Cannabis Varieties Research Articles

The Neurocognitive Effects of Cannabis Across the Lifespan.

This review examines the neurocognitive effects of cannabis and relevant developmental factors across adolescence (age 13-21), adulthood (21-65), and older adulthood (65+). Cannabis use is robustly associated with poorer neurocognitive functioning; however, studies that carefully control for confounds have often not found any evidence for impairment. Notably, the endocannabinoid system may underly how cannabis use affects neurocognitive functions, including heightened vulnerability during adolescence. In contrast, the endocannabinoid system may underlie protective neurocognitive effects of cannabis in older adults. Notably, older adults have reported sharp increases in recent cannabis use. As legalization increases the accessibility, variety, and potency of cannabis, strong empirical evidence is needed to understand its neurocognitive effects across the lifespan. In particular, rigorous study designs are needed to investigate the neurocognitive effects of cannabis, including among vulnerable populations (adolescents, older adults) and mediating (e.g., endocannabinoid system) and moderating factors (e.g., alcohol use).

Genomic Evidence That Governmentally Produced Cannabis sativa Poorly Represents Genetic Variation Available in State Markets.

The National Institute on Drug Abuse (NIDA) is the sole producer of Cannabis for research purposes in the United States, including medical investigation. Previous research established that cannabinoid profiles in the NIDA varieties lacked diversity and potency relative to the Cannabis produced commercially. Additionally, microsatellite marker analyses have established that the NIDA varieties are genetically divergent form varieties produced in the private legal market. Here, we analyzed the genomes of multiple Cannabis varieties from diverse lineages including two produced by NIDA, and we provide further support that NIDA’s varieties differ from widely available medical, recreational, or industrial Cannabis. Furthermore, our results suggest that NIDA’s varieties lack diversity in the single-copy portion of the genome, the maternally inherited genomes, the cannabinoid genes, and in the repetitive content of the genome. Therefore, results based on NIDA’s varieties are not generalizable regarding the effects of Cannabis after consumption. For medical research to be relevant, material that is more widely used would have to be studied. Clearly, having research to date dominated by a single, non-representative source of Cannabis has hindered scientific investigation.

Cannabinoids vs. whole metabolome: Relevance of cannabinomics in analyzing Cannabis varieties

Cannabis sativa has a long history of domestication both for its bioactive compounds and its fibers. This has produced hundreds of varieties, usually characterized in the literature by chemotypes, with Δ9-THC and CBD content as the main markers. However, chemotyping could also be done based on minor compounds (phytocannabinoids and others). In this work, a workflow, which we propose to name cannabinomics, combines mass spectrometry of the whole metabolome and statistical analysis to help differentiate C. sativa varieties and deciphering their characteristic markers. By applying this cannabinomics approach to the data obtained from 20 varieties of C. sativa (classically classified as chemotype I, II, or III), we compared the results with those obtained by a targeted quantification of 11 phytocannabinoids. Cannabinomics can be considered as a complementary tool for phenotyping and genotyping, allowing the identification of minor compounds playing a key role as markers of differentiation.

Separation and non-separation methods for the analysis of cannabinoids in Cannabis sativa L.

Cannabis sativa L. is a plant known all over the world, due to its history, bioactivity and also social impact. It is chemically complex with an astonishing ability in the biosynthesis of many secondary metabolites belonging to different chemical classes. Among them, cannabinoids are the most investigated ones, given their pharmacological relevance. In order to monitor the composition of the plant material and ensure the efficacy and safety of its derived products, extraction and analysis of cannabinoids play a crucial role. In this context, in addition to a conventional separation method based on HPLC with UV/DAD detection, a new strategy based on a non-separation procedure, such as 13C-qNMR, may offer several advantages, such as reduced solvent consumption and simultaneous acquisition of the quali/quantitative data related to many analytes. In the light of all the above, the aim of this work is to compare the efficiency of the above-mentioned analytical techniques for the study of the main cannabinoids in different samples of cannabis inflorescences, belonging to fibre-type, recreational and medical varieties. The 13C-qNMR method here proposed for the first time for the quantification of both psychoactive and non-psychoactive cannabinoids in different cannabis varieties provided reliable results in comparison to the more common and consolidated HPLC technique.

Δ9-cis-Tetrahydrocannabinol: Natural Occurrence, Chirality, and Pharmacology.

The cis-stereoisomers of Δ9-THC [(-)-3 and (+)-3] were identified and quantified in a series of low-THC-containing varieties of Cannabis sativa registered in Europe as fiber hemp and in research accessions of cannabis. While Δ9-cis-THC (3) occurs in cannabis fiber hemp in the concentration range of (-)-Δ9-trans-THC [(-)-1], it was undetectable in a sample of high-THC-containing medicinal cannabis. Natural Δ9-cis-THC (3) is scalemic (ca. 80-90% enantiomeric purity), and the absolute configuration of the major enantiomer was established as 6aS,10aR [(-)-3] by chiral chromatographic comparison with a sample available by asymmetric synthesis. The major enantiomer, (-)-Δ9-cis-THC [(-)-3], was characterized as a partial cannabinoid agonist in vitro and elicited a full tetrad response in mice at 50 mg/kg doses. The current legal discrimination between narcotic and non-narcotic cannabis varieties centers on the contents of "Δ9-THC and isomers" and needs therefore revision, or at least a more specific wording, to account for the presence of Δ9-cis-THCs [(+)-3 and (-)-3] in cannabis fiber hemp varieties.

Identification of Chemotypic Markers in Three Chemotype Categories of Cannabis Using Secondary Metabolites Profiled in Inflorescences, Leaves, Stem Bark, and Roots.

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.

Seed composition of non‐industrial hemp (Cannabis sativa L.) varieties from four regions in northern Morocco

SummaryThis study aims to determine the chemical composition of hemp (Cannabis sativa L.) seeds of two varieties cultivated in four different regions of northern Morocco. The content of protein, insoluble fibre, ash, total phenols and flavonoids was found to be 19.25%–24.18%, 26.40%–37.40%, 3.72%–5.39%, 134.57–199.90 mg 100g−1 and 39.40–69.54 mg 100g−1 respectively. Oil contents ranged from 26.42 to 35.19 g 100 g−1 of seeds in the four collection sites with variable oxidative stability at 100°C from 9.73 to 15.42 h. Total tocopherol content was between 376.46 and 796.06 mg kg−1 of oil, with a dominance of γ‐tocopherol. Fatty acids consisted mainly of unsaturated fatty acids (87.30‐88.96%) with a predominance of linoleic acid (48.26%–55.39%). Triacylglycerols (TAGs) composition analysis revealed the presence of 18 TAGs with an equivalent carbon number of 38–48. The composition of TAGs was predominated by LLL (14.34%–22.62%), OLL (12.77%–18.78%) and LLLn (12.32%–18.59%). Furthermore, the majority of the evaluated parameters were strongly influenced by the varietal factor, growing area and their interaction. The knowledge of this composition variability in seeds of non‐industrial Cannabis sativa varieties cultivated in northern Morocco could be useful in new Cannabis variety breeding programmes potentially valuable as non‐conventional oil in cosmetic and agro‐industry.

Cannabis Varieties Can Be Distinguished by Achene Shape Using Geometric Morphometrics.

Introduction: Cannabis plant uses are widespread across human cultures. The current tendency is to classify Cannabis varieties into chemovars upon their chemical fingerprint, mainly cannabinoids and terpenoids content. The identification of chemovars has important medical implications; however, their pharmacological characterization is costly and time consuming. The goal of this study was to assess whether achene shape variation could be related to Cannabis varieties with contrasting cannabinoid concentrations, as a first approach to chemovar identification. Methods: We used two-dimensional geometric morphometrics (GM) of the achenes and multivariate statistical analysis. We used achenes from five varieties, two from Type II chemotype (expressing both tetrahydrocannabinol [THC] and cannabidiol [CBD]), two Type I (THC-only), and one Type III (CBD-only). Results: The achenes from the different chemotypes were clearly distinguishable. No significant differences between varieties from the same chemotype were observed. The varieties with high THC concentration (Type I) were rounded and bigger, whereas achene from varieties containing only CBD (Type III) had a slender shape with smaller size. Conclusion: Achene shape variation is a potential biomarker of cannabinoid content in the plant flowers. Further studies are needed to confirm the suitability of GM methods for high-throughput screening of Cannabis cultivars, including larger diversity of varieties, and taking into account growth conditions, which can also influence plant chemical fingerprint.

Primary seed production of drug-free varieties of cannabis in the Middle Volga region

The article discusses the practical aspects of seed production of monoecious Ko noble planting. It is shown that the production of varietal seeds, maintenance of economically valuable traits, properties and preservation of biological purity of hemp varieties at a high level is possible only with a well-established seed production system. Long-term experimental data on the assessment of plants from 1-year breeding nurseries for the content of tetrahydrocannabinol (THC) by gas-liquid chromatography (GC) are presented. Techniques that ensure the formation of high varietal and sowing qualities as well as yield properties of seed material are considered.

Cannabinoid Content in Cannabis Flowers and Homemade Cannabis-Based Products Used for Therapeutic Purposes in Argentina.

Introduction: A recent law (DCTO-2020-883-APN-PTE-Law No. 27,350. Regulation) passed in Argentina put an end to the ban imposed for the last 60 years on cannabis cultivation within the country. The law permits restricted access to cannabis derivatives for medicinal, therapeutic, and palliative use by individuals and communities, allowing self- and community-based cannabis production. This is cause for concern in view of the lack of quality controls for cannabis derivatives. The several varieties of cannabis grown in Argentina have different chemical profiles and are processed in a variety of ways-mostly by alcohol extraction or maceration at different temperatures and for different amounts of times-making the cannabinoid content of these preparations highly variable. Determining the characteristics of home- and community-grown cannabis products will facilitate the implementation of public policies conducive to their safety and improvement. Objective: The aim of this study was to determine the cannabinoid chemotypes used for therapeutic purposes in Argentina and evaluate whether the cannabinoids present in homemade derivatives are comparable to those in commercially available products. Materials and Methods: High performance liquid chromatography with ultraviolet and diode array detector (HPLC/UV-DAD) analysis of 436 samples (oils, resins, and inflorescences) was carried out to determine the identity and concentration of five cannabinoids: tetrahydrocannabinolic acid (THCA), tetrahydrocannabinol (THC), cannabidiolic acid (CBDA), cannabidiol (CBD), and cannabinol (CBN). From three different sources, the samples represent the type of medical cannabis preparations to which patients have access. Results: The results indicate that the medium-to-low cannabinoid concentration in a significant number of homemade oil samples is similar to that found in commercial products. Most of the samples have a THC/CBD ratio >1 or only contain THC. Acidic cannabinoids were detected in homemade preparations, but were not reported in package inserts of commercial products. Conclusions: Our results indicate that despite their considerable variability, homemade preparations as a whole show cannabinoid levels and profiles equivalent to the commercially available products commonly used for medicinal, therapeutic, and palliative purposes in Argentina.

Phytocannabinomics: Untargeted metabolomics as a tool for cannabis chemovar differentiation

Cannabis sativa is traditionally classified according to five chemotypes based on the concentration of the main phytocannabinoids tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabigerol (CBG). However, cannabis chemovars and varieties very often present similar concentrations of such phytocannabinoids but different chemical profiles, which is unavoidably translated into different pharmacological effects when used for therapeutic purposes. For this reason, a more refined approach is needed for chemovar distinction, which is described in this study and named phytocannabinomics. The classification was achieved by a comprehensive characterization of the phytocannabinoid composition, by liquid chromatography coupled to high-resolution mass spectrometry untargeted metabolomics for the detection of over a hundred phytocannabinoids, and data analysis by chemometrics for chemovars differentiation. The method was developed on fifty cannabis varieties, grown under the same conditions, and was validated to discriminate between the standard chemotypes by partial least squares discriminant analysis. Then, the method was extended to consider the entire chemical variety of the cannabis accessions, by an unsupervised approach based on the principal component analysis. The latter approach clearly indicated several new subgroups within the traditional classifications, which arise from a unique composition of the minor phytocannabinoids. The existence of these subgroups, which were never described before, is of critical importance for evaluating the pharmacological effects of cannabis chemovars.

Palmitoylethanolamide and hemp oil extract exert synergistic anti-nociceptive effects in mouse models of acute and chronic pain

The use of products derived from hemp – i.e., cannabis varieties with low Δ9-tetrahydrocannabinol (Δ9-THC) content – as self-medication for pain and other health conditions is gaining in popularity but preclinical and clinical evidence for their effectiveness remains very limited. In the present study, we assessed the efficacy of a full-spectrum hemp oil extract (HOE; 10, 50 and 100 mg-kg-1; oral route), alone or in combination with the anti-inflammatory and analgesic agent palmitoylethanolamide (PEA; 10, 30, 100 and 300 mg-kg-1; oral route), in the formalin and chronic constriction injury (CCI) tests. We found that HOE exerts modest antinociceptive effects when administered alone, whereas the combination of sub-effective oral doses of HOE and PEA produces a substantial greater-than-additive alleviation of pain-related behaviors. Transcription of interleukin (IL)−6 and IL-10 increased significantly in lumbar spinal cord tissue on day 7 after CCI surgery, an effect that was attenuated to the same extent by HOE alone or by the HOE/PEA combination. Pharmacokinetic experiments show that co-administration of HOE enhances and prolongs systemic exposure to PEA. Collectively, our studies lend support to possible beneficial effects of using HOE in combination with PEA to treat acute and chronic pain.

Influence of seeding rates on yield and technological qualities of hemp fiber

When solving problems aimed at increasing the yield of cannabis and the marketability of the industry, improving the quality of products and reducing its cost, and ultimately restoring the previous size of the sown area, seed seeding rates are of great importance. Research in our experiments was carried out with a hashish-free variety of monoecious cannabis of the Central Russian type Diana. Studies have shown that in thickened crops by the time of harvesting, the number of fallen plants reaches 5-19 pieces per 1 m2 or 3.79-10.67%. The dependence of plant height on the area of nutrition is most clearly shown in the flowering phase: with a decrease in seed seeding rates, the height of plants increases. In this phase, the greatest growth of plants occurs, which reaches 87.7-119.1 cm. With the expansion of the area of supply is greatly enhanced by the formation of items such as plant height, stem thickness at the root collar and the base of the inflorescence, the mass of the stem, which together determine the length of the inflorescence of the plant. With the thickening of crops, the yield of stems and fibers tends to increase. The highest yield of stems and fiber was obtained at a seeding rate of 3.2 million units per 1 ha. With an increase in the seeding rate, the yield of total and long fiber increases. The yield of short fiber decreases noticeably with an increase in the seeding rate. The highest yield of long fiber 26% was provided by thickened sowing with a seeding rate of 3.2 million units per 1 ha.

Can public online databases serve as a source of phenotypic information for Cannabis genetic association studies?

The use of Cannabis is gaining greater social acceptance for its beneficial medicinal and recreational uses. With this acceptance has come new opportunities for crop management, selective breeding, and the potential for targeted genetic manipulation. However, as an agricultural product Cannabis lags far behind other domesticated plants in knowledge of the genes and genetic variation that influence plant traits of interest such as growth form and chemical composition. Despite this lack of information, there are substantial publicly available resources that document phenotypic traits believed to be associated with particular Cannabis varieties. Such databases could be a valuable resource for developing a greater understanding of genes underlying phenotypic variation if combined with appropriate genetic information. To test this potential, we collated phenotypic data from information available through multiple online databases. We then produced a Cannabis SNP database from 845 strains to examine genome wide associations in conjunction with our assembled phenotypic traits. Our goal was not to locate Cannabis-specific genetic variation that correlates with phenotypic variation as such, but rather to examine the potential utility of these databases more broadly for future, explicit genome wide association studies (GWAS), either in stand-alone analyses or to complement other types of data. For this reason, we examined a very broad array of phenotypic traits. In total, we performed 201 distinct association tests using web-derived phenotype data appended to 290 uniquely named Cannabis strains. Our results indicated that chemical phenotypes, such as tetrahydrocannabinol (THC) and cannabidiol (CBD) content, may have sufficiently high-quality information available through web-based sources to allow for genetic association inferences. In many cases, variation in chemical traits correlated with genetic variation in or near biologically reasonable candidate genes, including several not previously implicated in Cannabis chemical variation. As with chemical phenotypes, we found that publicly available data on growth traits such as height, area of growth, and floral yield may be precise enough for use in future association studies. In contrast, phenotypic information for subjective traits such as taste, physiological affect, neurological affect, and medicinal use appeared less reliable. These results are consistent with the high degree of subjectivity for such trait data found on internet databases, and suggest that future work on these important but less easily quantifiable characteristics of Cannabis may require dedicated, controlled phenotyping.

Effect of Fertilization with Urea and Inhibitors on Growth, Yield and CBD Concentration of Hemp (Cannabis sativa L.)

Field experiments were conducted during 2019 in two different locations in Greece (Athens and Farsala) to evaluate the effect of urea and urea fertilization with inhibitors on the agronomic yield and quality characteristics of two cannabis varieties (Cannabis sativa L.), “Uso31”and “Fedora 17”. The experimental design was split-plot with four different fertilization treatments—control, Urea (U), urea with Urease Inhibitor (UI), and urea with Nitrification Inhibitor (NI) and urease inhibitor (UI). The significance of differences between treatments was estimated by using Tukey’s test with a significance level of p = 0.05. The plant height was significantly affected by the different fertilizations and different varieties as well as by the two locations. The maximum plant height was 197 cm for “Fedora 17”in Farsala. The seed yield was higher forthe urea with inhibitors treatment in both varieties. The Cannabidiol (CBD) content was significantly affected by the fertilization—it was higher in urea with inhibitors in “Uso31”and “Fedora 17” treatments. The lowest CBD content value was 1.29% (control) and the highest was 1.69% (urea NI + UI). In conclusion, in both varieties, it seems that urea with inhibitors has a positive effect on their growth, as well as on the increase in cannabidiol (CBD) content.

CannabisGDB: a comprehensive genomic database for Cannabis Sativa L.

Being one of the world's oldest crops with significant economic and medicinal importance, Cannabis Sativa L. (cannabis) has attracted an enormous amount of attention and become one of the most popularly cultivated plants worldwide (Gao et al., 2020). The content of Δ9-tetrahydrocannabinol (THC) determines the legal status of the cannabis varieties. Uncontrolled varieties called hemp are defined as those that have 0.3% or less THC, while marijuana is defined as varieties with higher than 3% (4%-35%).

Cannabinoids: a class of unique natural products with unique pharmacology

The full understanding of the real therapeutic potential of cannabis preparations and of the dangers that their recreational use pose can only stem from the assessment of the chemical composition of such preparations and the knowledge of the pharmacology of their main components, the cannabinoids. Much in the same way cannabis is not just one plant, cannabinoids were not made all equal, and profound differences exist between the pharmacological properties of Δ9-tetrahydrocannabinol (Δ9-THC), the most abundant euphoric and psychotropic cannabinoid, and the other over one hundred natural products belonging to the same class of compounds. So far, only the bioactivity of Δ9-THC, the main component of marijuana, and a handful of cannabinoids, including cannabidiol (CBD), the most abundant cannabinoid in industrial varieties of cannabis, has been thoroughly investigated. While pharmacological studies on Δ9-THC led to the discovery of cannabinoid receptors, endocannabinoids (eCBs) and the endocannabinoid (eCB) system, which underlie most of the effects of this compound in mammals, the mechanisms of action of CBD and other non-euphoric cannabinoids are much more complex and are based on several molecular targets, including, but not limited to, the proteins (receptors and enzymes) of the “expanded eCB system,” or “endocannabinoidome” (eCBome), and hence on their capability of modulating the actions or tissue concentrations of eCB-related lipid mediators. In this article, we discuss to what extent the pharmacology of cannabinoids, and of Δ9-THC and CBD in particular, can be explained by the existence of the eCB system and the eCBome.

Features of changes in geometric parameters of stems of modern varieties of non-drug cannabis for substantiation of technology for producing from them fiber

Features of changes in geometric parameters of stems of modern varieties of non-drug cannabis for substantiation of technology for producing from them fiber

ИЗУЧЕНИЕ ГЕНЕТИЧЕСКОГО ПОЛИМОРФИЗМА СОРТОВ КОНОПЛИ ПОСЕВНОЙ РОССИЙСКОЙ СЕЛЕКЦИИ С ПРИМЕНЕНИЕМ ISSR–МАРКЕРОВ

Nowadays, hemp production places high demands on the efficiency of breeding and the reliability of seed multiplication of hemp (Cannabis sativa L. subsp. Sativa). Therefore, a detailed study of genetic polymorphism, intervarietal and intravarietal variability, and the peculiarities of the formation of the fund of available genetic diversity of culture becomes urgent. The aim was to study the genetic polymorphism of some modern Russian species of industrial hemp using molecular ISSR markers. Experimental data were obtained in 2021 in the laboratory of molecular-genetic researching and cell selection of the Federal Research Center of Bast Crops. We studied ten varieties of hemp of Russian breeding from the collection of Penza Unit of Federal Research Center for Fiber Crops: ‘Milena’, ‘Vera’, ‘Surskaya’, ‘Nadezhda’, ‘Ingreda’, ‘Diana’, ‘Juliana’, ‘Gentus’, ‘Dimra’, ‘Margo’. DNA was isolated from individual seeds by the CTAB method. When setting up PCR, 20 ISSR primers were used; amplification products were separated in agarose gel. A total of 99 alleles of 430–1500 bp were identified, the genetic profile of each studied sample turned out to be individual. Cluster analysis and genetic similarity dendrogram construction made it possible to reveal the presence of intra- and intervarietal genetic polymorphism, visualize the phylogenetic relationships of the studied species. The studied samples formed dense intravarietal groups and were divided into three intervarietal clusters. The presence of similar components of crossing in the pedigrees of the studied varieties is associated with the peculiarity of the identified clustering and determines the prerequisites for the formation of the genetic narrowness of modern varieties of cannabis. The set of 20 used ISSR-markers is distinguished by good resolution for studying the genetic polymorphism of hemp. Intravarietal polymorphism with an average genetic distance of 0.14 and intervarietal polymorphism with a distance of 0.61 were revealed in 10 cultivars of hemp of the Central Russian type. According to the results of cluster and factor analyzes, which showed the similarity of conclusions, the samples were divided into three main clusters, mainly differing in the originator of the variety. The study of DNA polymorphism of seeds of the ‘Diana’, obtained after reproduction of two years, indicates its genetic stability.

Identification of a new cannabidiol n-hexyl homolog in a medicinal cannabis variety with an antinociceptive activity in mice: cannabidihexol

The two most important and studied phytocannabinoids present in Cannabis sativa L. are undoubtedly cannabidiol (CBD), a non-psychotropic compound, but with other pharmacological properties, and Δ9-tetrahydrocannabinol (Δ9-THC), which instead possesses psychotropic activity and is responsible for the recreative use of hemp. Recently, the homolog series of both CBDs and THCs has been expanded by the isolation in a medicinal cannabis variety of four new phytocannabinoids possessing on the resorcinyl moiety a butyl-(in CBDB and Δ9-THCB) and a heptyl-(in CBDP and Δ9-THCP) aliphatic chain. In this work we report a new series of phytocannabinoids that fills the gap between the pentyl and heptyl homologs of CBD and Δ9-THC, bearing a n-hexyl side chain on the resorcinyl moiety that we named cannabidihexol (CBDH) and Δ9-tetrahydrocannabihexol (Δ9-THCH), respectively. However, some cannabinoids with the same molecular formula and molecular weight of CBDH and Δ9-THCH have been already identified and reported as monomethyl ether derivatives of the canonical phytocannabinoids, namely cannabigerol monomethyl ether (CBGM), cannabidiol monomethyl ether (CBDM) and Δ9-tetrahydrocannabinol monomethyl ether (Δ9-THCM). The unambiguously identification in cannabis extract of the n-hexyl homologues of CBD and Δ9-THC different from the corresponding methylated isomers (CBDM, CBGM and Δ9-THCM) was achieved by comparison of the retention time, molecular ion, and fragmentation spectra with those of the authentic standards obtained via stereoselective synthesis, and a semi-quantification of these cannabinoids in the FM2 medical cannabis variety was provided. Conversely, no trace of Δ9-THCM was detected. Moreover, CBDH was isolated by semipreparative HPLC and its identity was confirmed by comparison with the spectroscopic data of the corresponding synthetic standard. Thus, the proper recognition of CBDH, CBDM and Δ9-THCH closes the loop and might serve in the future for researchers to distinguish between these phytocannabinoids isomers that show a very similar analytical behaviour. Lastly, CBDH was assessed for biological tests in vivo showing interesting analgesic activity at low doses in mice.