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Tea is a recommended way of administration of prescribed cannabis plant products in Denmark. We aimed to investigate the cannabinoid and terpene doses contained in different teas. We analysed tetrahydrocannabinol (THC), tetrahydrocannabinolic acid (THCA), cannabidiol (CBD), cannabidiolic acid (CBDA), and terpene concentrations in three repeated preparations of each type of tea, and in plant material. In standard tea, concentrations of THC were [median (min‐max)] 9.5 (2.3–15), 19 (13–34), and 36 (26–57) μg/mL for products with a labelled content of 6.3%, 14%, and 22% total THC (THC + THCA), respectively. The CBD concentration in tea from a product labelled with 8% total CBD (CBD + CBDA) was 7.5 (1.9–10) μg/mL. Based on this, the recommended starting amount of 0.2 L of the different teas would contain between 0.46 and 11.3 mg THC, and 0.38 to 2.0 mg CBD. Adding creamer before, but not after boiling, increased the THC and CBD concentration 2.3–4.4 and 2.1‐fold, respectively. Terpenes were detected in plant material, but not in tea. The study elucidates THC and CBD doses in different teas, which may assist the clinician's choice of cannabis product. Moreover, it underscores the need for caution as administration as tea can result in exposure to different doses, even when the same cannabis product is used. [ABSTRACT FROM AUTHOR]

Interest in cultivating cannabis for medical and recreational purposes is increasing due to a dramatic shift in cannabis legislation worldwide. Therefore, a comprehensive understanding of the composition of secondary metabolites, cannabinoids, and terpenes grown in different environmental conditions is of primary importance for the medical and recreational use of cannabis. We compared the terpene and cannabinoid profiles using gas/liquid chromatography and mass spectrometry for commercial cannabis from genetically identical plants grown indoors using artificial light and artificially grown media or outdoors grown in living soil and natural sunlight. By analyzing the cannabinoids, we found significant variations in the metabolomic profile of cannabis for the different environments. Overall, for both cultivars, there were significantly greater oxidized and degraded cannabinoids in the indoor-grown samples. Moreover, the outdoor-grown samples had significantly more unusual cannabinoids, such as C4- and C6-THCA. There were also significant differences in the terpene profiles between indoor- and outdoor-grown cannabis. The outdoor samples had a greater preponderance of sesquiterpenes including β-caryophyllene, α-humulene, α-bergamotene, α-guaiene, and germacrene B relative to the indoor samples. [ABSTRACT FROM AUTHOR]

We have demonstrated in previous studies that the essential macro-nutrients nitrogen (N) and phosporous (P) have profound effects on the production of cannabinoids and terpenoids in the cannabis plant. The present study was undertaken to evaluate the hypothesis that potassium (K) supply, which is known to substantially affect plant development and function, affects the secondary metabolism of the cannabis plant. Two cultivars of medical cannabis were grown in controlled environment conditions, under five levels of K supply: 15, 60, 100, 175, and 240 mg L−1 K. The results revealed that the development and function of plants that received the low K supply of 15 mg L−1 K were impaired, as the plants suffered from visual chlorosis, and the inflorescence yield was reduced in both cultivars. Plants that received higher K inputs in the range of −175 mg L−1 K demonstrated optimal plant function and high yield, and one cultivar demonstrated over-supply symptoms under the high K level of 240 mg L−1. The concentrations of most cannabinoids and terpenoids declined with the elevation of K supply, thus supporting the hypothesis. As secondary metabolite concentrations decreased with the increase in K supply, and higher K levels had no positive effects, 60 mg L−1 K is the suggested application level to maintain high function and yield combined with high secondary metabolism. [ABSTRACT FROM AUTHOR]

Conventional cannabis drying is time-consuming and energy intensive. A quick and dependable drying of cannabis is essential to ensure high-quality products to meet increasing demands. This study explored a combined microwave and infrared (MI) drying on cannabis comparing with control environmental (CE) drying. MI was very efficient with a short drying time of 16–200 min, high moisture diffusivity of 7.95×10−09-8.70×10−08 m2/s, and low energy consumption of 390.49–1611.42 kJ. The Page and Modified Page drying models fitted well to describe and predict the MI drying characteristics of cannabis. Microstructural images identified shrinkage in glandular trichomes of cannabis, whereas colorimetric assessment resulted in alteration of color attributes due to MI drying. It also facilitated the conversion of acidic cannabinoids to their neutral forms by decreasing tetrahydrocannabinolic acid (THCA) in g/g of dry matter from 20.15% to 7.57% and increasing tetrahydrocannabinol (THC) from 6.31% to 16.65% that insignificantly (p ≥0.05) affected the total THC level. MI drying resulted in a total terpenes concentration (%g/g of dry matters) of 0.541–0.730, insignificantly lower than CE drying (0.768). Overall, the study highlights MI as a rapid and energy-efficient drying for obtaining high quality cannabis, particularly for medicinal applications. [Display omitted] • The effects of microwave-infrared drying on medicinal cannabis were investigated. • Microwave-infrared drying was quite faster and saved 95.54–98.92% energy. • Scanning electron microscopy shows shrinkage and damage of cannabis trichomes. • Tetrahydrocannabinol content increased from 6.31% to 16.65%. • Total terpenes of dried samples were statistically similar but lower than fresh. [ABSTRACT FROM AUTHOR]

Cannabis indica obtained from the Office of the Narcotics Control Board (ONCB) of Thailand was examined for medical purposes. The ONCB cannabis was seized and distributed to our research group for investigation of chemical compositions and purification of major cannabinoids. Its essential oil was analysed and aromadendrene was identified as the major terpene. Small amounts of cannabinoids were also observed. The components of the seized cannabis were extracted using ethanolic extraction and supercritical CO2 extraction. All crude extracts were analysed using high-performance liquid chromatography coupled with a cannabinoid analyser. All extracts contained cannabidiol (CBD), cannabinol (CBN), and Δ99-tetrahydrocannabinol (Δ9-THC) as the major constituents. A preliminary study showed notable anticancer activities of CBD, CBN and Δ99-THC against cervix carcinoma cell and epithelial human breast cancer cell. [ABSTRACT FROM AUTHOR]

Aims: Soils represent the natural habitat of entomopathogenic nematodes (EPNs). When moving in soil, EPNs are oriented to follow a chemical signal (chemotaxis). Cannabis sativa L. is known to secrete a very wide spectrum of secondary metabolites. Ethanol extracts (EE) of different C. sativa L. organs were used to study the effect on EPN chemotaxis. Methods: The root, leaf and inflorescence EE of two hemp varieties and two medical cannabis breeding lines were used in laboratory chemotaxis assays with three EPN species (Steinernema carpocapsae, S. feltiae and Heterorhabditis bacteriophora) at 20 and 25 °C. The content of terpenes and cannabinoids in C. sativa L. inflorescences was measured as well. Results: Overall, EPNs were most attracted or repelled by inflorescence extracts, followed by leaves and roots. The most abundant terpene in hemp inflorescences was trans caryophyllene (38.2% on average), the highest contents of total cannabidiol, total cannabigerol and total tetrahydrocannabinol were 9.65%, 0.89% and 0.62%, respectively. Conclusions: The attraction effect of S. sativa L. EE is a highly interesting outcome and could lead to the development of attractant compounds in EPN biological control. Cannabinoids and terpenes may be responsible for pronounced effects on EPN chemotaxis. [ABSTRACT FROM AUTHOR]

Summary: The cannabis leaf is iconic, but it is the flowers of cannabis that are consumed for the psychoactive and medicinal effects of their specialized metabolites. Cannabinoid metabolites, together with terpenes, are produced in glandular trichomes. Superficially, stalked and sessile trichomes in cannabis only differ in size and whether they have a stalk. The objectives of this study were: to define each trichome type using patterns of autofluorescence and secretory cell numbers, to test the hypothesis that stalked trichomes develop from sessile‐like precursors, and to test whether metabolic specialization occurs in cannabis glandular trichomes. A two‐photon microscopy technique using glandular trichome intrinsic autofluorescence was developed which demonstrated that stalked glandular trichomes possessed blue autofluorescence correlated with high cannabinoid levels. These stalked trichomes had 12–16 secretory disc cells and strongly monoterpene‐dominant terpene profiles. In contrast, sessile trichomes on mature flowers and vegetative leaves possessed red‐shifted autofluorescence, eight secretory disc cells and less monoterpene‐dominant terpene profiles. Moreover, intrinsic autofluorescence patterns and disc cell numbers supported a developmental model where stalked trichomes develop from apparently sessile trichomes. Transcriptomes of isolated floral trichomes revealed strong expression of cannabinoid and terpene biosynthetic genes, as well as uncharacterized genes highly co‐expressed with CBDA synthase. Identification and characterization of two previously unknown and highly expressed monoterpene synthases highlighted the metabolic specialization of stalked trichomes for monoterpene production. These unique properties and highly expressed genes of cannabis trichomes determine the medicinal, psychoactive and sensory properties of cannabis products. Significance Statement: The unique monoterpene‐rich cannabis stalked trichomes that determine the medicinal, psychoactive and sensory properties of cannabis flowers develop from sessile‐like intermediates. [ABSTRACT FROM AUTHOR]

The author reflects on the differences between the cannabinoids tetrahydrocannabinol (THC) and cannabidiol (CBD) that are oppositional in clinical and pharmadynamic effects. Topics discussed include the insufficient knowledge on cannabinoid components in cannabis that restricts their U.S. Food and Drug Administration (FDA) approval, and the need for federal regulation and taxation of cannabis similar to tobacco products and alcohol. Also noted are recommendations on cannabis regulation.

Perhaps no topic is more controversial than the use of marijuana in clinical practice. Within the United States, there are an estimated 55 million recent active users, defined as one to two uses within the previous year, and 35 million regular users, defined as one to two uses per month. [ABSTRACT FROM AUTHOR]

Cannabis sativa L. is used to treat a wide variety of medical conditions, in light of its beneficial pharmacological properties of its cannabinoids and terpenes. At present, the quantitative chemical analysis of these active compounds is achieved through the use of laborious, expensive, and time-consuming technologies, such as high-pressure liquid-chromatography- photodiode arrays, mass spectrometer detectors (HPLC-PDA or MS), or gas chromatography-mass spectroscopy (GC-MS). Hence, we aimed to develop a simple, accurate, fast, and cheap technique for the quantification of major cannabinoids and terpenes using Fourier transform near infra-red spectroscopy (FT-NIRS). FT-NIRS was coupled with multivariate classification and regression models, namely partial least square-discriminant analysis (PLS-DA) and partial least squares regression (PLS-R) models. The PLS-DA model yielded an absolute major class separation (high-THC, high-CBD, hybrid, and high-CBG) and perfect class prediction. Using only three latent variables (LVs), the cross-validation and prediction model errors indicated a low probability of over-fitting the data. In addition, the PLS-DA model enabled the classification of chemovars with genetic-chemical similarities. The classification of high-THCA chemovars was more sensitive and more specific than the classifications of the remaining chemovars. The prediction of cannabinoid and terpene concentrations by PLS-R yielded 11 robust models with high predictive capabilities (R 2 CV and R 2 pred > 0.8, RPD >2.5 and RPIQ >3, RMSECV/RMSEC ratio <1.2) and additional 15 models whose performance was acceptable for initial screening purposes (R 2 CV > 0.7 and R 2 pred < 0.8, RPD >2 and RPIQ <3, 1.2 < RMSECV/RMSEC ratio <2). Our results confirm that there is sufficient information in the FT-NIRS to develop cannabinoid and terpene prediction models and major-cultivar classification models. Dried and homogenously ground cannabis inflorescence samples were directly analyzed using NIR spectrometer and then extracted with ethanol for the determination of cannabinoids and terpenes using HPLC-PDA and GC-MS, respectively. Finally, NIR spectra and the chemical composition of the samples were used to build PLS-R and PLS-DA models. [Display omitted] • FT-NIR based PLS-R model enables accurate quantification of cannabinoid and terpene. • FT-NIR based PLS-DA model enables accurate classification of cannabis chemovars. • FT-NIR has been proven as a promising fast alternative to chromatographic analysis. [ABSTRACT FROM AUTHOR]

Cannabis is used to treat various medical conditions, and lines are commonly classified according to their total concentrations of Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD). Based on their ratio of total THC to total CBD, cannabis cultivars are commonly classified into high-THC, high-CBD, and hybrid classes. While cultivars from the same class have similar compositions of major cannabinoids, their levels of other cannabinoids and their terpene compositions may differ substantially. Therefore, a more comprehensive and accurate classification of medicinal cannabis cultivars, based on a large number of cannabinoids and terpenes is needed. For this purpose, three different chemometric-based classification models were constructed using three sets of chemical profiles. We examined those models to determine which provides the most accurate "chemovar" classification. This was done by analyzing profiles of cannabinoids, terpenes, and the combination of these substances using the partial least square-discriminant analysis multivariate (PLS-DA) technique. The chemical profiles were selected from the three major classes of medicinal cannabis that are most commonly prescribed to patients in Israel: high-THC, high-cannabigerol (CBG), and hybrid. We studied the correlations between cannabinoids and terpenes to identify major bio-indicators representing the plant's terpene and cannabinoid content. All three PLS-DA models provided highly accurate classifications, utilizing six to nine latent variables with an overall accuracy ranging from 2 to 11% CV. The PLS-DA model applied to the combined cannabinoid-and-terpene profile did the best job of differentiating between the chemovars in terms of misclassification error, sensitivity, specificity, and accuracy. The combined cannabinoid-and-terpene PLS-DA profile had cross-validation and prediction misclassification errors of 4% and 0%, respectively. This is the first study to demonstrate the highly accurate classification of samples of medicinal cannabis based on their cannabinoid and terpene profiles, as compared to cannabinoid profiles alone. Furthermore, our correlation analysis indicated that 11 cannabinoids and terpenes might serve as bio-indicators for 32 different active compounds. These findings suggest that the use of multivariate statistics could assist in breeding studies and serve as a tool for minimizing the mislabeling of cannabis inflorescences. [Display omitted] • Classification of major cannabis classes was good for all data sets. • Terpene-cannabinoid–based PLS-DA provided the best chemovar classification. • Eleven potential bio-indicators predicted 32 cannabinoids and terpenes. [ABSTRACT FROM AUTHOR]

Cannabis cultivation for medical purposes in Brazil has been increased in the last years. While cannabis crops are prohibited, hundreds patients have been granted with judicial authorizations and there is little information about the cultivation conditions, yields and chemical profiles of the plants. Cannabis plants contain hundreds of compounds, with cannabinoids and terpenes the main drivers of their toxicological and pharmacological properties. Besides the cannabinoids, terpene contents are useful for the chemotaxonomic classification of different varieties, and their role in forensic analyses should be further delineated. The present study monitored cannabis crops of fifteen participants who were granted special licenses by the Brazilian Courts in Rio de Janeiro and São Paulo. The cultivation conditions were monitored and five cannabinoids (tetrahydrocannabinol acid-THCA, tetrahydrocannabinol-THC, cannabidiolic acid-CBDA, cannabidiol-CBD and cannabinol-CBN) and nineteen terpenes were quantified in cannabis flowers. The total grow cycle of thirty-five cannabis plants ranged from 10 to 24 weeks. The dry flower yields ranged 22-90 g per plant. Most cannabis specimens were CBD-rich varieties (CBD levels from 1.6% to 16.7%, and THC levels from 0.0% to 2.6%, n = 22) used to treat epileptic patients. The THC-rich varieties contained CBD levels ranging from 0.03% to 0.8%, and THC levels from 0.7% to 20.1%, n = 11. Fewer of the samples contained THC:CBD ratios of approximately 1:1 (CBD levels of 3.3-3.8% and THC levels of 2.2-3.7%, n = 2). The most abundant terpenes in the cannabis flowers were beta-caryophyllene, alpha-humulene, guaiol and alpha-bisabolol. CBD-rich varieties showed significant higher levels of beta-caryophyllene and alpha-humulene in comparison with THC-rich varieties. Overall, the study herein provides data concerning medical cannabis crops grown in a region of Brazil that not only guide individual medical cannabis cultivation methods but also aid forensic analyses. [ABSTRACT FROM AUTHOR]

The purpose of this analytical study was to develop an advanced formulation of medical Cannabis oil (MCO) comparing the chemical profile of different extracts obtained with two existing methods (SIFAP and CALVI) and one original upgraded (CERFIT) method. Preparation methods were applied with varying solvent, temperature, and duration of the decarboxylation and extraction steps. HPLC-MS/MS TSQ and GC/FID-HS analyses were performed to investigate cannabinoid and terpene contents in the three oil extracts. Cannabinoids profile remained comparable between the formulations. CERFIT extracts exhibited a superior quantity of total terpene hydrocarbon forms (e.g., limonene and α-pinene) with no degradation occurrence (i.e., oxidized terpenes not quantifiable). Thus, this new method optimized the phytochemical profile of the MCO presenting a value opportunity to obtain a standardized high-level therapeutic product. [ABSTRACT FROM AUTHOR]

Cannabis (Cannabis sativa L.) for medical purposes has been legalized again in many countries in recent years. Currently, only two major cannabinoids (Δ9-THC and CBD) are considered in the legislation and medication, which is not sufficient in case of dried plant material or resulting extract. Other substances (mainly terpenes/terpenoids), or their specific combinations, could influence the resulting therapeutic effect for specific oncology diagnosis and specific patients. Six different genotypes (Conspiracy Kush, Jilly Bean, Jack Cleaner 2, Jack Skellington, Nordle and Nurse Jackie) were cultivated indoor at the Czech University of Life Sciences Prague. Ethanol extracts taken from the inflorescences were assayed for their content of main cannabinoids and terpenes/terpenoids. The extracts were used for in vitro cytotoxicity studies on hepatocarcinoma human cell lines Hep-G2 and colorectal carcinoma human cell lines Caco-2 and Ht-29. Healthy lung fibroblast MRC-5 and healthy intestinal cells FHs 74 Int were used to compare selectivity of cytotoxicity. The average content of Δ9-THC in extracts was 59.1 ± 2.43%, and of CBD 1.84 ± 0.17%. The content of main cannabinoids in the Nurse Jackie genotype extract was significantly greater than that of the other genotypes. Overall, more than 60 different terpenes/terpenoids were identified in the extracts. The major terpenes/terpenoids detected in most genotypes were limonene, linalool, α-terpineol, β-caryophyllene, trans -α-bergamotene, α-humulene, β-caryophyllene oxide, guaiol, γ-eudesmol, β-eudesmol and α-bisabolol. Differences in the terpene composition of individual genotypes were caused by minor terpenoids, such as β-ocimene, isopulegol acetate, β-elemene, β-selinene and spathulenol. All extracts were highly cytotoxic to Ht-29 colorectal carcinoma cells and showed positive selectivity compared to healthy FHs 74 Int colon cells. The Jack Cleaner 2 extract was cytotoxic to all cell lines tested at the lowest concentrations (8.48 ± 2.4–16.14 ± 0,07 μg/mL), but was positively selective only for colorectal cancer cells, especially Ht-29 and to a lesser extent for Caco-2. Similarly, the Nordle extract showed positive selectivity for Ht-29 and Caco-2 only. Jilly Bean was unique in this study, in that its extract functioned on all cell lines at the highest concentrations (20.13 ± 3.05–49.88 ± 1.5 μg/mL), whilst also being highly positively selective in all carcinoma lines (Ht-29, Caco-2 and Hep-G2 hepatocarcinoma) compared to healthy cell lines (FHs 74 Int and MRC-5). The results suggest that Δ9-THC and CBD are responsible for the in vitro cytotoxicity of the extracts, but observed differences in selectivity reveal their synergies with other substances. According to chemical analysis, higher concentrations of myrcene, β-elemene, β-selinene and α-bisabolol oxide found in the Jilly Bean genotype may positively affect the selectivity of cytotoxic activity. It is therefore vital that similar studies are performed on other cell lines, in order to be able to recommend these cannabis genotypes for preclinical and clinical studies, which are still lacking. • Two major cannabinoids and more than 60 terpenes/terpenoids have been identified in extracts. • Composition of cannabis extracts modifies cytotoxicity on various human cell lines. • All tested extracts were selectively cytotoxic to colorectal cancer cells Ht-29. • One of the six extracts tested was selectively cytotoxic for all cancer lines tested. [ABSTRACT FROM AUTHOR]