Industrial Crops & Products 145 (2020) 112082

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Industrial Crops & Products

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Cannabinoid content in industrial hemp (Cannabis sativa L.) varieties grown T

in Slovenia
Taja Glivara,*, Jan Erženb, Samo Kreftc, Marjeta Zagožend, Andreja Čerenakd, Barbara Čehd,
Eva Tavčar Benkovićb,c
a
Biotechnical Faculty, University of Ljubljana, Jamnikarjeva ulica 101, 1000, Ljubljana, Slovenia
b
Freyherr d.o.o., Kersnikova 10, 1000, Ljubljana, Slovenia
c
Faculty of Pharmacy, University of Ljubljana, Aškerčeva cesta 7, 1000, Ljubljana, Slovenia
d
Slovenian Institute of Hop Research and Brewing, Cesta Žalskega tabora 2, 3310, Žalec, Slovenia

A R T I C LE I N FO A B S T R A C T

Keywords: Cannabinoid content in different hemp varieties from the Common catalogue of varieties of agricultural plant species
Cannabis sativa L. is not well known. Hemp (Cannabis sativa L., subsp. sativa) contains a wide range of cannabinoids, where can-
Hemp nabidiol (CBD) and (−)-trans-Δ9-tetrahydrocannabinol (Δ9-THC) are the constituents with known therapeutic
Cannabinoids activity. Also, Δ9-THC is recognized as an illicit drug; therefore, cultivation of hemp is restricted to a 0.2 % limit
THC
of THC content in many European countries. In this study, the cannabinoid profiles of 15 hemp varieties, ac-
CBD
cessible to our research group, were analysed. The content of 13 cannabinoids was determined with HLPC (high-
Varieties
performance liquid chromatography) analysis. Large variations in cannabinoid content among varieties that
grew in uniform conditions (ANOVA p < 0.05) and also within a single variety were found, which shows on un-
uniform genetic profiles of the seed material. The varieties Fedora 17, USO 31, Tisza, Tiborszallasi, and Antal all
displayed good response to growth conditions, related to cannabinoid content, in Slovenia.

1. Introduction tetrahydrocannabinolic acid (Δ9-THC-A), cannabidiolic acid (CBD-A),

Hemp (Cannabis sativa L. subsp. sativa) is a subspecies of the genus
Cannabis (Cannabaceae), which also includes medicinal cannabis
(Cannabis sativa L. subsp. indica). It is an annual plant that occurs in
versatile forms, depending on the variety and availability of space for
growth. It is mostly dioecious, meaning that the male and female
flowers are located on separate plants. Seeds can only be found on fe-
male plants.
Cannabis plants contains two main cannabinoids; one is a psy-
choactive cannabinoid substance (−)-trans-Δ9-tetrahydrocannabinol,
better known as THC, which earns medicinal cannabis its classification
as an illicit drug. The second is cannabidiol (CBD), represented pri-
marily in hemp. Cannabidiol is known as a pharmacologically active
substance and is becoming more and more important for use in med-
icinal applications.
A broad chemical profile of cannabinoids, that can be found in
cannabis, exhibits the typical C21 terpenophenolic skeleton, including
their derivatives and transformation products (Pertwee, 2014). Can-
nabinoids are initially formed as carboxylic acids (eg, Δ9-
cannabichromenic acid (CBC-A), and tetrahydrocannabivarin car-
boxylic acid (Δ9-THCV-A)) that, when heated, are decarboxylated to
their corresponding neutral forms. Another process caused by heating,
light exposure or ageing, is oxidative degradation, wherein the con-
version of Δ9-THC to cannabinol (CBN) and isomerization of Δ9-THC to
Δ8-tetrahydrocannabinol (Δ8-THC) occurs. The concentrations of can-
nabinolic acid (CBN-A) and CBN, primary chemical degradants of Δ9-
THC-A and Δ9-THC, increase during storage, if exposed to light at 22 °C
as compared with storage in darkness at 4 °C. The concentration of CBD-
A, when exposed to light and 22 °C, decreases with cyclization from
CBD-A to Δ9-THC-A, followed by the degradation of Δ9-THC-A to CBN-A
(Thomas and Elsohly, 2016).
Cannabinoid production in hemp can be affected by numerous
biotic and abiotic factors such as the sex and maturity of the plant, daily
light integral, ambient temperature, nutrient availability, and intensity
of ultraviolet light (Hillig and Mahlberg, 2004). The content of can-
nabinoids was found to be differently distributed in plant parts with the
highest percentage in secretory cells inside glandular trichomes (up to
60 %), highly concentrated in unpollinated female flowers prior to

⁎
Corresponding author.
E-mail addresses: tajcyka@gmail.com (T. Glivar), jan@freyherr.com (J. Eržen), samo.kreft@ffa.uni-lj.si (S. Kreft), marjeta.zagozen@ihps.si (M. Zagožen),
andreja.cerenak@ihps.si (A. Čerenak), barbara.ceh@ihps.si (B. Čeh), eva.tavcar.benkovic@freyherr.com (E. Tavčar Benković).

https://doi.org/10.1016/j.indcrop.2019.112082
Received 22 August 2019; Received in revised form 22 December 2019; Accepted 29 December 2019
0926-6690/ © 2019 Elsevier B.V. All rights reserved.
T. Glivar, et al.
senescence (up to 30 %), pollinated flowers (up to 13 %), leaves (0.05
%), and least in the stem (0.02 %). No cannabinoids have been found in
the roots or seeds (Russo and Marcu, 2017) (Russo, 2011). The highest
cannabinoid concentration (in % of dry weight plant material) can be
found in the bracts of the flowers that contain abundant glandular tri-
chomes (Turner et al., 1981). Better knowledge of the cannabinoid
production in glandular trichomes would help the breeder to identify
the optimum time for harvest (Potter and D., 2009). Mature female
plants are preferred for cannabis extract production, since they produce
higher amounts of cannabinoids. The highest yield of cannabinoids are
achieved if grown without male plants to prevent pollination and the
formation of seeds (Thomas and Elsohly, 2016).
Hemp is becoming well-known for its versatile use and great eco-
nomic importance. Recently, there has been a dramatic increase in
CBD-rich supplementation in the food supplement and cosmetic in-
dustries. Even more potential is reported on the pharmaceutical use of
cannabinoids. Since the pharmaceutical industry requires the highest
quality of materials, it is of the upmost importance to ensure con-
sistency in the chemical profiles of plants intended for such use
(Thomas and Elsohly, 2016). Chemotypes are highly dependent on the
genetics, and in addition the plants are exposed to many natural factors,
which can be very variable among seeds within a single variety. As a
result, a constant profile of cannabinoids is not easy to achieve with
sowing seed. The best way for the propagation to produce uniform
plants is with cuttings (clones).
Current legislation in Slovenia allows cultivation of hemp for the
purpose of seeds production and further propagation, production of
food and beverages, the production of substances for cosmetic purposes,
production of fibres, animal feed, and for other industrial purposes.
This applies to all varieties listed in the Common catalogue of varieties of
agricultural plant species, whose THC content does not exceed 0.2 %
(“Rules on conditions for obtaining a permit for hemp and poppy cul-
tivation,” 2018) in the upper third part of the dried plant or in a 30 cm
dried plant part, containing at least one female inflorescence
(“Regulation (EU) No 809/2014,” 2014Regulation (EU) No 809/,
2014Regulation (EU) No 809/2014,” 2014).
The aim of this study was to determine the chemical profile of 13
cannabinoids in 15 different hemp varieties, grown in Slovenia.
2. Materials and methods
2.1. Plant material
A total of 15 industrial hemp varieties (Cannabis sativa L., subsp.
sativa) from the Common catalogue of varieties of agricultural plant species
were grown on the fields of the Slovenian Institute of Hop Research and
Brewing (IHPS) in Žalec in the year 2017 and in Gornja Radgona (ap-
proximately 100 km from Žalec) in 2018. The plants were collected at
the phase of maturity at which most Slovenian hemp farmers usually
harvest. The seeds of the same producer were used for each variety in
both years. Properties of investigated varieties, obtained from produ-
cers, web pages (“Zadruga konopko, 2019),(“Ihempfarms, 2019). and
data source (Pacifico et al., 2008), are summarized in Table 1.
On the 3 May 2017, 13 varieties of hemp (Table 1), were sown on
9 m2 large parcels, with each variety in a separate plot, but with no
physical obstacles between them. The plants were not irrigated during
the growth phase. During the growth period, the weather was warmer
than usual; this was one of the warmest summers in Žalec since 1961
(Fig. 1A). The vegetation phase lasted for 135 days. Sampling of the
female plants was carried out on the 15 September 2017. Plants were
dried for 24 h at 35 °C and transferred to the analytical laboratory at the
Faculty of Pharmacy, University of Ljubljana, to be stored in a cold
room in paper bags, protected from light until preparation for analytical
testing.
The second study began on the 29 May 2018, when 14 varieties of
hemp (Table 1) were sown. The parcels were 2 m2 large. The plants
2
Industrial Crops & Products 145 (2020) 112082
were not irrigated during the growth phase. Compared with the year
2017, 2018 was cooler with more precipitation (Fig. 1B). Sampling of
the female flowers was carried out on the 30 August 2018. The vege-
tation period lasted for 93 days. Plants were dried, transferred, and
stored with the same procedure as in 2017.
In this study, only few plants were sampled (see Section 2.2), while
50 plants/ha should be sampled according to the legislation directive
and the average content should not exceed 0.2 % (“Rules concerning
the requirements for obtaining permission to cultivate hemp,” 2004).
2.2. Preparation of plant material for chemical characterization
In 2017, three inflorescences, each from different female plants of
each variety were studied. Table 2 illustrates the experimental proce-
dure of the sample preparation. The preparation of the samples began
with careful removal of the seeds from each inflorescence. The bracts
were separated from the stems, leaflets, and supporting leaves. Stems,
leaflets, and supporting leaves of the inflorescence were ground with a
laboratory grinder (30 s, 20.000 rpm) to a size of ≤ 0.5 mm. Samples
from the first and second plants were prepared identically, while the
third inflorescence was additionally separated into upper and lower
parts. The third flower of the varieties Finola and Ferimon was not
separated into an upper and lower part; it was analysed as a whole
flower since not enough material was available.
Developing seeds were present in most plants, so if the seeds would
not be removed the mass ratio of seeds to other parts of the in-
florescences would differ from plant to plant, which would influence
the representability of the results of cannabinoid content. By removal of
the seeds representative results were achieved. For additional com-
parison, bracts were used to obtain the most accurate and comparable
results on cannabinoid profiles among the plants. Also, glandular tri-
chomes contained the highest content of cannabinoids and were mostly
located on the bracts (Turner et al., 1981).
In 2018, one inflorescence of each variety was prepared according
to the procedure described above, but only bracts were analysed. For
the second sample, inflorescences from another plant of each variety
were used with prior removal of seeds and ground with a laboratory
grinder (30 s, 20.000 rpm) to a size of ≤ 0.5 mm as shown in Table 2.
2.3. Cannabinoid content evaluation
The following cannabinoids were analysed: Cannabidivarinic acid
(CBDV-A), Cannabidivarin (CBDV), Tetrahydrocannabivarin (THCV),
Cannabidiolic acid (CBD-A), Cannabidiol (CBD), Cannabigerolic acid
(CBG-A), Cannabigerol (CBG), Cannabinol (CBN), (−)trans-Δ9-tetra-
hydrocannabinol (Δ9-THC), Δ8-tetrahydrocannabinol (Δ8-THC),
Cannabicyclol (CBL), Δ9-tetrahydrocannabinolic acid (Δ9-THC-A), and
Cannabichromene (CBC).
2.4. Cannabinoid extraction
The samples were extracted by adding 10 mL of ethanol (96 %, Kefo
Slovenia) to the 100−200 mg of the accurately weighted ground or
non-ground material in closed plastic tubes. Samples were incubated for
30 min in an ultrasonic bath at 50 °C. The supernatant was removed and
filtered through a 25 μm filter in the HPLC (high-performance liquid
chromatography) vial.
2.5. Chromatographic analysis
Samples were analysed using HPLC as described in one of our other
articles (submitted for publication). The amount of each cannabinoid
(%w/w [%mg of cannabinoid/100 mg herbal drug]) in the original
sample was calculated by using calibration curves of standard com-
pounds.
Additionally, to show the total concentration of Δ9-THC, CBD, and
T. Glivar, et al. Industrial Crops & Products 145 (2020) 112082

Table 1
Basic characteristics of hemp varieties.
Variety Grown in research study in Country origin Genotypic expression Climate adaptation Vegetative cycle CBD content Δ9-THC content
years

Fedora 17 2017 and 2018 France Monoecious Atlantic Early < 125 days 1.5 – 2.0 % < 0.06 %
KC Dora 2017 and 2018 Hungary Dioecious Continental Late < 145 days 1.5 – 2.0 % < 0.12 %
Monoica 2017 and 2018 Hungary Dioecious Continental Medium < 135 days 1.5 – 2.0 % < 0.12 %
USO 31 2017 and 2018 Germany Monoecious Atlantic Early < 125 days 0.5 – 1.0 % < 0.06 %
Helenaa 2017 and 2018 Serbia Dioecious No data available Early < 125 days No data No data available
available
Santhica 27 2017 and 2018 France Monoecious Atlantic Medium < 135 days 1.0–1.5 % < 0.02 %
Tisza 2017 and 2018 Hungary Dioecious Continental Late < 145 days 2.0–3.0 % < 0.12 %
Tiborszallasi 2017 and 2018 Hungary Dioecious Continental Late < 145 days 2.0–3.0 % < 0.20 %
Antalb 2017 and 2018 Hungary Dioecious Continental Late < 145 days 2.0–3.0 % < 0.12 %
Carmagnolac 2017 and 2018 Italy Dioecious Mediterranean Very late < 160 days No data < 0.30 %
available
Finola 2017 and 2018 Finland Dioecious Continental Very early < 110 No data No data available
days available
Kompolti hibrid TC 2017 and 2018 Hungary Dioecious Continental Medium < 135 days 2.0–3.0 % < 0.12 %
Ferimon 2017 France Monoecious Atlantic Early < 125 days 1.0–1.5 % < 0.06 %
Novosadskaa 2018 Serbia No data available No data available Very late < 160 days No data No data available
available
Marinaa 2018 Serbia Dioecious No data available Late < 145 days No data No data available
available

Cannabidiol (CBD), (−)-trans-Δ9-tetrahydrocannabinol (Δ9-THC).

a
Varieties Novosadska, Marina, and Helena are new and have never been included into the Common catalogue of varieties of agricultural plant species. Data
regarding those varieties was gained at producers.
b
Antal was removed from the Common catalogue of varieties of agricultural plant species after experiments were completed.
c
Carmagnola was deleted from Common catalogue of varieties of agricultural plant species in 2019, with market extension until 30 June 2021.

CBG in %w/w [%mg of cannabinoid/100 mg herbal drug], the fol-
lowing equations were used:
% of Total Δ9-THC = % Δ9-THC + (% Δ9-THC-A x 0.877)
% of Total CBD = % CBD + (% CBD-A x 0.877)
% of Total CBG = % CBG + (% CBG-A x 0.878)
The conversion factors 0.877 and 0.878 represent a loss of mole-
cular mass during the decarboxylation reaction (transformation of
carboxylic acid derivatives [Δ9-THC-A, CBD-A, CBG-A] to non-car-
boxylic acid forms [Δ9-THC, CBD, CBG]).
2.6. Statistical analysis
All statistical analyses were performed by using Excel software
Office Home and GraphPad Prism 6. Concentrations, mean values, and
standard deviations were calculated with Excel. In addition, for results
from study in 2017 (number of repetitions: N = 3), statistical analysis
ANOVA and t-tests, were performed with IBM SPSS. For study from
2018, results have not been statistically proceeded since no repetitions
were done (N = 1).
3. Results and discussion
The cannabinoid content of varieties grown in 2017 and 2018 are
summarized in Tables 3,4, and 5. Additionally, total CBD, total Δ9-THC,
total CBG, and the ratio of total Δ9-THC/total CBD content is presented.
The differences in the content of CBD-A, CBG-A, CBN, Δ9-THC-A, CBC,
total CBD, total CBG, and total Δ9-THC between the varieties were
significant (ANOVA p < 0.05). The content of remaining cannabinoids
were not significantly different between the varieties (ANOVA
p > 0.05).
3.1. Total CBD and total Δ9-THC content
It is well known that Cannabis sativa L. is a highly variable species in
terms of botany, genetics, and its chemical profile (Thomas and Elsohly,
2016). Considering that higher parts of the inflorescences are more
exposed to sunlight and exhibit differential hormonal distribution than
lower parts, it is expected that cannabinoid concentration in upper
parts would be higher compared with lower parts. Variability between
the upper part and the lower part of the inflorescence was examined in
the year 2017, as summarized in Table 4. Evaluation was carried out

Fig. 1. Weather conditions of precipitation and temperature (average of the Tmin and Tmax of the daily temperature) during the growth period presented in decades
(data source: ARSO - Slovenian Environment Agency): A.) from May to September in 2017 in Žalec, compared to the average of 30 years, B.) from May to August in
2018 in Gornja Radgona, compared to the average of 49 years.

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T. Glivar, et al. Industrial Crops & Products 145 (2020) 112082

Table 2
Experimental design in 2017 and 2018 for inflorescences presenting year of sowing, sequential flower, part of the flower, and sample condition.
Year Inflorescence Part of flower Grinded/non-grinded sample

2017 first Bracts from whole inflorescence non-grinded

first Rest of the inflorescence grinded
second Bracts from whole inflorescence non-grinded
second Rest of the inflorescence grinded
third Bracts from upper inflorescence part non-grinded
third Bracts from lower inflorescence part non-grinded
third Rest of the upper inflorescence part grinded
third Rest of the lower inflorescence part grinded
2018 first Bracts from whole inflorescence non-grinded
second Whole inflorescence grinded

with comparison of the total CBD and total Δ9-THC content, as their
ratios are the most interesting of all the analyzed cannabinoids.
The distribution of total CBD in bracts from the upper and lower
inflorescence parts was found to be equal. A considerable difference
was observed only with the variety Antal (upper part 5.970 %, lower
part 13.460 %, mean 9.715 % ± 3.745). If compared with the results
from bracts of whole inflorescence (of first and second flower) analysis
(mean value was found to be 9.513 % ± 1.572), results are similar. The
difference could be explained by ununiformed quality of the in-
florescences. It could be theorized that the upper parts stopped ma-
turing during growth because of excessive sunlight exposure.
Concentration of total Δ9-THC had shown no important differences
between parts of the inflorescences for any variety. With two varieties,
Tiborszallasi (mean value 3.304 % ± 0.097) and Carmagnola (mean
value 2.457 % ± 0.089), extreme values of high total Δ9-THC content
were detected. High values are presumably due to the non-uniform
genetic material, possibly due to cross-pollination in a previous gen-
eration between seed-producing female plants and male plants from
neighboring fields with contrasting genetics, causing higher Δ9-THC
content.
According to the obtained results, it can be concluded that no sig-
nificant differences in the total Δ9-THC/total CBD content between the
upper and lower bracts of the uppermost inflorescences was observed
(paired Student t-test: p = 0.44). Additionally, in a research study by
Namdar et al. where cannabinoids were analysed in the uppermost,
middle, and lowermost inflorescences of hemp, it has been found that
the uppermost inflorescences contained a higher content of cannabi-
noids compared with the lowermost inflorescences (Namdar et al.,
2018). In this study, the uppermost inflorescences were used. The
highest content of total CBD (> 5 % reported as %w/w) produced in
both years (Table 4 and Table 5) was found to be in the bracts of the
varieties Antal (9.580 % ± 1.287 in 2017 and 6.159 % in 2018), Car-
magnola (7.069 % ± 3.339 in 2017 and 5.726 % in 2018), Helena
(5.873 % ± 1.816 in 2017 and 8.108 % in 2018), and Tiborszallasi
(4.666 % ± 1.931 and 4.876 % in 2018). The lowest total CBD content
was detected with the varieties USO 31 (1.207 % ± 0.356 in 2017 and
0.735 % in 2018) and Santhica 27 (0.038 % ± 0.042 in 2017 and 0.010
% in 2018) (Fig. 2A).
Important information about a variety is its uniformity in terms of
cannabinoid content (chemotype), which can be evaluated with the
standard deviation in total CBD, in our case from inflorescences from
three different plants in experiment from 2017. High uniformity within
the population (standard deviation < 0.5) was found in Fedora 17, USO
31, and Santhica 27 (Table 4). The varieties Carmagnola, Monoica,
Helena, Tisza, and Tiborszallasi displayed low uniformity of the seeds
with high standard deviation (sd > 2) within the population (Table 4).
In hemp cultivation, special focus must also be given to the allowed
concentration of Δ9-THC as different countries have different regula-
tions. The limit of 0.2 % Δ9-THC is prescribed by Slovenian legislation
(“Rules on conditions for obtaining a permit for hemp and poppy cul-
tivation,” 2018). The highest content of total Δ9-THC (> 0.2 %) was
detected in bracts of the following varieties in 2017: Tiborszallasi
(2.509 % ± 1.627), Carmagnola (1.015 % ± 1.020), Tisza (0.895
% ± 1.004), KC Dora (0.812 % ± 0.991), and Antal (0.317 % ± 0.042).
Other varieties showed high uniformity (sd < 0.07) and lower total
THC content (< 0.2 %) (Fig. 2B) (Table 4). Results from 2018 confirm
that levels of total Δ9-THC in bracts for all listed varieties except for
Tiborszallasi (0.172 %) exceeded suitable quantities, with the addition
of the variety Helena (0.280 %) with a value slightly above the limit
(Fig. 2B) (Table 5).
It was shown that total Δ9-THC and total CBD reached higher con-
centrations in 2017 for the majority of the varieties. In 2017, en-
vironmental conditions were warmer and dryer, compared with year
2018. From this study it can be concluded that these environmental
conditions are more suitable for CBD production, but it could not be
summarized, which of the above mentioned weather parameters con-
tributed most to the higher content of total CBD. The data of total Δ9-
THC content in the bracts are higher than they would be with me-
chanical harvesting since with manual harvesting only inflorescences
were removed and agitation of the inflorescences is minimized. With
mechanical harvesting of bigger stems, their contribution to the final
weight of the sample is added.
In order to confirm that total Δ9-THC values do not exceed the al-
lowed levels, we verified the content of total CBD and total Δ9-THC in
the remaining parts of the inflorescences (stems, leaflets, and sup-
porting leaves) compared with bracts from inflorescences in the study
from 2017, as shown in Figs. 3A and 3B. Information gathered from
publications indicate that it is expected that cannabinoid ratios between
isolated glandular trichomes and whole pollinated flowers is approxi-
mately 1:4 (Russo and Marcu, 2017) in favor of glandular trichomes.
From our results, as shown by evidence in Figs. 3A and 3B, it can be
proven that the content of total CBD and total Δ9-THC in the bracts is
between two to eight times (on average four times) higher than in other
parts of the inflorescence (without bracts). It can be stated that the
remaining parts of the inflorescence present an additional mass with
lower cannabinoid concentration. In the case of analysing the whole
inflorescence, leaflets and stems present a contributing factor to de-
tection of lower cannabinoid concentration than the ones detected in
bracts.
To further confirm this finding, in 2018, cannabinoid comparisons
between bracts and whole inflorescence (with bracts, stems, leaflets,
and supporting leaves) had been performed (Table 5). Results showed
up to four times lower values detected in whole inflorescences (with the
exception of USO 31). Also, Turner et al. reported about the highest
levels of cannabinoids being found in bracts (Turner et al., 1981). The
content of total Δ9-THC in case of whole inflorescences would exceed
the limit of < 0.2 % only with KC Dora (0.878 %). According to pre-
vious findings regarding the KC Dora uniformity, this phenomenon is
probably caused due to a different chemotype of the variety.
Relating to the above facts it can be assumed that, according to
current Slovenian legislation, varieties wouldn’t presumably exceed the
required limit, especially when plants are harvested mechanically, since

4
T. Glivar, et al. Industrial Crops & Products 145 (2020) 112082

Table 3
Cannabinoid content of bracts from the first, second, and third flower from a study in the year 2017, with results of statistical analysis performed by ANOVA.
Cannabinoids CBG (Cannabigerol), CBDV (Cannabidivarin), THCV (Tetrahydrocannabivarin), CBL (Cannabicyclol), and Δ8-THC (Δ8-tetrahydrocannabinol) were not
detected in any of the varieties. The values of the third flower was previously averaged (up and low) before the total averaging and the calculation of the standard
deviation was performed.
Variety Flower CBDV-A CBD-A CBD CBG-A CBN Δ9-THC Δ9-THC-A CBC
(%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w)

Fedora 17 first 0.112 1.587 0.071 0.021 ND ND ND 0.020

second 0.016 2.281 0.416 0.040 ND ND 0.070 0.022
thirdup 0.016 2.040 0.071 0.029 ND ND 0.060 0.018
thirdlow 0.020 3.520 0.132 0.044 ND ND 0.092 0.016
Mean ± sd 0.049 ± 0.045 2.216 ± 0.489 0.196 ± 0.156 0.032 ± 0.008 ND ND 0.049 ± 0.034 0.020 ± 0.002
KC Dora first 0.016 2.232 0.407 0.039 ND 0.027 0.068 0.021
second 0.007 2.699 0.440 0.095 ND 0.528 1.922 0.017
thirdup 0.051 5.618 0.136 0.217 ND ND 0.176 ND
thirdlow 0.040 4.351 0.088 0.114 ND ND 0.133 ND
Mean ± sd 0.023 ± 0.017 3.305 ± 1.203 0.320 ± 0.147 0.100 ± 0.052 ND 0.185 ± 0.243 0.715 ± 0.854 0.013 ± 0.009
Monoica first 0.050 5.113 0.168 0.173 ND 0.025 0.137 0.020
second 0.020 9.819 0.566 0.343 ND 0.043 0.320 0.026
thirdup 0.018 6.993 0.340 0.145 ND 0.026 0.224 0.018
thirdlow 0.015 6.685 0.279 0.107 ND 0.030 0.210 0.019
Mean ± sd 0.029 ± 0.015 7.257 ± 1.944 0.348 ± 0.165 0.214 ± 0.093 ND 0.032 ± 0.008 0.225 ± 0.075 0.022 ± 0.003
USO 31 first 0.026 0.785 0.027 0.012 ND ND 0.021 0.040
second 0.009 1.224 0.472 0.029 ND ND 0.058 0.054
thirdup 0.021 1.452 0.008 0.033 ND ND 0.047 0.051
thirdlow 0.018 1.563 0.071 0.038 ND ND 0.048 0.052
Mean ± sd 0.018 ± 0.007 1.172 ± 0.297 0.180 ± 0.207 0.025 ± 0.010 ND ND 0.042 ± 0.015 0.048 ± 0.006
Helena first 0.032 8.618 0.259 0.184 ND 0.024 0.263 0.010
second 0.032 6.863 0.335 0.244 ND 0.028 0.197 0.013
thirdup 0.045 4.190 0.147 0.212 ND 0.010 0.126 0.011
thirdlow 0.034 3.392 0.100 0.124 ND 0.022 0.097 0.012
Mean ± sd 0.034 ± 0.004 6.424 ± 1.995 0.239 ± 0.088 0.199 ± 0.033 ND 0.022 ± 0.005 0.190 ± 0.062 0.012 ± 0.001
Santhica 27 first 0.002 0.009 ND 0.090 ND ND ND ND
second ND 0.110 ND 2.585 ND ND 0.006 0.005
thirdup 0.003 0.006 ND 2.371 ND ND ND ND
thirdlow 0.003 0.014 ND 3.424 ND ND ND ND
Mean ± sd 0.002 ± 0.001 0.043 ± 0.047 ND 1.857 ± 1.256 ND ND 0.002 ± 0.003 0.002 ± 0.002
Tisza first 0.024 3.466 0.081 0.185 ND 0.083 2.543 ND
second 0.017 8.398 0.472 0.188 ND 0.028 0.254 ND
thirdup 0.020 3.480 0.112 0.189 ND 0.013 0.128 ND
thirdlow 0.020 3.421 0.109 0.183 ND 0.016 0.118 ND
Mean ± sd 0.020 ± 0.003 5.105 ± 2.329 0.221 ± 0.178 0.186 ± 0.001 ND 0.042 ± 0.030 0.973 ± 1.111 ND
Tiborszallasi first 0.026 4.241 0.156 0.417 ND 0.172 4.345 ND
second 0.024 7.667 0.601 0.210 ND 0.036 0.235 0.033
thirdup 0.023 3.182 0.132 0.208 ND 0.182 3.671 0.013
thirdlow 0.017 2.925 0.105 0.190 ND 0.145 3.491 0.012
Mean ± sd 0.023 ± 0.003 4.987 ± 1.956 0.292 ± 0.219 0.275 ± 0.100 ND 0.124 ± 0.062 2.720 ± 1.785 0.015 ± 0.013
Antal first 0.193 8.747 0.270 0.598 ND 0.024 0.274 ND
second 0.058 12.119 0.456 0.512 ND 0.034 0.380 ND
thirdup 0.110 6.420 0.340 0.263 ND 0.022 0.199 ND
thirdlow 0.098 14.743 0.530 0.503 ND 0.042 0.458 ND
Mean ± sd 0.118 ± 0.056 10.483 ± 1.378 0.387 ± 0.083 0.497 ± 0.088 ND 0.030 ± 0.004 0.327 ± 0.043 ND
Carmagnola first 0.052 8.459 0.789 0.154 ND 0.058 0.237 0.038
second 0.050 10.918 0.893 0.211 ND 0.068 0.292 0.049
thirdup 0.528 2.840 0.085 0.054 ND 0.138 2.745 0.008
thirdlow 0.498 2.739 0.082 0.076 ND 0.105 2.581 0.011
Mean ± sd 0.205 ± 0.218 7.389 ± 3.404 0.589 ± 0.360 0.143 ± 0.060 ND 0.082 ± 0.028 1.064 ± 1.131 0.032 ± 0.016
Kompolti hibrid first 0.017 3.882 0.094 0.072 ND ND 0.123 0.009
TC second 0.015 4.251 0.242 0.438 ND ND 0.147 0.013
thirdup 0.036 5.884 0.189 0.473 ND ND 0.179 0.020
thirdlow 0.042 6.433 0.209 0.334 ND ND 0.195 0.021
Mean ± sd 0.024 ± 0.011 4.764 ± 0.998 0.178 ± 0.062 0.305 ± 0.165 ND ND 0.152 ± 0.026 0.014 ± 0.005
Finola first 0.052 1.126 0.670 0.009 0.003 0.034 0.031 0.022
second 0.115 1.727 1.413 0.011 0.005 0.059 0.039 0.067
thirda 0.050 1.659 0.187 0.007 ND 0.017 0.045 0.055
Mean ± sd 0.072 ± 0.030 1.504 ± 0.269 0.757 ± 0.504 0.009 ± 0.002 0.003 ± 0.002 0.037 ± 0.017 0.038 ± 0.006 0.048 ± 0.019
Ferimon first 0.024 4.455 0.250 0.045 ND 0.009 0.116 0.003
second 0.017 6.096 0.521 0.023 ND 0.033 0.181 0.020
thirda 0.018 4.719 0.275 0.064 ND 0.014 0.118 0.004
Mean ± sd 0.020 ± 0.003 5.090 ± 0.719 0.349 ± 0.122 0.044 ± 0.017 ND 0.019 ± 0.010 0.138 ± 0.030 0.009 ± 0.008
p (ANOVA of the differences 0.192 0.001 0.164 0.002 / 0.315 0.037 0.001
between varieties)

Cannabidivarinic acid (CBDV-A), Cannabidiolic acid (CBD-A), Cannabidiol (CBD), Cannabigerolic acid (CBG-A), Cannabinol (CBN), (−) trans-Δ9-tetra-
hydrocannabinol (Δ9-THC), Δ9-tetrahydrocannabinolic acid (Δ9-THC-A), Cannabichromene (CBC), ND (not detected), up (upper), low (lower).
a
in the Finola and Ferimon varieties the third flower was not separated into upper and lower parts and was analysed as a whole flower (see Section 2.2).

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T. Glivar, et al. Industrial Crops & Products 145 (2020) 112082

Table 4
Total CBG, total CBD, total Δ9-THC content, and the ratio of total Δ9-THC/total CBD in the bracts of inflorescence from the first, second, and third flowers from a
study in the year 2017, with results of statistical analysis performed by ANOVA. The values of the third flower was previously averaged (up and low) before the total
averaging and the calculation of the standard deviation was performed.
Variety Flower Total CBG Total CBD Total Δ9-THC Ratio of total Δ9-THC/ total CBD
(%w/w) (%w/w) (%w/w)

Fedora 17 first 0.018 1.463 ND N/A

second 0.035 2.416 0.061 1:39
thirdup 0.026 1.861 0.052 1:35
thirdlow 0.038 3.220 0.081 1:40
Mean ± sd 0.028 ± 0.007 2.140 ± 0.481 0.043 ± 0.030 1:50
Monoica first 0.152 4.652 0.145 1:32
second 0.301 9.177 0.324 1:28
thirdup 0.128 6.473 0.222 1:29
thirdlow 0.094 6.141 0.214 1:29
Mean ± sd 0.188 ± 0.082 6.712 ± 1.870 0.229 ± 0.073 1:29
Helena first 0.162 7.817 0.254 1:31
second 0.214 6.354 0.201 1:32
thirdup 0.186 3.821 0.120 1:32
thirdlow 0.109 3.075 0.107 1:29
Mean ± sd 0.175 ± 0.029 5.873 ± 1.816 0.189 ± 0.058 1:31
Tisza first 0.163 3.121 2.313 1:1
second 0.165 7.837 0.251 1:31
thirdup 0.166 3.165 0.125 1:25
thirdlow 0.160 3.109 0.119 1:26
Mean ± sd 0.164 ± 0.001 4.698 ± 2.219 0.895 ± 1.004 1:5
Antal first 0.525 7.941 0.264 1:30
second 0.449 11.085 0.367 1:30
thirdup 0.231 5.970 0.196 1:30
thirdlow 0.442 13.460 0.444 1:30
Mean ± sd 0.437 ± 0.078 9.580 ± 1.287 0.317 ± 0.042 1:30
Finola first 0.008 1.658 0.061 1:27
second 0.009 2.928 0.093 1:31
thirda 0.006 1.643 0.057 1:29
Mean ± sd 0.008 ± 0.002 2.076 ± 0.602 0.071 ± 0.016 1:29
Kompolti hibrid TC first 0.064 3.498 0.108 1:32
second 0.385 3.970 0.129 1:31
thirdup 0.415 5.350 0.157 1:34
thirdlow 0.293 5.850 0.171 1:34
Mean ± sd 0.268 ± 0.145 4.356 ± 0.900 0.133 ± 0.023 1:33
(%w/w) (%w/w) (%w/w)
KC Dora first 0.034 2.365 0.087 1:27
second 0.083 2.807 2.213 1:1
thirdup 0.191 5.063 0.155 1:33
thirdlow 0.100 3.904 0.117 1:33
Mean ± sd 0.088 ± 0.045 3.219 ± 0.913 0.812 ± 0.991 1:4
USO 31 first 0.010 0.716 0.019 1:38
second 0.026 1.545 0.051 1:30
thirdup 0.029 1.282 0.041 1:31
thirdlow 0.033 1.442 0.042 1:34
Mean ± sd 0.022 ± 0.009 1.207 ± 0.356 0.037 ± 0.014 1:33
Santhica 27 first 0.079 0.008 ND N/A
second 2.269 0.096 0.005 1:20
thirdup 2.082 0.005 ND N/A
thirdlow 3.006 0.012 ND N/A
Mean ± sd 1.631 ± 1.103 0.038 ± 0.042 0.002 ± 0.002 1:23
Tiborszallasi first 0.366 3.876 3.983 1:1
second 0.184 7.325 0.241 1:30
thirdup 0.182 2.923 3.401 1:1
thirdlow 0.167 2.671 3.207 1:1
Mean ± sd 0.241 ± 0.088 4.666 ± 1.931 2.509 ± 1.627 1:2
Carmagnola first 0.135 8.208 0.265 1:31
second 0.185 10.468 0.324 1:32
thirdup 0.047 2.576 2.545 1:1
thirdlow 0.067 2.484 2.368 1:1
Mean ± sd 0.126 ± 0.053 7.069 ± 3.339 1.015 ± 1.020 1:7
Ferimon first 0.040 4.158 0.111 1:38
second 0.020 5.867 0.191 1:31
thirda 0.056 4.413 0.117 1:38
Mean ± sd 0.039 ± 0.015 4.812 ± 0.753 0.140 ± 0.037 1:34
p (ANOVA of the differences between varieties) 0.002 0.001 0.042 /

Cannabidiol (CBD), (−)-trans-Δ9-tetrahydrocannabinol (Δ9-THC), Cannabigerol (CBG), ND (not detected), up (upper), low (lower).
a
in the Finola and Ferimon varieties the third flower was not separated into upper and lower part and it was analysed as a whole flower (see Section 2.2).

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T. Glivar, et al. Industrial Crops & Products 145 (2020) 112082

Table 5
Cannabinoid content and the ratio of total Δ9-THC/total CBD in bracts from inflorescences and whole inflorescences from a study in the year 2018. Cannabinoids CBG
(Cannabigerol), Δ8-THC (Δ8-tetrahydrocannabinol), and CBL (Cannabicyclol) were not detected in any of the varieties.
Variety B/WI CBDV-A CBDV THCV CBD-A CBD CBG-A CBN Δ9-THC Δ9-THC-A CBC Total CBG Total CBD Total Ratio total
Δ9 - Δ9−THC/
THC total CBD
(%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/w) (%w/
w)

Fedora 17 B 0.230 0.120 ND 3.158 1.754 0.104 0.005 0.090 0.056 0.099 0.091 4.523 0.139 1:33
WI 0.065 ND ND 1.301 0.492 0.041 0.001 0.028 0.025 0.031 0.036 1.633 0.050 1:33
KC Dora B ND ND ND 3.343 2.582 0.219 0.016 0.723 0.532 0.173 0.193 5.513 1.190 1:5
WI 0.004 ND ND 0.439 0.311 0.030 0.012 0.545 0.379 0.038 0.026 0.696 0.878 1:1
Monoica B 0.030 ND ND 1.490 1.156 0.049 ND 0.057 0.021 0.085 0.043 2.462 0.075 1:33
WI 0.005 ND ND 0.497 0.530 0.018 0.001 0.024 0.006 0.042 0.016 0.966 0.029 1:33
USO 31 B 0.008 ND ND 0.635 0.178 0.012 0.001 0.009 0.015 0.009 0.010 0.735 0.022 1:34
WI 0.007 ND ND 0.813 0.176 0.029 ND 0.010 0.020 0.014 0.025 0.889 0.027 1:32
Helena B 0.045 ND ND 8.825 0.368 0.283 ND 0.035 0.280 0.026 0.249 8.108 0.280 1:29
WI 0.009 ND ND 2.504 0.178 0.057 ND 0.016 0.075 0.015 0.050 2.374 0.082 1:29
Santhica 27 B ND ND ND 0.011 ND 3.566 ND ND ND 0.020 3.131 0.010 ND N/A
WI ND ND ND ND ND 1.423 ND ND ND 0.021 1.249 ND ND N/A
Tisza B 0.017 ND ND 3.152 1.235 0.109 0.005 0.432 0.413 0.094 0.095 4.000 0.794 1:5
WI 0.007 ND ND 1.489 0.606 0.066 0.001 0.040 0.029 0.044 0.058 1.912 0.066 1:29
Tiborszallasi B 0.033 ND ND 3.625 1.697 0.362 0.003 0.112 0.068 0.102 0.318 4.876 0.172 1:28
WI 0.006 ND ND 1.073 0.681 0.095 ND 0.040 0.020 0.038 0.083 1.622 0.058 1:28
Antal B 0.030 ND ND 5.153 1.639 0.491 0.006 0.769 1.314 0.159 0.431 6.159 1.922 1:3
WI 0.007 ND ND 1.234 0.744 0.078 ND 0.037 0.032 0.042 0.068 1.827 0.065 1:28
Carmagnola B 0.073 ND ND 4.120 2.113 0.184 0.003 0.150 0.120 0.140 0.162 5.726 0.255 1:22
WI 0.137 0.061 0.152 0.161 0.112 0.007 0.003 0.105 0.063 0.026 0.006 0.254 0.161 1:2
Kompolti B 0.024 ND ND 6.012 0.858 0.201 ND 0.071 0.140 0.064 0.177 6.131 0.193 1:32
hibrid TC WI 0.004 ND ND 1.047 0.459 0.076 0.002 0.027 0.018 0.031 0.067 1.377 0.043 1:32
Finola B 0.047 ND ND 1.409 1.545 0.017 0.004 0.067 0.022 0.068 0.015 2.781 0.086 1:32
WI 0.021 0.010 ND 0.573 0.474 0.005 0.001 0.020 0.011 0.024 0.004 0.977 0.030 1:33
Novosadska B 0.076 ND ND 6.485 0.292 0.146 ND 0.026 0.191 0.020 0.128 5.979 0.193 1:31
WI 0.019 ND ND 1.434 0.122 0.022 ND 0.011 0.038 0.012 0.019 1.380 0.044 1:31
Marina B 0.008 ND ND 1.460 0.890 0.081 ND 0.052 0.027 0.057 0.071 2.170 0.075 1:29
WI ND ND ND 0.332 0.389 0.019 0.002 0.016 0.004 0.026 0.017 0.680 0.020 1:34

Cannabidivarinic acid (CBDV-A), Cannabidivarin (CBDV), Tetrahydrocannabivarin (THCV), Cannabidiolic acid (CBD-A), Cannabidiol (CBD), Cannabigerolic acid
(CBG-A), Cannabinol (CBN), (−)-trans-Δ9-tetrahydrocannabinol (Δ9-THC), Δ9-tetrahydrocannabinolic acid (Δ9-THC-A), Cannabichromene (CBC), B (bracts), WI
(whole inflorescence), ND (not detected).

due to the diverse nature of cannabis varieties, more plants within the
sample population represent reliable results.
Influence of Slovenian soil and climate on selected varieties was not
studied up to now, which makes comparisons of the content of CBD and
Δ9-THC from whole inflorescences in year 2018 with declared values
published in publications (summarized in Table 1) interesting. For the
varieties Fedora 17, USO 31, Tisza, Tiborszallasi, and Antal comparable
values of cannabinoid concentrations were found. From this it can be
concluded that varieties reacted to the cultivation environment as
expected. Based on the obtained total CBD and total Δ9-THC values in
whole inflorescences, the variety Santhica 27 is a less appropriate
variety for CBD production since both cannabinoids were present in
minimal amounts.
3.2. Ratio of total Δ9-THC/total CBD
The ratio of total Δ9-THC/total CBD can primarily be used for the
identification of fiber or the indica type of the hemp. A ratio of total Δ9-

Fig. 2. Cannabinoid content from bracts: average value of three inflorescences from 2017 (mean value %w/w with standard deviation) and value in one inflorescence
from 2018 (single value %w/w): A.) total CBD (Cannabidiol), B.) total Δ9-THC ((−)-trans-Δ9-tetrahydrocannabinol).

7
T. Glivar, et al.
Fig. 3. Content of cannabinoids in bracts (average value of inflorescences from three plants) and in parts from the rest of inflorescences, study from 2017 (mean value
%w/w with standard deviation): A.) total CBD (Cannabidiol), B.) total Δ9-THC ((−)-trans-Δ9-tetrahydrocannabinol).
THC/total CBD lower than 1.0 is attributed to “fiber-type’’ plants (Hillig
and Mahlberg, 2004), which applies to all varieties considered in our
study. This ratio can also be used to measure seed uniformity. This is
important information for breeders, who must know if genetic sowing
material is consistent. When compared, the ratio of total Δ9-THC /total
CBD in bracts from the first, second, third upper, and third lower flower as
shown in Table 4 had high uniformity for the varieties Antal (with
standard deviation ± 0), Helena ( ± 1), Monoica ( ± 1), Kompolti hi-
brid TC ( ± 1), Finola ( ± 2), Ferimon ( ± 3), and USO 31 ( ± 3) has
been recognized. Varieties with low uniformity were found to be Car-
magnola ( ± 15), KC Dora ( ± 13), Tiborszallasi ( ± 13), and Tisza
( ± 12).
In order to confirm these findings, the ratio of total Δ9-THC/total
CBD from bracts and from whole inflorescences from 2018 (Table 5)
was added for comparison. When averaging all values, it was found that
the varieties Helena ( ± 1), Kompolti hibrid TC ( ± 1), Finola ( ± 2),
Monoica ( ± 2), and USO 31 ( ± 3) are highly uniform between both
seasons. For the mentioned varieties we could assume that homogenous
plant genetics are advantageous for farmers that are cultivating hemp
for production of CBD, since consistent content was shown between
both years. Low uniformity was detected with the same varieties as
listed above when compared inside year 2017. For the varieties No-
vosadska, Marina, and Ferimon ratios could not be compared since
these varieties were sown in only one season. The varieties Novosadska
and Marina showed high uniformity between the ratio of total Δ9-THC/
total CBD in bracts and whole inflorescence within year 2018.
3.3. Other cannabinoids in bracts
The following section describes the content of cannabinoids in 2017
and 2018 found in bracts (Tables 3–5).
Besides the total CBD and total Δ9-THC in 2017, the most re-
presented cannabinoid was found to be CBG-A with the highest content
in Santhica 27 (1.857 % ± 1.256), which did not contain significant
amounts of CBD, CBD-A, THC, or Δ9-THC-A. It is known the THCA- and
CBDA-synthases that metabolize cannabinoids from CBG-A are not well
expressed. This could be described and confirmed with results from
whole inflorescence and bracts in 2018, where the concentration of
CBG-A in Santhica 27 was found to be 3.566 % and levels of CBD-A and
Δ9-THC-A were minimal.
Cannabinoid CBG was not detected in any varieties in either year.
Relatively high CBG-A content in Antal (0.497 % ± 0.088 in 2017,
0.491 % in 2018) and in Tiborszallasi (0.362 % in 2018) could show
immaturity of the flowers. Concentration levels of CBG-A lower than
0.3 % correspond to others varieties for both years.
8
Industrial Crops & Products 145 (2020) 112082
The concentration of CBDV-A in 2017 was represented in all vari-
eties. In Carmagnola (0.205 % ± 0.218) and Antal (0.118 % ± 0.056)
levels were higher since in other varieties concentrations were found to
be lower than 0.07 %. In 2018 the highest levels of CBDV-A was found
in Fedora 17 (0.230 %). In other varieties, concentrations were found to
be under < 0.07 % or even nondetectable (Santhica 27 and KC Dora).
Only in Fedora 17, CBDV was detected (0.120 % in 2018); in others
varieties it was undetectable during both years.
In trace amounts CBC was detected (< 0.05 %) or even not detected
in 2017. Interestingly, in 2018 CBC was detected in higher quantities in
the varieties KC Dora (0.173 %), Antal (0.159 %), Carmagnola (0.140
%) and Tiborszallasi (0.102 %), while in other varieties the levels were
less than < 0.1 %. From the results it can be concluded that CBC con-
tent is higher in less mature flowers since flowers in 2018 were har-
vested earlier than those in 2017.
Trace amounts of CBN were detected in both years (< 0.02 %) or
were undetectable. Since CBN is an oxidative product of Δ9-THC, which
is more prominent in aged cannabis samples (Russo, 2011), (Andre
et al., 2016), results suggest that plants were collected at their maturity.
The content of the cannabinoid CBL rises by exposure of CBC to UV-
radiation (Hazekamp, 2007). Results from both years indicate that
storage of flowers after harvesting was appropriate (with no exposure to
light), since the concentration of CBL was found to be undetectable in
2017 and 2018.
Cannabinoids Δ8-THC and THCV were not detected at any of the
varieties in either year.
4. Conclusion
The response of hemp varieties used in research regarding growth
conditions in Slovenia are not well known since they are produced in
different European countries or breeding programs in geographical
areas. The purpose of this study was to compare and to show the che-
mical profile of cannabinoids in varieties that were accessible to our
research group. The major focus of the study was on the total con-
centration of CBD and Δ9-THC in the bracts of fertilized female flowers.
Some of them were already over the phase of their vegetative cycle and
were therefore treated as the cannabinoid production reached the
plateau phase.
From the results presented in this article it can be concluded that:
• The response of hemp varieties to growth conditions in Slovenia was
good in the sense of CBD production in the varieties Fedora 17, USO
31, Tisza, Tiborszallasi, and Antal since the values of CBD and Δ9-
THC in whole inflorescences were found to be comparable to
T. Glivar, et al.
declared values.
• The variety Santhica 27 was found to not be appropriate for CBD
production since only trace amounts of CBD and Δ9-THC were de-
tected.
• High uniformity of the seeds, according to preserved ratio of total
Δ9-THC/total CBD between years, was found with the varieties
Kompolti hibrid TC, USO 31, Finola, Helena, and Monoica. Low
uniformity was found in the varieties Carmagnola, Tiborszallasi,
Tisza, and KC Dora. For these varieties, it is possible that infinite
phenotypic change occurred.
• All tested varieties belong to the ‘fiber-type’’ plants according to the
ratio of total Δ9-THC/total CBD.
• No significant differences in cannabinoid concentration were found
between bracts from upper and lower inflorescence parts of hemp.
• It can be assumed that for all varieties the < 0.2 % limit of total Δ9-
THC will not be exceeded with appropriate sampling.
• The concentration of total CBD and total Δ9-THC proved to be
highest in bracts from inflorescence. It was found that the con-
centration was on average four times higher compared with the rest
parts of the inflorescence (without bracts) and up to four times
higher compared with the concentration of the whole inflorescence.
• High ratios (> 1:27) of total Δ9-THC/total CBD in bracts from both
years were found in the varieties USO 31, Monoica, Helena, Finola,
and Kompolti hibrid TC.
The use of genetically identical plant material from representative
clones under strictly controlled environmental conditions could ensure
reproducible cannabis material by reaching the same growth stages and
by controlling glandular trichomes profiles. Chemical differences can be
seen within the growth cycle, harvesting, and storage due to environ-
mental conditions. The major focus should be genotype consistency of a
specific chemical profile of cannabis to produce a reproducible and
stable chemical composition (Fischedick et al., 2010). With this in-
formation, the farmers and breeders will know more precisely what the
risks and benefits of cultivating a particular variety are in terms of
compliance with Slovenian legislation, depending on the production
purpose. This could enable a better understanding and an in-depth
knowledge of cannabis cultivation in Slovenia for increased yields of
dioecious and monoecious varieties.
CRediT authorship contribution statement
Taja Glivar: Conceptualization, Formal analysis, Data curation,
Writing - original draft, Visualization. Jan Eržen: Methodology,
Investigation. Samo Kreft: Formal analysis, Resources, Writing - review
& editing. Marjeta Zagožen: Investigation. Andreja Čerenak: Writing
- review & editing. Barbara Čeh: Resources, Writing - review & editing,
Project administration, Funding acquisition. Eva Tavčar Benković:
Conceptualization, Methodology, Validation, Writing - review &
editing, Supervision.
Industrial Crops & Products 145 (2020) 112082
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
Research was supported by the Slovenian Research Agency and
Slovenian Ministry of Agriculture, Forestry and Food (research project
reference: V4-1611). We would like to offer special thanks to Matevž
Štefančič, for his laboratory work.
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