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Evaluation of Protein Concentration, Amino Acid Profile and Antinutritional Compounds in Hempseed Meal from Dioecious and Monoecious Varieties

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Hempseed meal from three dioecious and three monoecious varieties has been evaluated for content and quality of the protein and for the concentration of antinutritional compounds. Hemp seeds were obtained from plants grown in two experimental fields for two consecutive years (2011-2012). For all the varieties, hempseed meal resulted in a rich source of protein (34% mean content) with an amino acid profile extremely rich in arginine and slightly poor in lysine. Differences between dioecious and monoecious varieties were observed in the content of antinutritional compounds. They were more concentrated in monoecious varieties in comparison with those dioecious. The concentration of phytic acid in hempseed meal deserves attention in both groups, being 63 and 75.4 g • kg −1 of dry matter in dioecious and monocieous varieties, respectively. The results show that, besides the recognized value of hemp oil, also the hempseed cake could find application in animal feed as a substitute of other cakes (soybean, rapeseed). From this point of view, the dioecious varieties showing lower contents of antinutritional compounds with respect to the monoecious varieties would be preferred.
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American Journal of Plant Sciences, 2015, 6, 14-22
Published Online January 2015 in SciRes. http://www.scirp.org/journal/ajps
http://dx.doi.org/10.4236/ajps.2015.61003
How to cite this paper: Russo, R. and Reggiani, R. (2015) Evaluation of Protein Concentration, Amino Acid Profile and Anti-
nutritional Compounds in Hempseed Meal from Dioecious and Monoecious Varieties. American Journal of Plant Sciences, 6,
14-22. http://dx.doi.org/10.4236/ajps.2015.61003
Evaluation of Protein Concentration, Amino
Acid Profile and Antinutritional Compounds
in Hempseed Meal from Dioecious and
Monoecious Varieties
Roberto Russo, Remo Reggiani
Istituto di Biologia e Biotecnologia Agraria, CNR, Via Bassini 15, Milano, Italy
Email: reggiani@ibba.cnr.it
Received 2 October 2014; revised 5 November 2014; accepted 12 December 2014
Copyright © 2015 by authors and Scientific Research Publishing Inc.
This work is licensed under the Creative Commons Attribution International License (CC BY).
http://creativecommons.org/licenses/by/4.0/
Abstract
Hempseed meal from three dioecious and three monoecious varieties has been evaluated for con-
tent and quality of the protein and for the concentration of antinutritional compounds. Hemp
seeds were obtained from plants grown in two experimental fields for two consecutive years
(2011-2012). For all the varieties, hempseed meal resulted in a rich source of protein (34% mean
content) with an amino acid profile extremely rich in arginine and slightly poor in lysine. Differ-
ences between dioecious and monoecious varieties were observed in the content of antinutri-
tional compounds. They were more concentrated in monoecious varieties in comparison with
those dioecious. The concentration of phytic acid in hempseed meal deserves attention in both
groups, being 63 and 75.4 gkg1 of dry matter in dioecious and monocieous varieties, respectively.
The results show that, besides the recognized value of hemp oil, also the hempseed cake could find
application in animal feed as a substitute of other cakes (soybean, rapeseed). From this point of
view, the dioecious varieties showing lower contents of antinutritional compounds with respect to
the monoecious varieties would be preferred.
Keywords
Antinutritional Compounds, Amino acid Profile, Dioecious, Monoecious, Protein
1. Introduction
Hemp (Cannabis sativa L.) is an annual herbaceous plant belonging to Cannabinaceae family known to have
R. Russo, R. Reggiani
15
played a historically important role in food, fiber and medicine production. For centuries, it has been considered
as one of the most important agricultural crops by providing necessities such as cordage, cloth, food, lighting oil
and medicines. In Europe, it was mainly utilized as a source of fiber and seed. In 1999, the EU produced about
27,000 t of hemp fiber and 6200 t of hemp seed, mostly in France, and 90% of this was used as animal feed. In
particular, the seeds have traditionally been employed as feed for bird and poultry [1]. In other parts of the world,
it was primarily used as a source of psychoactive drug [2]. Hemp has the greatest economical potential if grown
both for seeds and stems (fibers) as residual agricultural products [3].
Hempseed contains more than 30% oil of which more than 80% is polyunsaturated fatty acids [4]. Hempseed
oil is especially rich in two essential fatty acids-linoleic acid (18:2, omega-6) and alpha-linolenic acid (18:3,
omega-3). They are present in a ratio of about 3:1, considered optimal in healthy human adipose tissue [5], and
apparently unique among common plant oils [6]. This preferred omega-6/omega-3 ratio was successfully uti-
lized in animal feed to manipulate the fatty acid pattern in bovine adipose tissue and eggs [7] [8]. There are var-
ious benefits attributed to omega-3 and include anti-cancer, anti-inflammatory and anti-thrombosis properties,
stimulation of general metabolism and promotion of burning fat [5] [9].
Hempseed oil is commonly extracted by solvents and, the resulting by-product, is known as hempseed meal
(HSM) which has good protein level ranging from 30% to 40% in dry matter (DM) of flour depending upon the
variety of hemp used [10]. HSM is a rich source of protein and energy and can represent a nutritious feed sup-
plement for livestock or used for production of a high-protein flour.
Identification and characterization of hempseed proteins showed that edestin, rich in valuable amino acids,
constituted the main protein component in isolate hempseed protein [11]. Another protein structure, rich in me-
thionine and cystine, was found in hemp seeds and subsequently characterized as an albumin protein family
member [12]. Numerous factors are known to influence the nutritional quality of plant proteins, as measured by
their amino acid composition and digestibility. The amino acid composition may be affected by genotypic varia-
bility or agronomic conditions such as soil fertility and postharvest processing that alters the ratio of seed com-
ponents (e.g. shelling). The digestibility of proteins may be affected by protein structure, the presence of antinu-
tritional compounds and high temperature processing [13].
HSM contains antinutritional compounds that need to be considered when feeding this product. In hempseed,
among a vast number of antinutritional compounds, phytate has raised more attention [14]. Phytic acid (inositol
exaphosphate) is the main organic form of phosphorus present in plant seeds. Its presence reduces protein diges-
tibility and increases the excretion of endogenous nitrogen, amino acids and minerals, in particular bivalent ca-
tions [15]. Another nutritionally important group of compounds are the condensed tannins. Tannins (flavan-3-ol
based biopolymers) are known to negatively affect nitrogenous compounds uptake, absorption of minerals, and
reduce weight gain and feed consumption intake in broiler chicks [16]. In the case of sorghum, 1% of increased
tannin content reduced the dietary energy value by 10% [17]. Being protein precipitants, the tannins form com-
plexes with feed proteins and endogenous enzymes. Hence, the weight of the pancreas increases if the feed con-
tains high levels of tannins [18]. Other deleterious compounds considered by animal nutritionists are the sapo-
nins. These substances consist of a sugar moiety usually containing one sugar (glucose, galactose, glucuronic
acid, xylose, rhamnose or methylpentose), glycosidically linked to a hydrophobic aglycone (sapogenin) which
may be in nature triterpenoid or steroid [19]. Dietary saponins depressed growth, feed consumption and egg
production in poultry [20]. These negative effects have been ascribed to several properties of saponins such as
reduced feed intake caused by the astringent and irritating taste of saponins, reduction in intestinal motility and
reduction in protein digestibility, probably due to the formation of sparingly digestible saponin-protein com-
plexes [19]. Moreover, saponins have pronounced haemolytic properties and are responsible for bloat in rumi-
nats [21]. Trypsin inhibitors are considered one of the most important antinutritional factors and are found in
many species of graminaceous, cruciferous and leguminosae. In literature, there are few data about the trypsin
inhibitor activities in HSM [12]. Lastly, the metabolism of some substances contained in flour may lead to the
release of toxic products (e.g. cyanogenic glycosides to hydrogen cyanide) [22].
In the present study, HSM was evaluated as protein source (content and quality) and for the presence of the
main antinutritional compounds (phytic acid, condensed tannins, cyanogenic glycosides, trypsin inhibitors and
saponins). Two groups of hemp varieties were used. The first group was consisting of dioecious varieties (Car-
magnola, Carmagnola Selezionata [CS], Fibranova) and the second group of monoecious varieties (Fedora 17,
Felina 32, Ferimon). The dioecious and monoecious hemp varieties have strong differences in the flowering
time and seed filling [23]. The characterization of these biochemical features will help to better understand the
nutritional quality of HSM and its use as feed ingredient.
R. Russo, R. Reggiani
16
2. Materials and Methods
2.1. Plant Materials and Field Experiment
The experimental field trials on hemp were carried out for two consecutive years (2011-2012) in Italy in two
different locations of Lombardy region: Treviglio (45˚31'N, 9˚35'E; 125 m AMSL) and Cavriana (45˚20'N,
10˚35'E; 170 AMSL). The growing degree units were 579 and 645 for the two years (2011 and 2012, respec-
tively) in Treviglio, and 667 and 721 in Cavriana for the same years. The total rainfall during the growth period
of the hemp plants was 428 and 482 mm in the two years in Treviglio, and 498 and 344 for the same period in
Cavriana. In both sites, the experimental design included the evaluation of 3 dioecious (Carmagnola, CS and Fi-
branova) and 3 monoecious varieties (Futura 75, Felina 32 and Ferimon). The experimental design was a ran-
domized complete block with 3 replicate plots (10 m2/block) of each variety. Hemp plots were harvested at full
maturity stage. Seeds were forced-air dried (30˚C) in a ventilated chamber until 8% seed moisture was reached.
2.2. Oil Extraction and Protein Assay
Seeds were ground in a mortar and mixed with hexane (1:10, w/v). The solution was vigorously shaken for 30
min. After centrifugation, the upper-liquid was collected and the extraction procedure repeated. The dried flour
was used for analyses.
Protein was extracted from defatted flours by Plant Total Protein Extraction Kit (Sigma-Aldrich, Milan, Italy).
The kit includes two reagents, a plant specific protease inhibitor cocktail and a chaotropic reagent with increased
solubilizing power to extract more hydrophobic proteins. Protein content was determined by Quantum Protein
Kit (Euroclone, Milan, Italy) using Bovine Serum Albumin (BSA) as standard.
2.3. Amino Acid Composition
In order to obtain the amino acid profile of the protein of the six varieties of hemp, the protein of each variety
was extracted from a mixture of flours from the various plots. Fifty µL of protein extract was subjected to acid
hydrolysis in 3 M mercaptoethanesulfonic acid containing 2 g∙kg1 NaN3 at 110˚C for 16 h. Hydrolysed samples
were then diluted ten times with HPLC quality water before amino acid analysis. The amino acid composition
was determined by HPLC analysis of the 9-fluorenyl-methoxycarbonyl chloride/1-aminoadamantane (FMOC/
ADAM) derivatives. An aliquot (150 μL) of sample was added to 150 μL of a 200 mM Borate buffer (pH 10.0).
Then, 300 μL of FMOC-Cl (in acetonitrile) was added and derivatization occurred. After 5 min, the reaction was
stopped by the addition of 300 μL of 300 mM ADAM in water-acetonitrile (1:1, v/v) and the reaction lasted for
1 min to block the excess of FMOC through the formation of FMOC-ADAM complex. Then, the sample was
filtered and analyzed by HPLC. The separation of FMOC/ADAM derivatives was performed at a flow rate of
0.9 mLmin1 on a 150 × 4.6 mm Phenomenex Kinetex C18 2.6 µ reverse phase column. Two mobile phases
were used: a) 50 mM Na-acetate (pH 4.2); b) acetonitrile. Phase B was maintained at 28% (v/v) for 3 min, in-
creased linearly to 45% (v/v) over 24 min, increased linearly to 100% (v/v) over 5 min, remained to 100% for 5
min and then returned to 28% (v/v) in 2 min to regenerate the system. Fluorescence labelled amino acids were
monitored using a Jasco fluorescence detector (model FP-920) at an excitation wavelength of 263 nm and emis-
sion wavelength of 313 nm.
2.4. Analysis of Antinutritional Compounds
Phytic acid, condensed tannins, cyanogenic glycosides and trypsin inhibitors were extracted and assayed ac-
cording to Russo and Reggiani [24].
Saponins were extracted overnight from defatted flour with absolute methanol (1:20, w/v). The samples were
centrifuged for 15 min at 15,000 rpm and the methanol extract evaporated to dryness. Then the samples were
resuspended in water: n-butanol (1:2, v/v). After 1 h of agitation, the samples were centrifuged and the higher
alcoholic layer recovered. Saponin content was estimated according to Goel et al. [25] using Quillaja saponin
(Sigma-Aldrich, Milan, Italy) as a reference standard.
2.5. Statistical Analyses
All analyses were carried out in triplicate. Analysis of Variance (ANOVA) was applied to establish significant
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differences (P ≤ 0.01) between hemp varieties in the levels of protein and antinutritional compounds using SPSS
version 16.0 software. Mean separation was performed using Duncan’s test and referring to P ≤ 0.05 probability
level. Pearson’s correlations between antinutritional compounds were also calculated.
3. Results and Discussion
3.1. Hempseed Meal Protein Content
Figure 1 shows the protein content in six hemp varieties in the two years of cultivation (2011 and 2012) and in
the two experimental fields (Treviglio and Cavriana). The protein content in HSM was, on average, 340 g∙kg1
dry matter (DM). Substantial differences are evident between years of cultivation. In 2012, protein concentration
was higher respect to first year for all hemp varieties and in the two experimental fields. This was evidenced
also by ANOVA analysis that showed highly significant difference for year of cultivation (Table 1). In 2012,
the temperatures recorded during the seed filling were higher in comparison with 2011 (resulting in > growing
degree units in both sites), while rainfall resulted more abundant in the first year in Cavriana. As described in
literature, during seed formation hemp prefers high temperatures (27˚C) and mild climate [26]. Moreover, high
temperatures during seed filling intensify the protein synthesis with an increased protein content in meal due to a
better translocation of mineral salts (nitrogen and sulphur) into xylematic tubes [27]. Instead, no difference was
observed between dioecious and monoecious hemp groups in protein concentration (Table 2).
Table 1. F values from Analysis of Variance (ANOVA) for biochemical compound contents in six varieties of hemp.
Source of variance d.f.a Protein Phytic acid Condensed tannins Cyanogenic glycosides Trypsin inhibitors Saponins
Variety (V) 5 12.37** 57.02** 1130.35** 169.80** 445.84** 202.13**
Field (F) 1 6.60* 2.94 10.18** 18.73** 0.48 0.01
Year (Y) 1 269.41** 83.31** 118.88** 140.50** 4.47* 22.07**
VxF 5 0.80 5.52** 0.79 7.83** 2.30 1.51
VxY 5 6.02** 5.08** 14.98** 5.48** 5.08** 2.55*
FxY 1 1.38 8.85** 23.68** 2.87 0.07 0.11
VxFxY 5 4.89** 5.12** 0.12 1.78 3.55** 1.44
aDegrees of freedom; *Significant at P ≤ 0.05; **Significant at P ≤ 0.01.
Table 2. Protein and antinutritional compounds in three dioecious and three monoecious hemp varieties.
Variety Protein1 Phytic acid1 Condensed tannins1 Cyanogenic glycosides1 Trypsin inhibitors2 Saponins1
Carmagnola 337 (cd) 63.6 (cd) 2.14 (d) 0.09 (d) 22.6 (c) 0.47 (d)
CS 348 (ab) 64.8 (c) 2.49 (c) 0.08 (d) 21.7 (d) 0.53 (c)
Fibranova 351 (a) 61.5 (d) 2.21 (d) 0.05 (e) 10.8 (e) 0.49 (d)
Futura 75 342 (bc) 75.7 (ab) 4.56 (a) 0.12 (c) 27.3 (a) 0.65 (b)
Felina 32 351 (a) 76.7 (a) 4.42 (b) 0.17 (a) 26.2 (b) 0.68 (a)
Ferimon 331 (d) 73.9 (b) 4.40 (b) 0.15 (b) 27.7 (a) 0.70 (a)
Mean ± SE 343 ± 3.3 69.4 ± 2.8 3.50 ± 0.48 0.11 ± 0.02 22.7 ± 2.6 0.59 ± 0.04
P group 0.40 <0.01 <0.01 <0.01 <0.01 <0.01
1Data expressed as g∙kg1; 2Data expressed as unitmg1; Means with different letters in parentheses within the same row differ significantly by Dun-
can’s range test (P ≤ 0.05).
R. Russo, R. Reggiani
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Figure 1. Protein content in flours of six varieties of hemp in the two experi-
mental fields (Treviglio and Cavriana) and two years of cultivation (2011 and
2012).
3.2. Amino Acid Profile in Hemp Protein
The Table 3 shows the amino acid (AA) concentration in hydrolyzed protein of six varieties of hemp. The nutri-
tional value of protein is determined by the profile of essential AAs (threonine, methionine, valine, phenylanine,
isoleucine, leucine, tryptophan, histidine and lysine). Among AAs, glutamate and arginine were predominant in
all hemp varieties with a mean of 17.2 and 12.0 g 100 g1 protein, respectively. Arginine in hemp protein results
higher respect to soybean and almost double than in rapeseed protein [28] [29]. This datum could be very inter-
esting for feed industries, since young mammals have a high dietary requirement for arginine due to its role as a
nitrogen carrier in tissue proteins and its key role in gastrointestinal growth and development. Some researchers
consider arginine as a key essential AA for maximal growth of young pigs [30], especially for piglet plasma
concentrations decline throughout a sow’s lactation [31].
Among the essential AAs, threonine content was higher in the group of monoecious varieties (Futura 75, Fe-
lina 32, Ferimon) than in dioecious varieties (Carmagnola, CS, Fibranova). Plant storage protein is often poor in
lysine (cereals) and solforated AAs (legumes). The mean content of lysine was 4.2 g 100 g1 protein, while the
content of methionine and cysteine was 2.4 and 1.7 g 100 g1 protein, respectively. Fibranova showed a methio-
nine content (2.8 g 100 g1 protein) significantly above the mean. Lysine and methionine have been identified
frequently as first limiting essential AAs in feedstuffs of growing cattle and lactating cows. Hemp protein results
deficient in lysine respect to soybean and rapeseed proteins [28] [29]. Instead, methionine content is higher than
in soybean and rapeseed proteins [28] [29]. Feedstuffs rich in methionine are very important for feed industries
that often use methionine additive analogues to ensure animal requirements. In fact, methionine is clearly recog-
nized as first limiting AA in poultry, high-yielding cows and third limiting AA in pigs [32].
As described by House et al. [33], the PDCAAS (protein digestibility corrected amino acid score) value for
hemp protein sources is about 0.5 - 0.6 and is positioned in the same range as the major pulse protein sources
(e.g. beans, lentils), and above cereal grain products, such as whole wheat. The PDCAAS value for hemp protein
will continue to remain in this range due to the limitation in lysine content. Future breeding efforts to increase
lysine content in hemp may be guaranteed if the value of the protein component for animal consumption will
lead to the development of a market for this crop. Therefore, HSM could partly replace soybean meal, in Euro-
pean feed industries, to reduce the expensive import costs of soy from South America. Hempseed derived-
products are already used as feed material for all species [32]. In Mustafa et al. [34], each sheep was feed up to
200 g∙kg1 of HSM and concluded that HSM is an excellent source of RUP (rumen undegraded protein), with
high post-ruminal availability, and may replace canola and soybean meal with no detrimental effects on nutrient
utilization by sheep. Data from feeding trials indicate that HSM could be used up to 20% in laying hens diets,
although Gakhar et al. [35] reported a reduction of body weight of hen after a 4 weeks diet.
R. Russo, R. Reggiani
19
Table 3. Amino acid content (g 100 g1 protein) in hydrolyzed proteins of six varieties of hemp.
Variety Arg Ser Asx Glx Thr Gly Ala Tyr Pro Met Val Phe Ile Leu Cys Trp His Lys
Carmagnola 12.6 5.6 11.3 17.5 2.6 4.6 5.3 3.9 4.7 2.5 5.0 4.9 3.9 6.5 1.4 0.7 2.8 4.3
CS 11.6 5.4 11.3 17.2 2.6 4.8 5.1 3.4 4.8 2.3 5.5 4.7 4.2 6.9 1.8 0.8 2.8 4.0
Fibranova 11.5 4.9 10.3 17.3 3.1 5.0 5.2 4.0 5.1 2.8 5.0 4.6 3.9 7.1 1.9 0.9 3.1 4.4
Futura 75 11.9 5.4 11.1 17.7 3.6 5.0 5.3 2.9 4.9 2.1 5.1 4.8 4.0 7.1 1.4 0.8 2.8 4.0
Felina 32 12.5 5.3 11.0 16.7 3.7 5.0 5.1 3.0 4.1 2.4 5.5 4.6 4.4 7.1 1.8 0.9 2.9 4.0
Ferimon 11.6 4.9 10.5 17.0 3.9 4.9 5.0 3.8 4.5 2.1 5.7 4.6 4.1 6.9 1.9 0.8 3.2 4.6
Mean 12.0 5.3 10.9 17.2 3.3 4.9 5.2 3.5 4.7 2.4 5.3 4.7 4.1 6.9 1.7 0.8 2.9 4.2
SE 0.3 0.2 0.2 0.2 0.3 0.1 0.1 0.3 0.2 0.2 0.2 0.1 0.1 0.1 0.1 <0.1 0.1 0.2
3.3. Antinutritional Compounds in Hempseed Meal
In Table 1 it shows ANOVA for phytic acid, condensed tannins, cyanogenic glycosides, trypsin inhibitors and
saponins for six varieties of hemp. ANOVA showed that is present a great significant variability for varieties,
year of cultivation and their interaction for all the antinutritionals. Instead, variability attributable to experimen-
tal fields was significant only for condensed tannins and cyanogenic glycosides. In Table 2 it shows the levels
of antinutritional compounds for the three dioecious varieties and the three monocieous varieties. For each group
is shown the significance of variability between groups (P). As can be seen, phytic acid was lower in dioecious
varieties than in monocieous varieties. This huge difference between the two groups was put in evidence by the
P value and the Duncan’s test. These elevated contents of phytic acid observed in defatted hemp flour are defi-
nitely higher than soybean (2%) or rapeseed (2.2%) [29] [36]. The main effect of phytic acid is the binding of
important mineral nutrients (iron, zinc, calcium, phosphorus) which is a major concern when HSM is fed to
monogastric animals lacking of the phytase enzyme. As a consequence of this, it is advisable to mix HSM with
other plant flours exhibiting lower levels of phytic acid (cereals) for its use in monogastric animal feed formula-
tions.
In Table 2 it shows the content of condensed tannins. The monocieous and dioecious groups of plants showed
great significant differences between them evidenced by the P value. The monocieous varieties showed a mean
content of condensed tannins (4.46 g∙kg1 DM) about double respect to dioecious varieties (2.28 g∙kg1 DM).
Despite the total amount of condensed tannins in HSM resulted higher respect to soybean (0.1 g∙kg1) [37], it
can be considered relatively low if compared with other feed sources like forage [38] and, therefore, a negligible
nutritional interference can be expected [39]. Moreover, the tannin level necessary for rejection by grazing ani-
mals is about 20 g∙kg1 DM [40]. Since Colombini et al. [41] found that low levels of dietary tannins were re-
lated to increased yields of fat-corrected milk and protein (possibly due to lower protein degradation in the ru-
men and lower N excretion for ruminants), feeding of varieties with the lowest content of condensed tannins
(dioecious) would be desirable.
As can be seen, the mean content of cyanogenic glycosides was low even if different between dioecious and
monocioeus varieties (Table 2). At levels above 100 ppm, the cyanogenic glycosides are dangerous for animal
and human health [42]. However, having to dilute HSM with other flours to reduce the content of protein and
other antinutritionals, the cyanogenic glycosides content falls below the threshold of 100 ppm.
The range of variation for trypsin inhibitor activities in hemp varieties was quite high (Table 2). Trypsin in-
hibitors are responsible for reduced digestibility of seed proteins, and, for this reason, constitute the main
antinutritional factors of plant seed [43]. In Table 2, the activities varied from 10.8 to 27.7 unitmg1 of defatted
flour. However the levels of trypsin inhibitor activities result lower in comparison with those observed in some
cereals and soybean [44] [45]. Among hemp varieties, fibranova exhibited the lowest trypsin inhibitor activity
(letter “e” by Duncan’s range test). Generally plant grains require heat processing to reduce trypsin inhibitors
before their use as food or animal feed [44]. However, the excessive heat may decrease protein solubility and
amino acid availability. Therefore, flours low in trypsin inhibitor activity could be interesting for feed industries
to avoid expensive heat treatments and reduced nutritional quality of feed. The content of saponins showed sig-
nificant difference among hemp varieties (Table 2). The dioecious group showed a mean content of saponins
(0.50 g∙kg1) lower than that observed in monocioeus varieties (0.68 g∙kg1). These levels of saponins are lower
than that observed in flax and soybean [24] [45].
In Table 4 it shows the correlation matrix calculated for the different antinutritional compounds. The Pearson
R. Russo, R. Reggiani
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Table 4. Pearson correlation coefficient (r) among antinutritional compounds in six varieties of hemp.
Compounds Phytic acid Condensed tannins Cyanogenic glycosides Trypsin inhibitors Saponins
Phytic acid 1
Condensed tannins 0.670** 1
Cyanogenic glycosides 0.442** 0.791** 1
Trypsin inhibitors 0.624** 0.719** 0.660** 1
Saponins 0.653** 0.933** 0.796** 0.680** 1
**Correlation is significant at the 0.01 level.
correlation coefficients are given as a measure of linearity between two class of compounds. From Table 4 it is
clear that all the antinutritional compounds considered in the six varieties of hemp are positively correlated.
Probably, the biosynthetic pathways of these compounds in hemp are expressed simultaneously during the de-
velopment of the seed.
4. Conclusions
Feed industries need new rich-protein fodder for animal, to increase meat, milk and egg productions. The re-
maining hemp meal after extraction of oil (which has its own commercial value), could be a new rich protein
source available for use as animal feed. HSM shows a good protein content (34%), an interesting AA profile
with elevated levels of arginine (but slightly poor in lysine) and a discrete digestibility of protein. In ruminants,
heat treatment of hemp was shown to partially increase protein digestibility although such treatment leads to an
increase of costs [46]. Such costs for HSM need to be comparable to other protein feeds to become a viable al-
ternative on commercial farms [46].
The evaluation of antinutritional compounds in HSM varieties, put in evidence the extremely high levels of
phytic acid. From this point of view, HSM from dioecious varieties is better than that from monoecious varieties.
As mentioned above, the restricted limit of 20% of HSM in animal diet allows lowering the concentration of
antinutritionals and in particular of phytic acid. However, it is important to monitor phytate content in monogas-
tric animal, since exposure of animals to high level of phytate over the long term could cause important nutri-
tionally deficiencies, especially for iron and zinc [47]. To increase the use of HSM in feed would be desirable
breeding for low content of phytate. Otherwise, a research has to be undertaken to identify mutants with low-
phytic acid occurred in soybean [48].
Acknowledgements
The authors thank Regione Lombardia for financial support through project “VeLiCa” (no. 14840/RCC) and
“FilAgro” (No. 18093/RCC).
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... However, protein and other nutrients might vary based on various factors such as climate, cultivar, maturity, and oil extraction process. On the other hand, antinutritional (phytic acid, cyanogenic glycosides, trypsin inhibitors, condensed tannins, saponins) and psychoactive cannabinoid substances especially Delta-9-tetrahydrocannabinol (THC) (Russo and Reggiani, 2015) which is found in cannabis seeds should be used in limited levels. A cannabinoid is not produced in cannabis seeds but it can be transferred from the owers and leaves to the seeds and oil. ...
... We assumed that the hemp variety utilized in this study had a higher antinutrient content which exceeded the hens' tolerable limits. As reported by Russo and Reggiani, (2015) that the antinutrient content is in uenced by hemp varieties, where monoecious varieties are higher than dioecious varieties. When at a level of 8% and 12% hemp seed meal, accumulation of antinutrients might decrease protein digestibility and other nutrient absorption required for follicle development and ovulation consequently decreasing egg production which was shown clearly from the 4th week. ...
... However, dietary of 4% hemp seed meal might be still tolerated and had no harmful effects on laying hens, in addition, the high content of methionine, cysteine, and arginine amino acids as well as omega 3 and 6 fatty acids of hemp seed meal are favorable for egg formation (Darmawan et al. 2013;Konca et al. 2019 According to Pojic et al. (2014), the highest antinutrient in hemp seed meal is phytic acid (22.5mg/g) followed by condensed tannin (0.23mg/g), cyanogenic glycosides (3.80µmol/g), and trypsin inhibitors ( 2.88TIU/mg protein). Furthermore, Russo and Reggiani, (2015) discovered that the tannin content of monoecious and dioecious varieties (4.46g/kg; 2.28g/kg, respectively) was higher than that of soybean (0.1g/kg), but not in trypsin inhibitors and saponins. Antinutrients cause physiological and functional effects in animals when consumed above tolerance levels. ...
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Hemp seed meals contain high protein and unsaturated fatty acids which have the potential as a substitute for soybean meal and yield eggs rich in unsaturated fatty acids. Therefore, the study aimed to evaluate the impact of dietary hemp seed meal as a substitute for soybean meal protein on productive performances, egg quality, and yolk fatty acid composition. A total of 120 Lohmann Brown laying hens aged 50 weeks were separated into 4 groups and 10 repetitions with 3 birds each. Birds received treatment without hemp seed meal (control group), or soybean meal substituted with 4%, 8%, and 12% hemp seed meal. Dietary 4% hemp seed meal significantly increased (p < 0.05) feed efficiency and egg production compared to the 8% and 12% hemp seed meal group. Meanwhile, feed intake, egg weight, body weight loss, egg shape index, albumen index, albumen weight, Haugh unit, yolk weight, yolk index, and eggshell thickness had no significant influence (p > 0.05). However, dietary 8% and 12% hemp seed meal significantly increased (p < 0.05) eggshell weight and yolk color score compared to the control and 4% hemp seed meal group. There was a significant increase (p < 0.05) in omega-3 fatty acid levels and a significant decrease in yolk omega-6 to omega-3 fatty acids ratio with an increase in dietary hemp seed meal. It concluded that dietary 4% hemp seed meal as soybean meal substitution was able to increase egg production, and feed efficiency compared to the level of 8% and 12% hemp seed meal. Increasing dietary levels of hemp seed meal increased eggshell weight, yolk color, and omega-3 fatty acids content, and decreased the omega-6 to omega-3 fatty acids ratio.
... Hemp has six major anti-nutrients: phytic acid, condensed tannins, cyanogenic glycosides, trypsin inhibitors, and saponins. These compounds have been shown to reduce feed intake and lead to an enlarged animal pancreas (Russo and Reggiani 2015). Reports have also demonstrated the deleterious effects of these compounds; hence, they are considered unwanted components of hemp even though some of them, such as saponins, possess positive health impacts (Russo and Reggiani 2015). ...
... These compounds have been shown to reduce feed intake and lead to an enlarged animal pancreas (Russo and Reggiani 2015). Reports have also demonstrated the deleterious effects of these compounds; hence, they are considered unwanted components of hemp even though some of them, such as saponins, possess positive health impacts (Russo and Reggiani 2015). Also even though hemp is considered as low-allergenic food source, there are reports that its derived products, especially proteins induce certain mild symptoms when consumed (Shen et al. 2021). ...
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Hemp is a valuable crop with a wide range of use, from applications in foods and textiles to pharmaceuticals. Over recent years, the use of hemp as food and food ingredients has drastically increased. The growth is driven by numerous health benefits hemp possesses and its wide range of applications in the food industry. This review provides the scientific literature concerning the benefits of industrial hemp in the food industry. The relevant historical context of use, recent applications in the food industry, health benefits, various development challenges, and the global market outlook for hemp-based food products have been analyzed. Evidence suggests that today hemp is widely consumed as food or an ingredient in the food. Hemp-based foods are marketed as having various health benefits, although their reception by target consumers and success varies. Besides, scientific research on hemp-derived foods has dramatically increased over recent years. Numerous in vitro and in vivo studies have investigated the health benefits of hemp-based foods. Therefore, there is a promising growth trend in producing novel foods from industrial hemp. Nevertheless, due to health concerns related to THC, there is a general need for regulatory compliance when integrating hemp into foods to ensure product safety before use.
... Hemp (Cannabis sativa L.) is an annual herbaceous plant belonging to Cannabinaceae family known to have played a historically important role in food, fiber and medicine production (Russo and Reggiani, 2015). It commonly referred to as hemp, is a widely cultivated plant of industrial importance, as a source of whole seed, hulled seed, seed meal, oil and fibre (Callaway, 2004). ...
... Antinutrients in the hemp seed that have been isolated are tannins, cyanogenic glycosides, Saponins (Russo and Reggiani, 2015) and cyanogenic glycosides (Ernesto et al., 2002). However, diluting hemp seed meals with another type of flour reduces protein content and decreases cyanogenic glycosides' concentration below 100 ppm. ...
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Edible hemp products or superfood refers to Cannabis sativa or industrial hemp. In general, hemp is a rich source of functional metabolites, such as Tetrahydrocannabinol (THC), Cannabidiol (CBD), and other cannabinoids. Hemp has been widely used in food products, such as bread, cookies, meatballs, energy bars, cooking oil, snacks, and crackers. Hemp has been used for development of cosmetics and supplements. However, the use of hemp is far below its potential because of major challenges such as non‐cost‐effective extraction and isolation, stability and toxicity of the extracts, and legislation related to the use of the extracts. This narrative review comprehensively analyses major phytochemicals in hemp and hemp extracts, and also discusses the most common challenges in applications of hemp derived phytochemicals and hemp extracts in food and pharmaceutical products like stability, toxicity, legal limitations, isolation/extraction, and purification. In addition, this review outlines current applications of hemp extracts and proposes future trends for utilising hemp phytochemicals and extracts in food, cosmetic and pharmaceutical products to increase applications of hemp extracts and their phytochemicals for health benefits.
... (DM basis), which is the primary psychoactive substance [29]. Additionally, hempseed contains anti-nutritional compounds (e.g., phytate) that reduce the absorption of protein and micronutrients [58,59]. However, Reggiani and Russo (2016) observed that replacing 6.4% (on DM basis) of corn and soybean with hempseed or flax seed while maintaining isonitrogenous status can increase iron availability in Alpine lactating goats. ...
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This study aimed to determine whether increasing the levels of Hemp-seed Meal (HSM) Supplementation would affect Dry Matter Intake (DMI), Average Daily Gain (ADG), and rumen microbial community diversity of growing meat goats over a 60-d feeding trial. Forty, 5-month-old castrated Boer-cross goats with an average body weight (BW) of 25.65 ± 0.33 kg were randomly assigned to one of the four treatments (n=10/treatment): control , 10%, 20%, and 30% HSM of the total diets. The forage to concentrate ratio was adjusted to 50:50. Diets were pelleted as total mixed rations. The DMI was lower for 20 and 30% HSM supplementation (P>0.05) than for the control and 10% HSM diet. While, ADG (g/d) and live weight changes (kg/d) decreased (P<0.03) with increasing inclusion rate of HSM. Based on the bacterial clustering diversity, both the control and 10% treatment groups did display a significantly less (P<0.04) diverse environment when compared to the 20% and 30% groups. The fungi diversity detected within the control treatment group is significantly less (P<0.05) than that detected within the 10% and 20% groups. The relative abundance of Bacteroidetes (47.7%), Firmicutes (32.4%), and Proteobacteria (14.9%) were the most abundant bacterial phylum. In the present study, the relative abundance of Bacteroidetes phylum was lower (P<0.01) in animals fed 10 and 30% HSM diets. In comparison, the abundance of Firmicutes phylum (P<0.05) and Methanobrevibacter species were greater (P<0.01) in the rumen of goats fed 20 and 30% HSM than those consuming the control and 10% HSM diets. These findings provide the optimal level of HSM supplementation (<20% as-fed). Furthermore, more work needs to be completed to determine the best feeding strategy to improve the DMI, rumen fermentation, and animal performance. Citation: Abrahamsen F, Min BR, Gurung R, Gurung N, Abebe W. Altering the gut microbiome of meat goats: Feeding varying levels of hempseed meal on animal performance, rumen microbiome abundance, and methanogen community changes of meat goats. BAOJ Microbiol. 2022; 6(2): 1006.
... Hempseeds are an emerging protein-rich plant material and are becoming an important alternative protein source in the food and nutraceutical industry (Wang and Xiong, 2019). A study by Russo and Reggiani (2015) noted that the dioecious varieties of hempseeds are to be preferred as they show lower contents of antinutritional compounds with respect to the monoecious varieties. Data from a study by Tang et al. (2006) suggests that hemp protein isolates (HPI) from the species Cannabis sativa L. have poor functional properties when compared with soy protein isolates. ...
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Note by Note (NbN) cooking and 3D food printing individually and in combination allow for the creation of customized foods. A NbN recipe was developed and printed using a Procusini 3.0, 3D food printer. The recipe was adapted from a commercial powdered pastry dough recipe, to create two versions of a prototype food, containing in each case a different plant-based protein, i.e., soya (Glycine max L.) or hemp (Cannabis sativa). The printed foods were identical in shape and had a crisp consistency following cooking. They had different colours and contained different proteins in the amounts of 10.3 % of soya or 5.7 % of hemp before cooking, which increased to 17.5 % and 9.7 % respectively following evaporation of water during cooking. The main challenge was optimizing the consistency of each recipe mixture before adding it to the syringe prior to printing. The development of the prototype foods which were created in this study shows that recipes of pure compounds and/ or mixtures of compounds could be prepared by innovative chefs who want to surprise their diners. Food product developers can create customised foods for consumers, who wish to eat for example more plant-based proteins, those with allergies or intolerances to certain proteins and/or sports athletes.
... It is possible that phytochemical inhibitors in the hempseed diets interfered with mineral absorption [54], resulting in lower bone mineralization. Hempseed contains phytic acid, which forms insoluble complexes with minerals in the gastrointestinal tract [55][56][57][58]. Polyphenols, such as tannins, in hempseed may also inhibit the absorption of calcium and other minerals, although effects on bone structure are not consistent [59,60] and some studies show increases in calcium absorption with polyphenol supplementation [61]. ...
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Optimizing peak bone mass is critical to healthy aging. Beyond the established roles of dietary minerals and protein on bone integrity, fatty acids and polyphenols modify bone structure. This study investigated the effect of a diet containing hempseeds (HS), which are rich in polyunsaturated fatty acids and polyphenols, on bone mineral density, bone cell populations and body composition. Groups (n = 8 each) of female C57BL/6 mice were fed one of three diets (15% HS by weight; 5% HS; 0% HS (control)) from age 5 to 30 weeks. In vivo whole-body composition and bone mineral density and content were measured every 4 weeks using dual-energy X-ray absorptiometry. Ex vivo humeri cell populations in the epiphyseal plate region were determined by sectioning the bone longitudinally, mounting the sections on slides and staining with tartrate-resistant acid phosphatase and alkaline phosphatase stain to identify osteoclasts and osteoblasts, respectively. Mixed models with repeated measures across experimental weeks showed that neither body weight nor body weight gain across weeks differed among groups yet mice fed the 15% HS diet consumed significantly more food and more kilocalories per g body weight gained than those fed the 5% HS and control diets (p < 0.0001). Across weeks, fat mass was significantly higher in the 5% HS versus the control group (p = 0.02). At the end point, whole-body bone mineral content was significantly higher in the control compared to the 5% HS group (p = 0.02). Humeri from both HS groups displayed significantly lower osteoblast densities compared to the control group (p < 0.0001). No relationship was seen between osteoblast density and body composition measurements. These data invite closer examination of bone cell activity and microarchitecture to determine the effect of habitual HS consumption on bone integrity.
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Cannabis sativa L., has gained increasing attention due to its potential applications in various fields. However, the compositions of hemp seeds from different producing regions are quite different, significantly affects resource utilization. This work conducted the regional differentiation of hemp seeds from Dongbei, Guangxi, Inner Mongolia and Shanxi in China based on the headspace solid phase micro-extraction coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS) and multivariate analysis. Moreover, the chemical composition, amino acid and fatty acid profiles of hemp seeds and antioxidant activities of hemp protein isolate were evaluated from above four producing regions. The results showed that hemp seeds had large amounts of fat (24.30-34.65%) and protein (21.87-23.40%) with ideal amino acid and fatty acid profiles and large amounts of volatiles. More than 300 volatile compounds were detected, and 35 key contributors to the unique aroma were finally identified out of 110 volatile compounds with odor activity values (OAVs), including 14 aldehydes, 2 ketones, 4 alcohols, 2 esters, 1 acid, 3 aromatics, 5 terpenes and 4 other compounds. The content of alcohols in all hemp seeds was highest and 16 unique volatiles only found in hemp seeds from Shanxi. Meanwhile, the VIP scores indicated that acetic acid, hexanal, hexan-1-ol and eucalyptol contributed the most to the difference among groups. According to the results of fuzzy comprehensive evaluation, the hemp seeds from Shanxi seemed to have the better quality. Therefore, multivariate analysis combined with HS-SPME/GC-MS could effectively distinguish hemp seeds from different producing regions, and thus decipher the geographical characteristics of raw materials.
Chapter
Hemp is an ancient crop whose production has been curtailed until recently due to restriction imposed because of phytocannabinoids presence. Industrial hempseed low in tetrahydrocannabinol and cannabidiol is increasingly being grown in many countries, and its acreage is expected to increase further. Commercial hempseed processing generates two major products, oils and proteins and hempseed hulls as a by‐product. The polyunsaturated fatty acid rich oil has 3 : 1 n −6/ n− 3 ratio considered to benefit human health along with γ‐linolenic and stearidonic acids rarely encountered in commercial plant oils. Edestin one of the earliest and easily extractable hempseed proteins can be hydrolyzed along with other hempseed proteins to obtain hydrolysates and bioactive peptides with vascular tone modulation and other beneficial functionalities. The hull is a rich source of lignanamides with demonstrated neuroprotective effects. This chapter reviews the composition of industrial hempseed and processed products, as well as their characteristics, functionality, health benefits, and mode of action. Hempseed components, oil, protein, and lignanamides share common mechanism of action – NF‐κB signaling pathway and apoptosis despite their different mode of action.
Chapter
The hemp plant (Cannabis sativa L.) is one of the most cultivated plants in all of history. Hemp seeds (HSs), flowers, and leaves are an important source of food, fiber, textiles, medical products, cosmetics, and many others. HS protein is tasty (it has a nutty flavor) and is wholesome as it contains all the essential amino acids (which are often lacking in plant products). HSs have been called superfoods due to their ideal ratio of omega-6 to omega-3 fatty acids for human consumption (3:1) and unique amino acid profile, making them an excellent source of food. As a source of plant food, hemps, HS oil, and HS meal are a rich source of protein, polyunsaturated fatty acids, vitamins, and minerals. The valorization of food waste and secondary agroindustrial byproducts is an important part of the economy. In the case of multifunctional industrial crops such as hemp, this can contribute to providing a livelihood for people, meeting their needs without degrading natural resources.
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