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Natural Product Research
Formerly Natural Product Letters
ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/gnpl20
Chemical composition, antioxidant and
antibacterial activity of Crataegus monogyna
leaves’ extracts
Fouzia Belabdelli, Nabila Bekhti, Alessandra Piras, Fouad Mekhalef
Benhafsa, Mokhtari Ilham, Selka Adil & Lazzaz Anes
To cite this article: Fouzia Belabdelli, Nabila Bekhti, Alessandra Piras, Fouad Mekhalef Benhafsa,
Mokhtari Ilham, Selka Adil & Lazzaz Anes (2021): Chemical composition, antioxidant and
antibacterial activity of Crataegus�monogyna leaves’ extracts, Natural Product Research, DOI:
10.1080/14786419.2021.1958215
To link to this article: https://doi.org/10.1080/14786419.2021.1958215
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Published online: 02 Aug 2021.
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SHORT COMMUNICATION
Chemical composition, antioxidant and antibacterial
activity of Crataegus monogyna leaves’extracts
Fouzia Belabdelli
a,b
, Nabila Bekhti
a,c
, Alessandra Piras
d
,
Fouad Mekhalef Benhafsa
e
, Mokhtari Ilham
a,b
, Selka Adil
f
and Lazzaz Anes
b
a
Laboratory of Theoretical Chemistry of Bio & Nano-systems, Faculty of Exact Sciences, Djillali Liabes
University of Sidi Bel Abbes, Sidi Bel Abb
es, Algeria;
b
Department of Pharmacy, Faculty of Medicine,
Djillali Liabes University of Sidi Bel Abbes, Sidi Bel Abbes, Algeria;
c
Department of Process
Engineering, Faculty of Science and Technology, Mustapha Stambouli University of Mascara, Route
Mamounia, Mascara, Algeria;
d
Department of Chemical and Geological Sciences, University of
Cagliari, Cittadella Universitaria, Monserrato, Cagliari, Italy;
e
Center for Scientific and Technical
Research in Physico-Chemical Analysis (CRAPC), Algiers, Algeria;
f
Department of Pharmacy, Faculty of
Medicine, Abou Bakr Belkaïd University of Tlemcen, Tlemcen, Algeria
ABSTRACT
Crataegus monogyna is an important plant of the Rosaceae family,
widely used in traditional medicine to treat various conditions
such as cardiovascular diseases, cancer, diabetes, asthma, and
nephritis. The aim of the current study was to assess the chemical
composition, antioxidant, and antibacterial activity of leaves’
extracts against Bacillus cereus, Staphylococcus aureus, Escherichia
coli, and Pseudomonas aeruginosa. The total amounts of polyphe-
nols and flavonoids contained in the dry extracts of plants were
estimated by colorimetric methods. DPPH assay was utilized to
measure the antioxidant activity of C. monogyna. The phytochem-
ical compounds were determined through HPLC technique, and
the minimum inhibitory concentration (MIC) of ethanol extract
was performed using the broth dilution method. The ethanol
extract represented the richest extract in polyphenol with
473.4 mg GAE g
1
and flavonoids 80.9 mg CE g
1
and showed
considerable antioxidant potential IC
50
¼22.50 mg/ml. The
antibacterial susceptibility test against Staphylococcus aureus
0.512 mg/mL.
ARTICLE HISTORY
Received 10 April 2021
Accepted 10 July 2021
KEYWORDS
Crataegus monogyna; HPLC;
DPPH assay; MIC
CONTACT Nabila Bekhti nabilachimie@yahoo.fr
Supplemental data for this article can be accessed online at https://doi.org/10.1080/14786419.2021.1958215.
ß2021 Informa UK Limited, trading as Taylor & Francis Group
NATURAL PRODUCT RESEARCH
https://doi.org/10.1080/14786419.2021.1958215
1. Introduction
Crataegus monogyna species are commonly used in traditional medicine. This medi-
cinal plant constitutes a valuable source of bioactive phytochemicals or bio-nutrients.
Different hawthorn species have been found to exert various effects in all populations.
Those species are usually used in association with other drugs as antispasmodic, diur-
etic, febrifuge, hypertensive, analgesic, sedative, and antidepressant agents (Kirakosyan
et al. 2003; Guo et al. 2008; Zhao et al. 2012; Ku et al. 2015; Li et al. 2015). As shown
by Popovic-Milenkovic et al. 2014,Crataegus monogyna leaves and flowers contained a
high abundance of flavonoids and displayed a significant biological effect (Mudge
et al. 2016). C. monogyna is a species recognized by the European Pharmacopoeia
(2007). Whereas Crataegus monogyna has demonstrated a high antioxidant activity
attributed to flavonoids and procyanidins (Ozyurek et al., 2012).
The investigation of medicinal plants from temperate regions grasped the interest
of both botanists and pharmacologists. In addition to the scientific value, such investi-
gation can provide valuable information on general trends in research on medicinal
plants and on the practical possibilities of their use in modern pharmacopeia. The
therapeutic properties of these plants are due to the presence of natural bioactive
compounds, hawthorn fruits, and leave derivatives (NNHPD 2013).
This plant can grow in several types of soil, especially in glades and forests.
Although it prefers acidic and dry soils, the plant can also grow in neutral and alkaline
soils. This species is well represented in the global temperate zones. In Algeria, the
plant is more frequently found in the forests and the maquis of the Tellian Atlas.
Several studies have investigated the chemical composition and the antioxidant activ-
ity of Crataegus species and different organs of the plant (leaves, flowers, and fruits) of
the fruit Crataegus monogyna from different origins (Bernatonien _
e et al. 2008; Bouzi
et al. 2011). The flavonoids identified in flowers of Mexican hawthorns were the
2 F. BELABDELLI ET AL.
quercetin 3-O-glucoside, quercetin 3-0-rhamnoside, quercetin 3-O-rhamnosyl-(1!6)-
glucoside, and quercetin 3-O-rhamnosyl-(1!2)-[rhamnosyl-(1!6)] glucoside (Garc
ıa-
Mateos et al. 2013). Fifteen Crataegus L. species of leaves and flowers in Eurasia and
North America identified chlorogenic acid, hyperoside, Vitexin 2-rhamnoside, vitexin 2-
O-rhamnoside, vitexin, caffeic acid, and triterpenes oleanolic acid, ursolic acid
(Kateryna Khokhlova et al. 2020). Four flavonoids were quantified (naringenin, hypero-
side, rutin, and vitexin-20’-O-rhamnoside) and chlorogenic acid in leaf extracts of four
Crataegus species (C. monogyna,C. laevigata,C. douglasti, and C. okanaganensis) (Lund
et al. 2020). Five flavonoids quantified in hawthorn (C. douglasii and C. laevigata
leaves; C. monogyna leaves and flowers) vitexin2-O-rhamnoside, vitexin, isovitexin,
rutin and hyperoside (Mudge et al. 2016).epicatechin] and flavonoid (vitexin 20’-O-
rhamnoside, acetylvitexin 20’-O-rhamnoside, and hyperoside) (Kirakosyan et al. 2003).
In Turkey, the major compound is rutin in the ethanol extract of the flowers of
Crataegus monogyna Jacq as 2.68 mg/ml and provides considerable antioxidant activity
(78.80 ± 4.09) (Konyalioglu et al. 2017).
However, few studies investigated the Algerian species to reach sufficient data on
the chemical composition and the biological activity of Crataegus monogyna leaves. The
aim of the current study was, on one hand, the determination, with HPLC, of the chem-
ical composition of extracts plants of Crataegus monogyna, leaves growing in Sidi-Bel-
Abbes region (Northwest of Algeria). On the other hand, the assessment of the antioxi-
dant and the antibacterial activity of leaves’ethanol extract against common pathogens
such as Staphylococcus aureus,Escherichia coli,andPseudomonas aeruginosa.
2. Results and discussion
As the leaves of Crataegus monogyna, in Algeria are not phytochemically common, a
screening was carried out aiming to obtain a first indication of the main available
metabolites. Our results showed that flavonoids, phenol carboxylic acids, tannins, ster-
ols, and triterpene acids were present in the leaves of Crataegus monogyna. However,
saponosides, anthraquinones, alkaloids, anthocyanins, and coumarins were absent as
outlined in Table S1.
The ethanolic extract is considered the richest extract in polyphenol with 473.4 mg
GAE g
1
extract and flavonoids 80.9 mg CE g
1
. The chloroform extract contains a low
content of polyphenols with 17.7 mg GAE g
1
as shown in Table S2. Results obtained
showed that the total phenolic content and total flavonoid content of C. monogyna
leaves’extract were higher than those found in other species of different regions
worldwide. The variation of polyphenols and flavonoids of Crataegus leaves species of
different origins has previously been investigated by other researchers. In 2011, Bouzi
et al. reported in the eastern region of Algeria, that C. monogyna fruit polyphenol and
flavonoids values were 43.92 mg EAG/mg extract and 5.41 ug EQ/mg extract respect-
ively. Coimbra et al. (2020) have obtained in C. monogyna leaves 110.41 mg GAE g
1
extract and mg 29.94 QE g
1
extract. However, the extracts of C. monogyna leaves
tested by Iranian researchers Alirezalu et al. 2018, displayed values of 32.34 mg GAEg-
1 and 6.87 mg/g. Several studies have reported that the level of polyphenols varied
from 117.729 to 204.286 mg GAE/g in different regions of aerial parts of nine samples
of C. monogyna (Abuashwashi et al. 2016). Whereas the levels of flavonoids varied
NATURAL PRODUCT RESEARCH 3
from 2.12-32.62 mg/g in the same previous extracts. The highest phenolic (391.97GAE/
g) and flavonoid (17.60QE/g extract) rates were identified in the fruit C. monogyna
(Bardakci et al. 2019). Regarding the C. monogyna leaves’extract, an elevated content
was observed in comparison to those in some studies. These differences in the com-
position of hawthorn vary among species and geographic locations (Jones et al. 2001;
Abuashwashi et al. 2016).
After comparing the retention times of the extracts with those of the controls, an
HPLC separation was performed allowing the identification of nine phenolic com-
pounds: procyanidin (peak1), epicatechin (peak2), catechin (peak3), chlorogenic acid
(peak4), caffeic acid (peak5), vitexin-2-O-rhamnoside (peak6), vitexin (peak7), rutin
(peak8) and hyperoside (peak9), in ethanolic extract from C.monogyna leaves (Figure
S1) and Syringic acid (peak10), myricetin (peak11), quercetin (peak12), naringenin
(peak13), kaempferol (peak14), using chloroformic leaves extract as shown in Figure
S2.C. monogyna of chloroform extract hat the highest level of kaempferol. After
revealing a TLC chromatogram of the chloroform and ethanol extracts with vanillin-
sulfuric acid, two purplish-blue spots one below the other appeared in the middle
part which correspond according to the literature to oleanolic and ursolic acid
(Kateryna Khokhlova et al. 2020) and Caligiani et al. (2013) identified ursolic acid in C.
monogyna leaves.
Bouzi et al. showed that quercetin was present in the fruit of C. monogyna however
rutin and catechin were absent in the extracts. Ziouche et al. (2020) found that the
extracts of leaves of eastern Algeria contained chlorogenic acid, rutin, isoquercetin,
hesperidin, and catechin. Unlike the methanolic fruit extract that contained caffeic
acid and quercetin. Concerning flavonoids, including rutin, apigenin, myricetin, quer-
cetin, naringenin, and kaempferol, they were identified in the leaf hawthorn (Keser
et al. 2014). The NMR technique quantified vitexin-2-O-rhamnoside, hyperoside, and
chlorogenic acid contents and HPLC separated vitexin, vitexin-2-O-rhamnoside, isovi-
texin, rutin, hyperoside in the leaf extract of C. monogyna Lund et al. (2017/2020).
Vitexin-2-O-rhrmnoside, vitexin, isovitexin, rutin, and hyperoside were quantified in
hawthorn leaf and flower by Mudge et al. (2016). Kateryna Khokhlova in 2020 identi-
fied the flavonoids and two triterpenes of 15 various Hawthorn of leaves and flowers
such vitexin2- rhamnoside, hyperoside, rutin, chlorogenic acid, caffeic acid oleanolic,
and ursolic acids. According to Alirezalu et al. 2018, the most abundant phenolic
compounds in the extracts of hawthorn leaves were chlorogenic acid, vitexin, and
vitexin-2-O-rhamnoside. Interestingly, the contents of rutin, vitexin-2-O-rhamnoside,
kaempferol, and chlorogenic acid in our study were the highest concentration ever
quantified in the leaf of C. monogyna. Moreover, the diversity of compounds found in
the current investigation agrees with several previously published studies (Keser et al.
2014; Mudge et al. 2016, Lund et al. 2020).
Furthermore, a very weak anti-radical activity was expressed by the chloroform
extract. Our test showed that ethanolic extract of C. monogyna leaves recorded higher
radical scavenging activity with an IC
50
value of 22.5 mg/mL than that found by
Ziouche et al. (2020) in leaves’extract IC
50
; 24.52 mg/ml. In 2015, Loubna et al. found
an IC
50
of the aqueous extract of C. monogyna leaves at different stages of vegetation
ranging from 45 to 725 mg/mL. In 2021,
€
Ozy
€
Urek et al. have shown among the differ-
ent species studied in Turkey, that the C. monogyna sample exhibited significant
4 F. BELABDELLI ET AL.
antioxidant activity. However, the extracts of C. monogyna recently tested by Coimbra
et al. 2020 have presented an IC
50
value of 36.11 lg/mL. In our study, the antioxidant
activity in the leaves extract of C. monogyna displayed considerable antioxidant poten-
tial due to the phenolic content of this species extract. Flavonoids possess a much
higher concentration of rutin, vitexin-2-O-rhamnoside, kaempferol, chlorogenic acid,
and other phenolic derivatives.
Concerning the antibacterial activity assessment, the minimum inhibitory concentra-
tion (MIC) extract ethanol of leaves was observed at an interval of 0.512-1.024 mg/mL
against S. aureus. However, this was not active against E. coli and P. aeruginosa. The
extract leaves of C. monogyna of Algeria showed better activity compared to that C.
monogyna leaves observed by Coimbra et al. (2020) with 1 2 mg/mL and MIC of 1 g/
mL identified by Bouzi et al. (2011). Salmanian et al. (2014) found an antibacterial
activity of C. monogyna fruit between 5-10 mg/ml. Our results are in agreement with
those of Yi
git et al. (2014) who found the same MIC of alcoholic and aqueous extracts
against E. coli and S. aureus (1.25 and 0.625 mg/mL respectively).
3. Conclusion
Our results showed that variations were detected in C. monogyna leaves content and
composition compared to other authors’findings. Leaves’extract had strong antioxi-
dant and antibacterial activity, which were correlated with their high level of polyphe-
nols. Most of the substances found in C. monogyna extract of leaves were chlorogenic
acid, rutin, vitexin-2-O-rhamnoside and kaempferol. The variation in the chemical com-
position of C. monogyna might be attributed to the varied agroclimatic conditions of
the regions where it grows. High levels of polyphenolics found in C. monogyna leaves
allow this plant to exhibit its phyto and pharmaceutical potentials.
Acknowledgments
The authors would like to thank Professor Belhassaini Hachemi from the laboratory of
Biodiversity vegetable Faculty of Biology, Djillali Liabes University, Sidi Bel-Abbes for their scien-
tific collaboration, and Professor Meghit Boumediene Khaled for providing necessary corrections
to the last version of the article.
Disclosure statement
No potential conflict of interest was reported by the author(s).
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