1. No category

International Conference on Agricultural, Ecological and Medical Sciences (AEMS-2015) April 7-8, 2015 Phuket (Thailand)
Evaluation of Parietal Polysaccharides and
Polyphenols of Crataegus monogyna Jacq. and
Valorization Prospects
Zoheir Mehdadi1, Mokhtaria Hamdaoui2, and Ali Latreche3

catecholamines were identified in the genus Crataegus and
much of these substances are evaluated for their biological
activities [3].
Polysaccharides are macromolecules derived from various
living organisms, including the plants, they have the anti
tumoral, immunostimulant and antioxydant activity, they
characterized by a broad therapeutic spectrum and a low
toxicity [5]. They play a big role in the industry of materials
and health human and animal as a dietetic fiber [6].
C. oxyacantha, C. laciniata, C. monogyna and C. azarolus
are the most species used in the Maghreb countries; their
sheets, flowers and fruits are employed in form decoction or
infusion like anti spasmodic, cardiotonic, anti-diarrheal and for
the treatment of the calculous affections and the diabetes [7].
In Algeria, the fruit of C. monogyna is used in the solid mass
of Aurès like food of misery. On the other hand in Morocco
and in the Average Atlas, the ripe fruits of C. monogyna or C.
laciniata are eaten by the shepherds. They also constitute a
substantial nutritive contribution for birds [8]. Other studies
show that the extracts of the fruits of the European hawthorn
(C. monogyna or C. laevigata) are used as dietetic supplements
and in grass medicine to treat the heart, the light forms of the
arrhythmia [9] and angina pectoris [10]. The fruits of other
species like C. pinnatifida and C. scabrifolia were used
traditionally like peptic agent in Eastern medicine [11], in
jams, juice, foods and like basic ingredient to make the wines
and various soft foods [12].
C. monogyna is common in Algeria, localized in the forests
and the maquis of the Tellian Atlas [13]. The biochemical
studies undertaken on this species indicate its wealth of
polyphenolic compounds; heterosides of quercetol,
leucoanthocyanidines, oligomer procyanidines, acids phenolic
(chlorogenic and caffeic) and acids triterpenic (crataegolic,
ursolic, oleanolic) [7]. These molecules are regarded as
phytochimic substances with effects prebiotic, antioxydant and
anti-inflammatory [14].
Complementary to the work undertaken, our study consists
to valorize biochemically C. monogyna developing
spontaneously in the mount of Tessala (Western Algeria)
where is sheltered a flora diversified but subjected
unfortunately to a pressure anthropozoogene [15]. On the one
hand, evaluating quantitatively total phenols, the condensed
tannins, flavonoïdes, and hydrolysables tannins contained in
the sheets, the flowers and the fruits, and on the other hand
parietal polysaccharides (cellulose, hemicelluloses, pectins) in
Abstract—The aim of this study was to evaluate quantitatively
some parietal polysaccharides (cellulose, hemicelluloses and pectins)
and some polyphenols (total phenols, flavonoïdes, hydrolysables
tannins and condensed tannins) on methanolic extracts prepared with
the sheets, flowers and pericarp of the fruit of Crataegus monogyna
Jacq., of the Mount of Tessala (Western Algeria).
The results obtained show a quantitative variability of the
substances between the analyzed vegetable bodies (P < 0.05).
Considerable quantities of cellulose, hemicelluloses and pectins
characterize the pericarp of the fruit; on the other hand the
polyphenols are represented better in the sheets and the flowers.
The results indicate that the studied species is a plant potential that
can be exploited in various fields, especially in therapy.
Keywords—Mount of Tessala, Crataegus monogyna Jacq.,
polyphenols, analyzes quantitative, parietal polysaccharides.
I. INTRODUCTION
H
AWTHORN (Crataegus spp.) is a member of the
Rosaceae family, belonging to the Spiraeoideae subfamily
in the Pyroidae supertribe in the Pyreae subtribe [1].
There are approximately 280 species in the genus Crataegus,
which are widely distributed across the Mediterranean, North
Africa, Europe, Central and Eastern Asia and North America
[2]. They were employed a long time in folk medicine for the
treatment of various diseases: heart, central nervous system,
immune system, eyes, reproductive system, liver, kidneys, etc.
They present also antimicrobial activity, cytotoxic, gastroprotective, anti-HIV and anti-inflammatory [3].
All species of the genus Crataegus are recommended to treat
the
myocardial
weakness,
paroxysmal
tachycardia,
hypertension and arteriosclerosis [2]. The most species
employed in medicine are C. monogyna and C. laevigata, and
less frequently other species such as C. pentagyna, C. nigra
and C. azarolus L. [4]. The phytochimic substances like
oligomers procyanidines, flavonoïdes, triterpenes and
Zoheir Mehdadi1, Laboratory of Plant Biodiversity: conservation and
enhancement, faculty of Natural and Life Sciences, Djillali Liabes University,
Sidi Bel Abbes, 22000, Algeria (corresponding author’s phone:
00213774751626 ; e-mail: mehdadiz@yahoo.fr).
Mokhtaria Hamdaoui2, Laboratory of Plant Biodiversity: conservation and
enhancement, faculty of Natural and Life Sciences, Djillali Liabes University,
Sidi Bel Abbés, 22000, Algeria ( e-mail: hamdaouidoc@yahoo.fr).
Ali Latreche3, Laboratory of Plant Biodiversity: conservation and
enhancement, faculty of Natural and Life Sciences, Djillali Liabes University,
Sidi Bel Abbés, 22000, Algeria ( e-mail: ali_latreche@yahoo.fr).
http://dx.doi.org/10.15242/IICBE.C0415012
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International Conference on Agricultural, Ecological and Medical Sciences (AEMS-2015) April 7-8, 2015 Phuket (Thailand)
mixture, the filtrate is preserved at -20 °C for proportioning.
The quantitative analysis of hydrolysables tannins is carried
by spectrophotometry [17]. For that, 1 ml of the filtrate is
added to 3, 5 ml with a reagent prepared containing ferric
trichloride (FeCl3) with 0,01M in hydrochloric acid (HCl)
with 0,001M. The absorptance of the mixture is read to 660
Nm.
The contents of hydrolysables tannins are expressed by the
following formula:
the fruits.
II. BIOLOGICAL MATERIAL AND METHODS OF STUDY
A. Biological material
Previous research focused on biochemical dosages on C.
monogyna evolving in the mount of Tessala (Western Algeria).
The polyphenols are quantified on the sheets, the flowers and
the pericarp of the fruit; parietal polysaccharides are
proportioned only on the pericarp. The sheets and the flowers
were collected in April, the fruits in December of the year
2011.
The parameters stationers relating to the site of the
collection sample are noted: altitude: 1015 m; North latitude:
35° 16. 57 "; Western longitude: 0°46. 55 "; exposure: SouthEast.
Figures
T (%) = (D.O × M × V)/E mole × P
T: content of tannins; D.O: optical density; M: mass = 300; V: volume of
extract used; E mole: 2169 of the ellagic acid (constant expressed in mole); P:
weight of the sample.
For the estimate of condensed tannins, 1 ml of filtrate is
added to 2 ml of a solution prepared containing vanillin with
1%, in the sulphuric acid with 70%. The mixture thus obtained
is placed in a bain-marie during 15 mn with 20 °C safe from
the light in order to determine its optical density with 500 nm
[18].
The quantities of condensed tannins are calculated
according to the following formula:
B. Extraction and proportioning of the total phenols and
the flavonoïdes
The samples are harvested fresh and dried at the shade, in a
dry and aired place, then reduced powders by a knife crusher.
0, 2 g of vegetable powder are put in 10 ml of methanol at
80%; after agitation with the vortex, the mixture is centrifuged
at a speed of 4000 turn / minute during 10 minute then the
supernatant is recovered. This operation is repeated 3 times to
exhaust the contents of soluble phenolics compounds of the
sample. The supernatants obtained are gathered and constitute
the extract hydro-alcoholic which is preserved at - 20 °C for
the continuation of the analyzes.
The proportioning spectrophotometric of total phenols and
the flavonoïdes is carried by the technique of El Hadrami and
al. [16]. In parallel, curves standards are established by the
gallic acid and the catéchine respectively for total phenols and
the flavonoïdes.
For the proportioning of the total phenols, 50 μl of
methanolic extract is diluted in 2, 5 ml of distilled water, to
which 250 μl of Folin-Ciocalteu reagent are added. The
mixture is then subjected to an agitation with the vortex
followed by a rest of 5 minutes to room temperature, then 500
μl of sodium carbonate to 20% are added . After agitation and
incubation with 40 °C during 30 minutes, the mixture is kept
with the room temperature and the darkness during 60 minutes
for reading its optical density with 765 nm.
For the proportioning of the flavonoïdes, 500 μl of extract
methanolic are mixed with 1500 μl distilled water and 150 μl
of sodium nitrate with 5%; after a rest of 5 mn to the room
temperature and the darkness, 150 μl of aluminum trichlorure
at 10 % and 500 μl of sodium hydroxide (NaOH) with 1 M
are added. After agitation, the optical density of the mixture is
read to 510 nm.
The corresponding quantities of total phenols and
flavonoïdes are reported in equivalent mg of the standard used.
T (%) = (5, 2×10-2×D.O×V)/P.
T: content of tannins; 5, 2 X 10-2: constant expressed in equivalent of
cyanidines; D.O: Optical density; V: volume of extract used; P: weight of the
sample.
D. Extraction
and
quantification
of
cellulose,
hemicelluloses and pectin
The quantification of cellulose, hemicelluloses and pectin is
carried out on the parietal residue of the pericarp of the fruit
according to the modified method of Harche and al. [19].
The preparation of the parietal residue consists in putting
under agitation, during 14 hours and under hood, 50g of
vegetable powder in a mixture ethanol-toluene (1V-1V). After
filtration on fabric to be bolted, the parietal residue is put in
ethanol under agitation during 2 hours to eliminate the traces
from toluene; it then is rinsed with the water distilled then with
acetone and finally dried with the drying oven with 60 °C
during 24 hours. The residue obtained contains lignins,
cellulose, hemicelluloses and pectins.
Before extracting cellulose and hemicelluloses, the parietal
residue is delignified to break the bonds lignin-polysaccharides
and consequently to extract the maximum of polysaccharides.
For this purpose, 5g of parietal residue is put under agitation at
80 °C during 2 hours in 100 ml of ethanol at 70% container 1g
of soda (NaOH). After filtration on fabric to be bolted, the
residue and washed with the water distilled then dried with the
drying oven with 60 °C during 24 hours [20].
For the extraction of cellulose and hemicelluloses, 5g of
delignified parietal residue is put under agitation during 14
hours in Erlenmeyer containing 100 ml of soda (NaOH) at 4%.
After filtration on bolting cloth, this operation is renewed on
the residue to recover the maximum of hemicelluloses. The
residue obtained is washed with the water distilled then with
acetone, then dried with the drying oven with 60 °C during 14
C. Extraction and proportioning of hydrolysables tannins
and condensed tannins
0, 2 g of vegetable powder are macerated in 10 ml of
methanol with 80% during 18 hours. After filtration of the
http://dx.doi.org/10.15242/IICBE.C0415012
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International Conference on Agricultural, Ecological and Medical Sciences (AEMS-2015) April 7-8, 2015 Phuket (Thailand)
TABLE I
MEAN LEVELS OF TOTAL PHENOLS AND FLAVONOIDS IN THE LEAVES,
FLOWERS AND FRUIT PERICARP
Total phenols
Flavonoïdes
(mg EAG/g)
(mg EC/g)
hours and finally weighed. This part represents the cellulose
fraction [21].
Both filtrates obtained are neutralized by the pure acetic
acid, then precipitated in the ethanol (1V-3V) during 14 hours.
After centrifugation with 3600 tr/min during 30 minutes, the
base is washed by distilled water then acetone, then dried with
the drying oven with 60 °C during 14 hours and finally
weighed. This part represents the hemicellulosic fraction.
The extraction of highly methylated pectin (HMP) and
Weakly methylated pectin (WMP) are carried out according to
the technique Monties (1980) [22].
For HMP, 5g of residue is introduced into Erlenmayer
containing 100 ml of water distilled and put under agitation
during 14 hours, then put at boiling under backward flow twice
two hours. After filtration on fabric to be bolted, the filtrate is
concentrated with the rotavapor and precipitate in cold acetone
(1V-2V) during 14 hours. After centrifugation, the base is
washed with water distilled then with alcohol and put to dry in
the drying oven with 50 °C.
For SMP, the residue obtained is treated with 100 ml of
diamino-tetra-acetic acid ethylene (EDTA) with 1% with pH
6,5 adjusted with a solution of sodium hydroxide (NaOH),
then put under agitation during 6 hours at a temperature of 80
°C.
After filtration, the solution obtained is dialyzed against
running water during 14 hours, then against water distilled 4
times 1 hour in order to eliminate the traces from EDTA by
using a membrane of dialysis of the type SPECTRA POR 6
MWCO 1000. Dialyzate is concentrated with the rotavapor.
Pectins slightly methylated are recovered by precipitation of
the volume of dialyzate remaining in cold acetone (1V-2V)
during 14 hours. After centrifugation, the base is washed with
distilled then with the alcohol and put water dried in the drying
oven with 50 °C.
Leafs
Flowers
17,79 ± 0,55a
b
05,07 ± 0,10b
c
03,72 ± 0,34c
25,72 ± 0,14
Pericarp of fruit
16,62 ± 0,23
Variance analysis
+
+
EAG: gallic acid equivalent, EC: equity catechin, +: Significant difference
(P <0.05). a, b, c: Means in the same column followed by the same letter are
not significant
B. Evaluation of hydrolysable tannins and condensed
tannins
The most important contents of hydrolysable tannins are
recorded on the sheets (0,99 ± 0,01%). In the flowers and the
fruits, we obtained a low average contents even weak with
respectively 0, 39 ± 0,03% and 0,04 ± 0,01%.
The analysis of the variance and the test of Tukey confirm
this variability in the quantitative distribution of tannins
hydrolysables between the three bodies (P < 0,05).
This is not the case for condensed tannins which are present
with comparable proportions (P > 0.05) (table II).
TABLE II
AVERAGE PERCENTAGES OF HYDROLYSABLE AND CONDENSED TANNINS IN
LEAVES, FLOWERS AND PERICARP OF FRUIT
Hydrolysable tannins
Condensed tannins
E. Statistical analysis
The contents of the various biochemical compounds
correspond to averages evaluated on five samples.
The comparison of these contents is carried out by the test
of the analysis of the variance (comparison of several
averages) and the test of Tukey (comparison of the averages
two to two) by using software XLSTAT 2013
Leafs
0,99 ± 0,01a
0,22 ± 0,02a
Flowers
0,39 ± 0,03b
0,20 ± 0,04a
Pericarp of fruit
0,04 ± 0,01c
0,21 ± 0,01a
Variance analysis
+
C. Evaluation of parietal polysaccharides
The walls of the pericarp of the fruit offers 60, 86% of
parietal residue (RP), that is to say the three quarters of the dry
matter.
In 5g of delignified parietal residue, the cellulose present
with a rate of 2,22 ± 0,08 g either 44,40 ± 1,60%, is followed
by hemicelluloses with a rate of 1,77 ± 0,03 g or 35,40 ±
0,60%. With regard to pectins, the same weight in parietal
residue provided 0,57 ± 0,02 g or 11,40 ± 0,56% of highly
methylated pectins. However pectins slightly methylated do
not represent that 0,43 ± 0,02 g or 8,60 ± 0,42%. Compared to
cellulose and hemicelluloses, slightly and highly methylated
pectins are slightly represented in the pericarp (table III).
III. RESULTS
A. Evaluation of total phenols and the flavonoïdes
The results illustrated in table I show a variability in the
average content of total phenols and flavonoïdes in the various
studied bodies, confirmed by the analysis of the variance and
the test of Tukey (P < 0,05).
The sheets have the highest rate out of total phenols (34,82
± 0,63 mg/g) and flavonoïdes (17,79 ± 0,55 mg/g); however
these substances are slightly represented in the fruits: 16,62 ±
0,23 of total phenol mg/g and 3,72 ± 0,34 mg/g of flavonoïdes.
Intermediate rates are noted in the flowers: 25,72 ± 0,14 of
total phenol mg/g and 5,07 ± 0,10 mg/g of flavonoïdes.
http://dx.doi.org/10.15242/IICBE.C0415012
34,82 ± 0,63a
TABLE III
AVERAGE COMPOSITION OF PARIETAL POLYSACCHARIDES OF PERICARP OF
FRUIT
Percentage
Composition
Quantity in gr
(%)
Parietal residue (quantity in 50 gr of
30,31 ± 1,94
60,62 ± 3,75
plant powder)
Cellulose (quantity in 5gr of RPD)
02,22 ± 0,08
44,40 ± 1,60
Hemicelluloses (quantity in 5gr of
01,77 ± 0,03
35,40 ± 0,60
RPD)
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International Conference on Agricultural, Ecological and Medical Sciences (AEMS-2015) April 7-8, 2015 Phuket (Thailand)
Highly
methylated
00,57 ± 0,02
11,40 ± 0,56
Pectins (quantity
pectins (HMP)
in 5gr of RDP)
Weakly
methylated
00,43 ± 0,02
8,60 ± 0,42
pectins (WMP)
RP: parietal residue, RPD: parietal residue delignified
Generally, it arises that compared to the fruit, the sheets and
the flowers of the studied species are the bodies richest in
polyphenols (total phenols, flavonoïdes, condensed tannins
and hydrolysables tannins). This is in agreement with works of
Froehlicher and al. (2009) [34].
The synthesis of polyphenols is influenced by the genetic
factors and the environmental conditions. Other factors, such
the degree of maturity, the variety, the treatment and storage
also have an influence on the content of the phenolic
compounds of plants [35]. However, it is difficult to compare
these results with those of the bibliography because the use of
various methods of extraction reduces the reliability of a
comparison between the studies [36].
The average contents of cellulose, hemicelluloses and
pectins evaluated in the pericarp of the fruit of C. monogyna
are respectively 44,4%,35,40% and 20%. These contents are
quite higher than those found by Saadoudi (2007) [37], that is
to say 11,40 % of cellulose and 1,60 % of pectin at C.
monogyna of the area of Batna (Algerian East). These contents
are also more important with those obtained by Ozcan and al.
(2005) [38], is cellulose 4,67% at the fruits of species of the
genus of Crataegus evolving/moving in Turkey.
These quantitative variability noted within the same species
can be explained by the method adopted of extraction, solvents
used, the geographical location, the period of collection and
the degree of maturity of the fruit.
Pectins are represented in the fruit of C. monogyna with a
rate of 20%. A comparable content of this substance, is 21%
was reported at the fruit of C. pubescens [39]. Pectins are the
main component of the middle lamella of the cells of plant
tissues, they contribute significantly to the softening of the
texture of the bodies of the plants [40]. In China, the fruits of
hawthorns are usually transformed into jam and/or gel,
because of their high percentage of pectins [41]. Whereas in
Mexico, the fruits of the genus Crataegus are employed to
work the local cakes, jams and drinks containing fruits for the
person which follow a mode, and in food industry like a source
of pectin and vitamin C and other components dietetic [42].
On the other hand, the polysaccharides isolated from the
plants like hemicelluloses and pectin have an anticoagulant
activity which had with the presence of the hexuronic acids
residues and its derivatives (pectins are richest an acids
hexuronic) [43]. Many bioactives substances such as cellulose
and bio-cellulose of the plants and the oligosaccharides of
pectins derived from the hawthorns were shown for their effect
hypo-cholesterolemic [44], [45]. Many studies showed the
beneficial effects of the extracts of bays of hawthorns on the
system of circulation of blood [46].
The fruits of C. monogyna, represent an excellent source of
these macromolecules which influence on health human and
prevent the occurrence of many diseases, because the fibers
resulting from the plants have various biological activities: to
reduce cholesterol level, to promote regularity of défécations,
to improve rate of sugar blood at diabetics, to deal disease
diverticulaire (ignition of part of the intestines) and syndrome
with colon irritable (abdominal pain, diarrhea and constipation
which is presented by intervals), to help to prevent certain
cancers, like the cancer of the colon (cancer colorectal) and
IV. DISCUSSION
The average contents of total phenols, flavonoïdes and
tannins condensed in the pericarp of the fruit of C. monogyna
are estimated respectively at 16.62 mg/g, 3.72 mg/g and
0.20%. These results are comparable with those obtained on
the same species evolving/moving in the area of Batna
(Algerian East), that is to say 21.72 mg/g for total phenols,
3.20 mg/g for the flavonoïdes and 0.21% for condensed
tannins [23]. A low content of total phenol 8.17 mg/g is noted
in the pericarp of the fruit of the same species [24].
At the flowers of C. monogyna of the north of France, the
total phenols and the flavonoïdes are present with respective
rates of 15.63 mg/g and 10.26 mg/g [25]. These rates differ
from those recorded in our species, that is to say 25.72 mg/g of
total phenols and 5.07mg/g of flavonoïdes.
On C. oxyacantha of the area of Oum el Alou de Tlemcen
(Western Algerian), it was noted 4.60 mg/g of polyphenols and
0.74 mg/m of flavonoïdes [7]. These contents are rather weak
compared to those which were recorded on C. monogyna.
The evaluation of phenolic compound on extracts
methanolic of the fresh fruits of C. oxyacantha L. evolvin in
Serbia, shows the presence of the contents total phenols (18.21
mg/g) comparable with those evaluated on the fruit of C.
monogyna; as for the flavonoïdes, they are present there with
low contents (0.99 mg/g) [26].
The contents of polyphenols in the fruit of C. pinnatifida are
approximately 96.90 ± 4.30 mg/g [27], value largely
exceeding that which we obtained on the fruit of C. monogyna.
In other work, the total contents in phenolic compounds in
the fruits of the species of the genus Crataegus are estimated at
3.54% [28].
The average content of flavonoïdes in the sheets and the
flowers is respectively 1,77% and 0,50%. These percentages
with those are found comparable by Rose and Treadway
(1999) [29] is 1% in the same bodies. The hawthorns have
several composed flavonoïques, present qualitative and
quantitative differences within the sheets, flowers and fruits of
the same plant [30].
Our species is rich in flavonoïdes compared to other species
with knowing C. azarolus which contains between 0,06 and
0,08% [31].
The contents of procyanidines indicated by the European
pharmacopeia for the fruits of C. monogyna are comparable
with the contents evaluated on the fruit of C. azarolus [31].
The fruits of other species contain high percentages of
tannins compared to our species, like fruits of C. pinnatifida
Bge. which contains 3,74% of condensed tannin [32].
Condensed tannins are found in the fruits, barks, sheets and
seeds of much of the plants including the genus Crataegus [2],
[33]. Their function is defense against herbivores and
resistance to the abiotic effects [33].
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International Conference on Agricultural, Ecological and Medical Sciences (AEMS-2015) April 7-8, 2015 Phuket (Thailand)
breast cancer [47].
V. CONCLUSION
[14]
In the light of the results obtained, it arises that sheets and
flowers of C. monogyna are the bodies richest in polyphenols.
The pericarp of the fruit contains considerable quantities of
fibers (cellulose, hemicelluloses and pectins) which can be
developed in various fields (medicine, industry, dietetics, etc).
This biochemical evaluation shows that the studied specie is
a vegetable resource in phytochimic substances, being able to
contribute in écodéveloppement of our country. This
encourages us to preserve this species which unfortunately is
subjected to a pressure anthropozoogene.
These results remain preliminary, it would be desirable to
deepen it by the characterization of the substances quantified
by using advanced techniques like the HPLC.
[15]
[16]
[17]
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