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By , Potter could find hardly any traces of CBD at all in a batch of seized cannabis samples given to him for analysis by the Home Office. The exact effect this change has had on the mental health of cannabis users is hard to say — the link between cannabis and schizophrenia remains complex and controversial. Researchers have struggled to prove whether cannabis causes psychosis, or whether people predisposed to psychosis are just more likely to smoke cannabis. The best evidence currently suggests that in people who are genetically at risk of schizophrenia , regular cannabis use doubles their risk of experiencing psychotic symptoms.
But no long-term studies of people with schizophrenia have chemically analysed the type of cannabis the subjects were smoking. Potter has seen the extreme effects of both substance abuse and schizophrenia in his role as a magistrate for his local court. He recounts a recent case where a seemingly pleasant teenager, suffering from acute and sudden psychotic illness, had turned to alcohol and become violent.
The defendant, now diagnosed and medicated, will still be found guilty despite his temporary insanity. The need for new antipsychotic drugs is pressing. Existing medication does little to treat the other disabling symptoms of the disease — chronic inflammation, low mood, anxiety, and cognitive impairment — and often has worrying side-effects including weight gain, involuntary movements and drowsiness.
Results from double-blind, phase II clinical trials for CBD as a schizophrenia treatment will report next year. For now hopes rest on a small trial involving 39 patients with schizophrenia, 19 of whom were given the antipsychotic drug amisulpride, the rest CBD. At the end of the four-week trial, both groups showed significant improvement in their symptoms, but the CBD cohort reported far fewer side effects.
Potter and his team remain cautious, as it is not yet fully understood how CBD actually works — it is simplistic to say CBD just does the opposite of the psychosis-inducing THC. One theory is that CBD boosts the activity of other cannabinoids produced naturally by the brain. Potter is passionate about cannabis, but he is not a politician. All of the medicines GW develops are administered in traditional routes such as throat sprays or liquids, and none get users high.
He now boasts that his plants have as much CBD as there is THC in the strongest skunk, after years of selective breeding in the opposite direction by illegal producers. It has taken him more than a decade to get to a point where he can grow both CBD- and THC-rich cannabis of consistent strength on a wide scale. The surreal, secret glasshouse in which Potter works understandably involved extensive discussions around security before the government would grant a licence.
So far, says Potter, no attempts have been made on the bounty inside. The two varieties are harvested; the cannabinoids extracted and blended in different ratios for different therapeutic effects.
Perhaps his latest crop, rich in CBD and low in THC, will be the next cannabis plant of significant importance to society. Cannabinoids from the cannabis plant are being explored for a range of therapeutic uses. Medicinal cannabis extracts are currently prescribed in a number of countries to alleviate pain, treat muscle spasticity, and reduce nausea during chemotherapy.
CBD cannabidiol is being investigated as a potential treatment for epilepsy, diabetes, appetite-loss, a range of inflammatory diseases such as arthritis, and psychosis and schizophrenia. This much simpler protocol recovers equivalent levels of terpenoids and cannabinoids as demonstrated below.
There is a limitation to this method; we cannot detect the free acid forms of the cannabinoids, only the neutral forms of the cannabinoids. The heat in the protocol converts all free acid forms to neutral forms. In order to quantify the free acid forms of cannabinoids, the samples must be derivatized prior to GC separation [ 37 ]. The cannabinoids present in leaf and floral samples of 16 different medicinal marijuana plants were determined.
The levels of six different cannabinoids in floral samples are listed in Table 3 and the levels in leaf samples are listed in Table 4. In both tables the levels are reported as percent of dry weight of the respective organ, these organs were collected from plants at 50 to 65 d post-light induction. As expected the levels of cannabinoids are variable between strains, and the levels in floral tissues are much higher than in leaf tissues.
The other important medical cannabinoid CBD, was barely detectable in leaf or floral tissue of most of the strains except Alien Blues, Thunderstruck, Love Lace and Juanita. A third cannabinoid, CBC, was present at similar levels in leaves and floral tissues, and varied between strains. Platinum Buffalo had the highest leaf levels, 0.
Thunderstruck had the highest CBC levels in floral tissues, 0. CBG levels in floral samples ranged between 2.
The strain Platinum Buffalo had the highest levels in floral samples at 1. The higher levels of cannabinoids in floral versus leaf tissue is expected and has been described by many other investigators [ 2 , 9 , 22 ].
The induction of flowering in the clonally propagated plants is initiated by a shift to a short-day photoperiod. Floral and leaf samples were collected from two strains, Sour Willie and Bohdi Tree at multiple intervals following the floral induction. This plant material was collected from greenhouse grown plants.
These are the averages of multiple flowers collected from different parts of the plant. When those flowers are separated into their locations on the plant, high, middle and low, a pattern of accumulation is detected.
This was observed in both Sour Willie and Bohdi Tree and is anecdotal knowledge among producers of this crop. This same pattern of accumulation was observed in two additional strains, increasing the biological replication of this observation. The observation that the cannabinoid content increases during floral development has been described by other investigators for plants cultivated in vitro as well as in greenhouse settings [ 22 , 39 ].
The quantification of the variability within the plant for floral content of cannabinoids has not been described in the literature previously.
For example, does the cannabinoid content of vegetative leaves, those on the plant prior to flower induction predict the cannabinoid content of the mature flowers that develop later on that plant.
A comparison of the levels of CBD in vegetative leaves and mature flowers is plotted for samples from 16 strains Fig 4. There is a positive correlation between the CBD content of leaf and flower samples, with an R 2 value of 0. Conversely, if a strain is going to have appreciable levels of CBD in its flowers, then the vegetative leaves will have at least 0.
Testing vegetative leaves to determine if a medical marijuana strain predicted to have high CBD floral content will provide valuable information early on in the management of the plant. Flowers and leaves were collected from 16 Cannabis strains. A detailed analysis of the terpenoid and cannabinoid content of commercially generated medical Cannabis floral samples was conducted.
The growth conditions of the plant, the harvest of the samples and their curing process were all under the purview of the producer. The strain names for these samples are provided in the supporting information S1 Table , but we have somewhat limited confidence in the provenance of the strain name.
However, the chemical composition of this material used medicinally in New Mexico has been accurately determined. In the supplemental material we provide the abundances of 19 terpenoids and 6 cannabinoids determined on 72 strains of medical Cannabis. The totals of these values were plotted to look for correlations in levels Fig 5.
Cured trimmed flowers were extracted and the terpenoid and cannabinoid composition determined by GC-FID. The total values for cannabinoids and terpenoids in each sample were plotted.
The total terpenoid content ranged between 0. There are limited biosynthetic interactions between these two biosynthetic pathways, so a strong correlation based on shared biochemical pathways is not predicted. One group has reported a positive correlation between the terpenoid and cannabinoid content [ 1 ] of selected strains developed in the Netherlands for medicinal uses. Those authors do not consider the abundance of these metabolites to be linked metabolically either.
The positive association between the abundance of terpenoids and cannabinoids in Fig 5 probably reflects increased production of all metabolites, including oils in larger healthier floral buds. We also tested for chemotypes within the medical cannabis population based on terpenoid profiles. For that analysis we clustered the Cannabis strains using the relative abundance of 19 unique terpenoids in 72 different strains.
The dendrogram that resulted from this analysis is presented in Fig 6. The specific composition and strain identifications are presented in supporting information S1 Table. Pie charts for selected strains are presented along with the color code for the terpenoid composition in S1 Table. As demonstrated in Fig 6 , there were three major clades or chemotypes: The biosynthetic pathway for the monoterpenoids and sesquiterpenoids present in the Cannabis samples are highly interrelated and positive and negative correlations between the accumulations of specific compounds are expected.
Terpenoid types are identified: Several other groups around the world have used terpenoid profiles to categorize Cannabis strains [ 1 , 22 , 26 , 40 ]. In these cases the authors used a principal components analysis approach to identify metabolites important in either clustering or discriminating specific strains.
Recently, a PCA and hierarchical analysis of 30 cultivars from a medical Cannabis dispensary in California also identified five major groups based on the abundance of 16 terpenoids in these samples [ 40 ]. Our approach of agglomerative clustering and then inspection of pie chart displays of terpenoid composition, allowed us to identify the clades in the trees with their most abundant terpenoid.
Plant samples for these studies were collected from greenhouse grown plants. Transcript levels for these three genes were quantified in leaves Fig 7 and three different development stages in floral tissues Fig 8A—8C. RNA was isolated from flowers collected from the indicated strains at early blue , middle orange or late purple after the floral induction phase.
Transcripts for all three genes were detected in leaves of all four strains. The levels of transcripts for prenyl transferase and THCA synthase were similar within a strain. Transcripts for all three genes were detected in the flowers of all four strains with notable variations. Maximal accumulation of a transcript occurred early in floral development and transcript levels decreased over time. The absolute levels of transcripts in leaves for these enzymes were 10 to fold lower than in the floral RNA samples.
Also, transcripts accumulated early in floral development with decreased transcript abundance as the flower matures, suggests that the enzymes translated from these transcripts persist in the flowers and continue to support cannabinoid biosynthesis. The genes and enzymes for capsaicinoid biosynthesis in fruit of Capsicum sp.
A few other groups have investigated the transcription of cannabinoid biosynthetic genes using qRT-PCR approaches, however in all of those cases, the levels of transcript are reported in relative terms not absolute amounts as we do here. This method difference may be the basis for some of the different results reported by these groups. The increase in CBDAS transcripts in flowers of hemp strains relative to THCAS transcripts in flowers of marijuana strains has been reported in a transcriptomic study [ 18 ] and in a gene mapping study [ 19 ].
The difference in this case may again be due to the quantification of transcripts, as we determine absolute mass amounts and other groups report relative expression that has been normalized by a range of factors. In summary we report a simple, robust and reliable method for the chemical characterization of two classes of bioactive compounds in medical Cannabis samples, terpenoids and cannabinoids. Given the myriad of health conditions treated by medical marijuana, detailed and complete compositional analyses are essential to determine the efficacy of this material for those conditions, and the optimal strain for specific conditions.
The opportunity to predict high CBD floral samples from analyses on vegetative leaves should be explored further as a way to help producers increase the availability of this medically important cannabinoid. Cold Creek Kush, or D. Blue Dream at the days post-induction are represented. Samples are shown as flowers collected from the upper third of the plant blue , middle third of the plant red or bottom third of the plant green. Flowers upper portion of the plant and leaves prior to floral induction were collected from 21 Cannabis strains.
The results are organized by terpenoid type detected following the hierarchical clustering. Representative pie charts for the terpenoid composition are provided for each of the four terpenoid types: A key to the color code for the pie chart is provided. In addition, Rio Grande Analytics, a commercial licensed medical marijuana testing laboratory provided access to research space licensed for analysis of the plant material in this report.
All material costs and salaries of staff working on this project were paid through the Agricultural Experiment Station at New Mexico State University.
RDR is an owner of Rio Grande Analytics, in addition to his full-time position at NMSU, however, Rio Grande Analytics did not provide salary support or have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. National Center for Biotechnology Information , U.
Published online Jul Author information Article notes Copyright and License information Disclaimer. This work was possible because research scientists employed at a public university collaborated with two licensed medical marijuana industry members in NM: Rio Grande Analytics and Ultra Health.
These commercial entities did not play a role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript; they provided no financial support for supplies or salaries. They did provide analysis space and access to medical marijuana plants during their growth cycles.
Received May 21; Accepted Jul 9. This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
This article has been cited by other articles in PMC. Cannabinoid and terpenoid composition in flower samples as cured commercially available medicinal marijuana. Abstract There has been an increased use of medical Cannabis in the United States of America as more states legalize its use.
Introduction Cannabinoids are the class of terpenophenolic secondary metabolites commonly produced by members of the Cannabis genus [ 1 ]. Open in a separate window. Biosynthetic schema for cannabinoids. Materials and methods Plant material All of the plant material characterized in this report will be referred to as Cannabis , without a species level description. Table 1 Linear calibration parameters for quantification of terpenoids and cannabinoids.
RNA isolation and analysis Flowers and leaves were harvested and immediately frozen in liquid nitrogen; RNA was isolated using methods described earlier [ 35 ]; the quality of the RNA was confirmed by formaldehyde agarose gel electrophoresis. Results and discussion Detection of terpenoids and cannabinoids in a single GC-FID run Gas chromatography is a useful approach for quantifying terpenoids and the abundant cannabinoids; however, achieving base line separation of some of the minor cannabinoids, CBC and CBD, can be difficult.
GC-FID separation of terpenoids and cannabinoids. Comparison of cannabinoid content in leaf and floral samples from medicinal marijuana plants The cannabinoids present in leaf and floral samples of 16 different medicinal marijuana plants were determined.
Table 3 Inflorescence cannabinoid content of Cannabis plants cultivated in a greenhouse. Table 4 Leaf cannabinoid content of Cannabis plants cultivated in a greenhouse. Paired comparisons of CBD levels in leaf and floral samples of Cannabis plants. Terpenoid content and composition in medical cannabis A detailed analysis of the terpenoid and cannabinoid content of commercially generated medical Cannabis floral samples was conducted.
Paired comparisons of total cannabinoid and total terpenoid levels. Hierarchical clustering of medical Cannabis strains based on floral terpenoid composition.
Transcript accumulation in leaf samples of cannabinoid biosynthetic genes. Transcript accumulation of cannabinoid biosynthetic genes during floral development. TIF Click here for additional data file. S1 Table Cannabinoid and terpenoid composition in flower samples as cured commercially available medicinal marijuana.
XLSX Click here for additional data file. Data Availability All relevant data are within the paper and its Supporting Information files. Metabolic fingerprinting of Cannabis sativa L. The Plant of the Thousand and One Molecules.
History of cannabis and its preparations in saga, science and sobriquet. The Genetic Structure of Marijuana and Hemp. Hillig K, Mahlberg P. A chemotaxonomic analysis of cannabinoid variation in Cannabis Cannabaceae. Thomas B, Elsohly MA.
The Botany of Cannabis sativa L In:
How ancient viruses got cannabis high
We know sativa plants are tall, feature slim leaves and are usually Ruderalis is often lower in THC in its resin and is known for its high cannabidiol (CBD) content . Ruderalis genetics are often bred into strains to control certain factors All of these fantastic autoflowering strains, and more, have won well. All autoflowering strains contain ruderalis genetics. Plants can deliver high doses of CBD-rich buds in just several weeks And even though THC is already a great painkiller, CBD makes it even better. Stress Killer won't grow as fast. Still, it's not all bad for hemp: The plant produces another lucrative compound These genetic maps showed that the THC and CBD genes are.