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Cannabis - from cultivar to chemovar.

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Cannabis - from cultivar to chemovar
A. Hazekamp
* and J. T. Fischedick
The medicinal use of Cannabis is increasing as countries worldwide are setting up ofcial programs to provide patients with access to
safe sources of medicinal-grade Cannabis. An important question that remains to be answered is which of the many varieties of
Cannabis should be made available for medicinal use. Drug varieties of Cannabis are commonly distinguished through the use of
popular names, with a major distinction being made between Indica and Sativa types. Although more than 700 different cultivars
have already been described, it is unclear whether such classication reects any relevant differences in chemical composition. Some
attempts have been made to classify Cannabis varieties based on chemical composition, but they have mainly been useful for
forensic applications, distinguishing drug varieties, with high THC content, from the non-drug hemp varieties. The biologically active
terpenoids have not been included in these approaches. For a clearer understanding of the medicinal properties of the Cannabis
plant, a better classication system, based on a range of potentially active constituents, is needed. The cannabinoids and terpenoids,
present in high concentrations in Cannabis owers, are the main candidates. In this study, we compared cultivars obtained from
multiple sources. Based on the analysis of 28 major compounds present in these samples, followed by principal component
analysis (PCA) of the quantitative data, we were able to identify the Cannabis constituents that dened the samples into distinct
chemovar groups. The study indicates the usefulness of a PCA approach for chemotaxonomic classication of Cannabis varieties.
Copyright © 2012 John Wiley & Sons, Ltd.
Keywords: cannabis; cannabinoids; terpenoids; chemical proling; cultivar; principle component analysis
Cannabis as a medicine
Although there are an estimated 165 million frequent users of
Cannabis worldwide,
it is presently unclear how many of these
are medicinal users. Nevertheless, through persistent lobbying by
patients as well as through mounting scientic evidence, the
medicinal use of Cannabis is slowly gaining acceptance from
authorities. Over the last decade, both the Netherlands
have implemented state-run medicinal Cannabis
programmes, and other countries are considering a similar move
(Israel, Brazil) or are importing herbal material from the Dutch
programme (Italy, Finland, Germany; pers. comm. Ofce of
Medicinal Cannabis, the Netherlands).
In the USA, a growing
number of states have adopted medical marijuana laws to
provide safer access of Cannabis for medicinal use to patients,
despite the fact that this is vehemently opposed by the Federal
Besides obvious legal implications,Cannabisasanherbalmedicine
poses serious challenges to modern medicine, which operates
according to the single compound, single targetparadigm of
pharmacology. An obvious question therefore is how the chemical
constituents found in Cannabis reect different medicinal properties,
and what types of Cannabis should consequently be made available
to patients. In fact, the Canadian programme is currently under
review, after increasing complaints from patients that the single
variety of Cannabis that is currently available is not effective for a
large proportion of patients.
It is now widely accepted that Cannabis is monotypic and consists
only of a single species Cannabis sativa, as described by Leonard
Fuchs in the sixteenth century.
But as a result of centuries of
breeding and selection, a large variation of cultivated varieties
(cultivars) have been developed. These are commonly distinguished,
by plant breeders, recreational users, and medical Cannabis patients
alike, through the use of popular names such as White Widow,
Northern Lights, Amnesia, or Haze. Already, over 700 different
varieties have been described
and many more are thought to exist.
However, it is unclear whether or not these names reect any
relevant differences in chemical composition.
Classication systems
Some attempts have been made to classify Cannabis varieties based
on chemical composition. A rst study was done by Grlic,
recognized different ripening stages. Later, Fettermann
different phenotypes based on quantitative differences in the
content of main cannabinoids and he was the rst to distinguish
the drug- and bre-type. Further extension of this approach was
done by Small and Beckstead,
and Brenneisen.
However, it was found that a single plant could be classied into
different phenotypes, according to plant age. More recently, a
classication system was developed by de Meijer,
who recognized
ve different Cannabis types based on the (relative) content of three
major cannabinoids.
For forensic and legislative purposes, the most important classica-
tion of Cannabis types is that into the drug-type and the bre-type
(hemp). The main difference between these two is found in the
content of the psychotropic component delta-9-tetrahydrocannabinol
(THC): a high content of THC classies as drug-type Cannabis, while a
low content is found in bre-type Cannabis (max. 0.20.3% THC on
basis of dry matter in the upper reproductive part of the plants), which
* Correspondence to: A. Hazekamp, R&D Department, Bedrocan BV, P.O. Box
2009 Veendam, the Netherlands. E-mail:
aR&D Department, Bedrocan BV, Veendam, the Netherlands
bNatural Products Laboratory, Institute of Biology, Leiden University, the
Drug Test. Analysis (2012) Copyright © 2012 John Wiley & Sons, Ltd.
Research article
Drug Testin
and Anal
Received: 18 November 2011 Revised: 29 November 2011 Accepted: 29 November 2011 Published online in Wiley Online Library
( DOI 10.1002/dta.407
may also be cultivated for its seeds for human or animal consumption.
The content of the closely related but psychotropically inactive canna-
bidiol (CBD) is not regulated by law, and its levels tend to be higher in
Cannabis cultivated for seed or bre.
With almost no exception, Cannabis varieties presently used for
medicinal purposes in ofcial programs belong to the drug-type,
because of their high content of the biologically active THC. But
although bre-type Cannabis is not commonly used for either
medicinal or recreational purposes, it does contain components that
were found to be biologically active such as CBD,
indicating that
the distinction between the two types has limited relevance for
medicinal research into Cannabis. Moreover, it is becoming increas-
ingly clear that components in Cannabis beyond THC and CBD, such
as other minor plant-cannabinoids and volatile secondary metabo-
lites called terpenoids are involved in the drugsoveralleffect.
All major terpenoids present in Cannabis (including e.g. myrcene,
alpha-pinene, beta-caryophyllene) can be found ubiquitously in
For this reason these components did not receive much
scientic attention, until it was suggested that the terpenoid prole
of Cannabis products may help in determining the origin of Cannabis
in custom seizures.
However, no further reports on this approach
were subsequently made in the scientic literature, most likely
because of limited accuracy for forensic use.
The most common way currently used to classify Cannabis
cultivars is through plant morphology (phenotype) with two types
typically recognized: Sativa and Indica. Cannabis cultivars of the
Indica type are smaller in height with broader leaves, while Sativa
types are taller with long, thin-ngered leaves.
Indica plants typ-
ically mature faster than Sativa types under similar conditions, and
the types tend to have a different smell, perhaps reecting a different
prole of terpenoids.
Bridging the gap between culture and science
As a result of limited understanding and support from the medical
community, medicinal users of Cannabis generally adopt the termi-
nology derived from recreational users to describe the therapeutic
effects they experience. Although it is hard to study the popular
Cannabis literature and come to a single clear conclusion, the
following general picture emerges about the difference between
typical Sativa and Indica effects upon smoking.
The Sativa high is often characterized as uplifting and energetic.
The effects are mostly cerebral (head-high), also described as spacey
or hallucinogenic. This type gives a feeling of optimism and well-
being, as well as providing a good measure of pain relief for certain
symptoms. Although Indicas are generally said to contain more
THC, a few pure Sativa types are also very high in THC content.
Sativas are considered a good choice for daytime smoking. In con-
trast, the Indica high is most often described as a pleasant body buzz
(body-high). Indicas are primarily enjoyed for relaxation, stress relief,
and for an overall sense of calm and serenity. Indicas are supposedly
effective for overall body pain relief, and often used in the treatment
of insomnia; they are the late-evening choice of many smokers as an
aid for uninterrupted sleep. A few pure Indica strains are very potent
in THC, and may cause the couchlockeffect, enabling the smoker to
simply sit still and enjoy the experience of the Cannabis.
It has not been properly conrmed whether subjective descriptions
such as these are correlated in any way to the morphological distinc-
tions between Indica and Sativa cultivars, or to any other chemical
classication described above. It is obvious that a better understand-
ing of chemical differences between Cannabis cultivars could help
to bridge the gap between the vast knowledge on Cannabis that
exists within the community of recreational users, and the information
needed by medicinal users and health professionals. However, the
high number of (potential) active components present in Cannabis
signicantly complicates a conventional reductionist approach using
analytical chemistry, animal studies, and clinical trials, where an active
ingredient needs to be identied before further study is possible.
From cultivar to chemovar
An alternative approach to this multiple component problem may be
to simultaneously identify and quantify all major components pres-
ent in various Cannabis types, and use a multi-variant data analysis
tool such as principal component analysis (PCA) to classify cultivars
in a small number of chemically distinct groups. With well-designed
animal studies and/or clinical trials, and using a range of distinct
chemical varieties, correlations may then be observed between spe-
cic chemical characteristics, and potentially benecial biological
effects. Of course, such an approach ts exactly within the paradigm
of Systems Biology, which is recognized as a way to better under-
stand the complex interactions that can be involved in the effects
of medicinal plants with multiple active ingredients.
With this ap-
often vague and unsubstantiated characteristics, towards a new clas-
sication using chemovars with a complex, but nevertheless well-
dened chemical composition (also known as a chemical ngerprint).
In this study we attempt for the rst time to directly compare
cultivars obtained from ofcial as well as illicit sources through a
comprehensive chemical prole, including cannabinoids such as
THC and CBD, but also minor cannabinoids and a range of mono-
and sesqui-terpenoids. Two popular and widely available Cannabis
cultivars were obtained from a number of Dutch coffee shops
(highly regulated outlets of small amounts of Cannabis for recrea-
tional use, tolerated under Dutch law).
All samples were chemi-
cally proled using gas chromatography with ame ionization
detection (GC/FID). Because major components are more likely to
be involved in medicinal effects, only compounds present above
a 0.5 mg/gram threshold level were selected for further analysis,
in order to determine the chemical variation present in both
varieties. This approach resulted in the quantication of 28 different
sample components. The results were then compared to those
obtained by analyzing three medicinal-grade Cannabis varieties
currently available from Dutch pharmacies.
The study indicated
the usefulness of PCA analysis for chemotaxonomic classication of
Cannabis varieties, and may assist in a better identication of
Cannabis cultivars with a potential for medicinal use.
Materials and methods
Sample collection
In order to evaluate the chemical variation found in some major
Cannabis cultivars, ten coffee shops in four major, geographically
dispersed, cities in the Netherlands (Amsterdam, Utrecht, Groningen,
and Maastricht) were selected for sample collection. All locations
were visited a second time about two months later to obtain a larger
variation (different batches) of Cannabis samples. Two popular strains
were selected for analysis, because they were generally available at
all locations throughout the year: Amnesia (a Sativa-dominant type
Cannabis) and White Widow (an Indica-dominant type).
One gram
of each sample was purchased. All samples were delivered in small
zip-lock bags, as typically provided by Dutch coffee shops.
A. Hazekamp and J. T. Fischedick
Drug Testin
and Anal
sis Copyright © 2012 John Wiley & Sons, Ltd. Drug Test. Analysis (2012)
Three varieties of pharmaceutical-grade Cannabis (ofcial product
name: Bedrocan
directly from the ofcial Dutch cultivator, Bedrocan BV (Veendam,
the Netherlands). Plants were grown from clones under standardized
indoor conditions, and quality-controlled for chemical composition,
water content, and the absence of adulterants.
Flowertops were
harvested and air-dried for one week at controlled temperature
and humidity. Samples were delivered in airtight, triple-laminate
bags. All samples were kept in a freezer until analyzed.
Solvents and chemicals
Authentic standards for alpha-bisabolol, myrcene, alpha-pinene,
beta-pinene, gamma-terpineol and beta-caryophyllene were pur-
chased from Sigma-Aldrich (Steinheim, Germany). alpha-Humulene
was from Fluka (Steinheim, Germany). Terpinolene, gamma-
cadinene, cis-ocimene and beta-phellandrene were from a chemical
database of the authors.
Calibrated standards for THC, CBD, cannabigerol (CBG), cannabi-
chromene (CBC), trans-()-delta-9-tetrahydrocannabivarin (THCV),
and cannabinol (CBN) were puried and quantied as previously
All cannabinoid references were >98% pure in
ethanol. For the structures of the cannabinoids(Figure1).Astandard
for cannabigerol-monomethylether (CBGM) was not available.
All organic solvents used were of analytical reagent (AR) grade
(Biosolve BV, Valkenswaard, the Netherlands).
Extracts were prepared as described recently by Fischedick et al.
In short, 500 mg (+/2 mg) of each sample was extracted with
40 ml of absolute ethanol in plastic serum tubes (total volume
50 ml) while mechanically shaking for 10 min. Tubes were
centrifuged and clear supernatant was transferred to a 100-ml
volumetric ask. For exhaustive extraction, the procedure was
repeated twice more with 25 ml of ethanol, and supernatants were
combined. Volumes were adjusted to 100 ml and mixed well. Finally,
the solution was ltered through a 0.45 mm PTFE syringe lter.
Filtrates were stored at 20 C until analysis.
An Agilent GC 6890 series (Agilent Technologies Inc., Santa Clara, CA,
USA) equipped with a 7683 autosampler and a ame ionization
detector (FID) was used for the simultaneous analysis of monoterpe-
noids, sesquiterpenoids, and cannabinoids, as previously described
by Fischedick.
The instrument was equipped with a DB5 capillary
column (30 m length, 0.25 mm internal diameter, lm thickness
0.25 mm; J&W Scientic Inc., Folsom, CA, USA). The injector
temperature was 230 C, with an injection volume of 4 ml, a split ratio
of 1:20 and a carrier gas (N
)ow rate of 1.2 ml/min. The temperature
gradient started at 60 C and increased at a rate of 3 C/min until
240 C which was held for 5 min making a total run time of
65 min/sample. The FID detector temperature was set to 250C.
The GC-FID was controlled by Agilent GC Chemstation software
version B.04.01.
All terpenoids were quantied from a 4-point standard curve of
gamma-terpinene while all cannabinoids were quantied from a
4-point standard curve of THC, which was previously validated to give
adequate quantitative results.
The nal result, expressing the
concentration of each component in mg/gram of plant material,
was used for PCA data analysis. Values were not corrected for water
content of the samples.
In order to conrm peak identication, samples and available
standards were further analyzed by gas chromatographymass
spectrometry (GC-MS) as recently described
using a single
quadrupole MS in Total Ion Count (TIC) mode. Compounds were
identied by comparing their mass spectra and retention times with
authentic references as well as literature reports.
library was also used to assist in compound identication (version
2.0f; Standard Reference Data Program of the National Institute of
Standards and Technology, as distributed by Agilent Technologies).
PCA data analysis
All compounds present at a mean concentration of 0.5 mg/gr in at
least one of the Cannabis varieties were identied and quantied
as described above. Resulting data was subjected to principal
component analysis (PCA ) with the aid of SIMCAP + version 12.0
software (Umetrics, Umeå, Sweden). Unit variance scaling was used.
The rst two principal components (PC1 and PC2) were visualized
in a scatter-plot, in combination with its accompanying loading-plot.
For more information on interpreting PCA data, you are referred to
the tutorials section of the Umetrics website.
Figure 1. Structures of main cannabinoids.
Towards a better denition of Cannabis potency
Drug Testin
and Anal
Drug Test. Analysis (2012) Copyright © 2012 John Wiley & Sons, Ltd.
Results and discussion
Sampling and analysis
In total, we intended to obtain 40 Cannabis samples for our study
(10coffeeshopsx2visitsx2varieties).However,inve cases the
desired product was not available from the coffee shop visited (2x
Amnesia, 3x White Widow), which resulted in a total of 18 Amnesia
samples and 17 White Widow samples available for analysis. In total,
28 different components were found to be present in one or more
Cannabis samples at a mean level of 0.5 mg/gr. These are indicated
in Table 1 (coffee shop samples) and Table 2 (pharmacy samples).
Values were not corrected for water content of the samples as
this may require more sample (cost) and it may induce the loss
of volatile constituents. All components could be positively iden-
tied with the help of retention time, mass-data and authentic
standards, except one single compound (m/z: 356 [M+], 313,
297 [base], 243, 231). Because we expected this component to
be a cannabinoid, it was used in the nal data analysis (com-
pound 24 in Tables 1 and 2).
The use of only one terpenoid (gamma-terpinene) and one
cannabinoid (THC) for quantication of all sample components
may demand some further explanation. In a previously published
report, we showed that the differences in FID-detector response
between different types of cannabinoids, or between a range of
mono- and sesqui-terpenoids, were relatively small.
quantifying all 28 sample components with their own standard
curves would be preferred, but this is a tremendous and costly task.
Our current approach may give small deviations from actual
concentrations, but the method is easy to perform and it does not
invalidate our ndings. In our case, the method is used for selecting
the main sample components (above ca. 0.5mg/gr) and to subse-
quently compare the relative differencesbetween samples. Because
our intention is to keep extending our database of cultivars on a
regular basis, and therefore would need a rapid and simple method-
ology, we believe the limitations of our method are acceptable.
The data presented in Table 1 prompt some intriguing ques-
tions about the popular name used for these cultivars. For exam-
ple, Amnesia has a mean titer of alpha-pinene that is less than
half that of White Widow. If Amnesia actually has more amnesic
effects, this distinction is noteworthy, since that component is
an anticholinesterase inhibitor, and may reduce associated
short-term memory decits engendered by THC.
the myrcene titre of White Widow is more than four times that
in Amnesia. If White Widow, being of the Indica chemotype, is
indeed noted for its sedating properties (see Introduction), this
could be correlated, since myrcene has been associated with
Table 1. GC quantitative data of coffee shop samples.
white widow amnesia
ID# Compound RRT
1alpha-pinene 0.257 1.1 0.24 21.1 0.5 0.04 6.9
2beta-pinene 0.306 0.5 0.08 16.3 0.7 0.16 22.7
3myrcene 0.319 2.9 1.16 39.5 0.7 0.16 23.4
4beta-phellandrene 0.371 0.7 0.13 17.9 0.7 0.13 19.9
5cis-ocimene 0.397 ——0.7 0.14 19.3
6terpinolene 0.458 ——1.9 0.77 39.6
7terpineol 0.622 0.6 * 0.0 —— —
8beta-caryophyllene 1.000 1.2 0.39 31.1 2.1 0.59 27.6
9alpha-guaiene 1.030 ——0.5 * 0.0
10 humulene 1.054 0.6 0.11 19.2 0.8 0.21 25.2
11 beta-farnesene 1.057 0.5 0.06 11.2 0.5 0.00 0.7
12 gamma-selinene 1.136 0.7 0.25 34.6
13 delta-guaiene 1.136 ——
14 gamma-cadinene 1.180 0.6 0.22 34.3 0.6 0.08 14.2
15 eudesma-3,7(11)-diene 1.190 0.8 0.27 34.9 0.6 0.09 15.2
16 elemene 1.214 0.7 0.18 27.2 0.5 0.07 13.8
17 guaiol 1.274 0.6 0.12 20.0 —— —
18 gamma-eudesmol 1.307 0.6 0.21 35.7 —— —
19 beta-eudesmol 1.355 0.5 0.06 11.2 —— —
20 alpha-bisabolol 1.398 0.5 * 0.0 —— —
21 THCV 2.179 1.0 0.22 23.1 1.2 0.21 17.2
22 CBD 2.292 0.7 0.10 14.3 0.6 0.05 9.1
23 CBC 2.303 1.8 0.33 18.4 2.8 0.73 26.3
24 Unknown compound 2.348 0.7 0.26 36.2 0.7 0.12 17.4
25 CBGM 2.358 1.9 * 0.0 —— —
26 THC 2.399 159.5 26.76 16.8 167.5 21.88 13.1
27 CBG 2.456 5.4 2.34 43.2 12.5 2.95 23.6
28 CBN 2.461 0.9 0.19 21.0 1.4 0.41 29.6
All levels are expressed in mg of analyte per gram of cannabis sample.
* Analyte detected in only one sample, so standard deviation could not be calculated. RRT: relative retention time compared to beta-caryophyllene.
A. Hazekamp and J. T. Fischedick
Drug Testin
and Anal
sis Copyright © 2012 John Wiley & Sons, Ltd. Drug Test. Analysis (2012)
PCA and quantitative analysis of coffee shop samples
PCA is a mathematical way of identifying patterns in data, and
expressing the data in such a way as to highlight their similarities
and differences. Since patterns can be hard to nd in data of higher
dimension, where the luxury of graphical representation (i.e. a 2- or
3-dimensional diagram) is not available, PCA is a powerful tool for
analyzing data.
In this study, PCA was used to analyze our data set,
which consisted of 44 samples with up to 28 compounds each, equal-
ing 1232 data points. Each compound represented a variable while its
quantity represented the observation. Figure 2 shows an evaluation
of the rst two principal components of all samples in the form of
a scatter plot (Figure 2a) and its related loading plot (Figure 2b).
The data as represented in the PCA scatter plot clearly shows that
Cannabis cultivars Amnesia and White Widow cluster in two chem-
ically distinct groups. Principal component 1 (PC1; 27.2%) and PC2
(17.0%) together explained 44.2% of the total variance found in the
sample set, indicating a relatively high degree of condence. Anal-
ysis of PC3 added only another 11.4%. Interestingly, the mean THC
content found in Amnesia (167.5mg/gr, or 16.75 %) and White
Widow (159.5 mg/gr, or 15.95 %) samples showed no signicant
difference, providing no basis for distinguishing both popular
varieties. However, the loading plot (Figure 2b) helps us under-
stand how exactly the two cultivars differ from one another. Based
on our PCA analysis, it becomes clear that monoterpenoids,
sesquiterpenoids, and minor cannabinoids are important in distin-
guishing the varieties.
Amnesia samples were characterized mainly by the presence of
the terpenoids terpinolene, alpha-guaiene and gamma-selinene,
which were not detected (i.e. below threshold levels) in White
Widow. Levels of beta-caryophyllene, as well as cannabinoids THCV,
CBC and, particularly, CBG were higher in Amnesia than in White
Widow. In contrast, White Widow samples showed signicantly
higher content of myrcene and alpha-pinene. Samples of this variety
were further characterized by the presence of guaiol, beta-eudesmol,
gamma-eudesmol, and alpha-bisabolol, which were not detected in
Amnesia samples. Such sesquiterpene alcohols have previously been
reported to be important for the chemotaxonomic discrimination of
Indica Cannabis varieties originating from Afghanistan.
In three cases, chemical analysis showed that the obtained sample
was likely not of the desired cultivar type. More specically, two
samples that were purchased as White Widow were found to
resemble the typical chemical ngerprint of Amnesia, while for one
Amnesia sample the opposite was true. Although these samples
are shown in Figure 2, we assumedthatamix-uphadoccurredat
the coffee shop, and these outliers were not included in our nal
evaluation of chemical diversity. However, it is also possible that
these samples represented actual specimens of the desired cultivars,
which would even further increase the chemical diversity (i.e. STDEV)
found in both cultivar groups (Table 1). Naturally, a mix-up of samples
Table 2. GC quantitative data of pharmacy samples.
ID# Compound
1alpha-pinene 0.5 0.03 6.3 1.9 0.03 1.8 1.1 0.02 2.0
2beta-pinene 0.8 0.03 3.8 0.5 0.02 3.3 0.3 0.28 86.7
3myrcene 2.9 0.11 3.6 5.9 0.24 4.1 3.7 0.18 4.8
4beta-phellandrene 1.2 0.05 4.0 —— ——— —
5cis-ocimene 2.0 0.07 3.8 —— ——— —
6terpinolene 5.8 0.27 4.6 ——— —— —
7terpineol —— ——— ——— —
8beta-caryophyllene 1.5 0.05 3.3 0.9 0.04 4.2 1.3 0.01 0.6
9alpha-guaiene ——— —— ——— —
10 humulene 0.5 0.03 5.8 ——— —— —
11 beta-farnesene —— ——— ——— —
12 gamma-selinene ——— —— ——— —
13 delta-guaiene 0.7 0.02 3.5 ——— —— —
14 gamma-cadinene 0.6 0.03 5.2 ——— —— —
15 eudesma-3,7(11)-diene 0.8 0.00 0.6 —— —0.6 0.03 5.5
16 elemene 0.9 0.03 3.0 ——— —— —
17 guaiol —— ——— —0.6 0.03 5.3
18 gamma-eudesmol ——— —— —0.6 0.04 6.8
19 beta-eudesmol —— ——— ——— —
20 alpha-bisabolol —— ——— —0.7 0.03 3.6
21 THCV 1.3 0.04 3.2 0.9 0.08 8.4 1.0 0.09 8.5
22 CBD 0.8 0.03 4.3 0.6 0.01 ——1.7
23 CBC 2.8 0.02 0.8 2.5 0.02 0.6 2.6 0.11 4.2
24 Unknown compound ——— —— ——— —
25 CBGM —— ——— ——— —
26 THC 211.9 6.48 3.1 135.0 5.02 3.7 174.5 6.14 3.5
27 CBG 12.0 0.64 5.3 2.4 0.11 4.4 12.4 0.47 3.8
28 CBN 0.8 0.16 19.0 ———0.7 0.08 11.1
All levels are expressed in mg of analyte per gram of cannabis sample.
Towards a better denition of Cannabis potency
Drug Testin
and Anal
Drug Test. Analysis (2012) Copyright © 2012 John Wiley & Sons, Ltd.
during extraction or processing may also be considered as a possible
reason for the observed effect. However, this risk was minimized by
clearly marking all samples directly after purchase, while keeping
them in their original packaging. Samples were then extracted and
processed in groups of eight sequential samples by two people
checking each other.
It may be concluded that, based on the overall chemical prole
consisting of 28 major components, it is indeed possible to
distinguish between two cultivars that are popularly recognized by
recreational Cannabis users: Amnesia and White Widow. However,
an important issue when using Cannabis as a medicine is the repro-
ducibility of the chemical composition of the product that the patient
is trying to obtain. For both Cannabis varieties analyzed in this study,
chemical variability was considerable, as shown in Table 1 and visual-
ized in the PCA score plot. Deviations (relative standard deviation) of
more than 25% were found for 14 analyzed compounds. To put
this variability in perspective, it makes sense to compare these results
to the chemical variation commonly observed in Cannabis obtained
from standardized sources (see next section).
Comparing coffee shop to pharmacy samples
Three different types of medicinal-grade Cannabis, currently avail-
able on prescription in the Netherlands, were analyzed according
to the procedures mentioned. In order to obtain better statistical
results, these analyses were done on triplicate samples for each
variety (compared to obtaining samples in 20-fold for both coffee
shop varieties). Chemical proles were then compared to
determine similarities and differences between coffee shop and
pharmacy samples.
Although varieties Bedrobinol and Bedica have quite different
genetic backgrounds, PCA analysis indicated that both closely
resemble the typical ngerprint of the White Widow variety.
Indeed, variety Bedica is genetically derived from an Indica type,
so this would explain their similarity in chemical prole, such as
the presence of sesquiterpene alcohols. The fact that variety
Bedrobinol resembles White Widow is more surprising, as it was
genetically derived from a Haze variety, which has a large propor-
tion of Sativa genes in its background.
A potential explanation
is the fact that variety Bedrobinol has very low content of
terpenes in general, with only a few components detectable
above our 0.5 mg/gram threshold level. As a result, there may
not have been enough distinguishing features to clearly show
differences with the other analyzed samples. Finally, the differ-
ence between Bedrobinol and Bedica is mainly found in the
presence of sesquiterpenes such as guaiol, gamma-eudesmol,
and alpha-bisabolol in Bedica, while Bedrobinol was largely
devoid of these compounds.
Bedrocan is currently the most popular medicinal Cannabis
variety in the Netherlands. It is interesting to conclude that it does
not resemble either coffee shop type, or the two other pharmacy
varieties. In Figure 2, the data for Bedrocan (left-lower corner) even
falls outside the 95% condence interval region, underscoring the
fact that its composition is signicantly different from any of the
other four varieties analyzed. Bedrocan was originally developed
from a Jack Herer genetic background, which supposedly is a
slightly Sativa dominant hybrid.
Indeed, Bedrocan is closer to
Amnesia, which is also considered Sativa dominant, than to White
Widow. The main distinguishing feature of Bedrocan compared
to Amnesia is the markedly higher content of myrcene, cis-
ocimene and terpinolene by at least a factor of 2. In contrast, a
few components found in Amnesia were not detected (below
threshold levels) in Bedrocan (i.e. beta-farnesene, gamma-selinene,
gamma-cadinene and eudesma-3,7(11)-diene).
It is clear that plant-based products can never be perfectly
standardized for content of active components, as they are
dependent on too many environmental factors to completely predict
the chemical composition of the nal product. For that reason, a
variability of up to 15% is allowed for the THC and CBD levels in
the Dutch medicinal Cannabis. Although currently no regulations
exist for the levels of minor cannabinoids or terpenoids, any producer
of a medicinally used product should strive for a minimum of
variation in chemical composition. In our previous study on
medicinal-grade Cannabis
using the same analytical methodol-
ogy, an average variation (RSD) of about 7.6% was found for
cannabinoids, and 11.0% for terpenoids in a standard batch of
variety Bedrocan; for variety Bedica these numbers were 4.3%
(terpenoids) and 5.5% (cannabinoids). Incontrasttothecoffee
shop samples discussed above, none of the components analyzed
in the pharmacy varieties showed a variability of over 25%.
Obviously, this result may be expected based on the fact that the
Cannabis samples of pharmaceutical grade were cultivated under
-8 -7 -6 -5 -4 -3 -2 -1 0 1 2 3 4 5 6 7 8
PC2 [17.0%]
PC1 [27.2%]
White widow
Bedrocan Bedrobinol
-0.30 -0.25 -0.20 -0.15 -0.10 -0.05 0.00 0.05 0.10 0.15 0.20 0.25
Figure 2. PCA scatter plot. Solid ellipse indicates 95% condence inter-
val. Dotted lines were drawn by the authors to facilitate visual interpreta-
tion of this gure only; it does not signify signicance or condence
intervals. Note that 2 samples purchased as Amnesia (closed triangles)
were chemically similar to White Widow (closed circles), while for one
sample the opposite was observed. It was assumed these samples were
mixed up by the coffee shop, and they were not used for nal statistical
evaluations. Amnesia and White Widow data all represents individually
purchased samples from different batches. Data for Bedrocan, Bedrobinol
and Bedica represents triplicate analyses of the same batch.
A. Hazekamp and J. T. Fischedick
Drug Testin
and Anal
sis Copyright © 2012 John Wiley & Sons, Ltd. Drug Test. Analysis (2012)
strictly controlled indoor conditions, and each batch was ana-
lyzed for chemical content by validated laboratory methods.
The psychotropic compound THC is often considered to be the main
component responsible for (medicinal) effects of Cannabis products,
through activation of specic cannabinoid-binding receptors.
increasingly, other, non-psychotropic cannabinoids are showing their
medicinal potential,
sometimes even through mechanisms that
do not involve binding to the known cannabinoid receptors.
a growing number of medicinal Cannabis users worldwide, it there-
foremaybetimetodene the potencyof Cannabis products in a
more comprehensive manner than by THC content alone. With mod-
ern analytical techniques, a full- range analysis of cannabinoids and
terpenoids has become possible.
In previous studies, our group showed that a range of Cannabis
cultivars grown by clonal propagation under controlled environmen-
tal conditions could be accurately distinguished based on their
cannabinoid and terpenoid content. We also observed that the
chemical prole of cannabinoids and terpenoids in different batches
of the same cultivar (grown sequentially over the period of several
months) was highlyreproducible.
These data suggest it is possible
to fully standardize the chemical content of cannabinoids as well as
terpenoids in Cannabis, when cultivated in a controlled setting.
In this study we applied the same analytical methodology to eval-
uate the chemical variation of two popular Cannabis cultivars
obtained in Dutch coffee shops. With this study we intended to (1)
determine whether there is a chemical basis to consider the obtained
varietiesastrulydifferentproducts, and (2) evaluate the chemical
variability that exists within each cultivar type. In order to bridge
the gap between recreational and medicinal use of Cannabis, we
then compared the overall chemical composition of the obtained
coffee shop samples with three types of medicinal Cannabis from a
government-licensed facility in the Netherlands. The fourth variety
of Cannabis currently available on prescription in the Netherlands,
called Bediol
, was not included in this study because its high
content of CBD (7.5%) completely changes its pharmacological spec-
trum, compared to the varieties analyzed in this study (CBD <0.1%).
In a total of 40 visits to the coffee shop, the desired product was
not available on ve occasions, while in three cases a product was
obtained that signicantly differed from the average composition
of the requested variety. As we assumed these three samples had
been mixed up in the coffee shop (either intentionally or
accidentally), we chose not to include them in our nal analysis. If
our experiences can be considered as representative, it means that
in 20% of cases (8/40) a consumer is not able to receive a reliable
service when visiting an average coffee shop. The observed variabil-
ity in chemical composition may also be a cause of concern for
medicinalusers.Asitisoftenclaimed that the exact combination
of active constituents (synergy) gives each Cannabis variety its
unique (medicinal) properties, medicinal Cannabis users may
inadvertently purchase a product that has unexpected effects on
their health and/or psyche. It should be noted that changes in
chemical composition may not only be derived from genetic differ-
ences between batches of the same Cannabis variety, but could also
be caused by differences between coffee shops (or their suppliers) in,
for example, cultivation conditions, drying, processing, and storage.
However, no matter what the cause,Cannabisproductsfroma
standardized and quality-controlled source may be the safer choice
for medicinal users.
An important reason for patients to keep purchasing their
materials from illicit markets is the fact that, by trial and error,
they have found a strain that works optimally for treatment of
their specic symptoms. With the limited choice of Cannabis
varieties currently available from ofcial sources, it is hard to
deny the value of such choice. By making use of the comprehen-
sive chemical ngerprint of herbal Cannabis, our results may help
medicinal users of Cannabis and their doctors to switch from a
benecial variety obtained through the street market, to a strain
that is available through ofcial state-run programmes. It may
also help these ofcial programmes to narrow down the search
for future Cannabis varieties to be standardized and introduced
as an ofcial medicine.
As this study intended to compare chemical data about
Cannabis samples with knowledge available from Cannabis
popular culture, we decided to use information from the Sensi
Seed Bank (the Netherlands) website as a reference for the Sativa
or Indica background of cultivars. Sensi Seed Bank is perhaps the
largest supplier of Cannabis seeds in the world, and an authorita-
tive voice among producers and users of recreational Cannabis.
In a next phase of our ongoing studies, we would like to obtain
typical pureSativa and Indica strains to determine the two ex-
treme ends of the chemical diversity scale. Although this was
our intention for the present study, we were not able to identify
such strains that were available from all ten coffee shops we
visited for sample collection. According to Sensi Seed Bank, a
typical Sativa type Cannabis would be Durban (90% Sativa), while
some well-known examples of Indica types would be Hindu kush
(100% Indica), Afghani #1 (95%), or Northern lights (90%). As an
example of pure Sativa we could also select a non-drug variety
(i.e. hemp) available in the Netherlands, and grow it under
standardized conditions.
We consider this study a work in progress, and wish to add a
larger number of popular strains from the recreational scene,
as well as Cannabis varieties from ofcial programmes to our
database. Over time, we hope this will lead to a better under-
standing of the overlap between medicinal use of Cannabis,
and the street culture around the same plant. It is our personal
belief that, with such an approach, the endless number of popu-
lar cultivars could be reduced to perhaps two dozen chemically
distinct chemical varieties, or chemovars. Further study into the
chemical similarities between popular coffee shop (or other illicit
sources) Cannabis types, and available medicinal-grade strains
may help medical users to accurately and efciently nd a more
reliable source of medicine within the various national medicinal
Cannabis programmes. Exchange of cultivars and information
between the various national programmes, in countries including
the Netherlands, Canada, and the USA, may greatly facilitate such
a transition.
The authors would like to pay their nal respect to their former
colleague Gian van der Water, whose enthusiasm was a driving
force behind this study.
AH has been employed by Bedrocan BV as the head of Research
and Development since February 2011.
Towards a better denition of Cannabis potency
Drug Testin
and Anal
Drug Test. Analysis (2012) Copyright © 2012 John Wiley & Sons, Ltd.
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A. Hazekamp and J. T. Fischedick
Drug Testin
and Anal
sis Copyright © 2012 John Wiley & Sons, Ltd. Drug Test. Analysis (2012)
... Gas chromatography was used for the simultaneous qualitative analysis of monoterpenes, sesquiterpenes, and cannabinoids as previously reported (Hazekamp and Fischedick, 2012). An Agilent GC 6890 series (Agilent Technologies Inc., Santa Clara, CA, USA) equipped with a 7683 autosampler and a flame ionization detector (FID) was used. ...
... For each variety, control (non-irradiated) samples, and treated (irradiated) samples are shown side by side, using the same vertical scale to allow direct comparison. The main peaks in each variety were identified based on previously published data (Hazekamp and Fischedick, 2012). While the overall qualitative composition of the samples was unaltered, differences in several terpene components could be detected after irradiation in the cannabis varieties studied. ...
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In several countries with a National medicinal cannabis program, pharmaceutical regulations specify that herbal cannabis products must adhere to strict safety standards regarding microbial contamination. Treatment by gamma irradiation currently seems the only method available to meet these requirements. We evaluated the effects of irradiation treatment of four different cannabis varieties covering different chemical compositions. Samples were compared before and after standard gamma-irradiation treatment by performing quantitative HPLC analysis of major cannabinoids, as well as qualitative GC analysis of full cannabinoid and terpene profiles. In addition, water content and microscopic appearance of the cannabis flowers was evaluated. This study found that treatment did not cause changes in the content of THC and CBD, generally considered as the most important therapeutically active components of medicinal cannabis. Likewise, the water content and the microscopic structure of the dried cannabis flowers were not altered by standard irradiation protocol in the cannabis varieties studied. The effect of gamma-irradiation was limited to a reduction of some terpenes present in the cannabis, but keeping the terpene profile qualitatively the same. Based on the results presented in this report, gamma irradiation of herbal cannabis remains the recommended method of decontamination, at least until other more generally accepted methods have been developed and validated.
... For example we found that Jamaican Lambs Bread (100% reported C. sativa) was nearly identical (IBS = 0.98) to a reported 100% C. indica strain from Afghanistan. Sample mix-up cannot be excluded as a potential reason for these discrepancies , but a similar level of misclassification was found in strains obtained from Dutch coffee shops based on chemical composition [10]. The inaccuracy of reported ancestry in marijuana likely stems from the predominantly clandestine nature of Cannabis growing and breeding over the past century. ...
... The inaccuracy of reported ancestry in marijuana likely stems from the predominantly clandestine nature of Cannabis growing and breeding over the past century. Recognizing this, marijuana strains sold for medical use are often referred to as Sativa or Indica " dominant " to describe their morphological characteristics and therapeutic effects [10]. Our results suggest that the reported ancestry of some of the most common marijuana strains only partially captures their true ancestry. ...
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Despite its cultivation as a source of food, fibre and medicine, and its global status as the most used illicit drug, the genus Cannabis has an inconclusive taxonomic organization and evolutionary history. Drug types of Cannabis (marijuana), which contain high amounts of the psychoactive cannabinoid Δ9-tetrahydrocannabinol (THC), are used for medical purposes and as a recreational drug. Hemp types are grown for the production of seed and fibre, and contain low amounts of THC. Two species or gene pools (C. sativa and C. indica) are widely used in describing the pedigree or appearance of cultivated Cannabis plants. Using 14,031 single-nucleotide polymorphisms (SNPs) genotyped in 81 marijuana and 43 hemp samples, we show that marijuana and hemp are significantly differentiated at a genome-wide level, demonstrating that the distinction between these populations is not limited to genes underlying THC production. We find a moderate correlation between the genetic structure of marijuana strains and their reported C. sativa and C. indica ancestry and show that marijuana strain names often do not reflect a meaningful genetic identity. We also provide evidence that hemp is genetically more similar to C. indica type marijuana than to C. sativa strains.
... Additionally , Cannabis secondary metabolites such as terpenoids and flavonoids likely contribute to therapeutic or psychoactive effects (Russo, 2011 ). For example, b-myrcene , humulene, and linalool are proposed to produce sedative effects associated with specific varieties (Hazekamp and Fischedick, 2012). In this study, plants that produce low levels of total cannabinoids are herein referred to as hemp, while high cannabinoid producing varietals are described as drug-type plants. ...
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Plants of the Cannabis genus are the only prolific producers of phytocannabinoids, compounds that strongly interact with the evolutionarily ancient endocannabinoid receptors shared by most bilaterian taxa. For millennia, the plant has been cultivated not only for these compounds, but also for food, rope, paper, and clothing. Today, specialized varieties yielding high-quality textile fibers, nutritional seed oil, or high cannabinoid content are cultivated across the globe. However, the genetic identities and histories of these diverse populations remain largely obscured. We analyzed the nuclear genomic diversity among 340 Cannabis varieties, including fiber and seed oil hemp, high cannabinoid drug-types, and feral populations. These analyses demonstrate the existence of at least three major groups of diversity with European hemp varieties more closely related to narrow leaflet drug-types (NLDTs) than to broad leaflet drug-types (BLDTs). The BLDT group appears to encompass less diversity than the NLDT, which reflects the larger geographic range of NLDTs, and suggests a more recent origin of domestication of the BLDTs. As well as being genetically distinct, hemp, NLDT, and BLDT genetic groups produce unique cannabinoid and terpenoid content profiles. This combined analysis of population genomic and trait variation informs our understanding of the potential uses of different genetic variants for medicine and agriculture, providing valuable insights and tools for a rapidly emerging valuable industry.
... At higher concentrations, terpenoids produce psychoactive effects and possibly interact with cannabinoids, altering the effect of both chemicals (McPartland and Russo, 2001; Adams and Taylor, 2010). Recent studies have shown that some terpenoids have sedative effects (Hazekamp and Fischedick, 2012 ) and show potential as treatments for illnesses such as inflammation (Gil et al., 1989; Lorenzetti et al., 1991), pain (Russo, 2006; Huestis, 2007), depression (Komori et al., 1995), convulsions (Elisabetsky et al., 1995), cancer (De-Oliveira et al., 1997; Vigushin et al., 1998), and fungal infections (Langenheim, 1994). Without a clear way to define the taxonomy of Cannabis , previous research relied on terpenoid variation as a measure of divergence within the genus (Hillig, 2004). ...
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The Cannabis industry is currently one of the fastest growing industries in the United States. Given the changing legal status of the plant, and the rapidly advancing research, updated information on the advancement of Cannabis genomics is needed. This versatile plant is used as medicine and for food, fiber, and bioremediation. Insights from modern, high-throughput genomic technology are revolutionizing our understanding of the plant and are providing new tools to further improve our knowledge and utilization of this unique species. This review quantifies and evaluates the currently available genomic resources for Cannabis research, including six whole-genome assemblies, two transcriptomes, and 393 other substantial genomic resources, as well as other smaller publicly available genetic and genomic resources. The open-source approaches followed by many leading scientists in the field promote collaboration and facilitate these rapid advances.
... Unfortunately, the long-term effects of his publication would turn out to do the exact opposite, and well over two hundred years later we are still left in confusion. Modern research tools, involving detailed chemical [27] and genetic [28] analysis of a wide variety of cannabis types, may finally solve this sativa-indica dilemma: is it a single species, two species , or even more. If such scientific evaluation can take into account the vast knowledge that exists within current communities of cannabis users, we may finally decide on a definition of indica that is acceptable to all. ...
Cannabis sativa (hemp) as multifunctional crop have traditional application as fiber, food, paper, textile and pharmaceutical potential as inflorescences and seed as sources of exciting bioactive secondary metabolites. The Genus Cannabis is the only producer of phytocannabinoids. Extensive studied have been made to describe the origin history, geographical ranges and genetic identity of the Cannabis species but it remains obscured to date. Various high through put genetic marker have been studied to assess the genetic diversity in hemp varieties. Studies also indicated that domestication origin affects the genetic groups of hemp which further consequences on the chemical diversity of the cannabis. Chemotaxonomy using chemical markers also played a crucial role in differencing and allocating the Cannabis taxa. Cannabinoids ratio and terpene composition are the major marker to play an important role in studying chemical diversity of Cannabis sp. Cannabis genus is the only source of phytocannabinoids the dominant chemical class. Other than cannabinoids terpene and non-cannabinoid phenolic compounds also contribute in the chemical diversity of the species. The vast array of phytochemicals presents in the genus have potential application in pharmaceutical industries. However, due to its legalization status very limited study on its chemical and genetic diversity have been done. Therefore, the species needs attention to explore its commercial value.Keywords Cannabis sativa HempPhytocannabinoidsGenetic diversityChemotaxonomy
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For over a century, research on cannabis has been hampered by its legal status as a narcotic. The recent legalization of cannabis for medical purposes in North America requires rigorous standardization of its phytochemical composition in the interest of consumer safety and medicinal efficacy. To utilize medicinal cannabis as a predictable medicine, it is crucial to classify hundreds of cultivars with respect to dozens of therapeutic cannabinoids and terpenes, as opposed to the current industrial or forensic classifications that only consider the primary cannabinoids tetrahydrocannabinol (THC) and cannabidiol (CBD). We have recently developed and validated analytical methods using high-pressure liquid chromatography (HPLC-DAD) to quantify cannabinoids and gas chromatography with mass spectroscopy (GC-MS) to quantify terpenes in cannabis raw material currently marketed in Canada. We classified 32 cannabis samples from two licensed producers into four clusters based on the content of 10 cannabinoids and 14 terpenes. The classification results were confirmed by cluster analysis and principal component analysis in tandem, which were distinct from those using only THC and CBD. Cannabis classification using a full spectrum of compounds will more closely meet the practical needs of cannabis applications in clinical research, insdustrial production, and patients’ self-production in Canada. As such, this holistic classification methodology will contribute to the standardization of commercially-available cannabis cultivars in support of a continuously growing market.
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Introduction: There is a large disparity between the ''cultural'' language used by patients using cannabis for self-medication and the ''chemical'' language applied by scientists to get a deeper understanding of cannabis effects in laboratory and clinical studies. The distinction between Sativa and Indica types of cannabis, and the different biological effects associated with them, is a major example of this. Despite the widespread use of cannabis by self-medicating patients, scientific studies are yet to identify the biochemical markers that can sufficiently explain differences between cannabis varieties. Methods: A metabolomics approach, combining detailed chemical composition data with cultural information available for a wide range of cannabis samples, can help to bridge the existing gap between scientists and patients. Such an approach could be helpful for decision-making, for example, when identifying which varieties of cannabis should be made legally available under national medicinal cannabis programs. In our study, we analyzed 460 cannabis accessions obtained from multiple sources in The Netherlands, including hemp-and drug-type cannabis. Results: Based on gas chromatography analysis of 44 major terpenes and cannabinoids present in these samples , followed by Multivariate Data Analysis of the resulting chromatographic data, we were able to identify the cannabis constituents that may act as markers for distinction between Indica and Sativa. This information was subsequently used to map the current chemical diversity of cannabis products available within the Dutch medicinal cannabis program, and to introduce a new variety missing from the existing product range. Conclusion: This study represents the analysis of the widest range of cannabis constituents published to date. Our results indicate the usefulness of a metabolomics approach for chemotaxonomic mapping of cannabis varieties for medical use.
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Hemp is diploid organism (2n = 2x = 20, genome size 534 Mb) with nine pairs of autosomes plus XX (♀) or XY (♂) chromosomes. Cannabis sativa L. is an important economic plant for the production of food, fibre, oils, and intoxicants. Genotypes (varieties or chemovar) of hemp with low Δ9-tetrahydrocannabinol content are used for industrial applications. Varieties with high Δ9-tetrahydrocannabinol or high cannabidiol content are used for medicinal applications. Biochemical and molecular methods can be used for identification and classification. An important step for molecular biology methods is to obtain the matrix of the native and sufficiently pure DNA. We tested two different experimental variant of samples (20 mg and 100 mg) of seeds, oilcake and dried flowers for analysis of the Italian variety Carmagnola for analysis (harvested in 2014, Hempoint Ltd., Czech Republic). The DNeasy® Plant Mini Kit (Qiagen, GE) was used to isolate the DNA. The DNA concentration and purity was assessed by agarose electrophoresis and via a spectrophotometer. Samples of lower weight yielded lower values of DNA concentration (average 16.30 - 38.90 ng.μL-1), but with better purity than samples of higher weight (ratio A260nm/A280nm for low-weight samples was near 1.80). To test the applicability of DNA analysis, we used two SSR markers (CAN1347 and CAN2913). PCR products were separated on 1% agarose and on 8% polyacrylamide electrophoresis. DNA samples obtained from samples of higher weight exhibited less PCR amplification than samples of lower weight. We found no effect of sample weight on the formation of non-specific amplification products during the PCR reaction. Based on our results we can be recommended for practical isolation procedure using DNeasy® Plant Mini Kit with lower of sample weight (20 mg). In future work the procedure for DNA isolating from wheat-cannabis products, e. g. breads, rolls or pasta, will be optimized.
The recreational and medicinal desirability of marijuana (Cannabis sativa), fomented by widespread and generationally shifted social acceptance, has created a perfect storm for the medical community. The emergence of this drug highlights the lack of scientific and clinical understanding of its effects, including those that are health adverse as well as medicinal. This review illustrates established findings to guide psychiatric practice (and nonpractice) and outlines open questions to motivate further study. This work provides a general introduction on the properties of marijuana, briefly describes the cannabinoid receptors (cell surface proteins that recognize the plant’s psychoactive constituent, Δ9-tetrahydrocannabinol), and delineates the potential utility of agents that activate such receptors. Current theories on the workings of the endogenous neurotransmitter system that is hijacked by Δ9-tetrahydrocannabinol are considered. Finally, this review proposes possible ways in which growing knowledge of ...
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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.
Delta(9)-tetrahydrocannabinol binds cannabinoid (CB(1) and CB(2)) receptors, which are activated by endogenous compounds (endocannabinoids) and are involved in a wide range of physiopathological processes (e.g. modulation of neurotransmitter release, regulation of pain perception, and of cardiovascular, gastrointestinal and liver functions). The well-known psychotropic effects of Delta(9)-tetra hydrocannabinol, which are mediated by activation of brain CB(1) receptors, have greatly limited its clinical use. However, the plant Cannabis contains many cannabinoids with weak or no psychoactivity that, therapeutically, might be more promising than Delta(9)-tetra hydrocannabinol. Here, we provide an overview of the recent pharmacological advances, novel mechanisms of action, and potential therapeutic applications of such non-psychotropic plant-derived cannabinoids. Special emphasis is given to cannabidiol, the possible applications of which have recently emerged in inflammation, diabetes, cancer, affective and neurodegenerative diseases, and to Delta(9)-tetrahydrocannabivarin, a novel CB(1) antagonist which exerts potentially useful actions in the treatment of epilepsy and obesity.
A genetic factor that blocks the cannabinoid biosynthesis in Cannabis sativa has been investigated. Crosses between cannabinoid-free material and high content, pharmaceutical clones were performed. F1s were uniform and had cannabinoid contents much lower than the mean parental value. Inbred F2 progenies segregated into discrete groups: a cannabinoid-free chemotype, a chemotype with relatively low cannabinoid content and one with relatively high content, in a monogenic 1:2:1 ratio. In our model the cannabinoid knockout factor is indicated as a recessive allele o, situated at locus O, which segregates independently from previously presented chemotype loci. The genotype o/o underlies the cannabinoid-free chemotype, O/o is expressed as an intermediate, low content chemotype, and O/O is the genotype of the high content chemotype. The data suggests that locus O governs a reaction in the pathway towards the phenolic cannabinoid precursors. The composition of terpenoids and various other compound classes of cannabinoid-free segregants remains unaffected. Backcrossing produced cannabinoid-free homologues of pharmaceutical production clones with potential applications in pharmacological research. A new variant of the previously presented allele ‘B 0’, that almost completely obstructs the conversion of CBG into CBD, was also selected from the source population of the cannabinoid knockout factor.
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.