Cannabis was considered a narcotic and has been banned by lawmakers all over the world. In recent years, interest in this plant has increased due to its therapeutic potential, in addition to a very interesting chemical composition, characterized by the presence of an atypical family of molecules known as phytocannabinoids. With this emerging interest, it is very important to take stock of what research has been conducted so far on the chemistry and biology of Cannabis sativa. The aim of this review is to describe the traditional uses, chemical composition and biological activities of different parts of this plant, as well as the molecular docking studies. Information was collected from electronic databases, namely SciFinder, ScienceDirect, PubMed and Web of Science. Cannabis is mainly popular for its recreational use, but it is also traditionally used as remedy for the treatment of several diseases, including diabetes, digestive, circulatory, genital, nervous, urinary, skin and respiratory diseases. These biological proprieties are mainly due to the presence of bioactive metabolites represented by more than 550 different molecules. Molecular docking simulations proved the presence of affinities between Cannabis compounds and several enzymes responsible for anti-inflammatory, antidiabetic, antiepileptic and anticancer activities. Several biological activities have been evaluated on the metabolites of Cannabis sativa, and these works have shown the presence of antioxidant, antibacterial, anticoagulant, antifungal, anti-aflatoxigenic, insecticidal, anti-inflammatory, anticancer, neuroprotective and dermocosmetic activities. This paper presents the up-to-date reported investigations and opens many reflections and further research perspectives.
... Cannabis sativa L., like every plant, has cellulose polymeric chains linked by microfibrils in its fundamental plant structure [10]. The primary metabolites' structure and quantities are comparable to those of any other plant. ...
... So far, more than 90 cannabinoids have been identified. The most common are cannabinodiolic acid, cannabinolic acid, and cannabidiolic acid, followed by cannabidgerolic acid, cannabichromic acid, and tetra-hydro-cannabinolic acid [10,11]. Cannabinoids are typically found as carboxylic acids, which are decarboxylated by dehydrating or scorching the collected plant over the period. ...
Hemp is a high-value crop that originated in Central Asia and is a historic but emerging cultivated plant. It may be grown for fiber, food, paper making, textiles, and therapeutic reasons. In the 21st century, market interest in hemp and its products has notably increased because seed portions can be utilized in the agri-food business, the woody component of the stem can be used in green buildings, the outer layer of the stems can be used in the textile industry, and the extraction of bioactive components from roots can play a vital role in the pharmacological industries. Hemp has recently been demonstrated to be a viable alternative for economies built on synthetic materials by the food, pharmaceutical, textiles, paper, building, and energy industries, among others. As a result, the goal of this study is to assemble the significant advancements in hemp, as well as to identify research gaps and research direction opportunities. The hemp plant will be provided more encouragement to be grown and be used. Many applications of hemp may be pushed to the next level for both producing a green environment and profit. A strong vision and a well-defined plan will pave the path for the discovery of new technologies and concepts.
... The chemical profile of particular metabolites has mainly been studied as a function of the plant's genetics and environment. It stands to reason that the physiological effects and therapeutic benefit of different cannabis strains is linked to the diversity and the quantities of these secondary metabolites [7,8]. ...
... A common method in cannabis cultivation to avoid genetic variations is to grow genetically identical plants from clones. Moreover, by implementing biotechnological tools such as genetic engineering, it is possible to produce plants with overexpressed genes responsible for the biosynthesis of particular bioactive metabolites [7,9]. Environmental conditions such as mineral nutrition, temperature, humidity, soil bacteria, and light intensity/spectra are important factors affecting the chemical composition and secondary metabolism in cannabis plants [10][11][12][13]. ...
Interest in cultivating cannabis for medical and recreational purposes is increasing due to a dramatic shift in cannabis legislation worldwide. Therefore, a comprehensive understanding of the composition of secondary metabolites, cannabinoids, and terpenes grown in different environmental conditions is of primary importance for the medical and recreational use of cannabis. We compared the terpene and cannabinoid profiles using gas/liquid chromatography and mass spectrometry for commercial cannabis from genetically identical plants grown indoors using artificial light and artificially grown media or outdoors grown in living soil and natural sunlight. By analyzing the cannabinoids, we found significant variations in the metabolomic profile of cannabis for the different environments. Overall, for both cultivars, there were significantly greater oxidized and degraded cannabinoids in the indoor-grown samples. Moreover, the outdoor-grown samples had significantly more unusual cannabinoids, such as C4- and C6-THCA. There were also significant differences in the terpene profiles between indoor- and outdoor-grown cannabis. The outdoor samples had a greater preponderance of sesquiterpenes including β-caryophyllene, α-humulene, α-bergamotene, α-guaiene, and germacrene B relative to the indoor samples.
... Other studies [25] found also that β-caryophyllene was the main compound in cannabis. The results confirm that the terpenoids in the plant vary according to numerous parameters including the variety of cannabis, the plant part, the environmental conditions, and the maturity stage of the plant [51,52]. ...
... Moderate water stress, coupled with the use of biostimulants was reported to enhance specialized metabolites like EO components, but also yield [34]. Imposed water-limited stress has led to differential responses of pharmacologically useful diterpenoids for the obtention of the desired composition [35]. Finally, the results confirm that the interfaces in the production and synthesis of fatty acids in plants are influenced by variations in temperature, light, moisture amount, and farming conditions, as reported by [50][51][52][53][54][55]. ...
A field experiment was carried out in Lebanon to assess the agronomic and essential oil characteristics of cannabis as affected by sowing date and irrigation practice. The experiment consisted of a split-plot design with the water regime being the main factor (Iopt-irrigated when the readily available soil water is depleted; I50-receiving 50% of the irrigation amounts in Iopt treatments) and sowing date as the sub-plot factor (mid-April; end of April; mid-May). Biometric and seed quality parameters of the cannabis crop were determined. The essential oils (EO) of the inflorescence were subjected to a multivariate analysis such as principal component analysis (PCA) and hierarchical cluster analysis (HCA). The obtained results revealed that the aboveground fresh biomass, the dry matter, and the plant height were 55.08%, 59.62%, and 43.11% higher in Iopt than in I50, respectively. However, the EO content was neither statistically affected by the irrigation regime nor by the sowing date. Under early sowing, both the water-use efficiency (WUE) for biomass and the EO production reached their highest values. All treatments presented a similar seed composition except that the crude fat and crude protein content were more elevated in Iopt than in I50 treatments. The main extracted essential oils in cannabis inflorescence corresponded to twenty-six identified compounds representing 79.34% of the monoterpenes and 81.25% of the sesquiterpenes. The mon-oterpenes were highly correlated with the irrigation treatment and early-April sowing while the sesquiterpenes were better enhanced under I50 and end of April to mid-May sowing. The study reveals that agronomic practices lead to differential responses of pharmacologically useful plant compounds for improved health benefits. Further research is required to clarify the potential for cannabis cultivation in Lebanon.
... Cannabinoid detection, and primarily THC quantification, is well established in an array of separation techniques, mainly including highperformance liquid chromatography (HPLC), gas chromatography (GC), and thin-layer chromatography (TLC) (Leghissa et al., 2018). These methods may be coupled with either a flame ionization detector (FID), such as for GC-FID, or mass spectral detection methods (MS, tandem MS-MS, TOF, and others) (Fischedick et al., 2010). Chromatographic techniques are vehicles for the separation of mixtures of compounds extracted from any substrate. ...
... Despite these limitations, GC still remains a useful technique for the analysis of cannabis (Hazekamp et al., 2005). Nevertheless, for chemical diversity monitoring in cannabis, a comprehensive method is GC-FID-based metabolomics (Fischedick et al., 2010;Johnson, 2019). ...
Recent studies highlight the therapeutic virtues of cannabidiol (CBD). Furthermore, due to their molecular enriched profiles, cannabis inflorescences are biologically superior to a single cannabinoid for the treatment of various health conditions. Thus, there is flourishing demand for Cannabis sativa varieties containing high levels of CBD. Additionally, legal regulations around the world restrict the cultivation and consumption of tetrahydrocannabinol (THC)-rich cannabis plants for their psychotropic effects. Therefore, the use of cannabis varieties that are high in CBD is permitted as long as their THC content does not exceed a low threshold of 0.3%–0.5%, depending on the jurisdiction. These chemovars are legally termed ‘hemp’. This controlled cannabinoid requirement highlights the need to detect low levels of THC, already in the field. In this review, cannabis profiling and the existing methods used for the detection of cannabinoids are firstly evaluated. Then, selected valuable biosensor technologies are discussed, which suggest portable, rapid, sensitive, reproducible, and reliable methods for on-site identification of cannabinoids levels, mainly THC. Recent cutting-edge techniques of promising potential usage for both cannabis and hemp analysis are identified, as part of the future cultivation and agricultural improvement of this crop.
... In addition, previous studies have suggested that EtOH shows high affinity for phytocannabinoids, with absolute EtOH being the most commonly used organic solvent (Dussy et al., 2005;Bruci et al., 2012;Gunjević et al., 2021). In GC/FID, GC/EI-MS, and GC/MS, a multivariate analysis can also be employed to discriminate a large number of cannabis plant samples and extracts into different chemovars based on analyses of their ethanol extracts (Fischedick et al., 2010;Hazekamp and Fischedick, 2012;Elzinga et al., 2015;Giese et al., 2015;Hazekamp et al., 2016;Ciolino et al., 2018;Béres et al., 2019;Namdar et al., 2019). Consistent with previous research, we also propose the use of EtOH as an extraction solvent for use with cannabis THC, is synthesized in the plastid of the secretory glandular trichomes and exported via an unknown mechanism into the secretory reservoir of glandular trichomes. ...
... As a result, there is a growing market for minor cannabinoids other than D9-THC. Selective breeding techniques have enabled cannabis varieties enriched for minor cannabinoids, 3,4 and minor cannabinoids can be extracted for use as concentrates/isolates. Cannabinol (CBN), produced from the oxidation of D9-THC, is present in higher quantities in aged cannabis. ...
Introduction: Minor cannabinoids are increasingly being consumed in oral formulations (i.e., edibles, tinctures) for medical and nonmedical purposes. This study examined the pharmacokinetics (PKs) of cannabinoids tetrahydrocannabivarin (THCV), cannabichromene (CBC), cannabinol (CBN), and delta-8-tetrahydrocannabinol (D8-THC) after the first and last oral dose during a 14-day administration period.
Materials and Methods: Sprague-Dawley rats (N=6 animals/dose, 50% female) were given an assigned dose of one of four cannabinoids (THCV=3.2–100 mg/kg, CBC=3.2–100 mg/kg, CBN=1–100 mg/kg, or D8-THC=0.32–10 mg/kg) or vehicle (medium-chain triglyceride oil) through oral gavage once daily for 14 days. Blood was collected 45 min and 1.5, 3, and 24 h following the first dose (day 1) and the last dose (day 14) of repeated oral cannabinoid treatment for PK analysis. Outcomes of interest included time to maximum concentration (Tmax), maximum concentration (Cmax), and area under the concentration versus time curve (AUClast). Dose-normalized (DN) Cmax and DN AUClast were also calculated. Brain tissue was collected 24 h post-administration of the first (day 1) and the last (day 14) dose of each cannabinoid to determine concentrations in brain.
Results: All cannabinoids tested were detectable in plasma after single and 14-day repeated dosing. DN Cmax and DN AUClast were highest for D8-THC, followed by CBC, CBN, and THCV. There was no sex difference observed in cannabinoid kinetics. Accumulation of D8-THC in plasma was observed after 14 days of administration. THCV levels in plasma were lower on day 14 compared to day 1, indicating potential adaptation of metabolic pathways and increased drug elimination. Cannabinoids were detected in brain tissue 24 h post-administration of the first and the last dose of 17–100 mg/kg THCV, 3.2–100 mg/kg CBC, 10–100 mg/kg CBN, and 10 mg/kg D8-THC.
Conclusions: THCV, CBC, CBN, and D8-THC produced detectable levels in plasma and translocated to brain tissue after the first dose (day 1) and the last dose (day 14) of repeated oral dosing. Examination of PKs of these minor cannabinoids in blood and brain provides a critical step for informing target dose ranges and dosing schedules in future studies that evaluate the potential effects of these compounds.
... In 2020, Aliferis et al. introduced the term cannabinomics (cannabis metabolomics) for indicating the approaches that go beyond the analysis of the sole major constituents for cannabis research and development [21]. Hazekamp et al., in the early 2010s, pioneered the use of metabolomics-based workflows for discriminating cannabis cultivars and chemovars by focusing on both major and minor phytocannabinoids as well as terpenes by gas chromatography coupled to flame ionization detector (GC-FID) and mass spectrometry (GC-MS) [22][23][24]. Similarly, a GC-FID approach targeted to phytocannabinoids and terpenes was employed by Al Bakain et al. for discriminating the geographical field location of cannabis samples from 23 states of the United States [25]. ...
... In addition, β-elemene is also one of the common sesquiterpenes of several aromatic essential oils extracted from Proteus vulgaris [49]. β-elemene was also found in wild hops from Lithuania at levels up to 14% [50], and in notable amounts in the medical cannabis cultivar 'bedropuur' [51]. The same compound has notable antimicrobial activity against different pathogens, including Mycobacterium tuberculosis, as reported by Sieniawska et al. [52]. ...
The genus Beauveria includes important entomopathogenic and endophytic fungi; among them, Beauveria bassiana is the most studied species. However, there is little knowledge regarding their antimicrobial activity. The current research has been conducted to evaluate the in vitro antagonistic activity of B. bassiana and the antimicrobial efficacy of its Exo and Endo metabolites against Bacillus cereus, B. megaterium, Clavibacter michiganensis (Gram positive bacteria, G+ve), Xanthomonas campestris, Pseudomonas aeruginosa and P. fluorescence (Gram negative bacteria, G−ve). In addition, solid-phase microextraction (SPME) was coupled with Gas Chromatography-Mass Spectrometry (GC/MS) to qualitatively measure the volatile organic compounds’ (VOCs) metabolic profile of the most efficient studied isolate of B. bassiana. The obtained results showed that the isolate UniB2439-3 has a promising antibacterial effect against most of the studied target bacteria. An SPME-GC/MS analysis of VOCs revealed the presence of ethanol, butanal,2-methyl, 2,4-dimethyl-1-heptene, octane, 4-methyl and β-elemene as the dominant bioactive compounds. The results demonstrated that the efficient isolate of B. bassiana can be potentially used as a biocontrol agent against several bacteria, especially G+ve ones.
... The cannabis plant synthesizes a myriad of secondary metabolites, but the most valuable products from a medical and commercial standpoint are phytocannabinoids (Radwan et al. 2009;Fischedick et al. 2010). The most abundant cannabinoids in fresh plant material are 9-tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), and cannabichromenic acid (CBCA). ...
i>Cannabis sativa is one of the oldest cultivated crops, used for its fiber and medicinal properties. The cannabis plant synthesizes a myriad of secondary metabolites, but the most valuable products from a medical and commercial standpoint are cannabinoids. Despite significant advances in elucidating the biochemistry and genetics that govern cannabinoid accumulation, we still do not have conclusive evidence for the role of these secondary metabolites in the physiology of C. sativa . In line with known functions of other secondary metabolites, the protective functions of cannabinoids against temperature stress, poor micronutrient soil content, drought, UV-B radiation, and as anti-microbial agents have been suggested, but are yet to be conclusively demonstrated. Recent research suggests that the environment has a major effect on cannabis growth and productivity, but the relationship between stress, cannabinoid accumulation, and plant health is complex. Here, we summarize the current insights on how abiotic and biotic stress affect C. sativa biology. We also examine the available evidence to support the hypothesis for the protective function of cannabinoids against environmental stressors. Maintaining optimal growth and high cannabinoid synthesis is a balancing act, one that can only be achieved by better understanding of the effects on the environment on the cannabis plant.
... In addition, β-elemene is also one of the common constituents of several aromatic essential oils extracted from Proteus vulgaris [41]. β-elemene was also found in wild hops from Lithuania at levels up to 14% [42] and in notable amounts in the medical cannabis cultivar 'bedropuur' [43]. The same compound has notable antimicrobial activity against different pathogens including Mycobacterium tuberculosis as reported by Sieniawska et al. [44]. ...
The genus Beauveria include important entomopathogenic and endophytic fungi, among them, Beauveria bassiana is the most studied species. However, there are few knowledge regarding their antimicrobial activity. The current research has been conducted to evaluate in vitro antagonistic activity of B. bassiana and the antimicrobial efficacy of its Exo and Endo metabolites against Bacillus cereus, B. megaterium, Clavibacter michiganensis (gram positive bacteria, G+ve), Xanthomonas campestris, Pseudomonas aeruginosa and P. fluorescence (gram negative bacteria, G-ve). In addition, Solid-phase microextraction (SPME) was coupled to Gas Chromatography-Mass Spectrometry (GC/MS) to qualitatively measure the volatile organic compounds (VOCs) metabolic profile of the most efficient studied isolate of B. bassiana. The obtained results showed that, the isolate UniB2439-3 has promising antibacterial effect against most of studied target bacteria. SPME-GC/MS analysis of VOCs revealed the presence of ethanol; butanal,2-methyl; 2,4-dimethyl-1-heptene; octane, 4-methyl and β-elemene as the main dominant bioactive compounds. The outgoing results explicated that the efficient isolate of B. bassiana can be potentially used as a biocontrol agent against several bacteria especially G+ve ones.
... Moreover, monoterpenoids can help to distinguish accessions containing similar cannabinoid and sesquiterpenoid profiles. The cannabinoid and terpenoid concentrations are reproducible for cannabis clones grown at separate times under standardized environmental conditions [21]. ...
With the increasing accessibility of cannabis (Cannabis sativa L., also known as marijuana and hemp), its products are being developed as extracts for both recreational and therapeutic use. This has led to increased scrutiny by regulatory bodies, who aim to understand and regulate the complex chemistry of these products to ensure their safety and efficacy. Regulators use targeted analyses to track the concentration of key bioactive metabolites and potentially harmful contaminants, such as metals and other impurities. However, the metabolic complexity of cannabis metabolic pathways requires a more comprehensive approach. A non-targeted metabolomic analysis of cannabis products is necessary to generate data that can be used to determine their authenticity and efficacy. An authentomics approach, which involves combining the non-targeted analysis of new samples with big data comparisons to authenticated historic datasets, provides a robust method for verifying the quality of cannabis products. To meet International Organization for Standardization (ISO) standards, it is necessary to implement the authentomics platform technology and build an integrated database of cannabis analytical results. This study is the first to review the topic of the authentomics of cannabis and its potential to meet ISO standards.
... In addition, β-elemene is also one of the common constituents of several aromatic essential oils extracted from Proteus vulgaris [44]. β-elemene was also found in wild hops from Lithuania at levels up to 14% [45] and in notable amounts in the medical cannabis cultivar 'bedropuur' [46]. The same compound has notable antimicrobial activity against different pathogens including Mycobacterium tuberculosis as reported by Sieniawska et al. [47]. ...
The genus Beauveria include important entomopathogenic and endophytic fungi, among them, B. bassiana is the most studied species. However, there are few knowledge regarding their antimicrobial activity. The current research has been conducted to evaluate in vitro antibacterial efficacy of five isolates of B. bassiana against Bacillus cereus, B. megaterium, B. mojavensis, Clavibacter michiganensis (gram positive bacteria, G+ve), Xanthomonas campestris, X. vesicatoria, Escherichia coli, Pseudomonas aeruginosa and P. fluorescence (gram negative bacteria, G-ve). In addition, chemical composition of the principal diffusible metabolites and volatile organic compounds (VOCs) of the most efficient studied isolate of B. bassiana has been carried out using GC-MS analysis. The obtained results showed that, the isolate UniB2439-3 has promising antibacterial effect against most of studied target bacteria. GC-MS analysis of diffusible metabolites detected the presence of hexanedioic acid, bis(2-ethylhexyl) ester as the main compound in the cell-free culture filtrate. Furthermore, GC-MS analysis of VOCs revealed the presence of ethanol; butanal,2-methyl; 2,4-dimethyl-1-heptene; octane, 4-methyl and β-elemene as the main dominant bioactive compounds. The outgoing results explicated that, the isolates of B. bassiana have promising antibacterial activity which could be correlated to their diffusible and VOCs metabolites. Therefore, the selected isolate can be potentially used as a biocontrol agent against several bacteria especially G+ve ones. Taking in consideration that the antibiotics are forbidden in agriculture in many countries worldwide, search for possible natural alternatives as efficient antimicrobial agents are highly interesting.
... Additionally, cannabinoids cause cell death in plant tissue culture (Sirikantaramas et al., 2005). Substantial energy and carbon is devoted to cannabinoid biosynthesis; leaves and flowers are up to 6 and 20% cannabinoid, respectively (Pacifico et al., 2008) and unpollinated pistillate flowers are up to 20% cannabinoid by dry weight (Fischedick et al., 2010). The abundance and phytotoxicity of hemp secondary metabolites led us to ask if these chemicals could be allelopathic, defined as causing physiological inhibition of other plants through chemical release Duke, 2015). ...
Evolved resistance to herbicides necessitates alternative weed control strategies. Allelopathic crops show promise as an alternative to exogenous herbicides and could be used to reduce establishment, growth, and reproduction of weeds. Individual cannabinoids and terpenes found in hemp ( Cannabis sativa L.) show allelopathic potential, but allelopathic effects of plant residue have not been characterized. A laboratory assay found that crude, acetone-soluble hemp extracts diluted to 2.5 mg mL ⁻¹ reduced the germination of a bioindicator species. However, tested concentrations below 2.5 mg mL ⁻¹ were not more effective than the no-extract control treatment at reducing germination. A greenhouse study found that soil-incorporated hemp residue was not more effective than a maize ( Zea mays L.) residue treatment comparison in reducing plant growth. However, a simulated chaff line experiment in the greenhouse showed that the equivalent of 378 g m ⁻² hemp residue on the soil surface can effectively reduce and delay the germination of waterhemp [ Amaranthus tuberculatus (Moq.) Sauer] in comparison to bare-soil, or a maize residue treatment comparison. Together, these results show that incorporated hemp residue will likely not be an effective weed control practice. However, chaff lining hemp residue may be an effective practice for the control of certain weeds and warrants further research in a field setting.
... Terpene profile is a unique characteristic, that can be even used as chemotaxonomic markers, for determination and identification of different varieties (Fischedick et al., 2010). ...
... To conclude, the applied method was sensitive and suitable for the qualification of a broad range of Cannabis samples. It was already mentioned that in the literature cannabis quantification is commonly carried out by GC-FID (Borges et al., 2020;Fischedick et al., 2010;Ibrahim et al., 2017). However, GC-MS can also be applied as a quantification tool but expensive deuterated standards are necessary which are not commercially available for all cannabinoids (Citti et al., 2018). ...
Cannabis sativa L. is undoubtedly the most used recreational drug worldwide because of its desired acute psychotropic effects, like relaxation, euphoria and altered perceptions. In addition, promising medical properties of Cannabis components have gained a lot of attention, resulting in a debate to permit recreational Cannabis use in several countries. In recent years, this controversial plant was increasingly studied and a large number of scientific papers were published. Herbal Cannabis consists of a variable and complex matrix, which makes it challenging to properly seize and prepare the sample for qualitative and quantitative analysis. Moreover, both the adoption of legal cut‐off values in different countries for the Δ9‐tetrahydrocannabinol (THC) content in seizures, and the emergence of cannabidiol (CBD) based products, containing generally small but variable amounts of THC, urged the need for sensitive and reliable analytical techniques to accurately identify and quantify the components of interest. This review presents detailed information on the procedure prior to analysis and covers chromatographic and spectroscopic methods developed for the analysis of cannabinoids in seizures for different forensic purposes, that is, identification/quantification, potency testing, drug‐ and fiber‐type differentiation, age estimation, yield determination and Cannabis profiling. Advantages and drawbacks of existing methods, within a specific forensic context, are discussed. The application of chemometrics, which offers a powerful tool in interpreting complex data, is also explained. This article is categorized under: Toxicology > Cannabis Toxicology > Drug Analysis Forensic Chemistry and Trace Evidence > Presentation and Evaluation of Forensic Science Output Cannabis sativa L. is the most used and seized recreational drug worldwide. For forensic institutes/laboratories, it is important to obtain reliable and reproducible data about seized samples, as it is used in judicial investigations. A thorough consideration about the sampling, sample preparation, instrumental analysis and subsequent data handling is needed, and depends on the forensic purpose. Moreover, chemometrics, which is already applied in certain herbal cannabis studies, will become an important tool in forensics to interpret large and complex data.
... Table 1 illustrates the concentration range (mg/g) of terpenes and terpene derivatives reported in published research articles investigating the terpene content of specific cannabis chemotypes. Chemical profiles of common cannabis cultivars continue to show that myrcene, β-caryophyllene, limonene, α-terpinene, and α-pinene are the most prominent terpenes that can be found in the first three chemotype varieties [34,[45][46][47][48]. Terpene profiles of the remaining chemotypes are limited or have yet to be investigated. ...
Cannabis is a complex biosynthetic plant, with a long history of medicinal use. While cannabinoids have received the majority of the attention for their psychoactive and pharmacological activities, cannabis produces a diverse array of phytochemicals, such as terpenes. These compounds are known to play a role in the aroma and flavor of cannabis but are potent biologically active molecules that exert effects on infectious as well as chronic diseases. Furthermore, terpenes have the potential to play important roles, such as synergistic and/or entourage compounds that modulate the activity of the cannabinoids. This review highlights the diversity and bioactivities of terpenes in cannabis, especially minor or secondary terpenes that are less concentrated in cannabis on a by-mass basis. We also explore the question of the entourage effect in cannabis, which studies to date have supported or refuted the concept of synergy in cannabis, and where synergy experimentation is headed, to better understand the interplay between phytochemicals within Cannabis sativa L.
... The use of Cannabis sativa has recently received more attention: its main components, ∆9-tetrahydrocannabinol (∆9-THC), with psychotropic activity, and cannabidiol (CBD), non-psychoactive [1], can be used as symptomatic supportive treatment in many diseases, i.e., to reduce convulsions in cases of childhood and/or adolescent epilepsy [2], to manage neuropathic pain [3] and counteract the side effects of chemotherapy [4]. Other than two main components, up to 125 other structurally related compounds, called cannabinoids, [5] have been extracted and characterized to date [6]. Since the synergies existing in the phytocomplex can increase the activity of ∆9-THC and limit side effects, medicinal products should be based on the natural chemical complexity of Cannabis, rather than using a single [7] or synthetic cannabinoid [8]. ...
Over the past 20 years, the interest in Cannabis oily extracts for medicinal use compounded in pharmacy has consistently grown, along with the need to have preparations of adequate quality. Hot maceration (M) is the most frequently used method to compound oily solutions. In this work, we systematically studied the possibility of using an ultrasonic homogenizer and a sonotrode (US) as an alternative extraction method. Oily solutions were prepared using two available varieties of Cannabis for medicinal use, called FM2 and Bedrocan. All preparations resulted with an equivalent content in CBD and THC, with the advantage of a faster process using US. In particular, 10 min sonication at the amplitude optimized for the sonotrode used (2 or 7 mm) provides not statistically different total Δ9-tetrahydrocannabinol (M-FM2: 0.26 ± 0.02 % w/w; US-FM2: 0.19 ± 0.004 % w/w; M-Bedrocan: 1.83 ± 0.17 % w/w; US-Bedrocan: 1.98 ± 0.01 % w/w) and total cannabidiol (M-FM2: 0.59 ± 0.04 % w/w; US-FM2: 0.58 ± 0.01 % w/w) amounts extracted in refined olive oil. It can therefore be confirmed that sonotrode is an efficient and fast extraction technique and its use is without negative consequence on the solvent properties. Despite DSC evidencing that both maceration and sonication modify the Tonset and enthalpy of the event at about −10 °C, the qualitative characteristics of the oil remained constant for the two treatments and similar to the starting material.
... Terpenes contribute to the aroma (Andre et al., 2016;Brenneisen, 2007;Giese et al., 2015;Small, 2015). Geranyl pyrophosphate is the precursor for synthesising the terpenoids, leading to the creation of monoterpenoids in secretory cell plastids (Loza-Tavera, 1999;Bertoli et al., 2010), sesquiterpenoids and triterpenoids in the cytoplasm (Russo, 2011;Bertoli et al., 2010;Dewick, 2002;Fischedick et al., 2010). Other compounds isolated from the hemp include phenylpropanoids (Flores-Sanchez & Verpoorte, 2008;Docimo et al., 2013). ...
Edible hemp products or superfood refers to Cannabis sativa or industrial hemp. In general, hemp is a rich source of functional metabolites, such as Tetrahydrocannabinol (THC), Cannabidiol (CBD), and other cannabinoids. Hemp has been widely used in food products, such as bread, cookies, meatballs, energy bars, cooking oil, snacks, and crackers. Hemp has been used for development of cosmetics and supplements. However, the use of hemp is far below its potential because of major challenges such as non‐cost‐effective extraction and isolation, stability and toxicity of the extracts, and legislation related to the use of the extracts. This narrative review comprehensively analyses major phytochemicals in hemp and hemp extracts, and also discusses the most common challenges in applications of hemp derived phytochemicals and hemp extracts in food and pharmaceutical products like stability, toxicity, legal limitations, isolation/extraction, and purification. In addition, this review outlines current applications of hemp extracts and proposes future trends for utilising hemp phytochemicals and extracts in food, cosmetic and pharmaceutical products to increase applications of hemp extracts and their phytochemicals for health benefits.
... As hemp varieties become legal worldwide, their economic value as an agricultural product is developing into a multi-billion-dollar industry [50]. Although breeding of individuals for high CBD production has not been actioned in Korea yet, many varieties are emerging abroad [51][52][53][54][55]. ...
Cannabis breeders are combining several genes to develop economically valuable fiber, seed, and medicinal hemp. This study analyzed the characteristics and selection of traits based on cannabidiol production of medicinal cannabis lines successfully grown under artificial light and nutrient solution cultivation conditions in smart farm conditions. Sixteen female plants were selected by seeding medical hemp F1 hybrid specimens obtained by randomly crossing Cherry Wine and native hemp from each country. The F1 generation was treated with 12 h light to induce flower differentiation. CBD production peaked on day 50 of the treatment, and this was selected as the harvesting day. All F1 hybrids were separated by leaf and inflorescence after collecting morphological data, and fresh and dry weights were measured. The CBD production of leaf and inflorescence per cubic meter was calculated. The CW21-5 line produced a total of 53.002 ± 0.228 g of CBD per cubic meter, the highest CBD producer. In addition, heatmap correlation analysis showed that most morphological data were not related to cannabinoid content. Principal Component Analysis (PCA) and Self-Organizing Map (SOM) analysis showed that CW21-5 is an arbitrary line that does not cluster with other lines, and the reason for its excellent CBD yield per cubic meter is that it has a narrow plant diameter and a high CBD content at the same time.
... Tobacco and cannabis smoke contains different organic and inorganic chemical compounds that could have adverse effects to cannabis smokers (Lehrer and Rheinstein, 2022). The most used cannabis plant, Cannabis sativa, reportedly contains more than 60 different phytocannabinoids (Fischedick et al., 2010). ...
Objectives
This study aims to investigate the effects of cannabis smoke condensate (CSC) on the adhesion, growth, and signaling pathways of human gingival epithelial cells.
Design
The effects of CSC on cell shape and adhesion, and viability were evaluated after 30 min, 60 min, 2 h, and 24 h of exposure using microscopic observation, cell metabolic activity, and lactate dehydrogenase activity assays. The effects of CSC on cell apoptosis, necrosis, autophagy, and oxidative stress were determined through flow cytometry, while apoptotic and autophagic gene expression were identified via an RT²-PCR array. Phosphorylated signaling pathway proteins were measured using flow cytometry.
Results
CSC deregulated gingival epithelial cell shape and adhesion, decreased cell viability, and increased lactate dehydrogenase release. Its toxic effects included apoptosis, autophagy, and oxidative stress. Moreover, it modulated seven specific apoptotic and six autophagic genes. Furthermore, it decreased phosphorylation in signaling proteins, such as STAT5, ERK12, P38, and nuclear factor κB.
Conclusions
CSC has notable adverse effects on gingival epithelial cells. This finding indicates that cannabis smoke could impair gingival epithelial cell innate immune function, leading to gingivitis and periodontitis. Oral health professionals may need to document observed modifications in the oral cavity of patients who smoke cannabis and consider these potential changes during clinical care.
... June 2022 | Volume 13 | Article 906038 (Russo 2011;Radwan 2021). Typically found in cannabis trichromes at about 10%, and in flowers at about 1%, some chemotypes have been bred to contain as much as 3.5% terpene content or higher in the flowers (Potter, 2009;Fischedick et al., 2010). Although relatively modest in concentration to the cannabinoids, terpenes (mainly mono and sesquiterpenes) are another important class of compounds. ...
“Hemp” refers to non-intoxicating, low delta-9 tetrahydrocannabinol (Δ9-THC) cultivars of Cannabis sativa L. “Marijuana” refers to cultivars with high levels of Δ9-THC, the primary psychoactive cannabinoid found in the plant and a federally controlled substance used for both recreational and therapeutic purposes. Although marijuana and hemp belong to the same genus and species, they differ in terms of chemical and genetic composition, production practices, product uses, and regulatory status. Hemp seed and hemp seed oil have been shown to have valuable nutritional capacity. Cannabidiol (CBD), a non-intoxicating phytocannabinoid with a wide therapeutic index and acceptable side effect profile, has demonstrated high medicinal potential in some conditions. Several countries and states have facilitated the use of THC-dominant medical cannabis for certain conditions, while other countries continue to ban all forms of cannabis regardless of cannabinoid profile or low psychoactive potential. Today, differentiating between hemp and marijuana in the laboratory is no longer a difficult process. Certain thin layer chromatography (TLC) methods can rapidly screen for cannabinoids, and several gas and liquid chromatography techniques have been developed for precise quantification of phytocannabinoids in plant extracts and biological samples. Geographic regulations and testing guidelines for cannabis continue to evolve. As they are improved and clarified, we can better employ the appropriate applications of this uniquely versatile plant from an informed scientific perspective.
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.
Cannabinoids (CBDs) represent a group of C21 or C22 terpenophenolic compounds predominantly produced by Cannabis but have also been found in plants from the Radula and Helichrysum genera. There are about 100 different cannabinoids, although some of them are metabolites. They are generally classified into ten subclasses [1–3].
Hemp (Cannabis sativa L.) is a globally distributed plant with a complex phytochemistry. Secondary metabolism is extremely rich. Secondary metabolites are substances whose role is the interaction of the plant with the environment. Their synthesis depends on the presence of stress and is very energy-consuming. They protect plants from pests and pathogens, as well as from abiotic factors. Cannabinoids, terpenoids and flavonoids are the predominant substances concentrated in the inflorescences of female plants. Their ecological role and function are not yet fully understood. The cannabinoids Δ-9-trans-tetrahydrocannabinol (Δ9-THC), cannabidiolic acid (CBDA), and others probably act as antioxidants, and based on their spatially isolated synthesis and cytotoxic by-products, they are thought to be involved in the plant’s defense system. Under stress, their content increases and the chemical profile changes. Stress activates stress signalling molecules - jasmonic acid and its derivatives, which affect the release of terpenoids, and increases the expression of genes in the phenylpropanoid pathway, which is crucial for the synthesis of flavonoids. Secondary metabolites act synergistically and in conjunction with the microbiome - endophytic bacteria and fungi - have a negative effect on harmful organisms and protect the plant.
Industrial hemp, Cannabis sativa L., production has been negatively impacted by larvae of corn earworm, Helicoverpa zea (Boddie), which feed on developing inflorescences. Adult H. zea oviposit on hemp once flowers develop, and late-instar larvae can cause serious loss to both quality and yield. A 2-year study to assess the influence of hemp variety and fertilization practices on damage caused by H. zea was conducted. Differences in damage ratings among varieties were observed in both years; however, the rate of nitrogen applied did not influence biomass yield or damage rating. These results indicate that increasing nitrogen fertility may not be an effective means of cultural control for mitigating damage from H. zea. Floral maturity was very influential on damage caused by H. zea as late-maturing varieties had much less floral injury than those which matured early in outdoor field trials. Some cannabinoids were also correlated to damage rating, but this relationship was due to late-maturing plants with immature flowers low in cannabinoid concentrations receiving less floral injury. Based on these results, the selection of high-yielding varieties that flower when ovipositional activity of H. zea is expected to decline should be the first step in an integrated pest management program for hemp production. This research expanded our knowledge of the role of fertility rate, varietal characteristics, cannabinoid profile, and floral maturity on damage caused by H. zea to hemp. Findings from this research will allow growers to make more informed agronomic decisions before planting to improve hemp production.
Cannabis sativa L. is an ancient crop used for fiber and seed production and not least for its content of cannabinoids used for medicine and as an intoxicant drug. Due to the psychedelic effect of one of the compounds, tetrahydrocannabinol (THC), many countries had regulations or bands on Cannabis growing, also as fiber or seed crop. Recently, as many of these regulations are getting less tight, the interest for the many uses of this crop is increasing. Cannabis is dioecious and highly heterogenic, making traditional breeding costly and time consuming. Further, it might be difficult to introduce new traits without changing the cannabinoid profile. Genome editing using new breeding techniques might solve these problems. The successful use of genome editing requires sequence information on suitable target genes, a genome editing tool to be introduced into plant tissue and the ability to regenerate plants from transformed cells. This review summarizes the current status of Cannabis breeding, uncovers potentials and challenges of Cannabis in an era of new breeding techniques and finally suggests future focus areas that may help to improve our overall understanding of Cannabis and realize the potentials of the plant.
Fossil fuel supplies are becoming scarce as a result of the growing world’s population. The increasing use of fossil fuels also pose a threat to ecosystem. Renewable energy sources should be revived in order to meet future energy demands, and minimize the adverse effects on environment. Biofuels and biochemicals can be made from a wide variety of lignocellulosic biomass, which has been studied for decades. Several useful bioproducts can now be made from hemp's lignocellulosic biomass (cellulose, hemicellulose, lignin, protein, and others). Hemp has considerable commercial potential as it can be used in biorefinery applications for the production of bioethanol, biodiesel, biohydrogen, biogas, organic acids, biomaterials, bio-oil and various pharmaceutical and nutraceuticals compounds. Effective conversion of lignocellulosic biomass to value-added bioproducts for biorefinery and other potential applications relies on a wide range of pretreatment processes. This article aims to highlight recent developments in hemp lignocellulosic conversion technologies for biorefinery applications and other value-added products development with the goal of bolstering future industries in a manner that is more environmentally friendly and secure.
Background:
Hemp and marijuana are the two major varieties of Cannabis sativa. While both contain Δ9-tetrahydrocannabinol (THC), the primary psychoactive component of C. sativa, they differ in the amount of THC that they contain. Presently, U.S. federal laws stipulate that C. sativa containing greater than 0.3% THC is classified as marijuana, while plant material that contains less than or equal to 0.3% THC is hemp. Current methods to determine THC content are chromatography-based, which requires extensive sample preparation to render the materials into extracts suitable for sample injection, for complete separation and differentiation of THC from all other analytes present. This can create problems for forensic laboratories due to the increased workload associated with the need to analyze and quantify THC in all C. sativa materials.
Method:
The work presented herein combines direct analysis in real time-high-resolution mass spectrometry (DART-HRMS) and advanced chemometrics to differentiate hemp and marijuana plant materials. Samples were obtained from several sources (e.g., commercial vendors, DEA-registered suppliers, and the recreational Cannabis market). DART-HRMS enabled the interrogation of plant materials with no sample pretreatment. Advanced multivariate data analysis approaches, including random forest and principal component analysis (PCA), were used to optimally differentiate these two varieties with a high level of accuracy.
Results:
When PCA was applied to the hemp and marijuana data, distinct clustering that enabled their differentiation was observed. Furthermore, within the marijuana class, subclusters between recreational and DEA-supplied marijuana samples were observed. A separate investigation using the silhouette width index to determine the optimal number of clusters for the marijuana and hemp data revealed this number to be two. Internal validation of the model using random forest demonstrated an accuracy of 98%, while external validation samples were classified with 100% accuracy.
Discussion:
The results show that the developed approach would significantly aid in the analysis and differentiation of C. sativa plant materials prior to launching painstaking confirmatory testing using chromatography. However, to maintain and/or enhance the accuracy of the prediction model and keep it from becoming outdated, it will be necessary to continue to expand it to include mass spectral data representative of emerging hemp and marijuana strains/cultivars.
Cannabis sativa is a flowering plant belonging to the Cannabinaceae family. There have been numerous ongoing studies globally on C. sativa which include investigating the yield and quality of essential oils. However, this study is the first to assess the chemical composition and biological activities of Jamaican C. sativa essential oils (EOs) extracted via hydrodistillation as the plants mature from week 8 through to week 12. EOs from fresh landrace female plants were analysed by gas chromatography–mass spectrometry (GC–MS) and GC coupled to a flame ionization detector (GC‐FID). Cannabis sativa essential oils were screened for their in vitro antimicrobial activity using the disc diffusion method and further with tube dilution method. The antioxidant properties were investigated using the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical scavenging assay. In total, 36 constituents were identified in the C. sativa EOs by comparison of the Kováts retention indices (RI), the mass spectra data with those in the Natural Institute of Standards and Technology (NIST) library and by co‐elution with authentic samples where available. The major constituents were β‐caryophyllene (25.34%) and α‐humulene (α‐caryophyllene) (10.94%) both at the highest levels in week nine and were the most abundant constituents present at all stages of maturity. All extracted C. sativa essential oils did not exhibit antioxidant properties. However, the C. sativa essential oils exhibit significant to moderate antimicrobial property against the test gram‐positive microorganisms, Enterococcus faecalis, Streptococcus Group A and Streptococcus Group B with MIC values ranging from 16 to 125 μg/mL. Jamaican Cannabis sativa essential oils extracted via hydrodistillation as the plants mature from week 8 through to week 12 were assessed for its chemical composition and biological activities. A total of 36 compounds were identified with the major constituents being β‐caryophyllene (25.34%) and α‐humulene (10.94%) both at highest level in week nine and were the most abundant constituents present at all stages of maturity. The oils showed antimicrobial against three gram‐positive micro‐organisms assayed, however no significant antioxidant activities were observed.
In 2019, an air emissions field sampling study was conducted by the Colorado Department of Public Health and Environment's Air Pollution Control Division (APCD) at four commercial cannabis cultivation facilities. Measurements of ambient biogenic volatile organic compounds (VOC) concentrations were collected from various growing stages of cannabis (vegetative and flowering) and during post-harvest activities (drying and trimming). This data was then used to determine room-specific biogenic VOC emission rates for three of the facilities from the vegetative stage of the life cycle through post-harvest activities. This study shows that the magnitude of biogenic VOC emissions within a cannabis cultivation facility varies widely with the highest emission rates of up to 7.18E-1 kg/hr found during mechanical trimming and up to 2.33E-1 kg/hr in the drying rooms. These were up to an order of magnitude higher than emission rates found in the cultivation rooms. For example, Facility A vegetative room had an emissions rate of 1.46E-2 kg/hr. Normalized by the amount of biomass present, the drying rooms had the highest VOC emissions rates, with a maximum rate of 1.6E-3 kg/hr/kg biomass. The flowering room rates were found to be up to 3.25E-4 kg/hr/kg biomass and drying rooms up to 1.16E-3 kg/hr/kg biomass. When normalized by plant count, emission rates in the flower rooms ranged from 8.11E-6 to 3.62E-4 kg/hr/plant. The dominant monoterpenes from sampling were β-myrcene, terpinolene, and D-limonene. These data suggest that the variability in emission rates across cannabis production will create a challenge in establishing a generalized emission factor for all facilities. Across the industry, cannabis cultivation conditions and strategies can vary widely impacting the amount and type of VOC emissions. Minimizing uncertainties for VOC emission from cannabis facilities requires site specific information on air exchange rates, plant counts, cannabis strains, biomass, and if hand or mechanical processing is used.Implications This study found that the magnitude of biogenic VOC emissions within a cannabis cultivation varies widely throughout rooms found in the facility, with the highest emissions found during post-harvest activities (i.e. trimming) and the lowest rates in the vegetative room. These data suggest that the large emission sources of VOCs are found post-harvest and emission inventories based solely on cultivation emissions will underestimate total biogenic VOC emissions from indoor cannabis cultivation facilities. The dominant measured terpenes throughout all facilities from cultivation to post harvest were: β-myrcene, terpinolene, and D-limonene.
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Absolute content of terpenes in inflorescences of two strains of Cannabis sativa L., CAT 1 and CAT 3, has been determined. Twenty terpenes commonly present in these samples were quantified by solid phase microextraction combined with gas chromatography and flame ionization detection (SMPE/GC-FID). High amounts of β-myrcene, α-pinene, β-pinene, limonene, (E)-β-ocimene, β-caryophyllene, α-humulene, (E)-nerolidol, and linalool, were found in both strains. Lower concentrations (< 20 µg·g⁻¹) of other terpenes were also determined. Only (E)-β-ocimene was detected at 50 µg·g⁻¹ in CAT 3 whereas it was below the LOD in CAT 1. Concentrations of other compounds for which standards were not available, were estimated based on a response factor obtained from the calibration curves of compounds with similar chemical structures. Fingerprints of both CAT strains were obtained and the identities of most volatile compounds were assigned using gas chromatography coupled to mass spectrometer detector (GC-MS).
Additionally, an assessment of variability of terpenes was achieved by analyzing ten plants of each strain grown under controlled conditions and harvested at the same time. This variability was about 20 %, considering terpenes at concentration above 20 µg·g ⁻¹.
Cannabis sativa L. has been used as medicine for thousands of years. Since the early identification of tetrahydrocannabinol (THC) in 1960, pharmacological activities were attributed to a group of unique structures named cannabinoids. For decades, research and development were applied to determine different cannabinoids and their medicinal properties. Nowadays there is evidence that the therapeutic benefits of the plant are based on the synergy of cannabinoids and other secondary metabolites such as terpenes and flavonoids. Differences between the medical performance of isolated compounds like cannabidiol (CBD) or THC and full-spectrum plant extracts are notable. Indeed, the superiority of the last one is provoked by the synergy between various different compounds. This improved medicinal effect is called the entourage effect. Chromatography has become the method of choice for the determination of cannabinoids, terpenes, and flavonoids, so it represents an excellent tool for a proper characterization of the plant and plant derived products. The objective of characterization relies not only in analyzing the fingerprint of cannabis, but also to identify different chemotypes for medical purposes. To understand the contributions of each natural product to this “entourage effect”, this review presents an in-depth analysis of the utilization of High-performance liquid chromatography (HPLC), Gas chromatography (GC) and other methods for the analysis of phytocomponents of Cannabis sativa L. In this sense, a representative number of examples and advances made in the field together with limitations and future needs are provided. It can be concluded that standardized protocols and quality control policies and procedures are necessary for the comprehensive analysis of cannabis extracts and derivatives.
Design of Experiments (DoE) is a well-established tool used for analytical methods robustness studies, because of its ability to assess the effect of a great number of factors in a minimal number of experiments. However, when assessing the robustness of an analytical method the analysis of the individual effect of each factor is not sufficient on its own. Some factors may not influence the robustness of the method, but their effect combined with the effects of other factors may have a significant contribution on the robustness of the method, which is not given by conventional analysis of DoE results. The aim of this work is to propose, in addition to the analysis of the individual effects of the factors, to estimate the joint effect of the factors by means of the matrix experimental results prediction interval. This prediction interval is the interval in which, with a given probability, should fall the next results, therefore it is an interesting tool to estimate the variation limits of the method results during routine use. We also propose the use of two other prediction intervals which can help to analyze the DoE results and give a conclusion on the method robustness. The first one is based on the DoE experimental error information, and it gives an estimation of the experimental error component impact on the factors joint effect. The second one is based on the factors non-significance limits, and it provides the information regarding the factors impact on the responses in the case where the conditions are, by definition, robust. We applied these proposals to the robustness study of a UHPLC method for the separation of phytocannabinoids and we could demonstrate that, in addition to the calculated effects values and robustness information, the use of the prediction intervals information provided additional information that allowed a better interpretation of the method performance parameters.
The need to find a rapid and worthwhile technique for the in situ detection of the content of delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) in Cannabis sativa L. is an ever-increasing problem in the forensic field. Among all the techniques for the detection of cannabinoids, Raman spectroscopy can be identified as the most cost-effective, fast, noninvasive, and nondestructive. In this study, 42 different samples were analyzed using Raman spectroscopy with 1064 nm excitation wavelength. The use of an IR wavelength laser showed the possibility to clearly identify THC and CBD in fresh samples, without any further processing, knocking out the contribution of the fluorescence generated by visible and near-IR sources. The results allow assigning all the Raman features in THC- and CBD-rich natural samples. The multivariate analysis underlines the high reproducibility of the spectra and the possibility to distinguish immediately the Raman spectra of the two cannabinoid species. Furthermore, the ratio between the Raman bands at 1295/1440 and 1623/1663 cm-1 is identified as an immediate test parameter to evaluate the THC content in the samples.
Cannabis sativa (hemp) as multifunctional crop have traditional application as fiber, food, paper, textile and pharmaceutical potential as inflorescences and seed as sources of exciting bioactive secondary metabolites. The Genus Cannabis is the only producer of phytocannabinoids. Extensive studied have been made to describe the origin history, geographical ranges and genetic identity of the Cannabis species but it remains obscured to date. Various high through put genetic marker have been studied to assess the genetic diversity in hemp varieties. Studies also indicated that domestication origin affects the genetic groups of hemp which further consequences on the chemical diversity of the cannabis. Chemotaxonomy using chemical markers also played a crucial role in differencing and allocating the Cannabis taxa. Cannabinoids ratio and terpene composition are the major marker to play an important role in studying chemical diversity of Cannabis sp. Cannabis genus is the only source of phytocannabinoids the dominant chemical class. Other than cannabinoids terpene and non-cannabinoid phenolic compounds also contribute in the chemical diversity of the species. The vast array of phytochemicals presents in the genus have potential application in pharmaceutical industries. However, due to its legalization status very limited study on its chemical and genetic diversity have been done. Therefore, the species needs attention to explore its commercial value.Keywords
Cannabis sativa
HempPhytocannabinoidsGenetic diversityChemotaxonomy
To date, a large number of controlled clinical trials have been done evaluating the therapeutic ap- plications of cannabis and cannabis-based preparations. In 2006, an excellent review was pub- lished, discussing the clinical trials performed in the period 1975 to June 2005 (Ben Amar 2006). The current review reports on the more recent clinical data available. A systematic search was per- formed in the scientific database of PubMed, focused on clinical studies that were randomized, (double) blinded, and placebo-controlled. The period screened was from July 1, 2005 up to August 1, 2009. The key words used were: cannabis, marijuana, marihuana, hashish, cannabinoid(s), tetrahydro- cannabinol, THC, CBD, dronabinol, Marinol, nabilone, Cannador and Sativex. For the final selec- tion, only properly controlled clinical trials were retained. Open-label studies were excluded, ex- cept if they were a direct continuation of a study discussed here. Thirty-seven controlled studies evaluating the therapeutic effects of cannabinoids were identified. For each clinical trial, the country where the project was held, the number of patients assessed, the type of study and comparisons done, the products and the dosages used, their efficacy and their adverse effects are described. Based on the clinical results, cannabinoids present an interesting therapeutic potential mainly as analgesics in chronic neuropathic pain, appetite stimulants in de- bilitating diseases (cancer and AIDS), as well as in the treatment of multiple sclerosis.
A simple method is presented for the preparative isolation of seven major cannabinoids from Cannabis sativa plant material. Separation was performed by centrifugal partition chromatography (CPC), a technique that permits large‐scale preparative isolations. Using only two different solvent systems, it was possible to obtain pure samples of the cannabinoids; (−)‐Δ‐(trans)‐tetrahydrocannabinol (Δ‐THC), cannabidiol (CBD), cannabinol (CBN), cannabigerol (CBG), (−)‐Δ‐(trans)‐tetrahydrocannabinolic acid‐A (THCA), cannabigerolic acid (CBGA), and cannabidiolic acid (CBDA). A drug‐type and a fiber‐type cannabis cultivar were used for the isolation. All isolates were shown to be more than 90% pure by gas chromatography. This method makes acidic cannabinoids available on a large scale for biological testing. The method described in this report can also be used to isolate additional cannabinoids from cannabis plant material.
Since 2003 medicinal grade cannabis is provided in the Netherlands on prescription through phar- macies. Growing, processing and packaging of the plant material are performed according to pharmaceutical standards and are supervised by the official Office of Medicinal Cannabis (OMC). The quality is guaranteed through regular testing by certified laboratories. However, in the Nether- lands a tolerated illicit cannabis market exists in the form of so-called 'coffeeshops', which offers a wide variety of cannabis to the general public as well as to medicinal users of cannabis. Since cannabis has been available in the pharmacies, many patients have started to compare the price and quality of OMC and coffeeshop cannabis. As a result, the public debate on the success and neces- sity of the OMC program has been based more on personal experiences, rather than scientific data. The general opinion of consumers is that OMC cannabis is more expensive, without any clear dif- ference in the quality. This study was performed in order to show any differences in quality that might exist between the official and illicit sources of cannabis for medicinal use. Cannabis samples obtained from ran- domly selected coffeeshops were compared to medicinal grade cannabis obtained from the OMC in a variety of validated tests. Many coffeeshop samples were found to contain less weight than expected, and all were contaminated with bacteria and fungi. No obvious differences were found in either cannabinoid- or water-content of the samples. The obtained results show that medicinal cannabis offered through the pharmacies is more reliable and safer for the health of medical users of cannabis.
. A central tenet underlying the use of botanical remedies is that herbs contain many active ingredients. Primary active ingredients may be enhanced by secondary compounds, which act in beneficial syn-ergy. Other herbal constituents may mitigate the side effects of dominant active ingredients. We reviewed the literature concerning medical can-nabis and its primary active ingredient, ∆ 9 -tetrahydrocannabinol (THC). Good evidence shows that secondary compounds in cannabis may enhance the beneficial effects of THC. Other cannabinoid and non-cannabinoid compounds in herbal cannabis or its extracts may reduce THC-induced anxiety, cholinergic deficits, and immunosuppression. Cannabis terpenoids and flavonoids may also increase cerebral blood flow, enhance cortical activity, kill respiratory pathogens, and provide anti-inflammatory activ-ity. [Article copies available for a fee from The Haworth Document Delivery Service: and: Cannabis Therapeutics in HIV/AIDS (ed: Ethan Russo) The Haworth Integrative Healing Press, an imprint of The Haworth Press, Inc., 2001, pp. 103-132. Single or multiple copies of this arti-cle are available for a fee from The Haworth Document Delivery Service [1-800-342-9678, 9:00 a.m. -5:00 p.m. (EST). E-mail address: getinfo@haworthpressinc.com].
Cannabis sativa L. (cannabis) extracts, vapor produced by the Volcano vaporizer and smoke made from burning cannabis joints were analyzed by GC-flame ionization detecter (FID), GC-MS and HPLC. Three different medicinal cannabis varieties were investigated Bedrocan, Bedrobinol and Bediol. Cannabinoids plus other components such as terpenoids and pyrolytic by-products were identified and quantified in all samples. Cannabis vapor and smoke was tested for cannabinoid receptor 1 (CB1) binding activity and compared to pure Delta(9)-tetrahydrocannabinol (Delta(9)-THC). The top five major compounds in Bedrocan extracts were Delta(9)-THC, cannabigerol (CBG), terpinolene, myrcene, and cis-ocimene in Bedrobinol Delta(9)-THC, myrcene, CBG, cannabichromene (CBC), and camphene in Bediol cannabidiol (CBD), Delta(9)-THC, myrcene, CBC, and CBG. The major components in Bedrocan vapor (>1.0 mg/g) were Delta(9)-THC, terpinolene, myrcene, CBG, cis-ocimene and CBD in Bedrobinol Delta(9)-THC, myrcene and CBD in Bediol CBD, Delta(9)-THC, myrcene, CBC and terpinolene. The major components in Bedrocan smoke (>1.0 mg/g) were Delta(9)-THC, cannabinol (CBN), terpinolene, CBG, myrcene and cis-ocimene in Bedrobinol Delta(9)-THC, CBN and myrcene in Bediol CBD, Delta(9)-THC, CBN, myrcene, CBC and terpinolene. There was no statistically significant difference between CB1 binding of pure Delta(9)-THC compared to cannabis smoke and vapor at an equivalent concentration of Delta(9)-THC.
The cannabis plant (Cannabis sativa L.) has a long history as a recreational drug, but also as part of traditional medicine in many cultures. Nowadays, it is used by a large number of patients worldwide, to ameliorate the symptoms of diseases varying from cancer and AIDS to multiple sclerosis and migraine. The discovery of cannabinoid-receptors and the endocannabinoid system have opened up a new and exciting field of research. But despite the pharmaceutical potential of cannabis, its classification as a narcotic drug has prevented the successful development of cannabis into modern medicine. Although a huge number of scientific papers has been published on cannabis, there is currently no scientific consensus on the usefulness of medicinal cannabis. In 2004, The Netherlands became the first country to make herbal cannabis available as a prescription drug. The phytochemical research presented in this thesis has been possible because of the availability of these high-grade cannabis plants. This thesis has a specific focus on the cannabinoids and on analytical problems that currently obstruct advanced study of the cannabis plant. Furthermore, it deals with much needed methods for quality control and with administration forms of medicinal cannabis. In general, it may be considered a general guidebook, covering all the basic phytochemical aspects of medicinal cannabis.
Metabolic fingerprinting techniques have received a lot of attention in recent years and the annual amount of publications in this field has increased significantly over the past decade. This increase in publications is due to improvements in the analytical performance, most notably in the field of NMR and MS analysis, and the increased awareness of the different applications of this growing field. Metabolomic fingerprinting or profiling is continuously being applied to new areas of research such as drug discovery from natural resources, quality control of herbal material, and discovering lead compounds. In this review the current state of the art of metabolic fingerprinting, focussing on NMR and MS technologies will be discussed. The application of these two analytical tools in the quality control of herbal material and phytopharmaceuticals forms the major part of this review. Finally we will look at the future developments and perspectives of these two technologies in the quality control of herbal material.
There has been controversy about whether the subjective, behavioral or therapeutic effects of whole plant marijuana differ from the effects of its primary active ingredient, Delta(9)-tetrahydrocannabinol (THC). However, few studies have directly compared the effects of marijuana and THC using matched doses administered either by the smoked or the oral form.
Two studies were conducted to compare the subjective effects of pure THC to whole-plant marijuana containing an equivalent amount of THC in normal healthy volunteers. In one study the drugs were administered orally and in the other they were administered by smoking.
In each study, marijuana users (oral study: n=12, smoking study: n=13) participated in a double-blind, crossover design with five experimental conditions: a low and a high dose of THC-only, a low and a high dose of whole-plant marijuana, and placebo. In the oral study, the drugs were administered in brownies, in the smoking study the drugs were smoked. Dependent measures included the Addiction Research Center Inventory, the Profile of Mood States, visual analog items, vital signs, and plasma levels of THC and 11-nor-9-carboxy-THC.
In both studies, the active drug conditions resulted in dose-dependent increases in plasma THC levels, and the levels of THC were similar in THC-only and marijuana conditions (except that at the higher oral dose THC-only produced slightly higher levels than marijuana). In both the oral study and the smoking study, THC-only and whole plant marijuana produced similar subjective effects, with only minor differences.
These results support the idea that the psychoactive effects of marijuana in healthy volunteers are due primarily to THC.
A (1)H-NMR method has been developed for the quantitative analysis of pure cannabinoids and for cannabinoids present in Cannabis sativa plant material without any chromatographic purification. The experiment was performed by the analysis of singlets in the range of delta 4.0-7.0 in the (1)H-NMR spectrum, in which distinguishable signals of each cannabinoid are shown. Quantitation was performed by calculating the relative ratio of the peak area of selected proton signals of the target compounds to the known amount of the internal standard, anthracene. For this method no reference compounds are needed. It allows rapid and simple quantitation of cannabinoids with a final analysis time of only 5 min without the need for a pre-purification step.
The metabolomic analysis of 12 Cannabis sativa cultivars was carried out by 1H NMR spectroscopy and multivariate analysis techniques. Principal component analysis (PCA) of the 1H NMR spectra showed a clear discrimination between those samples by principal component 1 (PC1) and principal component 3 (PC3) in cannabinoid fraction. The loading plot of PC value obtained from all 1)H NMR signals shows that Delta9-tetrahydrocannabinolic acid (THCA) and cannabidiolic acid (CBDA) are important metabolites to differentiate the cultivars from each other. The discrimination of the cultivars could also be obtained from a water extract containing carbohydrates and amino acids. The level of sucrose, glucose, asparagine, and glutamic acid are found to be major discriminating metabolites of these cultivars. This method allows an efficient differentiation between cannabis cultivars without any prepurification steps.
This study investigated the contribution of different cannabinoids to the subjective, behavioral and neurophysiological effects of smoked marijuana. Healthy marijuana users (12 men, 11 women) participated in four sessions. They were randomly assigned to a low or a high delta9-tetrahydrocannabinol group (THC; 1.8% versus 3.6%). In the four sessions under blinded conditions subjects smoked marijuana cigarettes containing placebo (no active cannabinoids), or cigarettes containing THC with low or high levels of cannabichromene (CBC; 0.1% versus 0.5%) and low or high levels of cannabidiol (CBD; 0.2% versus 1.0%). Dependent measures included subjective reports, measures of cognitive task performance and neurophysiological measures [electroencephalographic (EEG) and event-related potential (ERP)]. Compared to placebo, active THC cigarettes produced expected effects on mood, behavior and brain activity. A decrease in performance, reduction in EEG power and attenuation of ERP components reflecting attentional processes were observed during tests of working memory and episodic memory. Most of these effects were not dose-dependent. Varying the concentrations of CBC and CBD did not change subjects' responses on any of the outcome measures. These findings are consistent with previous studies indicating that THC and its metabolites are the primary active constituents of marijuana. They also suggest that neurophysiological EEG and ERP measures are useful biomarkers of the effects of THC.
Cannabis sativa L. is possibly one of the oldest plants cultivated by man, but has remained a source of controversy throughout its history. Whether pariah or panacea, this most versatile botanical has provided a mirror to medicine and has pointed the way in the last two decades toward a host of medical challenges from analgesia to weight loss through the discovery of its myriad biochemical attributes and the endocannabinoid system wherein many of its components operate.
This study surveys the history of cannabis, its genetics and preparations. A review of cannabis usage in Ancient Egypt will serve as an archetype, while examining first mentions from various Old World cultures and their pertinence for contemporary scientific investigation. Cannabis historians of the past have provided promising clues to potential treatments for a wide array of currently puzzling medical syndromes including chronic pain, spasticity, cancer, seizure disorders, nausea, anorexia, and infectious disease that remain challenges for 21st century medicine. Information gleaned from the history of cannabis administration in its various forms may provide useful points of departure for research into novel delivery techniques and standardization of cannabis-based medicines that will allow their prescription for treatment of these intractable medical conditions.
Twelve new cannabinoid esters, together with three known cannabinoid acids and Δ9-THC, were isolated from a high potency variety of Cannabis sativa L [1,2]. The structures were determined by extensive spectral analysis to be: β-fenchyl-Δ9-THCA ester (1), α-fenchyl-Δ9-THCA ester (2), bornyl-Δ9-THCA ester (3), epi-bornyl-Δ9-THCA ester (4), α-terpenyl-Δ9-THCA ester (5), 4-terpenyl-Δ9-THCA ester (6), α-cadinyl-Δ9-THCA ester (7), γ-eudesmyl-Δ9-THCA ester (8), inseparable mixture of two sesquiterpenyl-Δ9-THCA esters (9), α-cadinyl-CBGA ester (10), γ-eudesmyl-CBGA ester (11), 4-terpenyl-CBNA ester (12), Δ9-tetrahydrocannabinol (Δ9-THC), Δ9-tetrahydrocannabinolic acid A (Δ9-THCA), cannabinolic acid A (CBNA) and cannabigerolic acid (CBGA). CB-1 receptor assays [3–6] indicated that these esters, as well as the parent Δ9-THCA, are not active compared to Δ9-THC. Acknowledgements: This work is supported by the National Institute on Drug Abuse (contract # N01DA-5-7746) and by the Center of Research Excellence in Natural Products Neuroscience, The University of Mississippi (contract # 1P20RR021929-01). We are grateful to Dr. Bharathi Avula for assistance with the HR-ESI-MS, and to Dr. Melissa Jacob and Ms. Marsha Wright for conducting the antimicrobial testing. References: [1] ElSohly MA, et al. (2000) Journal of Forensic Science 45: 24–30. [2] ElSohly MA, Slade D (2005) Life Sciences 78: 539–548. [3] Devane WA, (1988) Molecular Pharmacology 34: 605–613. [4] Munro S, et al. (1993) Nature 365: 61–65. [5] Barth F (2005) Annual Reports in Medicinal Chemistry 40: 103–118. [6] Ashton JC, Giass M (2007) Current Neuropharmacology 5: 73–80.
Nine strains of Cannabis sativa L. (marijuana) were grown for research by the University of Mississippi. The seeds for these strains were obtained from Iowa, Minnesota, Mexico, Turkey, Italy, France, and Sweden. The cannabinoid content was determined using GLC, and the material was divided into two chemical phenotypes according to cannabinoid content. These phenotype categories are used to differentiate between drug-type and fiber-type Cannabis sativa. In addition, the ( - )-δ9−trans-tetrahydrocannabinol content was determined for both male and female plants, various plant parts, and a Turkish variety during various stages in its growth.
IT has been suggested that ``drug'' strains and ``non-drug'' strains of Cannabis sativa L. comprise two comprehensive groups1,2, which can be identified on the basis of their relative content of two of the principal ``cannabinoids''. Drug strains have been thought to contain an excess, usually substantial, of (-)-Delta9-trans-tetrahydrocannabinol (Delta9-THC) in comparison with the amount of cannabidiol (CBD), including carboxylate forms of both compounds, and non-drug strains have been held to have the reverse ratio. The former compound is considered psychotomimetic (psychosis-imitating), whereas the latter is not3. In examining the above cannabinoid ratio to decide in which phenotypic group a strain belongs, some investigators2 add the amount of cannabinol (CBN) to the amount of THC. The former seems to be an oxidation product of Delta9-THC (ref. 4) and is not considered to be psychoactive.
Sixty-eight compounds were identified by coupled gas chromatography and mass spectrometry (GC-MS) in the chemosphere of Cannabis sativa L. pollen and entire male and female plants of two cultivated varieties, Northern Lights and Hawaian Indica. Twenty-one and 28 substances, respectively, were present in pollen of the two forms. To conserve the natural composition of volatiles a delicate headspace method was employed. The two varieties represent different chemotypes which distinguish themselves, in the main quantitatively, in the setup of volatiles from pollen and entire male and female plants. Twenty compounds were monoterpenes, including the five major components: β-myrcene (E)-β-ocimene, terpinolene, β-pinene and limonene; 25 were sesquiterpenes, and the other 23 were of mixed biogenetic origin, including 3-methyl-1-butanol and benzylalcohol which occurred only in pollen; two pyrazines occurred only in Northern Lights females. Besides being of interest in natural products chemistry, the results should have relevance for plant systematics and for the pharmaceutical and technical applications of Cannabis. We demonstrate that the pollen has a distinct chemical character in possessing two exclusive volatiles, while lacking seven compounds occurring in males and females of both variants. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society, 2005, 147, 387–397.
This article provides a critical overview of current methods to quantify essential oil components. The fields of application and limits of the most popular approaches, in particular relative percentage abundance, normalized percentage abundance, concentration and true amount determination via calibration curves, are discussed in detail. A specific paragraph is dedicated to the correct use of the most widely used detectors and to analyte response factors. A set of applications for each approach is also included to illustrate the considerations. Copyright (C) 2008 John Wiley & Sons, Ltd.
To determine whether the terpenoid composition of the essential oil of Cannabis is useful for chemotaxonomic discrimination, extracts of pistillate inflorescences of 162 greenhouse-grown plants of diverse origin were analyzed by gas chromatography. Peak area ratios of 48 compounds were subjected to multivariate analysis and the results interpreted with respect to geographic origin and taxonomic affiliation. A canonical analysis in which the plants were pre-assigned to C. sativa or C. indica based on previous genetic, morphological, and chemotaxonomic studies resulted in 91% correct assignment of the plants to their pre-assigned species. A scatterplot on the first two principal component axes shows that plants of accessions from Afghanistan assigned to the wide-leaflet drug biotype (an infraspecific taxon of unspecified rank) of C. indica group apart from the other putative taxa. The essential oil of these plants usually had relatively high ratios of guaiol, isomers of eudesmol, and other unidentified compounds. Plants assigned to the narrow-leaflet drug biotype of C. indica tended to have relatively high ratios of trans-β-farnesene. Cultivars of the two drug biotypes may exhibit distinctive medicinal properties due to significant differences in terpenoid composition.
Cannabinoids are important chemotaxonomic markers unique to Cannabis. Previous studies show that a plant's dry-weight ratio of Δ(9)-tetrahydrocannabinol (THC) to cannabidiol (CBD) can be assigned to one of three chemotypes and that alleles B(D) and B(T) encode alloenzymes that catalyze the conversion of cannabigerol to CBD and THC, respectively. In the present study, the frequencies of B(D) and B(T) in sample populations of 157 Cannabis accessions were determined from CBD and THC banding patterns, visualized by starch gel electrophoresis. Gas chromatography was used to quantify cannabinoid levels in 96 of the same accessions. The data were interpreted with respect to previous analyses of genetic and morphological variation in the same germplasm collection. Two biotypes (infraspecific taxa of unassigned rank) of C. sativa and four biotypes of C. indica were recognized. Mean THC levels and the frequency of B(T) were significantly higher in C. indica than C. sativa. The proportion of high THC/CBD chemotype plants in most accessions assigned to C. sativa was <25% and in most accessions assigned to C. indica was >25%. Plants with relatively high levels of tetrahydrocannabivarin (THCV) and/or cannabidivarin (CBDV) were common only in C. indica. This study supports a two-species concept of Cannabis.
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Nine new cannabinoids (1-9) were isolated from a high-potency variety of Cannabis sativa. Their structures were identified as (+/-)-4-acetoxycannabichromene (1), (+/-)-3''-hydroxy-Delta((4'',5''))-cannabichromene (2), (-)-7-hydroxycannabichromane (3), (-)-7R-cannabicoumarononic acid A (4), 5-acetyl-4-hydroxycannabigerol (5), 4-acetoxy-2-geranyl-5-hydroxy-3-n-pentylphenol (6), 8-hydroxycannabinol (7), 8-hydroxycannabinolic acid A (8), and 2-geranyl-5-hydroxy-3-n-pentyl-1,4-benzoquinone (9) through 1D and 2D NMR spectroscopy, GC-MS, and HRESIMS. The known sterol beta-sitosterol-3-O-beta-d-glucopyranosyl-6'-acetate was isolated for the first time from cannabis. Compounds 6 and 7 displayed significant antibacterial and antifungal activities, respectively, while 5 displayed strong antileishmanial activity.
Medicines that activate cannabinoid CB(1) and CB(2) receptor are already in the clinic. These are Cesamet (nabilone), Marinol (dronabinol; Delta(9)-tetrahydrocannabinol) and Sativex (Delta(9)-tetrahydrocannabinol with cannabidiol). The first two of these medicines can be prescribed to reduce chemotherapy-induced nausea and vomiting. Marinol can also be prescribed to stimulate appetite, while Sativex is prescribed for the symptomatic relief of neuropathic pain in adults with multiple sclerosis and as an adjunctive analgesic treatment for adult patients with advanced cancer. One challenge now is to identify additional therapeutic targets for cannabinoid receptor agonists, and a number of potential clinical applications for such agonists are mentioned in this review. A second challenge is to develop strategies that will improve the efficacy and/or the benefit-to-risk ratio of a cannabinoid receptor agonist. This review focuses on five strategies that have the potential to meet either or both of these objectives. These are strategies that involve: (i) targeting cannabinoid receptors located outside the blood-brain barrier; (ii) targeting cannabinoid receptors expressed by a particular tissue; (iii) targeting up-regulated cannabinoid receptors; (iv) targeting cannabinoid CB(2) receptors; or (v) 'multi-targeting'. Preclinical data that justify additional research directed at evaluating the clinical importance of each of these strategies are also discussed.
In previous reports the presence of cannabinoids in the distilled essential oil of Cannabis sativa L. was proved, besides the presence of mono– and sesquiterpene hydrocarbons.
In this paper the localization of the cannabinoids in the glandular hairs of the leaves and with that the possible biogenetic relation with the components of the essential oil are demonstrated by microscopic examination after colouring tests and gaschromatographic analysis of the isolated contents of individual glandular hairs. Quantitative data about the relation between essential oil and cannabinoids are obtained by comparing the extracts without and after preceding steam distillation.
On acount of the origin of the seed (birdseed), special attention was paid to the botanical description of the plant material and to the counting of chromosomes.
Three-hundred and fifty diverse seed acquisitions of Cannabis were grown outdoors under uniform conditions in Ottawa, Canada, and analysed for their content of CBD, Δ9-THC, Δ8-THC, CBGM and CBN by glc. Several patterns of association recurred frequently. Plants originating from countries north of latitude 30° N almost always had notably higher contents of cannabinoids in the females than in the males. Considerable amounts of CBD were present. Less frequently, moderate or high amounts of THC were also present in the females. In plants originating from countries south of latitude 30° N, high amounts of THC and low amounts of CBD were frequently present in both sexes. Plants probably conforming to this latter type frequently failed to reach the flowering stage in the relatively short growing season of Ottawa. CBN was rarely present in freshly harvested plants, and then only in trace amounts. Δ8-THC was usually present in trace amounts. Trace amounts of a compound having the same retention time as CBGM were consistently present in plants originating from northeastern Asia.
The aqueous extract (marijuana tea) and volatile oil prepared from marijuana were compared with (−)-trans-Δ9-tetrahydrocannabinol for their effect on hexobarbital sleeping time and analgesic action in mice. All three substances prolonged hexobarbital sleeping time with an order of potency of (−)-trans-Δ9-tetrahydrocannabinol > aqueous extract > volatile oil. Each agent produced significant analgesic activity. However, the potencies of the aqueous extract and the volatile oil were similar to each other but only 1/200 that of (−)-trans-Δ9-tetrahydrocannabinol.
1 Conditions have been worked out for a reliable estimation of the cataleptic activity of delta'-trans-tetrahydrocannabinol (THC) after oral administration to mice, using the ring test over a period of 6 h. 2 By this method, the activity of cannabis herb and 5 crude fractions were measured against THC; at the same time the THC contents were determined chemically. 3 The B/C ratio (biological activity divided by chemical assay) was calculated for each. With cannabis herb the value was 3.3 and with extracts prepared with ethanol or 70% ethanol the values ranged from 3.2 to 7.1, indicating that in all samples the activity was much higher than would be expected from their THC content. 4. The cannabinoids were completely extracted from a sample of herb using petroleum spirit and the marc examined for a possible synergist. Surprisingly, it contained a powerful inhibitor of the action of THC, which could be restored by intraperitoneal prostaglandin E2 (3 microgram/kg). Some crude fractions had inhibitory activities about 10 times that of aspirin. 5 In contrast, the petroleum spirit extract (referred to in 4) had a surprisingly high B/C ratio of 23, indicating that a powerful synergist of THC activity is present. 6 The net effect of the herb and ethanol extracts is probably due to a balance of synergist and inhibitor.
1. The oral sedative potencies of cannabis herb, crude ethanolic and petroleum-ether fractions, were assayed against delta'-trans-tetrahydrocannabinol (THC) administered orally to mice, by measuring spontaneous motor activity over 30 min periods, at selected times, up to 6 h. 2. The THC contents of the extracts were determined chemically by gas-liquid chromatography analysis and the B/C ratio (biological activity divided by chemical activity) calculated for each. The B/C values for cannabis herb, which contained THC but no CBD, was 4.47 and for ethanolic and petroleum-ether extracts, 5.26 and 4.39, respectively. 3. The sedative potency expressed as SDA50, the dose required to give 50% effect over 6 h, was 1.06 (0.98 to 1.15) mg/kg for THC; 4.72 (4.22 to 5.27) mg/kg for cannabidiol and 1.26 (1.22 to 1.80) mg/kg for chlorpromazine. 4. An infusion of cannabis herb made with boiling water was shown to have sedative activity of very low potency. 5. When the cannabinoids were completely extracted from a sample of herb with petroleum-ether the aqueous and ethanolic extracts of the marc had some sedative activity; but the 70% ethanolic fraction had none. 6. The sedative activity of THC, cannabis herb and a water soluble fraction is blocked by aspirin, a cyclo-oxygenase inhibitor, and restored by prostaglandin E2 (PGE2). 7. The sedative effect of chlorpromazine is not blocked by aspirin.
This basic fraction (BF) of marihuana whole smoke condensate was subjected to pharmacological testing in males, Swiss-Webster mice. In a general pharmacological activity screen looking at behavioral, neurologic, and autonomic parameters, BF, at iv doses of 5, 10, and 20 mg/kg, caused impairment of visual placing, increase in tail pinch response, decrease in tail elevation, and induction of piloerection. These effects, although statistically significant, were slight and not consistently dose dependent. In a second study with doses ranging from 10 to 29 mg/kg, BF caused a decrease in spatial locomotion, rearing behavior, and urination incidence. In a third study, body temperatures of mice were measured periodically for 2 hr following administration of BF (1.2, 2.4, and 4.8 mg/kg) alone or in combination with 1.0 mg/kg delta-9-tetrahydrocannabinol (THC). BF did not alter body temperature, nor did it affect THC-induced hypothermia. These results, although suggesting that the basic fraction of marihuana whole smoke condensate has pharmacological activity in mice, offers little evidence for the presence of highly active compounds.
The composition of the steam-distilled volatile oil of fresh and air-dried, indoor-grown marijuana was studied by GC/FID and GC/MS. In all, 68 components were detected of which 57 were fully identified. Drying of the plant material had no effect on the qualitative composition of the oil and did not affect the ability of individuals familiar with marijuana smell to recognize the odor.
Cannabis has a potential for clinical use often obscured by unreliable and purely anecdotal reports. The most important natural cannabinoid is the psychoactive tetrahydrocannabinol (Δ9-THC); others include cannabidiol (CBD) and cannabigerol (CBG). Not all the observed effects can be ascribed to THC, and the other constituents may also modulate its action; for example CBD reduces anxiety induced by THC. A standardised extract of the herb may be therefore be more beneficial in practice and clinical trial protocols have been drawn up to assess this. The mechanism of action is still not fully understood, although cannabinoid receptors have been cloned and natural ligands identified.
Cannabis is frequently used by patients with multiple sclerosis (MS) for muscle spasm and pain, and in an experimental model of MS low doses of cannabinoids alleviated tremor. Most of the controlled studies have been carried out with THC rather than cannabis herb and so do not mimic the usual clincal situation. Small clinical studies have confirmed the usefulness of THC as an analgesic; CBD and CBG also have analgesic and antiinflammatory effects, indicating that there is scope for developing drugs which do not have the psychoactive properties ofTHC. Patients taking the synthetic derivative nabilone for neurogenic pain actually preferred cannabis herb and reported that it relieved not only pain but the associated depression and anxiety. Cannabinoids are effective in chemotherapy-induced emesis and nabilone has been licensed for this use for several years. Currently, the synthetic cannabinoid HU211 is undergoing trials as a protective agent after brain trauma. Anecdotal reports of cannabis use include case studies in migraine and Tourette’s syndrome, and as a treatment for asthma and glaucoma.
Apart from the smoking aspect, the safety profile of cannabis is fairly good. However, adverse reactions include panic or anxiety attacks, which are worse in the elderly and in women, and less likely in children. Although psychosis has been cited as a consequence of cannabis use, an examination of psychiatric hospital admissions found no evidence of this, however, it may exacerbate existing symptoms. The relatively slow elimination from the body of the cannabinoids has safety implications for cognitive tasks, especially driving and operating machinery; although driving impairment with cannabis is only moderate, there is a significant interaction with alcohol.
Natural materials are highly variable and multiple components need to be standardised to ensure reproducible effects. Pure natural and synthetic compounds do not have these disadvantages but may not have the overall therapeutic effect of the herb.
Cannabis is under clinical investigation to assess its potential for medicinal use, but the question arises as to whether there is any advantage in using cannabis extracts compared with isolated Delta9-trans-tetrahydrocannabinol (Delta9THC), the major psychoactive component. We have compared the effect of a standardized cannabis extract (SCE) with pure Delta9THC, at matched concentrations of Delta9THC, and also with a Delta9THC-free extract (Delta9THC-free SCE), using two cannabinoid-sensitive models, a mouse model of multiple sclerosis (MS), and an in-vitro rat brain slice model of epilepsy. Whilst SCE inhibited spasticity in the mouse model of MS to a comparable level, it caused a more rapid onset of muscle relaxation, and a reduction in the time to maximum effect compared with Delta9THC alone. The Delta9THC-free extract or cannabidiol (CBD) caused no inhibition of spasticity. However, in the in-vitro epilepsy model, in which sustained epileptiform seizures were induced by the muscarinic receptor agonist oxotremorine-M in immature rat piriform cortical brain slices, SCE was a more potent and again more rapidly-acting anticonvulsant than isolated Delta9THC, but in this model, the Delta9THC-free extract also exhibited anticonvulsant activity. Cannabidiol did not inhibit seizures, nor did it modulate the activity of Delta9THC in this model. Therefore, as far as some actions of cannabis were concerned (e.g. antispasticity), Delta9THC was the active constituent, which might be modified by the presence of other components. However, for other effects (e.g. anticonvulsant properties) Delta9THC, although active, might not be necessary for the observed effect. Above all, these results demonstrated that not all of the therapeutic actions of cannabis herb might be due to the Delta9THC content.
Cannabis is a potential treatment for epilepsy, although the few human studies supporting this use have proved inconclusive. Previously, we showed that a standardized cannabis extract (SCE), isolated Delta9-tetrahydrocannabinol (Delta9-THC), and even Delta9-THC-free SCE inhibited muscarinic agonist-induced epileptiform bursting in rat olfactory cortical brain slices, acting via CB1 receptors. The present work demonstrates that although Delta9-THC (1 microM) significantly depressed evoked depolarizing postsynaptic potentials (PSPs) in rat olfactory cortex neurones, both SCE and Delta9-THC-free SCE significantly potentiated evoked PSPs (all results were fully reversed by the CB1 receptor antagonist SR141716A, 1 microM); interestingly, the potentiation by Delta9-THC-free SCE was greater than that produced by SCE. On comparing the effects of Delta9-THC-free SCE upon evoked PSPs and artificial PSPs (aPSPs; evoked electrotonically following brief intracellular current injection), PSPs were enhanced, whereas aPSPs were unaffected, suggesting that the effect was not due to changes in background input resistance. Similar recordings made using CB1 receptor-deficient knockout mice (CB1-/-) and wild-type littermate controls revealed cannabinoid or extract-induced changes in membrane resistance, cell excitability and synaptic transmission in wild-type mice that were similar to those seen in rat neurones, but no effect on these properties were seen in CB1-/- cells. It appears that the unknown extract constituent(s) effects over-rode the suppressive effects of Delta9-THC on excitatory neurotransmitter release, which may explain some patients' preference for herbal cannabis rather than isolated Delta9-THC (due to attenuation of some of the central Delta9-THC side effects) and possibly account for the rare incidence of seizures in some individuals taking cannabis recreationally.