Article

Metabolic fingerprinting of Cannabis sativa L., cannabinoids and terpenoids for chemotaxonomic and drug standardization purposes

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  • Hazekamp Herbal Consulting
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... 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. ...
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Citation: Trancoso, I.; de Souza, G.A.R.; dos Santos, P.R.; dos Santos, K.D.; de Miranda, R.M.d.S.N.; da Silva, A.L.P.M.; Santos, D.Z.; García-Tejero, I.F.; Campostrini, E. Cannabis sativa L.: Crop Management and Abiotic Factors That Affect Phytocannabinoid Production.
... The genus Cannabis contains different types of chemicals with a diverse phytocannabinoid profile and range of effects [1]. The differences in phytocannabinoids composition and quantities of cannabis chemotypes should be searched in the genetic background of their biosynthesis pathways and the environmental conditions where they have been evolved [6][7][8]. Precursor synthesis of cannabinoids occurs from two distinct biosynthesis pathways: the polyketide and the methylerythritol phosphate (MEP) pathways, which produce olivetolic acid (OLA) and geranyl diphosphate (GPP), respectively [5]. Geranylpyrophosphate:olivetolate geranyltransferase catalyse the alkylation of OLA with GPP, leading to formation of CBGA (cannabigerolic acid), the main precursor of various cannabinoids, ...
... 4 Average annual maximum temperat age annual relative humidity. 6 Average annual minimum humidity. 7 Average annual maximum humidity. ...
... 5 Average annual relative humidity. 6 Average annual minimum humidity. 7 Average annual maximum humidity. ...
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Cannabis (Cannabis sativa L.) has a rich history of human use, and the therapeutic importance of compounds produced by this species is recognized by the medical community. The active constituents of cannabis, collectively called cannabinoids, encompass hundreds of distinct molecules, the most well-characterized of which are tetrahydrocannabinol (THC) and cannabidiol (CBD), which have been used for centuries as recreational drugs and medicinal agents. As a first step to establish a cannabis breeding program, we initiated this study to describe the HPLC-measured quantity of THC and CBD biochemistry profiles of 161 feral pistillate cannabis plants from 20 geographical regions of Iran. Our data showed that Iran can be considered a new region of high potential for distribution of cannabis landraces with diverse THC and CBD content, predominantly falling into three groups, as Type I = THC-predominant, Type II = approximately equal proportions of THC and CBD (both CBD and THC in a ratio close to the unity), and Type III = CBD-predominant. Correlation analysis among two target cannabinoids and environmental and geographical variables indicated that both THC and CBD contents were strongly influenced by several environmental–geographical factors, such that THC and CBD contents were positively correlated with mean, min and max annual temperature and negatively correlated with latitude, elevation, and humidity. Additionally, a negative correlation was observed between THC and CBD concentrations, suggesting that further studies to unravel these genotype × environment interactions (G × E interactions) are warranted. The results of this study provide important pre-breeding information on a collection of cannabis that will underpin future breeding programs.
... The terpenes are among the main compounds of the essential oil (EO) of cannabis [76]. Cannabis' EO is a liquid with a yellow color more or less pronounced, composed of mono-and sesquiterpenes but also of other molecules such as terpene alcohols and cannabinoids [75,[77][78][79][80]. Diverse biological activities have been associated with C. sativa EO including insecticidal, nematicide, antimicrobial, fungicidal, anti-leishmanial, antioxidant, anti-acetylcholinesterase or neuroactive activities [81][82][83][84][85]. ...
... The main objective of studies dealing with the extraction of terpenes from cannabis using CSE is the identification of these compounds. Cannabis EO are well extracted with both polar [78,79] and nonpolar solvent [75,79,89], however Namdar et al. (2018) [79] determined that a mixture of polar and nonpolar solvents results in the most efficient extraction of terpenes and cannabinoids from inflorescences. Indeed, the authors showed that the solvent mixture hexane:EtOH (7:3 v/v) is the most efficient for the extraction in comparison with pure hexane and ethanol. ...
... The use of polar solvent benefits to the cannabinoids yield [79]. Although the operating conditions are not detailed, L/S ratios vary in the range of 5-50 mL/g, the extractions last up to 1 h and the application of heat does not seem necessary [75,78,79,89]. Besides, high temperature during extended extraction time may cause a loss of volatile compounds and thus lower the EO yield. ...
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Cannabis sativa L. is a controversial crop due to its high tetrahydrocannabinol content varieties; however, the hemp varieties get an increased interest. This paper describes (i) the main categories of phenolic compounds (flavonoids, stilbenoids and lignans) and terpenes (monoterpenes and sesquiterpenes) from C. sativa by-products and their biological activities and (ii) the main extraction techniques for their recovery. It includes not only common techniques such as conventional solvent extraction, and hydrodistillation, but also intensification and emerging techniques such as ultrasound-assisted extraction or supercritical CO2 extraction. The effect of the operating conditions on the yield and composition of these categories of phenolic compounds and terpenes was discussed. A thorough investigation of innovative extraction techniques is indeed crucial for the extraction of phenolic compounds and terpenes from cannabis toward a sustainable industrial valorization of the whole plant.
... Cannabis classification is a fundamental requirement for future medical research and applications, and it is best enabled through an overview of the class and content of potentially therapeutic secondary metabolites in each plant part. Currently, researchers attempted to discriminate and identify the chemical differences between the categories of "Sativa" (narrow-leaflet drug, NLD) and "Indica" (wide-leaflet drug, WLD) (Fischedick et al., 2010;Hazekamp and Fischedick, 2012;Hazekamp et al., 2016). Results of the chemotaxonomic separation of "Sativa" and "Indica" were mixed, and THC and CBD concentrations appeared to have no differentiation value. ...
... Results of the chemotaxonomic separation of "Sativa" and "Indica" were mixed, and THC and CBD concentrations appeared to have no differentiation value. However, certain terpenoids were more prominent in some strains than others (Hillig, 2005b;Fischedick et al., 2010;Hazekamp and Fischedick, 2012;Fischedick, 2015Fischedick, , 2017Hazekamp et al., 2016;Jin et al., 2017;McPartland and Guy, 2017). The mixed results in the current body of literature may be due to experimental design shortcomings. ...
... Firstly, the vernacular terminology ("Sativa" and "Indica") is inadequate for medical applications due to the misuse of the botanical nomenclature, extensive cross-breeding, and unreliable labeling during unrecorded hybridization (McPartland, 2017). Secondly, samples in most classification studies were collected from disparate sources (Fischedick et al., 2010;Hazekamp et al., 2016) and are subject to inconsistent environmental factors during the growth phases (Aizpurua-Olaizola, 2016) and post-harvest treatment (Jin et al., 2019). Additionally, inappropriate sample preparation and extraction procedures during laboratory analysis may affect classification results (Jin et al., 2020). ...
Article
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Previous chemotaxonomic studies of cannabis only focused on tetrahydrocannabinol (THC) dominant strains while excluded the cannabidiol (CBD) dominant strains and intermediate strains (THC ≈ CBD). This study investigated the utility of the full spectrum of secondary metabolites in different plant parts in three cannabis chemotypes (THC dominant, intermediate, and CBD dominant) for chemotaxonomic discrimination. Hierarchical clustering, principal component analysis (PCA), and canonical correlation analysis assigned 21 cannabis varieties into three chemotypes using the content and ratio of cannabinoids, terpenoids, flavonoids, sterols, and triterpenoids across inflorescences, leaves, stem bark, and roots. The same clustering results were obtained using secondary metabolites, omitting THC and CBD. Significant chemical differences were identified in these three chemotypes. Cannabinoids, terpenoids, flavonoids had differentiation power while sterols and triterpenoids had none. CBD dominant strains had higher amounts of total CBD, cannabidivarin (CBDV), cannabichromene (CBC), α-pinene, β-myrcene, (-)-guaiol, β-eudesmol, α-eudesmol, α-bisabolol, orientin, vitexin, and isovitexin, while THC dominant strains had higher total THC, total tetrahydrocannabivarin (THCV), total cannabigerol (CBG), camphene, limonene, ocimene, sabinene hydrate, terpinolene, linalool, fenchol, α-terpineol, β-caryophyllene, trans-β-farnesene, α-humulene, trans-nerolidol, quercetin, and kaempferol. Compound levels in intermediate strains were generally equal to or in between those in CBD dominant and THC dominant strains. Overall, with higher amounts of β-myrcene, (-)-guaiol, β-eudesmol, α-eudesmol, and α-bisabolol, intermediate strains more resemble CBD dominant strains than THC dominant strains. The results of this study provide a comprehensive profile of bioactive compounds in three chemotypes for medical purposes. The simultaneous presence of a predominant number of identified chemotype markers (with or without THC and CBD) could be used as chemical fingerprints for quality standardization or strain identification for research, clinical studies, and cannabis product manufacturing.
... Even though these varieties might differ in morphologic and organoleptic features and are commonly distinguished by names, it is inconclusive to which extent these varieties present true differences in chemical composition [18]. There are some studies [6,24,27,[37][38][39] addressing this specific question. Fischedick et al. [37] cultivated eleven different varieties under equal and controlled conditions and then analyzed 36 different plant ingredients, seven of which were cannabinoids. ...
... There are some studies [6,24,27,[37][38][39] addressing this specific question. Fischedick et al. [37] cultivated eleven different varieties under equal and controlled conditions and then analyzed 36 different plant ingredients, seven of which were cannabinoids. Ultimately, the authors were able to distinguish between the investigated varieties. ...
... Traditional classification based on THC and CBD contents [6,16,17] allowed differentiation of the investigated cannabis varieties into phenotypes I and II. As previously observed in other studies [6,18,24,25,27,37,39,55], comprehensive analytical methods combined with multivariate statistical analyses, e.g., PCA, enabled for further subgrouping of cannabis varieties. The presented data concerning the PCA complemented the traditionally applied classification into phenotypes I, II, and III. ...
Article
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Cannabis sativa ( C. sativa ) is commonly chemically classified based on its Δ ⁹ -tetrahydrocannabinol (THC) and cannabidiol (CBD) content ratios. However, the plant contains nearly 150 additional cannabinoids, referred to as minor cannabinoids. Minor cannabinoids are gaining interest for improved plant and product characterization, e.g., for medical use, and bioanalytical questions in the medico-legal field. This study describes the development and validation of an analytical method for the elucidation of minor cannabinoid fingerprints, employing liquid chromatography coupled to high-resolution mass spectrometry. The method was used to characterize inflorescences from 18 different varieties of C. sativa , which were cultivated under the same standardized conditions. Complementing the targeted detection of 15 cannabinoids, untargeted metabolomics employing in silico assisted data analysis was used to detect additional plant ingredients with focus on cannabinoids. Principal component analysis (PCA) was used to evaluate differences between varieties. The overall purpose of this study was to examine the ability of targeted and non-targeted metabolomics using the mentioned techniques to distinguish cannabis varieties from each other by their minor cannabinoid fingerprint. Quantitative determination of targeted cannabinoids already gave valuable information on cannabinoid fingerprints as well as inter- and intra-variety variability of cannabinoid contents. The untargeted workflow led to the detection of 19 additional compounds. PCA of the targeted and untargeted datasets revealed further subgroups extending commonly applied phenotype classification systems of cannabis. This study presents an analytical method for the comprehensive characterization of C. sativa varieties. Graphical abstract
... sativa originated from Europe and C. indica from Asia), and the composition of their secondary metabolites (secondary plant metabolites are large numbers of chemical compounds produced by plant cells, using biosynthetic enzymes and building blocks derived from primary plant metabolic pathways) all showed differences, but the dilemma about their taxonomical separation, whether they are separate species or subspecies, continues to the present day (Russo, 2007;Small, 2015;Pollio, 2016). Several review and research papers have provided detailed reports about the classification and nomenclature of cannabis (Fischedick et al., 2010;Pollio, 2016;McPartland, 2017McPartland, , 2018. The current consensus is that C. sativa and C. indica should not be considered different species. ...
... This fiber-type cannabis is characterized by a higher level of CBD, and the maximum D 9 -THC content of this type of cannabis was defined as 0.2%-0.3% of dry matter (McPartland, 2017). Chemotaxonomic evaluation has led to the recognition of an intermediate type with similar levels of D 9 -THC and CBD (Fischedick et al., 2010). ...
... The average terpene concentration in cannabis flowers were previously reported in the range of 1%-10%, but due to selective breeding, the lower end of the average terpene content has increased up to 3.5% or even higher in modern chemovars (Fischedick et al., 2010;Lewis et al., 2018). Review papers published in the last decade revealed considerable variation in the terpene profile of cannabis and its products (Hillig, 2004;Fischedick et al., 2010;Casano et al., 2011;Da Porto et al., 2014;Elzinga et al., 2015;Aizpurua-Olaizola et al., 2016;Hazekamp et al., 2016;Fischedick, 2017;Jin et al., 2017;Richins et al., 2018;Mudge et al., 2019). ...
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Pain prevalence among adults in the United States has increased 25% over the past two decades, resulting in high health-care costs and impacts to patient quality of life. In the last 30 years, our understanding of pain circuits and (intra)cellular mechanisms has grown exponentially, but this understanding has not yet resulted in improved therapies. Options for pain management are limited. Many analgesics have poor efficacy and are accompanied by severe side effects such as addiction, resulting in a devastating opioid abuse and overdose epidemic. These problems have encouraged scientists to identify novel molecular targets and develop alternative pain therapeutics. Increasing preclinical and clinical evidence suggests that cannabis has several beneficial pharmacological activities, including pain relief. Cannabis sativa contains more than 500 chemical compounds, with two principle phytocannabinoids, Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD). Beyond phytocannabinoids, more than 150 terpenes have been identified in different cannabis chemovars. Although the predominant cannabinoids, Δ9-THC and CBD, are thought to be the primary medicinal compounds, terpenes including the monoterpenes β-myrcene, α-pinene, limonene, and linalool, as well as the sesquiterpenes β-caryophyllene and α-humulene may contribute to many pharmacological properties of cannabis, including anti-inflammatory and antinociceptive effects. The aim of this review is to summarize our current knowledge about terpene compounds in cannabis and to analyze the available scientific evidence for a role of cannabis-derived terpenes in modern pain management. SIGNIFICANCE STATEMENT: Decades of research have improved our knowledge of cannabis polypharmacy and contributing phytochemicals, including terpenes. Reform of the legal status for cannabis possession and increased availability (medicinal and recreational) have resulted in cannabis use to combat the increasing prevalence of pain and may help to address the opioid crisis. Better understanding of the pharmacological effects of cannabis and its active components, including terpenes, may assist in identifying new therapeutic approaches and optimizing the use of cannabis and/or terpenes as analgesic agents.
... Contribution to cannabinoid profile and biological activity (structures in Fig. 3, properties in Table 3). CBG (3) (first described by Gaoni and Mechoulam 1964) is commonly detected in cannabis samples, regardless of their origin, and can reach concentrations of up to 20.8 mg/g (Fischedick et al. 2010;Aizpurua-Olaizola et al. 2014;Jin et al. 2017;Richins et al. 2018;Wang et al. 2018;Zager et al. 2019) (Table 1). Among the quantitative analysis publications considered for this review, only two reported on CBGA (4) (first reported by Mechoulam and Gaoni 1965a), with the highest reported concentration being 14 mg/g (Jin et al. 2017;Wang et al. 2018;). ...
... Among the quantitative analysis publications considered for this review, only two reported on CBGA (4) (first reported by Mechoulam and Gaoni 1965a), with the highest reported concentration being 14 mg/g (Jin et al. 2017;Wang et al. 2018;). A quantitative analysis of cannabis samples for cannabigerol monomethyl ether (5) indicated that this metabolite is generally present at low levels but can accumulate to 2.6 mg/g in some chemovars (Fischedick et al. 2010) (Table 1). Reports on the concentrations of the remaining members of the CBG-type cannabinoids across chemovars are currently unavailable. ...
... Contribution to cannabinoid profile and biological activity (structures in Fig. 4, properties in Table 4). The content of CBC (27) (first described by Claussen et al. 1966, andMechoulam 1966) in cannabis tends to be extremely low but, in hashish samples, high concentrations of up to 5.4 mg/g have been reported to occur in certain chemovars (Fischedick et al. 2010;Jin et al. 2017;Richins et al. 2018;Wang et al. 2018;Zager et al. 2019) (Table 1). Quantitative data for other CBC-type metabolites have not been reported. ...
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Following decades of tight restrictions, recent legislative adjustments have decriminalized the use of products derived from cannabis (Cannabis sativa L.) in many countries and jurisdictions. This has led to a renewed interest in better understanding the chemical basis of physiological effects attributed to cannabis use. The present review article summarizes our current knowledge regarding the 130 structures of cannabinoids that have been characterized from cannabis extracts to date. We are also providing information on the methods employed for structure determination to help the reader assess the quality of the original structural assignments. Cannabinoid chemical diversity is discussed in the context of current knowledge regarding the enzymes involved in cannabinoid biosynthesis. We briefly assess to what extent cannabinoid levels are determined by the genotype of a given chemovar and discuss the limits of enzymatic control over the cannabinoid profile.
... Solvent-based (both polar and nonpolar) conventional extraction methods have also been implemented to isolate terpenes from Cannabis [66,67,75,107,108]. For example, Ibrahim et al. [72] used an ethyl acetate, ethanol, methanol, and chloroform/methanol (1:9; v/v) mixture and reported that ethyl acetate was the best solvent to recover terpenes from Cannabis [72]. ...
... Fischedick et al. [66] used ethanol as the extraction solvent to recover both terpenes and cannabinoids, and 36 compounds were identified in 11 varieties of C. sativa [66]. Conventional ethanolic extraction was also used by A. Hazekamp and Fischedick [88] to isolate monoterpenes (α-pinene, myrcene, and terpinolene); sesquiterpenes ((E)-caryophyllene and α-humulene) [88]; and oxygenated terpenes (guaiol, γ_eudesmol, and α_bisabolol) from marijuana and medical Cannabis inflorescences [82]. ...
... Fischedick et al. [66] used ethanol as the extraction solvent to recover both terpenes and cannabinoids, and 36 compounds were identified in 11 varieties of C. sativa [66]. Conventional ethanolic extraction was also used by A. Hazekamp and Fischedick [88] to isolate monoterpenes (α-pinene, myrcene, and terpinolene); sesquiterpenes ((E)-caryophyllene and α-humulene) [88]; and oxygenated terpenes (guaiol, γ_eudesmol, and α_bisabolol) from marijuana and medical Cannabis inflorescences [82]. ...
Article
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Cannabis is well-known for its numerous therapeutic activities, as demonstrated in pre-clinical and clinical studies primarily due to its bioactive compounds. The Cannabis industry is rapidly growing; therefore, product development and extraction methods have become crucial aspects of Cannabis research. The evaluation of the current extraction methods implemented in the Cannabis industry and scientific literature to produce consistent, reliable, and potent medicinal Cannabis extracts is prudent. Furthermore, these processes must be subjected to higher levels of scientific stringency, as Cannabis has been increasingly used for various ailments, and the Cannabis industry is receiving acceptance in different countries. We comprehensively analysed the current literature and drew a critical summary of the extraction methods implemented thus far to recover bioactive compounds from medicinal Cannabis. Moreover, this review outlines the major bioactive compounds in Cannabis, discusses critical factors affecting extraction yields, and proposes future considerations for the effective extraction of bioactive compounds from Cannabis. Overall, research on medicinal marijuana is limited, with most reports on the industrial hemp variety of Cannabis or pure isolates. We also propose the development of sustainable Cannabis extraction methods through the implementation of mathematical prediction models in future studies.
... Terpenes are present in plant resins and essential oils and are responsible for the pharmacological and physiological effects of many medicinal plants, including Cannabis (Fischedick et al., 2010b). Terpenes are defined by the number of isoprene units and can be categorized based on the size of the carbon skeleton. ...
... Mono-and sesquiterpenes are primarily found in the essential oils of Cannabis. A significant and positive correlation was found between the level of terpenes and cannabinoids because mono-and sesquiterpenes are synthesized in the same areas where cannabinoids are produced (Fischedick et al., 2010b). However, studying terpenes is difficult because their yield can depend on the part of the plant, age, cultivation, storage and other factors (Andre et al., 2016). ...
Article
Cannabis has been used for centuries for its medicinal properties. Given the dangerous and unpleasant side effects of existing analgesics, the chemical constituents of Cannabis have garnered significant interest for their antinociceptive, anti-inflammatory and neuroprotective effects. To date, Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) remain the two most widely studied constituents of Cannabis in animals. These studies have led to formulations of THC and CBD for human use; however, chronic pain patients also use different strains of Cannabis (sativa, indica and ruderalis) to alleviate their pain. These strains contain major cannabinoids, such as THC and CBD, but they also contain a wide variety of cannabinoid and noncannabinoid constituents. Although the analgesic effects of Cannabis are attributed to major cannabinoids, evidence indicates other constituents such as minor cannabinoids, terpenes and flavonoids also produce antinociception against animal models of acute, inflammatory, neuropathic, muscle and orofacial pain. In some cases, these constituents produce antinociception that is equivalent or greater compared to that produced by traditional analgesics. Thus, a better understanding of the extent to which these constituents produce antinociception alone in animals is necessary. The purposes of this review are to (1) introduce the different minor cannabinoids, terpenes, and flavonoids found in Cannabis and (2) discuss evidence of their antinociceptive properties in animals.
... There is some empirical evidence supporting that these two species do indeed contain different amounts of certain cannabis constituents. For example, various studies have analyzed commercially available cannabis labeled as "indica" or "sativa" and found that indica samples contained greater concentrations of myrcene and hydroxylated terpenes while sativa samples contained greater terpinolene, 3-carene, and several sesquiterpenes (Hazekamp et al., 2016), but indica and sativa samples generally contained similar concentrations of major cannabinoids (i.e., THC and CBD; Elzinga et al., 2015;Fischedick et al., 2010;Hazekamp & Fischedick, 2012;Hazekamp et al., 2016). Though these few studies suggest some indica and sativa products may be distinguishable by terpene content, there is presently no consensus, operational definition of "indica" or "sativa" based on chemical composition. ...
... 6 sativa for "enhanced energy" and indica for "sleep" and "sedation" (Cohen et al., 2016;Pearce et al., 2014). Prior studies that have conducted analytical testing on commercial indica and sativa cannabis products for cannabinoid and terpene content (Elzinga et al., 2015;Fischedick et al., 2010;Hazekamp & Fischedick, 2012;Hazekamp et al., 2016) may provide insight into the differences in perceived effects of indica and sativa observed in the present study. These studies have shown that cannabis labeled as "indica" contains similar concentrations of major cannabinoids (i.e., THC and CBD) to cannabis labeled as "sativa" (Elzinga et al., 2015;Hazekamp et al., 2016). ...
Article
Cannabis products available for retail purchase are often marketed based on purported plant species (e.g., "indica" or "sativa"). The cannabis industry frequently claims that indica versus sativa cannabis elicits unique effects and/or is useful for different therapeutic indications. Few studies have evaluated use patterns, beliefs, subjective experiences, and situations in which individuals use indica versus sativa. A convenience sample of cannabis users (n = 179) was surveyed via Amazon Mechanical Turk (mTurk). Participants were asked about their prior use of, subjective experiences with, and opinions on indica versus sativa cannabis and completed hypothetical purchasing tasks for both cannabis subtypes. Participants reported a greater preference to use indica in the evening and sativa in the morning and afternoon. Participants were more likely to perceive feeling "sleepy/tired" or "relaxed" after using indica and "alert," "energized," and "motivated" after using sativa. Respondents were more likely to endorse wanting to use indica if they were going to sleep soon but more likely to use sativa at a party. Hypothetical purchasing patterns (i.e., grams of cannabis purchased as a function of escalating price) did not differ between indica and sativa, suggesting that demand was similar. Taken together, cannabis users retrospectively report feeling different effects from indica and sativa; however, demand generally did not differ between cannabis subtypes, suggesting situational factors could influence whether someone uses indica or sativa. Placebo-controlled, blinded studies are needed to characterize the pharmacodynamics and chemical composition of indica and sativa cannabis and to determine whether user expectancies contribute to differences in perceived indica/sativa effects. (PsycInfo Database Record (c) 2021 APA, all rights reserved).
... By standardizing the genetic plant material and using the same cultivation and processing methods, uniform plants with a uniform phytochemical profile can thus be bred (Fischedick et al., 2010). ...
... Through this cultivation technique and processing procedure, uniform plants with a uniform phytochemical profile can be produced (Fischedick et al., 2010). ...
Thesis
Cannabis sativa L. eignet sich aufgrund der Möglichkeit, die ganze Pflanze zu nutzen, hervorragend für die Kreislaufwirtschaft und ist daher ein Paradebeispiel für eine multifunktionale Nutzpflanze. Die Cannabispflanze erlebt derzeit einen Boom aufgrund ihres reichhaltigen Repertoires an sekundären Pflanzeninhaltsstoffen, ihrer Fasern und ihres wertvollen Öls in zahlreichen Industriezweigen sowie ihrer positiven landwirtschaftlichen Eigenschaften. Das Hauptaugenmerk liegt dabei im medizinischen Nutzen, basierend auf den in Blüten und Blättern vorhandenen Phytocannabinoiden. Es ist wichtig, zwischen Nutzhanf Genotypen und phytocannabinoid-reichen (PCR) Genotypen zu unterscheiden. Nutzhanf erfüllt den von der EU-Gesetzgebung vorgeschriebenen THC-Grenzwert von 0,2% und kann daher im Feldmaßstab legal angebaut werden. PCR Genotypen, enthalten hohe Mengen an nicht-psychoaktiven Cannabinoiden, wie CBD und Cannabigerol (CBG), in einem Bereich von 10–30%, während ihr THC-Gehalt unter 0,2% liegt. Diese Genotypen werden derzeit gezüchtet und sind noch kaum auf dem Markt erhältlich. Die Cannabinoid-Extraktion von aus Nutzhanf gewonnenen Rohstoffen, könnte einen entscheidenden Wettbewerbsvorteil bieten, da die geerntete Biomasse durch eine bessere Flächennutzung und mehr Kosteneffizienz im Vergleich zu einem Indoor-Produktionssystem deutlich erhöht werden könnte. Darüber hinaus kann die Multifunktionalität der Nutzhanfpflanze einen wirtschaftlichen Mehrwert bieten. Bestehende Anbausysteme für die Faser- und Ölsaatenproduktion müssen neu entwickelt werden, da sich der Erntezeitpunkt und Ernteorgan stark von den bisherigen Systemen unterscheiden dürften. Um dies zu erreichen, befasst sich Publikation I mit folgenden Zielen: Ermittlung des Ertragspotenzials verschiedener Nutzhanf Genotypen hinsichtlich Blütenstand- und Biomasseertrag sowie Cannabinoidgehalt in Abhängigkeit von Genotyp, Wachstumsstadium und Biomassefraktion in einem Freilandanbausystem. In einem zweijährigen Feldversuch wurden sieben Nutzhanf Genotypen (Finola, Fédora17, Ferimon, Félina32, Futura75, USO31 und Santhica27) angebaut. Die Beprobung von Blättern und Blütenständen erfolgte zu vier spezifischen Wachstumsstadien: vegetatives Blattstadium, Knospenstadium, Vollblütestadium und zur Samenreife. Die Trockensubstanz wurde erfasst sowie der Cannabinoidgehalt analysiert. Die Ergebnisse zeigten, dass der Gehalt an Cannabinoiden stark vom Genotyp und dem Wachstumsstadium abhängt. Daher müssen für ein optimales Ernteergebnis die Biomasse und der Ertrag der Blütenstände berücksichtigt werden. Der Genotyp Santhica27 wies den höchsten Gehalt an CBG/A auf. Die Genotypen Futura75, Fédora17, Félina32, Ferimon, Finola und Santhica27, welche die höchsten CBD/A- bzw. CBG/A Gehalte aufwiesen hatten zur Samenreife die höchsten Biomasseerträge an Druschrückständen und somit einen höheren CBD/A- und CBG/A-Ertrag pro Fläche. Folglich ist eine Ernte nach der Samenreife wirtschaftlich vorteilhaft. Diese Ergebnisse machen ausgewählte Nutzhanf Genotypen zu idealen Kandidaten für den Mehrzweckanbau in Bezug auf Biomasseproduktion und CBD/A- bzw. CBG/A-Gewinnung, um das volle Potenzial der Hanfpflanze auszuschöpfen. Zusätzlich befasste sich die Arbeit mit einer weiteren Standardisierung von PCR Genotypen in Indoor-Anbausystemen. Aufgrund der vorgeschriebenen hohen Qualitätsanforderungen für medizinisches Cannabismaterial rückt der Indoor-Anbau immer mehr in den Fokus, da alle Produktionsparameter standardisiert werden können. Die Produktion von Cannabinoiden unter Indoor-Bedingungen ist aufgrund von Verarbeitungskosten und regulatorischen Einschränkungen teur. Daher wird eine kosteneffektive Produktionskette angestrebt. In Publikation II wurde die Anpassung der Pflanzenarchitektur durch den gezielten Einsatz von synthetischen Phytohormonen evaluiert. Mit dem Ziel eine kleine und kompakte Pflanzenmorphologie mit hohen Blütenerträgen zu generieren. Dies umfasste folgende Zielsetzungen: den Einfluss exogen applizierter Pflanzenwachstumsregulatoren (PGRs), wie NAA, BAP und einer Mischung (NAA/BAP-Mix) aus beiden auf die Pflanzenarchitektur verschiedener PCR Genotypen zu prüfen. Darüber hinaus den Biomasseertrag sowie den CBD/A Gehalt zu bestimmen. In einem Gewächshausexperiment wurden die Genotypen mit synthetischen Phytohormonen in verschiedenen Konzentrationen behandelt. Als Ergebnis wurde ein genotyp-spezifischer Einfluss der applizierten PGRs auf die Pflanzenarchitektur festgestellt. NAA führte beim Genotyp KANADA zu einer kompakteren Pflanzenmorphologie mit einem konstant hohen Blütenertrag, während der CBD/A-Gehalt nicht beeinflusst wurde. Die Genotypen 0.2x und FED zeigten durch die Anwendungen reduzierte Blütenerträge. Publikation III befasste sich mit der Bewertung von Ertragsparametern und CBD Gehalt von PCR Genotypen, welche in verschiedenen Substratzusammensetzungen in einem Indoor-Topfanbausystem kultiviert wurden. In einem Gewächshausexperiment wurde der Einfluss folgender Substratzusammensetzungen: Torf-Mix (PM); Torf-Mix, substituiert mit 30% Grünfasern (G30), und Kokosfaser (CC), auf Wachstumsleistung, N-Gehalt, Wurzelwachstum sowie CBD/A-Gehalt untersucht. Die verschiedenen Substrate zeigten signifikante Auswirkungen auf die Wachstumsleistung und die Wurzelentwicklung der getesteten Genotypen. Es wurde eine genotyp-spezifische Reaktion auf den Blütenertrag untersucht wobei kein limitierender Effekt auf den CBD/A-Gehalt festgestellt wurde. Es lässt sich schlussfolgern, dass organische Torfalternativen wie Grünfasern, die Torf in Standardtopfsubstraten teilweise ersetzen, eine genotyp-spezifische Option bieten.
... The concept of 'Cannabis fingerprinting' has already been mentioned in some research papers. Fischedick et al. (2010) [28] demonstrated that the metabolic fingerprinting of Cannabis, subjected to chemometric techniques, was able to chemotaxonomically differentiate Cannabis varieties. The authors were interested in finding the most discriminating components of the plant when applying principal component analysis. ...
... The concept of 'Cannabis fingerprinting' has already been mentioned in some research papers. Fischedick et al. (2010) [28] demonstrated that the metabolic fingerprinting of Cannabis, subjected to chemometric techniques, was able to chemotaxonomically differentiate Cannabis varieties. The authors were interested in finding the most discriminating components of the plant when applying principal component analysis. ...
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Cannabis sativa L. is widely used as recreational illegal drugs. Illicit Cannabis profiling, comparing seized samples, is challenging due to natural Cannabis heterogeneity. The aim of this study was to use GC–FID and GC–MS herbal fingerprints for intra (within)- and inter (between)-location variability evaluation. This study focused on finding an acceptable threshold to link seized samples. Through Pearson correlation-coefficient calculations between intra-location samples, ‘linked’ thresholds were derived using 95% and 99% confidence limits. False negative (FN) and false positive (FP) error rate calculations, aiming at obtaining the lowest possible FP value, were performed for different data pre-treatments. Fingerprint-alignment parameters were optimized using Automated Correlation-Optimized Warping (ACOW) or Design of Experiments (DoE), which presented similar results. Hence, ACOW data, as reference, showed 54% and 65% FP values (95 and 99% confidence, respectively). An additional fourth root normalization pre-treatment provided the best results for both the GC–FID and GC–MS datasets. For GC–FID, which showed the best improved FP error rate, 54 and 65% FP for the reference data decreased to 24 and 32%, respectively, after fourth root transformation. Cross-validation showed FP values similar as the entire calibration set, indicating the representativeness of the thresholds. A noteworthy improvement in discrimination between seized Cannabis samples could be concluded.
... Products containing cannabinoids are made by adding cannabis flour derived from the seeds as well as cannabinoid oils and/or extracts. The proportion of a given addition affects the final content of these compounds in the finished product, but the content mainly of THC must not exceed permissible limits [44,[76][77][78][79]. ...
... A study by Hazekamp and Fischedick [9] showed that nominal cannabinoid concentrations in plants of the same type but in varied geographical locations differed by more than 25%. To mitigate such differences, we can implement strict control over the varieties and their growing methods to ensure greater homogeneity or mix the extracts to ensure the desired homogeneity [9,[77][78][79]. ...
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Scientific demonstrations of the beneficial effects of non-psychoactive cannabinoids on the human body have increased the interest in foods containing hemp components. This review systematizes the latest discoveries relating to the characteristics of cannabinoids from Cannabis sativa L. var. sativa, it also presents a characterization of the mentioned plant. In this review, we present data on the opportunities and limitations of cannabinoids in food production. This article systematizes the data on the legal aspects, mainly the limits of Δ9-THC in food, the most popular analytical techniques (LC-MS and GC-MS) applied to assay cannabinoids in finished products, and the available data on the stability of cannabinoids during heating, storage, and access to light and oxygen. This may constitute a major challenge to their common use in food processing, as well as the potential formation of undesirable degradation products. Hemp-containing foods have great potential to become commercially popular among functional foods, provided that our understanding of cannabinoid stability in different food matrices and cannabinoid interactions with particular food ingredients are expanded. There remains a need for more data on the effects of technological processes and storage on cannabinoid degradation.
... In this study, the manual collection of mature inflorescences allows obtaining a high-quality plant material with yields in essential oils ranging among 0.85 and 1.33 EO/kg fresh flowers for EC and CS, respectively. The order of magnitude agrees with previous published data, also if a direct comparison is not easy to perform, due to the different cultivars, variability of operative conditions, different plant parts used, distillation of fresh/dry rather than raw/shredded material, harvesting periods and others [24][25][26]. The performances of experimental cultivation are summarized in Table 2, whereas the Table 3 summarizes the intrinsic scavenger/reducing properties of the three EOs. ...
... On the other side, heptacosane (peak 39) is only present in the Eletta campana sample (23.9%). The latter contains more complex mixture characterized by the exclusive presence of other six compounds (peaks 16,17,25,26,30,33). The quantitative differences as for myrcene (peak 4, range 6.8-26.4%), ...
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Industrial hemp is a multiuse crop that has been widely cultivated to produce fibers and nutrients. The capability of the essential oil (EO) from inflorescences as antimicrobial agent has been reported. However, literature data are still lacking about the hemp EO antiprotozoal efficacy in vivo. The present study aims to unravel this concern through the evaluation of the efficacy of hemp EOs (2.5 mL/kg, intraperitoneally) of three different cultivars, namely Futura 75, Carmagnola selezionata and Eletta campana, in mice intraperitoneally infected with Leishmania tropica. A detailed description of EO composition and targets-components analysis is reported. Myrcene, α-pinene and E-caryophyllene were the main components of the EOs, as indicated by the gas-chromatographic analysis. However, a prominent position in the scenario of the theoretical interactions underlying the bio-pharmacological activity was also occupied by selina-3,7(11)-diene, which displayed affinities in the micromolar range (5.4–28.9) towards proliferator-activated receptor α, cannabinoid CB2 receptor and acetylcholinesterase. The content of this compound was higher in Futura 75 and Eletta campana, in accordance with their higher scavenging/reducing properties and efficacy against the tissue wound, induced by L. tropica. Overall, the present study recommends hemp female inflorescences, as sources of biomolecules with potential pharmacological applications, especially towards infective diseases.
... Cannabis or its resins are reduced to small pieces by a grater [5] or spatula [45], grinded or pulverized, while cannabis oils are directly proceeded to instrumental analysis. Manual pulverization and homogenization of the dried plant material can be performed using mortar and pestle [46][47][48][49][50], metal spoon [51] or glass rod [52], by cutting the plant material [53] or crushing and riddling (0.5 mm) [54] or by manual grinder [50,55]. According to the UNODC [5], dried herbal cannabis material and cannabis resins should be pulverized by a cutter (at high revolution speed, i.e., 100 rps) and sieved (mesh size 1 mm). ...
... For the purpose of phytocannabinoid profiling, NMR spectroscopy is used as (semi)quantitative method alone [20,209] or as an orthogonal technique to LC [216][217][218] or GC [219] for the purpose of qualitative peak assignment of major phytocannabinoids [20], chemical and morphological examination [220], chemotaxonomic classification [20,219,220], metabolomics-based chemovar distinction [51] or quantitative analysis of cannabis plant material without the need of pre-purification step [221], chromatographic separation or use of certified reference standards [219]. Cryogenic NMR spectroscopy combines improved sensitivity and noise reduction with a cryogenic cooling system for the receiver coil and preamplifiers. ...
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Cannabis is gaining increasing attention due to the high pharmacological potential and updated legislation authorizing multiple uses. The development of time- and cost-efficient analytical methods is of crucial importance for phytocannabinoid profiling. This review aims to capture the versatility of analytical methods for phytocannabinoid profiling of cannabis and cannabis-based products in the past four decades (1980–2021). The thorough overview of more than 220 scientific papers reporting different analytical techniques for phytocannabinoid profiling points out their respective advantages and drawbacks in terms of their complexity, duration, selectivity, sensitivity and robustness for their specific application, along with the most widely used sample preparation strategies. In particular, chromatographic and spectroscopic methods, are presented and discussed. Acquired knowledge of phytocannabinoid profile became extremely relevant and further enhanced chemotaxonomic classification, cultivation set-ups examination, association of medical and adverse health effects with potency and/or interplay of certain phytocannabinoids and other active constituents, quality control (QC), and stability studies, as well as development and harmonization of global quality standards. Further improvement in phytocannabinoid profiling should be focused on untargeted analysis using orthogonal analytical methods, which, joined with cheminformatics approaches for compound identification and MSLs, would lead to the identification of a multitude of new phytocannabinoids.
... Indica and sativa plants have also been found to differ in terpene and cannabinoid profiles. Thus, these chemotaxonomic markers are a promising tool for screening hybrids (Hillig 2004, Hillig and Mahlberg 2004, Fischedick et al. 2010, Elzinga et al. 2015. Zhang et al. (2018) are recommending that Cannabis should be recognised as a monotypic species typified by Cannabis sativa L., containing three subspecies: subsp. ...
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The use of cannabis for medicinal purposes dates back well before the era of modern medicine, but in recent years research into the use of medical cannabis in the medical and pharmaceutical sciences has grown significantly. In European countries, most cannabis plants have been and still are grown for industrial purposes. For this reason, hemp cultivation technology is relatively well researched, while little is known about the key factors affecting cannabis cultivation for medical purposes. The active substances of cannabis plant targeted by this review are called phytocannabinoids. The biosynthesis of phytocannabinoids is relatively well understood, but the specific environmental factors that influence the type and number of phytocannabinoids have been much less studied. Indoor or greenhouse cultivation, which uses automated lighting, ventilation, irrigation systems and complex plant nutrition has become much more sophisticated and appears to be the most effective method for producing medical cannabis. There are many different cultivation systems for cannabis plants, but one of the essential elements of the process is an optimal plant nutrition and selection of fertilisers to achieve it. This review summarises the existing knowledge about phytocannabinoid biosynthesis and the conditions suitable for growing plants as sources of medical cannabis. This review also attempts to delineate how nutrient type and bioavailability influences the synthesis and accumulation of specific phytocannabinoids based on contemporary knowledge of the topic.
... sesquiterpenes 19,[28][29][30] . In addition, the contrasting aromas that have been associated with Sativa (that is, sweet) and Indica (that is, earthy) were key discriminators in a sensory evaluation of Cannabis cultivars and mediated customers' perceptions of potency and quality 9 . ...
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Analysis of over 100 Cannabis samples quantified for terpene and cannabinoid content and genotyped for over 100,000 single nucleotide polymorphisms indicated that Sativa- and Indica-labelled samples were genetically indistinct on a genome-wide scale. Instead, we found that Cannabis labelling was associated with variation in a small number of terpenes whose concentrations are controlled by genetic variation at tandem arrays of terpene synthase genes. By quantifying over 100 Cannabis samples for terpene and cannabinoid content and genotyping them for over 100,000 single nucleotide polymorphisms, this study finds that Cannabis labelling is associated with genetic variants in terpene synthase genes.
... From these values, LOD and LOQ were calculated using the equations (LOD = 3 × SD/S; LOQ = 10 × SD/S). Recoveries were determined by the method described previously [17]. ...
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In the chemical characterization of medically valued Cannabis, the present work has used a gas chromatography (GC) method coupled with mass spectrometry (MS) for identification and quantification of cannabinoids. We have modified a GC–MS method for chemical analysis of cannabinoids extracted from dried flowers of different Cannabis varieties. The method allows for quantification of major cannabinoids, i.e. cannabidiol (CBD), cannabichromene (CBC), tetrahydrocannabinol (THC), cannabigerol (CBG), and cannabinol (CBN) simultaneously. We found that the Cannabis cultivar called Cannabis 5-CW had the highest amount of CBD. We used a modified GC–MS method for chemical profiling of cannabinoids extracted from a variety of Cannabis samples, especially the high CBD, CBC, THC, CBG and CBN-producing ones. The method was successfully applied for identification and quantification of cannabinoids in a short time with better separation resolution among the reported methods.
... However, further studies with larger case numbers and users of more different medical cannabis strains are required to explore differentiability in serum samples. Moreover, fluctuations in cannabinoid profiles among different medical cannabis batches should be regarded [15,27,28]. ...
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Increasing prescription numbers of cannabis-based medicines raise the question of whether uptake of these medicines can be distinguished from recreational cannabis use. In this pilot study, serum cannabinoid profiles after use of cannabis-based medicines were investigated, in order to identify potential distinguishing markers. Serum samples after use of Sativex®, Dronabinol or medical cannabis were collected and analyzed for 18 different cannabinoids, using a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Analytes included delta-9-tetrahydrocannabinol, 11-hydroxy-tetrahydrocannabinol, 11-nor-9-carboxy-tetrahydrocannabinol, cannabidiol, cannabinol, cannabigerol, cannabichromene, cannabicyclol, tetrahydrocannabivarin, cannabidivarin, tetrahydocannabinolic acid A, cannabidiolic acid, cannabinolic acid, cannabigerolic acid, cannabichromenic acid, cannabicyclolic acid, tetrahydrocannabivarinic acid and cannabidivarinic acid. Cannabinoid profiles of study samples were compared to profiles of street cannabis user samples via principal component analysis and Kruskal–Wallis test. Potential distinguishing markers for Dronabinol and Sativex® intake were identified, including 11-hydroxy-tetrahydrocannabinol/delta-9-tetrahydrocannabinol ratios ≥1 and increased concentrations of 11-nor-9-carboxy-tetrahydrocannabinol, cannabidiol or cannabichromene. Larger quantities of minor cannabinoids suggested use of cannabis. Use of medical and street cannabis could not be distinguished, except for use of a cannabidiol-rich strain with higher cannabidiol/delta-9-tetrahydrocannabinol and cannabichromene/delta-9-tetrahydrocannabinol ratios. Findings of the study were used to classify forensic serum samples with self-reported use of cannabis-based medicines.
... For Uso-31 (chemotype V), whose inflorescences are characterized by a negligible level of cannabinoids, the lowest EO extraction yield was found irrespective of the year. A positive correlation between the accumulation of total cannabinoids and total terpenes, both synthesized in glandular trichomes [22], was already reported [32][33][34], and explains the low EO yield obtained for Uso-31. Considering both the years of cultivation, all the 2020 samples were characterized by a significantly higher EO extraction yield than the 2019 ones (Tables 2 and 3), and this could be at least partly due to the different meteorological conditions occurring in the two subsequent growing seasons, particularly regarding rainfalls and average temperature ( Table 5). ...
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Cannabis sativa L. is an annual species cultivated since antiquity for different purposes. While, in the past, hemp inflorescences were considered crop residues, at present, they are regarded as valuable raw materials with different applications, among which extraction of the essential oil (EO) has gained increasing interest in many fields. The aim of the present study is the evaluation of the yield and the chemical composition of the EO obtained by hydrodistillation from eleven hemp genotypes, cultivated in the same location for two consecutive growing seasons. The composition of the EOs was analyzed by GC-MS, and then subjected to multivariate statistical analysis. Sesquit-erpenes represented the main class of compounds in all the EOs, both in their hydrocarbon and oxygenated forms, with relative abundances ranging from 47.1 to 78.5%; the only exception was the Felina 32 sample collected in 2019, in which cannabinoids predominated. Cannabinoids were the second most abundant class of compounds, of which cannabidiol was the main one, with relative abundances between 11.8 and 51.5%. The statistical distribution of the samples, performed on the complete chemical composition of the EOs, evidenced a partition based on the year of cultivation, rather than on the genotype, with the exception of Uso-31. Regarding the extraction yield, a significant variation was evidenced among both the genotypes and the years of cultivation.
... Chemical differences can be seen within the growth cycle, harvesting, and storage due to environmental conditions. The major focus should be genotype consistency of a specific chemical profile of cannabis to produce a reproducible and stable chemical composition (Fischedick et al., 2010). With this information, the farmers and breeders will know more precisely what the risks and benefits of cultivating a particular variety are in terms of compliance with Slovenian legislation, depending on the production purpose. ...
... Sir William B. O'Shaughnessy, an Irish physician was the first to assess the therapeutic value of Cannabis scientifically when he was working with the Indian hemp, in Calcutta, India and publicized in the western countries in the early 19th century [10,11]. Cannabinoids were found to possess various pharmacological properties such as analgesic, anticonvulsant, appetite stimulant in cancer and AIDS patients, treatment of asthma and multiple sclerosis [12][13][14][15][16]. It has been also reported to possess antitumour and anticancer activities [17][18][19][20]. ...
Article
Cannabinoids are the major chemical constituents of the plant Cannabis sativa L. and are known to exhibit a wide range of pharmacological effects viz., psychotropic, analgesic, anticancer, antiinflammatory, antidiabetic, anticonvulsive, antibacterial and antifungal etc. The use of cannabis, cannabinoids and their products is restricted in several countries due to the high risk of misuse. Recently, cannabinoids have regained the interest of the researchers due to their therapeutic applications. Ever since the discovery of the cannabinoids, most of the studies carried out on the evaluation of their biological activities were limited to only preclinical levels. The quality of the preclinical data still remains only low to moderate, thus, leaving behind an uncertainty in their use for therapeutic applications. Problems associated with the solubility, stability and bioavailability of the cannabinoid drugs are also a major concern in the quality of the study. Nanoparticle based drug delivery system could be a potential method to increase the reliability of the data. While considering the immense pharmacological properties of the cannabinoids, there is an urgency to perform intensive clinical trials and to know their mechanism of action in various disease conditions, evaluate their efficacy and safety, and register them as drug candidates. This review highlights the chemistry, types and biological activities of the cannabinoids such as THC, CBD and CBN in focus with their anticancer activity, neuroprotective effect and nanoformulating the cannabinoid drugs.
... have also been identified: C. indica Lam. and C. ruderalis Janisch. -although there exists some debate regarding whether or not the three are in fact unique species (Russo, 2007;Fischedick et al., 2010;Hillig and Mahlberg, 2004). Currently (a) 2-AG is synthesized on-demand in neurons from arachidonic acid-containing membrane inositol phospholipids (phosphatidylinositol; PI). ...
Article
The first aim of the present review is to provide an in-depth description of the cannabinoids and their known effects at various neuronal receptors. It reveals that cannabinoids are highly diverse, and recent work has highlighted that their effects on the central nervous system (CNS) are surprisingly more complex than previously recognized. Cannabinoid-sensitive receptors are widely distributed throughout the CNS where they act as primary modulators of neurotransmission. Secondly, we examine the role of cannabinoid receptors at key brain sites in the control of fear and anxiety. While our understanding of how cannabinoids specifically modulate these networks is mired by their complex interactions and diversity, a plausible framework(s) for their effects is proposed. Finally, we highlight some important knowledge gaps in our understanding of the mechanism(s) responsible for their effects on fear and anxiety in animal models and their use as therapeutic targets in humans. This is particularly important for our understanding of the phytocannabinoids used as novel clinical interventions.
... With the discovery of biochemically-analogous endogenous cannabinoid ligands and the discovery of synthetic cannabinoids, the term ‗phytocannabinoids' is now used for psychoactive components isolated from the plant [5] . So far, more than 90 of these phytocannabinoids have been identified and are composed of C21 or C22 terpenophenolic compounds (including their breakdown products) produced by the cannabis plant [3,6,7] . These are further divided into 10 subclasses; namely, (-)-D9-transtetrahydrocannabinol (D9-THC), (-)-D8-transtetrahydrocannabinol (D8-THC), cannabinol (CBN), cannabidiol (CBD), cannabinodiol (CBND), cannabigerol (CBGs), cannabichromene (CBC), cannabicyclol (CBL), cannabielsoin (CBE), and cannabitriol (CBT) [3] . Figure 1 provides further information regarding the chemical structure of cannabinoids. ...
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Millions of Americans use cannabis for medical purposes including but not limited to pain, nausea, mood changes and appetite stimulation. The use of cannabinoid in the palliative care setting is a relatively new trend. Given the fact that a patient receiving palliative care is not necessarily approaching death, the increasing need for palliative care as the American population ages, this literature review was compiled in order to examine the potential efficacy of cannabis in treating the mental health comorbidities of palliative care patients. We attempted to create the most comprehensive report on cannabinoid use in palliative psychiatry. It summarizes the most recently published science on cannabinoid use in palliative care patients and its impact on mood and anxiety symptoms. The mechanism of action of cannabinoids on their associated receptors was elucidated, as were the pharmacological roles that specific molecules in cannabinoids, like cannabidiolic acid and terpenes, play in cannabinoids’ overall efficacy. The legal impediments to widespread cannabis use were also explored. While the potential efficacy of cannabinoids has proven to be mixed, more research is necessary to ensure that a potentially vital resource in treating palliative care patients does not go underutilized.
... The heating of the sample in the injector port induces the decarboxylation of the cannabinoid acids converting them into the neutral form. Then, the determination of both acids and free cannabinoids requires a derivatization step, which is a time-consuming procedure and does not guarantee 100% of the yield [13,14]. ...
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Cannabis sativa L. is an herbaceous plant belonging to the family of Cannabaceae. It is classified into three different chemotypes based on the different cannabinoids profile. In particular, fiber-type cannabis (hemp) is rich in cannabidiol (CBD) content. In the present work, a rapid nano liquid chromatographic method (nano-LC) was proposed for the determination of the main cannabinoids in Cannabis sativa L. (hemp) inflorescences belonging to different varieties. The nano-LC experiments were carried out in a 100 µm internal diameter capillary column packed with a C18 stationary phase for 15 cm with a mobile phase composed of ACN/H2O/formic acid, 80/19/1% (v/v/v). The reverse-phase nano-LC method allowed the complete separation of four standard cannabinoids in less than 12 min under isocratic elution mode. The nano-LC method coupled to ultraviolet (UV) detection was validated and applied to the quantification of the target analytes in cannabis extracts. The nano-LC system was also coupled to an electrospray ionization–mass spectrometry (ESI-MS) detector to confirm the identity of the cannabinoids present in hemp samples. For the extraction of the cannabinoids, three different approaches, including dynamic maceration (DM), ultrasound-assisted extraction (UAE), and an extraction procedure adapted from the French Pharmacopeia’s protocol on medicinal plants, were carried out, and the results achieved were compared.
... Studies have reported that terpenoids are powerful metabolites that have an interactive effect (or an "entourage effect") with cannabinoid receptors (Gertsch et al., 2008). However, terpene composition in cannabis resin is dependent upon genetic, environmental, and developmental factors, and highly variable terpene profiles additionally exist between individual plants (Fischedick et al., 2010;Hazekamp and Fischedick, 2012;Booth et al., 2017). Terpene diversity in cannabis resin is responsible for scent and flavor qualities of cannabis flowers (Booth et al., 2017). ...
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Cannabis sativa L. is cultivated for its secondary metabolites, of which the cannabinoids have documented health benefits and growing pharmaceutical potential. Recent legal cannabis production in North America and Europe has been accompanied by an increase in reported findings for optimization of naturally occurring and synthetic cannabinoid production. Of the many environmental cues that can be manipulated during plant growth in controlled environments, cannabis cultivation with different lighting spectra indicates differential production and accumulation of medically important cannabinoids, including Δ ⁹ -tetrahydrocannabinol (Δ ⁹ -THC), cannabidiol (CBD), and cannabigerol (CBG), as well as terpenes and flavonoids. Ultraviolet (UV) radiation shows potential in stimulating cannabinoid biosynthesis in cannabis trichomes and pre-harvest or post-harvest UV treatment merits further exploration to determine if plant secondary metabolite accumulation could be enhanced in this manner. Visible LED light can augment THC and terpene accumulation, but not CBD. Well-designed experiments with light wavelengths other than blue and red light will provide more insight into light-dependent regulatory and molecular pathways in cannabis. Lighting strategies such as subcanopy lighting and varied light spectra at different developmental stages can lower energy consumption and optimize cannabis PSM production. Although evidence demonstrates that secondary metabolites in cannabis may be modulated by the light spectrum like other plant species, several questions remain for cannabinoid production pathways in this fast-paced and growing industry. In summarizing recent research progress on light spectra and secondary metabolites in cannabis, along with pertinent light responses in model plant species, future research directions are presented.
... Moreover, results from previous studies on tobacco smokers suggested that the sensations which smoking creates in the airways contribute to short-term satisfaction, the rewarding effect, and reduced craving (49)(50)(51). One can therefore suppose that smoking CBD-rich cannabis may be "beneficial" as part of a strategy to lower exposure to THC: by preserving the smoking-related airway sensation as well as the terpene-related taste (52)(53)(54), a minimal reduction in the satisfaction experienced from the act of smoking may be derived from THC-low cannabis as compared to THChigh cannabis (44). In reality, smoking cannabis exposes persons to harmful substances, including carcinogens (55)(56)(57). ...
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Background Although cannabis use is common in France, it is still criminalized. Cannabidiol (CBD) products, including CBD-rich cannabis, are legally available. Although previous results suggested that CBD may have benefits for people with cannabis use disorder, there is a lack of data on cannabis users who use CBD to reduce their cannabis consumption. We aimed to identify (i) correlates of this motive, and (ii) factors associated with successful attempts to reduce cannabis use. Methods A cross-sectional online survey among French-speaking CBD and cannabis users was conducted. Logistic regressions were performed to identify correlates of using CBD to reduce cannabis consumption and correlates of reporting a large reduction. Results Eleven percent ( n = 105) of our study sample reported they primarily used CBD to reduce cannabis consumption. Associated factors included smoking tobacco cigarettes (adjusted odds ratio (aOR) [95% confidence interval (CI)] 2.17 [1.3–3.62], p = 0.003) and drinking alcohol (aOR [95%CI] 1.8 [1.02–3.18], p = 0.042). Of these 105, 83% used CBD-rich cannabis to smoke, and 58.7% reported a large reduction in cannabis consumption. This large reduction was associated with non-daily cannabis use (aOR [95%CI] 7.14 [2.4–20.0], p < 0.001) and daily CBD use (aOR [95%CI] 5.87 [2.09–16.47], p = 0.001). A reduction in cannabis withdrawal symptoms thanks to CBD use was the most-cited effect at play in self-observed cannabis reduction. Conclusions Cannabis use reduction is a reported motive for CBD use—especially CBD-rich cannabis to smoke—in France. More studies are needed to explore practices associated with this motive and to accurately assess CBD effectiveness.
... Terpenes are widely distributed in higher plants such as citrus, conifers, and eucalyptus and are detected in their leaves, flowers, stems, and roots (Ninkuu et al., 2021). According to Bueno et al. (2020) and Fischedick et al. (2010), terpene profile is influenced by a variety of factors such as genotypes, age of inflorescences, and environmental, production, and harvesting conditions. Although terpenes were most abundant in chili pepper "Hangjiao No.2" after harvest, their levels were relatively low compared to aldehydes in the present study, and their levels increased or diminished after treatment during the storage period, especially after day 3 of storage in the 1-MCP treatments compared to the control. ...
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This study aimed to determine the effects of different concentrations of 1-methyl cyclopropene (1-MCP) on the nutritional quality, antioxidant enzyme activities, and volatile compounds of “Hangjiao No.2” chili pepper during 12 days of storage at ambient temperature. The chili fruit were randomly selected and divided into four groups corresponding to the four treatments, thus, 0.5, 1.0, and 1.5 μl L –1 1-MCP and a control. The analysis of the nutritional value, enzyme activities, and volatile compounds were determined at 3 days interval. The results showed that the malondialdehyde (MDA) content was lower in the fruit treated with 1-MCP compared to the control. The treatment with 1.5 μl L –1 and the control showed the lowest superoxide dismutase (SOD) activity compared to the other treatments. Peroxidase (POD) and Catalase (CAT) were highest in the fruit treated with 0.5 μl L –1 compared to the control and treatment with 1.0 μl L –1 . The 1.5 μl L –1 treatment delayed the decline in vitamin C and protein content compared to the control. Nitrate levels increased 1.34-fold at 0.5 μl L –1 and 2.01-fold in the control. Chlorophyll content degradation was delayed at 1.0 μl L –1 compared to the control. A total of 88 volatile compounds, including terpenes, aldehydes, alkanes, esters, alcohols, acids, phenolic derivatives, ketones, and other aromatic compounds, were detected in “Hangjiao No.2” pepper during the 12-day storage period and treatment concentrations. The production of volatile terpenes was higher in the control than in the 1-MCP treatments, while the 0.5 μl L –1 1-MCP treatment generally suppressed the production of volatile compounds during storage. Overall, the production of volatile compounds after treatment was higher in the “Hangjiao No.2” chili fruit treated with 1.0 μl L –1 1-MCP than in the other treatments throughout the storage period. The results indicate that 1-MCP treatment was more effective in maintaining fruit quality, enhancing the activities of SOD, POD, and CAT, retarding the accumulation of MDA and restoring volatile aromas, with 1.0 μl L –1 having the best preservative effect on “Hangjiao No.2” chili fruit during storage, which could be useful for future marketing and processing.
... A. Lewis et al., 2018;A. Hazekamp & Fischedick, 2012;Justin Thomas Fischedick et al., 2010;Justin T. Fischedick, 2017;E. B. Russo & Marcu, 2017;Jin et al., 2020;Mudge et al., 2019;Zager et al., 2019;Jin et al., 2020) as well as additional research. ...
... Mass spectral library matching is often used, which is valid as long as a close match is found; however, the quality of the library matches is rarely reported. An examination of analytical results recently obtained by the CSIRO (data not published) as well as a selection of recently published cannabis terpene and terpenoid profiles [39][40][41][42][43][44][45][46][47][48][49][50] found that the following monoterpenes, 36-44 ( Fig. 5), sesquiterpenes, 45-49 (Fig. 6), and terpenoids, 50-57 ( Fig. 7) are most often found in cannabis varieties. Over 50 other cannabis terpenes and terpenoids have been described, but these were usually only reported in a single or small number of publications, and detected at low to very low levels. ...
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The science of cannabis and cannabinoids encompasses a wide variety of scientific disciplines and can appear daunting to newcomers to the field. The encroachment of folklore and ‘cannabis culture’ into scientific discussions can cloud the situation further. This Primer Review is designed to give a succinct overview of the chemistry of cannabis and cannabinoids. It is hoped that it will provide a useful resource for chemistry undergraduates, postgraduates and their instructors, and experienced chemists who require a comprehensive and up to date summary of the field. The Review begins with a brief overview of the history and botany of cannabis, then goes on to detail important aspects of the chemistry of phytocannabinoids, endocannabinoids and synthetic cannabinomimetics. Other natural constituents of the cannabis plant are then described including terpenes and terpenoids, polyphenolics, alkaloids, waxes and triglycerides, and important toxic contaminants. A discussion of key aspects of the pharmacology associated with cannabinoids and the endocannabinoid system then follows, with a focus on the cannabinoid receptors, CB1 and CB2. The medicinal chemistry of cannabis and cannabinoids is covered, highlighting the range of diseases targeted with cannabis and phytocannabinoids, as well as key aspects of phytocannabinoid metabolism, distribution, and delivery. The modulation of endocannabinoid levels through the inhibition of key endocannabinoid-degrading enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) is then discussed. The Review concludes with an assessment of the much touted ‘entourage effect’. References to primary literature and more specialised reviews are provided throughout.
... Similarly, readers interested in biotechnological synthesis of cannabinoids are referred to Luo et al. (2019), and to Leahy and coworkers for a clever example of their chemical synthesis (Shultz et al. 2018). The biosynthesis of the cannabis terpenes, present in small amounts in C. sativa (Hillig 2004;Fischedick et al. 2010;Booth et al. 2017;Booth and Bohlmann 2019;Mudge et al. 2019;Zager et al. 2019) and proposed to synergistically accentuate the pharmacological effects of cannabis consumption (Livingston et al. 2020;Russo 2011), is described elsewhere (Flores-Sanchez and Verpoorte 2008; Page et al. 2006). ...
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Cannabis has been integral to Eurasian civilization for millennia, but a century of prohibition has limited investigation. With spreading legalization, science is pivoting to study the pharmacopeia of the cannabinoids, and a thorough understanding of their biosynthesis is required to engineer strains with specific cannabinoid profiles. This review surveys the biosynthesis and biochemistry of cannabinoids. The pathways and the enzymes’ mechanisms of action are discussed as is the non-enzymatic decarboxylation of the cannabinoic acids. There are still many gaps in our knowledge about the biosynthesis of the cannabinoids, especially for the minor components, and this review highlights the tools and approaches that will be applied to generate an improved understanding and consequent access to these potentially biomedically-relevant materials. Graphical abstract
... Plants with THC concentration of <0.3% are considered as hemp (fiber-type) while those containing THC of ≥0.3% are considered as marijuana (drugtype) [13,21]. Besides using phytocannabinoids, monoterpenoids and sesquiterpenoids content can also be used to distinguish hemp from marijuana [22]. Variations in terpene synthase genes is also associated with the differences between drug-type Cannabis cultivars [23]. ...
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Cannabis sativa L. is an illegal plant in many countries. The worldwide criminalization of the plant has for many years limited its research. Consequently, understanding the full scope of its benefits and harm became limited too. However, in recent years the world has witnessed an increased pace in legalization and decriminalization of C. sativa. This has prompted an increase in scientific studies on various aspects of the plant’s growth, development, and use. This review brings together the historical and current information about the plant’s relationship with mankind. We highlight the important aspects of C. sativa classification and identification, carefully analyzing the supporting arguments for both monotypic (single species) and polytypic (multiple species) perspectives. The review also identifies recent studies on suitable conditions and methods for C. sativa propagation as well as highlighting the diverse uses of the plant. Specifically, we describe the beneficial and harmful effects of the prominent phytocannabinoids and provide status of the studies on heterologous synthesis of phytocannabinoids in different biological systems. With a historical view on C. sativa legality, the review also provides an up-to-date worldwide standpoint on its regulation. Finally, we present a summary of the studies on genome editing and suggest areas for future research.
... This is underscored by the reported preference of many patients for the whole plant extract compared with purified THC, and especially for specific strains, for example, Cannabis sativa strains with mood elevating effects may be better suited for day use, while Cannabis indica strains with sedative effects being preferable for night use. These different subjective effects are likely to arise from variation in the ratios of major and minor cannabinoids, terpenes and other phytochemicals [70,72,73]. Information of cannabis-derived compounds other than THC and CBD, regarding their effects on pain in humans, their effective doses, pharmacokinetic profiles and bioavailability, is an important determinant in the development of cannabis-based analgesic therapeutic agents. ...
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The recent legalization of medicinal cannabis in several jurisdictions has spurred the development of therapeutic formulations for chronic pain. Unlike pure delta-9-tetrahydrocannabinol (THC), full-spectrum products contain naturally occurring cannabinoids and have been reported to show improved efficacy or tolerability, attributed to synergy between cannabinoids and other components in the cannabis plant. Although ‘synergy’ indicates that two or more active compounds may produce an additive or combined effect greater than their individual analgesic effect, potentiation of the biological effect of a compound by related but inactive compounds, in combination, was termed the ‘entourage effect’. Here, we review current evidence for potential synergistic and entourage effects of cannabinoids in pain relief. However, definitive clinical trials and in vitro functional studies are still required.
... This is achieved via the plastidial deoxyxylulose phosphate/methyl-erythritol phosphate (DOXP/MEP) pathway (monoterpenoids), and the cytoplasmic mevalonate (MVA) pathway (sesquiterpenoids, triterpenoids, and sterols) [2]. Along with cannabinoids, terpenes are considered a main physiological marker of secondary metabolites [15,16]. ...
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The cannabis plant (Cannabis sativa L.) produces an estimated 545 chemical compounds of different biogenetic classes. In addition to economic value, many of these phytochemicals have medicinal and physiological activity. The plant is most popularly known for its two most-prominent and most-studied secondary metabolites—Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD). Both Δ9-THC and CBD have a wide therapeutic window across many ailments and form part of a class of secondary metabolites called cannabinoids—of which approximately over 104 exist. This review will focus on non-cannabinoid metabolites of Cannabis sativa that also have therapeutic potential, some of which share medicinal properties similar to those of cannabinoids. The most notable of these non-cannabinoid phytochemicals are flavonoids and terpenes. We will also discuss future directions in cannabis research and development of cannabis-based pharmaceuticals. Caflanone, a flavonoid molecule with selective activity against the human viruses including the coronavirus OC43 (HCov-OC43) that is responsible for COVID-19, and certain cancers, is one of the most promising non-cannabinoid molecules that is being advanced into clinical trials. As validated by thousands of years of the use of cannabis for medicinal purposes, vast anecdotal evidence abounds on the medicinal benefits of the plant. These benefits are attributed to the many phytochemicals in this plant, including non-cannabinoids. The most promising non-cannabinoids with potential to alleviate global disease burdens are discussed.
... Studies have reported that terpenoids are powerful metabolites that have an interactive effect (or an "entourage effect") with cannabinoid receptors (Gertsch et al., 2008). However, terpene composition in cannabis resin is dependent upon genetic, environmental, and developmental factors, and highly variable terpene profiles additionally exist between individual plants (Fischedick et al., 2010;Hazekamp and Fischedick, 2012;Booth et al., 2017). Terpene diversity in cannabis resin is responsible for scent and flavor qualities of cannabis flowers (Booth et al., 2017). ...
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Differences in cannabis (Cannabis sativa L.) plant chemistry between strains are influenced by genetics, plant growth and development, in addition to environmental conditions such as abiotic and biotic stress. Resulting secondary metabolite profiles are further altered post-harvest by drying and extraction, all of which present sizable challenges to industrial scale producers of pharmaceutical grade products in Canada and elsewhere. Consistent quantity and quality of cannabis extracts, demonstrated by preferred cannabinoid ratios and other secondary metabolites present are important, particularly as the list of therapeutic uses for medical cannabis is expanding. As more countries contemplate the legalization and licensure of medical cannabis, the number of cannabis extraction and testing laboratories is increasing to keep up with demand. However, it is not always known what standards are adhered to, resulting in numerous non-validated methods. In this review, a summary of cannabis chemistry and biosynthesis of secondary compounds is provided, and post-harvest processing practices occurring along the cannabis product value chain that might affect cannabis phytochemistry, potency and volatility are presented. An emphasis is placed on improved drying and extraction methods for plant material suitable for the cannabis industry. Finally, new approaches to secondary metabolite profiling for cannabis products are compared.
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As the frequency of cannabis-based therapy increases, the ability to distinguish intake of cannabis-based medicines from recreational cannabis use becomes desirable. Minor cannabinoids have been suggested to indicate recreational cannabis use in biological matrices but are unreliable when presumably also present in directly plantderived medicines. Thus, for therapeutics such as medical cannabis, Sativex and Dronabinol, a more thorough investigation of cannabinoid profiles is required to identify possible distinguishing markers. In this study, 16 phytocannabinoids were quantified in samples of seized and medical cannabis, Sativex and Dronabinol from two different manufacturers, using a validated LC-MS/MS method. Analytes included delta-9- tetrahydrocannabinol, tetrahydocannabinolic acid A, cannabidiol, cannabidiolic acid, cannabigerol, cannabigerolic acid, cannabinol, cannabinolic acid, cannabichromene, canabichromenic acid, cannabicyclol, cannabicyclolic acid, tetrahydrocannabivarin, tetrahydrocannabivarinic acid, cannabidivarin and cannabidivarinic acid. Resultant cannabinoid profiles were compared, and markers were suggested. Characteristics of Sativex included a specific cannabidiol/tetrahydrocannabinol ratio and presence of cannabichromene, while acidic cannabinoids, cannabigerol and cannabinol occurred in only low amounts. As expected, the predominant ingredient in Dronabinol was tetrahydrocannabinol, but minor cannabinoids were quantified as well. Medical marihuana and seized cannabis were compared separately in a principal component analysis. Several medical marihuana varieties were found to significantly differ from seized cannabis, mostly regarding contents of tetrahydocannabinolic acid A and tetrahydrocannabivarinic acid and cannabidiolic and cannabidivarinic acid respectively.
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Background: Phytocannabinoids naturally occur in the cannabis plant (Cannabis sativa), and Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) predominate. There is a need for rapid inexpensive methods to quantify total THC (for statutory definition) and THC-CBD ratio (for classification into three chemotypes). This study explores the capabilities of a spectroscopic technique that combines ultraviolet-visible and fluorescence, absorbance-transmittance excitation emission matrix (A-TEEM). Methods: The A-TEEM technique classifies 49 dry flower extracts into three C. sativa chemotypes, and quantifies the total THC-CBD ratio, using validated gas chromatography (GC)-flame ionization (FID) and High-Performance Liquid Chromatography (HPLC) methods for reference. Multivariate methods used are principal components analysis for a chemotype classification, extreme gradient boost (XGB) discriminant analysis (DA) to classify unknown samples by chemotype, and XGB regression to quantify total THC and CBD content using GC-FID and HPLC data on the same samples. Results: The A-TEEM technique provides robust classification of C. sativa samples, predicting chemotype classification, defined by THC-CBD content, of unknown samples with 100% accuracy. In addition, A-TEEM can quantify total THC and CBD levels relevant to statutory determination, with limit of quantifications (LOQs) of 0.061% (THC) and 0.059% (CBD), and high cross-validation (>0.99) and prediction (>0.99), using a GC-FID method for reference data; and LOQs of 0.026% (THC) and 0.080% (CBD) with high cross-validation (>0.98) and prediction (>0.98), using an HPLC method for reference data. A-TEEM is highly predictive in separately quantifying acid and neutral forms of THC and CBD with HPLC reference data. Conclusions: The A-TEEM technique provides a sensitive method for the qualitative and quantitative characterization of the major cannabinoids in solution, with LOQs comparable with GC-FID and HPLC, and high values of cross-validation and prediction. As a spectroscopic technique, it is rapid, with data acquisition <45 sec per measurement; sample preparation is simple, requiring only solvent extraction. A-TEEM has the sensitivity to resolve and quantify cannabinoids in solution based on their unique spectral characteristics. Discrimination of legal and illegal chemotypes can be rapidly verified using XGB DA, and quantitation of statutory levels of total THC and total CBD comparable with GC-FID and HPLC can be obtained using XBD regression.
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Cannabis sativa L. is an industrial and medicinal crop that originated in Eurasia. Its female inflorescence is usually used as a drug because of its high content of Δ⁹-tetrahydrocannabinol (Δ⁹-THC) and cannabidiol (CBD), while the stem and leaf have great potential as sustainable sources of fiber and fuel. In this study, we used an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF MS)-based untargeted metabolomics strategy to profile the metabolome differences among those Cannabis leaves collected before flowering from Xinjiang, Qinghai and Sichuan Provinces. Kruskal–Wallis test, principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were applied to compare and discover the chemical characterization of the collected samples. Finally, potential high-contribution features were identified as 13 flavonoids (including C-glycoside and O-glycoside) and 1 cannabinoid based on high-resolution MS and MS/MS information, in which 4 flavonoids were unambiguously identified by reference standards. Cannabis leaves from Xinjiang and Qinghai accumulated higher flavonoid glucuronides, while flavonoid glycosides were more abundant in those from Sichuan. However, for the cannabinoids, such as Δ⁹-THC and CBD, there were no significant differences in leaves, even those samples shown difference genotypes. Our results indicating that flavonoids variation in Cannabis leaves has potential ability of chemotypic marker and geographical marker, and may improve the practical and economic value of Cannabis leaves.
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Medicinal cannabis (Cannabis sativa L.) is a growing agro-industrial sector with the end product required to be free of pesticides. C. sativa is covered by specialized hairs called trichomes, namely, two types of non-glandular and three types of glandular trichomes. Despite the great importance of biological pest control in medicinal cannabis little is known about the impact of cannabis trichomes on natural enemies´ mobility and their interaction with prey. In the current study, by employing a video recording set-up, we determined the mobility of Aphidoletes aphidimyza and Chrysoperla carnea larvae and their interaction with the aphid pests Phorodon cannabis and Aphis gossypii on C. sativa leaf disks and inflorescences. As benchmark, we tested the same parameters on sweet pepper leaf disks, a benign host plant for both predators, and without trichomes. It was found that A. aphidimyza females readily oviposited on P. cannabis colonies developing on C. sativa plants. On C. sativa leaves, covered by non-glandular trichomes, the larvae of both predators were able to move and prey upon aphids. On C. sativa inflorescences, where glandular trichomes prevailed, the larvae of A. aphidimyza were generally inactive, while the C. carnea larvae were still able to move and interact with prey albeit to a lesser degree compared to the cannabis leaves. We suggest that the use of A. aphidimyza and C. carnea in augmentative biological control programs in medicinal cannabis should depend on the crop´s growth stage; the former should be employed during the vegetative and the latter during vegetative and flowering crop stages.
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Introduction: As Cannabis sativa L. (Cannabaceae) ages, inflorescence phytochemicals are susceptible to oxidative degradation. Reduction of Δ9-tetrahydrocannabinol (Δ9-THC) content has the potential to impact the reliability and accuracy of dosing. Advances that improve cannabinoid stability during storage would have an important impact in medical cannabis markets. Reported here is the use of C. sativa terpenes with antioxidant properties that improve inflorescence cannabinoid stability. Materials and Methods: Killer Kush inflorescence samples were stored in a temperature-controlled environment, in opaque jars. To accelerate the rate of oxidate degradation, samples were stored with the oxidizing agent hydrogen peroxide. Vapor phase terpenes were added to inflorescence packaging. Two terpene blends and three different dosage amounts were evaluated. Inflorescence stability samples were prepared in triplicate for each sample type. Cannabinoid content was quantitatively assessed after 24, 81, and 127 days of storage using high-performance liquid chromatography. Terpene content was assessed using headspace gas chromatography mass spectrometry. Results from inflorescence stored with and without external terpenes were compared by analysis of variance (ANOVA) data processing. Results: After 127 days of storage, inflorescence in the accelerated study experienced a loss of 18.0% and 34.3% total Δ9-THC content for samples stored with and without external terpenes, respectively. The differences in cannabinoid content were found to be statistically significant at all timepoints using ANOVA processing. In the nonaccelerated study, only one of the six sample types investigated had a statistically significant greater total Δ9-THC content than control at all timepoints. Nevertheless, a dose-dependent relationship between the amount of external terpenes added to inflorescence and the preservation of total Δ9-THC content was observed. Discussion: In the accelerated study, exogenous terpenes reduced the degradation of inflorescence cannabinoid content by 47.4%. This represents the first reported addition of terpene antioxidants to inflorescence packaging for cannabinoid preservation. Of note, the antioxidants used in this system can be obtained from C. sativa. This is advantageous from a toxicological perspective as inhaling synthetic antioxidants presents unknown and unpredictable risks. When fully developed, the novel system has applications for inflorescence packaged for individual sale, as well as long-term storage of bulk biomass.
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Polymyxins have resurged as the last-resort antibiotics against multidrug-resistant Acinetobacter baumannii. As reports of polymyxin resistance in A. baumannii with monotherapy have become increasingly common, combination therapy is usually the only remaining treatment option. A novel and effective strategy is to combine polymyxins with non-antibiotic drugs. This study aimed to investigate, using untargeted metabolomics, the mechanisms of antibacterial killing synergy of the combination of polymyxin B with a synthetic cannabidiol against A. baumannii ATCC 19606. The antibacterial synergy of the combination against a panel of Gram-negative pathogens (Acinetobacter baumannii, Klebsiella pneumoniae and Pseudomonas aeruginosa) was also explored using checkerboard and static time-kill assays. The polymyxin B–cannabidiol combination showed synergistic antibacterial activity in checkerboard and static time-kill assays against both polymyxin-susceptible and polymyxin-resistant isolates. The metabolomics study at 1 h demonstrated that polymyxin B monotherapy and the combination (to the greatest extent) significantly perturbed the complex interrelated metabolic pathways involved in the bacterial cell envelope biogenesis (amino sugar and nucleotide sugar metabolism, peptidoglycan, and lipopolysaccharide (LPS) biosynthesis), nucleotides (purine and pyrimidine metabolism) and peptide metabolism; notably, these pathways are key regulators of bacterial DNA and RNA biosynthesis. Intriguingly, the combination caused a major perturbation in bacterial membrane lipids (glycerophospholipids and fatty acids) compared to very minimal changes induced by monotherapies. At 4 h, polymyxin B–cannabidiol induced more pronounced effects on the abovementioned pathways compared to the minimal impact of monotherapies. This metabolomics study for the first time showed that in disorganization of the bacterial envelope formation, the DNA and RNA biosynthetic pathways were the most likely molecular mechanisms for the synergy of the combination. The study suggests the possibility of cannabidiol repositioning, in combination with polymyxins, for treatment of MDR polymyxin-resistant Gram-negative infections.
Chapter
The eye, nose, and throat (ENT)-related diseases are a great problem in the pediatric population, but the mortality is low, whereas complication rates are increasing in spite of the improvements in health care facilities. In children, middle ear infection is the most common disease, the reason being alterations in the eustachian tube anatomy, being straighter in children than adults. Nearly 42 million people (age >3 years) are facing a hearing loss, mainly because of otitis media, second only to the common cold as a cause of infection in kids, also the commonest cause of mild-to-moderate hearing impairment in industrializing countries. In the population above 5 years of age, nearly 16% suffer from this disorder and more than 55% of these cases occur in school-going children, generally from the lower socioeconomic class (Idu et al. 2008; Baldry and Hind 2008; Zumbroich 2009; Nepali and Sigdel 2012). Respiratory tract symptoms such as cough, sore throat, and earache are also frequent in children. Upper respiratory tract infections predispose a child to complications such as otitis media, tonsillitis, and sinusitis. Tonsillitis most often occurs in children, a condition rarely appreciated in kids below 2 years. Viral tonsillitis is more common in younger children, while tonsillitis caused by Streptococcus species typically occurs in children aged 5–15 years (Nepali and Sigdel 2012).
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The popularity of vaping cannabis products has increased sharply in recent years. In 2019, a sudden onset of electronic cigarette/vaping-associated lung injury (EVALI) was reported, leading to thousands of cases of lung illness and dozens of deaths due to the vaping of tetrahydrocannabinol (THC)-containing e-liquids that were obtained on the black market. A potential cause of EVALI has been hypothesized due to the illicit use of vitamin E acetate (VEA) in cannabis vape cartridges. However, the chemistry that modifies VEA and THC oil, to potentially produce toxic byproducts, is not well understood under different scenarios of use. In this work, we quantified carbonyls, organic acids, cannabinoids, and terpenes in the vaping aerosol of pure VEA, purified THC oil, and an equal volume mixture of VEA and THC oil at various coil temperatures (100-300 °C). It was found under the conditions of our study that degradation of VEA and cannabinoids, including Δ9-THC and cannabigerol (CBG), occurred via radical oxidation and direct thermal decomposition pathways. Evidence of terpene degradation was also observed. The bond cleavage of aliphatic side chains in both VEA and cannabinoids formed a variety of smaller carbonyls. Oxidation at the ring positions of cannabinoids formed various functionalized products. We show that THC oil has a stronger tendency to aerosolize and degrade compared to VEA at a given temperature. The addition of VEA to the e-liquid nonlinearly suppressed the formation of vape aerosol compared to THC oil. At the same time, toxic carbonyls including formaldehyde, 4-methylpentanal, glyoxal, or diacetyl and its isomers were highly enhanced in VEA e-liquid when normalized to particle mass.
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Cannabis has been at the center of scientific attention for some years now. Since its pharmacological potential has been highlighted, cannabis has become a hot topic in research laboratories, leading to the publication of many scientific studies. Focusing on analytical chemistry, an enormous number of analytical methods for cannabinoid (CNB) determination have been published, involving various tech-niques. However, no globally accepted reference method for CNB determination has yet been chosen.This review aims to identify very recent analytical methods developed to analyze phytocannabinoids in cannabis herbal samples. For certain techniques, stagnation in terms of employed operational conditions can be observed. In this context, a reference method of analysis should be proposed and accepted worldwide to standardize CNB determination. In contrast, for other techniques, we are witnessing a scientific ferment, which is resulting in the development of new interesting analytical options. In this regard, particular focus has been given to these niche techniques, which are now emerging in the analytical panorama of cannabis analysis, offering new important perspectives for the future of cannabis testing. Supercritical fluid chromatography and infrared spectroscopy showed tangible advantages when applied to CNB determination in herbal samples.
Chapter
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Leonotis leonurusL. R. Br. (Lamiaceae) is a medicinal plant native to the South Africancontinent also employed as a recreational drug and a substitute to Cannabis sativaL. (Cannabaceae). Given the interest of the last mentioned species as a source of treatments for epilepsy among many other pathologies and its possible substitution for L. leonurus, the aim of this article is obtain anatomical and micrographical characters for its identification in chopped or powdered material and to survey the user ́s perceptions about this plant based in posts extracted from a recreational drug user Internet forum. L. leonurusleaves have pluricellular tector trichomes and two classes of pluricellular trichomes with unicellular and pluricellular heads, styloid crystals in its mesophyll among many other characters, while the flowers have wooly trichomes and characteristic pollen granules. Regarding the Internet forum survey, it was reported that L. leonurusleaves and flowers were the employed parts and that the mode of use was smoked. The reported effect was sedative. The anatomical data reported in this article may help to identify L. leonurusin pharmaceutical or forensic contexts.
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Coincident with the cannabis legalization and the increased interest in the medicinal use of the plant, the cannabis marketplace and farming have seen tremendous growth. It is reported that there are more than 2000 cannabis varieties available to customers. However, the data that is available to the growers and breeders regarding the cannabinoid contents of various varieties remains low. Here, a high-performance liquid chromatography (HPLC) method was developed and validated for the simultaneous separation and determination of 11 cannabinoids. A total of 104 hemp bud materials belonging to 20 varieties were collected from farms in the state of Maryland and analyzed with the HPLC method. The contents of the 11 cannabinoids in various varieties were compared and discussed, highlighting the varieties that showed a high yield of cannabinoids and good consistency that are more appropriate for cannabinoid production.
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In recent years, hemp oils have become ubiquitous in health products on the European market. As the trend continues to grow and more cannabinoids are researched for their therapeutic benefits, more academic and industrial interests are drawn to this direction. Cannabidiol, Δ9-tetrahydrocannabinol, and their acidic forms remain the most examined cannabinoids in hemp and cannabis oils, in the case of cannabidiol due to its proven health implications in numerous articles, and in the case of Δ9-tetrahydrocannabinol, due to the legislation in the European area. These oils sold on the internet contain a wide range of cannabinoids that could demonstrate their effects and benefits. As a result of these claims, we developed a robust and rapid method that can identify and quantify 10 of the most common cannabinoids found in hemp oils: cannabivarin, cannabidiolic acid, cannabigerolic acid, cannabigerol, cannabidiol, cannabinol, Δ9-tetrahydrocannabinol, Δ8-tetrahydrocannabinol, cannabichromene, and tetrahydrocannabinolic acid in less than 11 min, with reverse-phase–high-performance liquid chromatography–photodiode matrix system (RP–UHPLC–PDA) equipped with C18 column, eluting in a gradient using water and acetonitrile with formic acid as mobile phases. The quantification of 9 sample products presented in different matrixes was performed using a calibration curve obtained by analyzing standard solutions from a 10-cannabinoid-mix-certified reference standard. The developed method demonstrated the ability to identify and quantify the main cannabinoids in hemp oil and is a useful tool for pharmaceutical professionals.
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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.
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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.
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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.
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. 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].
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
Article
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.
Article
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.
Article
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.
Article
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.
Article
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.
Article
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.