Diversity of active constituents in Cichorium endivia and Cynara cornigera extracts

Abstract

The present study attempts to explore the phytochemical constituents of different extracts from Cynara cornigera and Cichorium endivia plant materials. The two species studied are native in Egypt. Five different solvents, viz., aqueous, methylene chloride, petroleum ether, ethyl acetate, and n-butanol were used. Phytochemical analysis revealed the presence of phenols, flavonoids, sterols (stigmasterol and beta-sitosterol), terpenes (α-amyrin, ursolic and oleanolic acid), and hydrocarbons (n-alkane), the latter found in low amount. The ethyl acetate and water extracts of C. cornigera root showed lower mass fractions of phenolic compounds ranged from 20 to 81 g/100 g, and higher amounts in ethyl acetate extract of the inflorescences and butanol extract of the root where values ranged from 195 to 399 g/100 g. The β-sitosterol and stigmasterol were present in all plant extracts. Oleanolic and ursolic acids were detected in roots, leaves and inflorescences of C. cornigera and in C. endivia shoot. The ethyl acetate extracts from C. cornigera leaf and inflorescence attained higher chemical diversity than the other extracts. Alternatively, sterols and triterpenes were the major constituents. The high chemical diversity of active constituents justifies the future potential use of the two species at commercial level.

Full text

Acta Biologica Hungarica 66(1), pp. 103–118 (2015)
DOI: 10.1556/ABiol.66.2015.1.9
0236-5383/$ 20.00 © 2015 Akadémiai Kiadó, Budapest
DIVERSITY OF ACTIVE CONSTITUENTS
IN CICHORIUM ENDIVIA AND CYNARA CORNIGERA
EXTRACTS
AhmAd K. hegAzy,1,2 * Shahira M. Ezzat,3 iMan B. QaSEM,4
MohaMEd S. ali-ShtayEh,5
MohaMMEd o. BaSalah,1 haySSaM M. ali1 and aShraf a. hataMlEh1
1Department of Botany and Microbiology, College of Science, King Saud University,
Riyadh 11451, Saudi Arabia
2Department of Botany, Faculty of Science, Cairo University, Giza 12613, Egypt
3Department of Pharmacognosy, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt
4Biodiversity and Environmental Research Center (BERC)/ Til, POB 696 Nablus, West Bank, Palestine
5Department of Biology, Faculty of Science, An-Najah University, West Bank, Palestine
(Received: February 25, 2014; accepted: June 16, 2014)
The present study attempts to explore the phytochemical constituents of different extracts from Cynara
cornigera and Cichorium endivia plant materials. The two species studied are native in Egypt. Five dif-
ferent solvents, viz., aqueous, methylene chloride, petroleum ether, ethyl acetate, and n-butanol were
used. Phytochemical analysis revealed the presence of phenols, avonoids, sterols (stigmasterol and beta-
sitosterol), terpenes (α-amyrin, ursolic and oleanolic acid), and hydrocarbons (n-alkane), the latter found
in low amount. The ethyl acetate and water extracts of C. cornigera root showed lower mass fractions of
phenolic compounds ranged from 20 to 81 g/100 g, and higher amounts in ethyl acetate extract of the
inorescences and butanol extract of the root where values ranged from 195 to 399 g/100 g. The
β-sitosterol and stigmasterol were present in all plant extracts. Oleanolic and ursolic acids were detected
in roots, leaves and inorescences of C. cornigera and in C. endivia shoot. The ethyl acetate extracts from
C. cornigera leaf and inorescence attained higher chemical diversity than the other extracts.
Alternatively, sterols and triterpenes were the major constituents. The high chemical diversity of active
constituents justies the future potential use of the two species at commercial level.
Keywords: Sterols – triterpenes – avonoids – phenolic acids – Egypt
INTRODUCTION
The increasing use of plant extracts in food, cosmetic and pharmaceutical industries
suggests a systematic study of medicinal plants around the world [49]. Biotic and
abiotic stresses exert a considerable inuence on the secondary metabolite pool espe-
cially the qualitative and quantitative phenolic composition [1]. Polyphenols when
associated with various carbohydrates and organic acids exhibit a wide range of
pharmaceutical properties including antiallergic, antiatherogenic, antiinammatory,
antimicrobial, antioxidant, antithrombotic, cardioprotective, antioxidants, and vasodi-
latory effects [2, 5, 36].
* Corresponding author; e-mail address: akhegazy@yahoo.com

104 AhmAd K. hegAzy et al.
Acta Biologica Hungarica 66, 2015
The wild cichory or endive, Cichorium endivia L. (Asteraceae) is a Mediterranean
perennial herb indigenous to Europe, Western Asia and North Africa [17]. The genus
Cichorium comprises seven species native to the Mediterranean basin [42]. Strong
antioxidant activities were found in the polyphenolic fraction of two Cichorium vari-
eties; Cichorium endivia var. crispum and var. latifolium [41], where both varieties
contain chlorogenic acid derivatives [14, 26]. Alternatively, different active constitu-
ents of other Cichorium spp. were reported [27, 36a], and few other studies reported
different active constituents and medicinal uses of Cichorium endivia [29, 45, 54].
The wild artichoke, Cynara cornigera Lindl. (Asteraceae) is a perennial herba-
ceous species, originating from the Mediterranean region. The species is widely cul-
tivated for economic purposes all over the world [15]. Cynaroside, apigenin-7-O-
glucoside, and scolymoside and two sesquiterpens, viz., grosheimin and solstitalin A
were identied from the leaf extracts [6, 15, 52, 57]. Extracts from C. scolymus are
used in folk medicine against liver complaints and to improve liver regeneration after
partial hepatectomy [9, 19]. Flower head of C. cornigera is eaten as a vegetable and
prepared for different value-added products such as salad, concentrate, and canned
beverages [56]. The avonoid kaempferol 3-O-(6-O-malonyl) was reported from the
root extracts of the two species [14].
Considering the reported chemical constituents in other species of Asteraceae, this
study decided to screen for additional secondary metabolites that might be present in
the wild cichory and artichoke native to Egypt. The aim is to explore the active con-
stituents in the two species, particularly the phenolic compounds, terpenes and steroid
contents.
MATERIALS AND METHODS
Plant extracts
Plant materials were collected from wadi Habis, about 30 km west of Matrouh city,
Mediterranean coast, Egypt. Individual plants were excavated and separated into dif-
ferent organ components, viz., roots, stems, leaves and inorescences (ower heads)
for wild artichoke and whole plant phytomasss for wild cichory. The plant materials
were cut into pieces and air-dried in shade at room temperature. The dry material (792
gm) of wild cichory; and leaves (675 gm), roots (864 gm) and ower heads (630 gm)
of wild artichoke were ground into powder. Extraction was performed in 90% ethyl
alcohol at room temperature till exhaustion, and was then concentrated under reduced
pressure to dry at 40 °C using a rotary evaporator. Materials were subjected to suc-
cessive extraction with petroleum ether, methylene chloride, ethyl acetate, and
n-butanol using separating funnel. The solvents were used in the order to increase
polarity. The residue obtained from each solvent was dried and weighed. The extracts
were kept at 4 °C till used for phytochemical screening.

Phytochemical components in Cichorium and Cynara 105
Acta Biologica Hungarica 66, 2015
Phenolic compounds
Qualitative and quantitative determinations of phenolic compounds were carried out
according to Dimitrios et al. [12]. The retention time (RT) of the compounds isolated
from C. endivia and C. cornigera was compared with that of corresponding standards
under the same conditions and further conrmed by co-injection with isolated stand-
ards. The compounds were then injected to a High Performance Liquid Chromatography
(HPLC) system at the same time as the solvent extracts and the chromatogram was
run with a gradient of CH3OH in acetic water (10 to 90% CH3OH in water in the
presence of 5% CH3COOH). Throughout the experiment each peak detected in the
range from 254 to 280 nm in 30 min at 20 °C corresponded, as a rule, to a single
molecule. Most compounds exhibited absorption peaks in the low-wavelength region
245 nm.
All the HPLC samples (one gram each) were prepared by extraction with 50 ml
methanol (Lab scan) and sub-coiled condensed for one hour. After evaporation, the
residue was dissolved in methanol to give a nal concentration of 0.0025–0.025%
(w/v). Filtration over acrodisc-CR removed the insoluble particles. The samples were
analysed at room temperature (20 °C) in a Brucker LC 42 (Brucker Franzen Analytic
Gmbh) whith UV diode array detection, LC 21 pump, LC 51 automatic sample ana-
lyzer, LC 211 oven, using a spherisorb C16 ODS column, 5 µm , 250 mm × 4.6 mm
(Bischoff, Leonberg). The ow rate was 1 ml/min, and the injection volume 5 µl. The
operating system was the Chromstar Data system of the Brucker LC 42 connected to
an Epson computer. Both the height and the area of a well-resolved peak in a chro-
matogram were proportional to the concentration of the compounds detected. For
quantication, either peak height or peak area was used.
Lipids and triterpenes
Plant extracts were used for the investigation of lipid and triterpene contents using
Unicam Pro-GC with [(Column: 3%OV-17(Methyl phenyl Silicone) on chromosorb-
WHP); (Mesh: 100–120) and (Dimensions: 1.5 × 4 mm)]. The percentage composi-
tion of the extracts was computed from GC peak areas without correction factors.
Qualitative analysis was based on a comparison of retention times, indices and mass
spectra with the corresponding data base. The condtions of GC used are shown in
Table 1.
Table 1
Separation conditions of unsaponiable matters during GC analysis
Temperature Programing Gases Flow Rate
Initial
temperature Rate Final
temperature Final
time Injector Detector N2H2Air
70 °C 10 °C/min 270 °C 50 min 250 °C 300 °C 30 ml/min 33 ml/min 330 ml/min

106 AhmAd K. hegAzy et al.
Acta Biologica Hungarica 66, 2015
The quantication of Oleanolic acid (OA) and Ursolic acid (UA) followed the
method described by Wang et al. [53]. Standard solutions of OA and UA were pre-
pared in nal concentration of 25 µg/ml of each compound. The petroleum ether
extracts were dissolved in 5 ml methanol then an aliquot of 50 µl extract was diluted
in 5 ml methanol. Sampels of 20 µl were injected in the HPLC apparatus. The peaks
in the chromatogram were identied in comparison to their retention time with the
corresponding standards under the same conditions.
RESULTS
Successive extracts yield
The yield of ethanolic extracts and the sucssive solvent extraction of C. endivia and
C. cornigera are presented in Table 2. The highest yield of C. endivia was obtained
from the ethanolic and water extracts with values 18.95 and 16.47 mg/100 g, respec-
tively. The highest yield from C. cornigera was obtained from the ethanolic extract of
roots (111.22 mg/100 g), followed by leaves (110.24 mg/100 g) and the least amounts
in the ower heads (86.38 mg/100 g). Considering the order of extract dependent yield
in C. cornigera, the order of root extract yield was ethanol > water > ethyl acetate >
n-butanol > methylene chloride > petroleum ether, while for leaf and ower the yields
were ethanol > water > n-butanol > ethyl acetate > methylene chloride > petroleum
ether.
Phenolic compounds and avonoids
Twenty phenolic compounds were detected in the two study species, eight avonoids,
viz., Catechin, apigenin, luteolin, luteolin-3-methoxy-7-rutinoside, rutin, quercetin,
chrysin and 5-7-dihydroxy-4-methisoavone and 12 phenolic acids (Tables 3 and 4).
Table 2
The yield (mg/100 g) of Cichorium endivia and Cynara cornigera extracts obtained
by the different solvents
Plants
Total extract of dry plant material
Dry weight
(g) Ethanol Petroleum
ether Methylene
chloride Ethyl
acetate n-Butanol Water
C. endivia 812 18.95 0.12 0.56 0.44 1.34 16.47
C. cornigera
roots 675 111.22 0.06 0.19 1.14 1.13 108.68
C. cornigera
leaves 864 110.24 0.05 0.59 1.13 1.96 106.48
C. cornigera
ower heads 630 86.38 0.05 0.40 1.48 1.85 84.44
Values rounded to the nearest two decimals.

Phytochemical components in Cichorium and Cynara 107
Acta Biologica Hungarica 66, 2015
Table 3
Retention time (min) and percentage (%) of phenolic compounds in Cynara cornigera extrcts
Compound
Ethyl acetate
(Flower head) Ethyl acetate
(Leaf) Ethyl acetate
(Root) n-Butanol
(Root) Water
(Root)
Retention time
(min) %Retention time
(min) %Retention time
(min) %Retention time
(min) %Retention time
(min) %
Gallic acid 14.38 0.09 ND ND ND ND 14.68 0.14 14.68 8.52
Pyrogallic ND ND ND ND 17.87 0.11
Chlorogenic acid 22.02 1.75 ND ND ND ND ND ND ND ND
Resorcinol ND ND ND ND ND ND 22.80 1.11 22.43 5.55
Catechin 23.59 0.42 23.87 2.10 ND ND 23.83 3.60
p-Hydroxy benzoic acid 26.39 12.41 26.27 0.69 ND ND ND ND 26.41 1.23
Luteolin-3-methoxy-7-
rutinoside ND ND 27.01 2.46 26.89 1.30 27.06 0.17 ND ND
Caffeic acid 29.40 4.40 29.34 2.25 29.49 8.86 29.6 12.65 29.51 1.81
Vanillin 29.88 0.42 30.00 26.37 30.04 23.03 29.83 16.74 30.15 0.80
Ferulic acid 30.96 2.87 30.98 4.82 ND ND 30.86 16.33 ND ND
Rutin ND ND 31.53 0.23 31.47 4.81 ND ND ND ND
Phenol 32.16 0.95 32.16 7.70 32.21 2.12 32.22 0.41 32.25 0.36
p-Coumaric acid 32.39 4.07 32.72 1.33 ND ND ND ND ND ND
Luteoline ND ND 34.30 0.15 34.88 0.30 34.42 0.05 ND ND
Quercetin 36.32 0.57 36.54 2.71 36.44 1.67 ND ND 36.63 0.16
Salicylic acid ND ND 37.53 0.43 ND ND 37.35 1.11 ND ND
Apigenin ND ND ND ND ND ND ND ND ND ND
Euganol ND ND ND ND ND ND 41.44 0.03 ND ND
5,7-Dihydroy 4-methoxy
isoavone 43.68 0.35 43.60 0.38 43.49 0.63 ND ND ND ND
Chrysin ND ND ND ND ND ND ND ND 44.24 1.80
Values rounded to the nearest two decimals. ND = Not detected.

108 AhmAd K. hegAzy et al.
Acta Biologica Hungarica 66, 2015
All plant organs contained phenolic compounds in different concentrations, ranging
between 0.01 and 135.63 mg/100 g (Table 5). Ethyl acetate extracts displayed more
phenolic content (0.476–135.63 mg/100 g), while that of the water and n-butanol
extracts, only approached 38.03 mg/100 g, and 84.89 mg/100 g, respectively.
Only in the butanol root extract of C. cornigera the presence of euganol and pyro-
gallic acid was observed. Chlorogenic acid, ferulic acid, phenol and p-coumaric acid
were detected in the organic solvent extracts, while were absent in the aqueous
extract. Signicant amounts of chlorogenic acid, p-hydroxyl benzoic acid, p-coumar-
ic acid, caffeic acid, and ferulic acid were detected in ethyl acetate extract of the
ower heads, with values 135.63, 121.17, 56.66, 28.22 and 22.76 mg/100 g, respec-
tively. Caffeic acid, vanillin, phenol, and quercetin were present in all C. cornigera
Table 4
Retention time (min) and percentage (%) of phenolic compounds of Cichorium endivia extracts
Compound
Ethanol Ethyl acetate Water
Retention time
(min) %Retention time
(min) %Retention time
(min) %
Gallic acid 14.657 4.04 ND ND 14.62 6.26
Pyrogallic 17.70 2.33 ND ND 17.72 0.41
Chlorogenic acid 22.03 1.90 ND ND 21.95 0.51
Resorcinol ND ND ND 22.61 2.24
Catechin 23.69 2.10 ND ND 23.94 16.63
p-Hydroxy benzoic
acid 26.71 18.66 26.83 7.62 26.65 0.89
Luteolin-3-methoxy-7-
rutinoside ND ND ND ND 26.94 0.58
Caffeic acid 29.40 1.50 ND ND 27.31 0.76
Vanillin ND ND 30.06 1.65 29.69 5.35
Ferulic acid 30.98 0.65 30.96 0.60 ND ND
Rutin 31.23 1.31 ND ND 31.21 0.55
Phenol ND ND 32.02 2.29 ND ND
p-Coumaric acid 32.46 1.15 32.50 0.90 32.41 0.01
Luteoline 34.85 0.09 ND ND 34.73 0.06
Quercetin 36.28 3.10 ND ND ND ND
Salicylic acid 36.98 0.56 ND ND 37.36 0.08
Apigenin ND ND ND ND 39.75 0.06
Euganol ND ND 41.37 0.49 ND ND
5,7-dihydroy
4-methoxy isoavone ND ND 43.71 0.50 43.37 0.04
Chrysin 44.050 0.7659 ND ND ND ND
Values rounded to the nearest two decimals. ND = Not detected.

Phytochemical components in Cichorium and Cynara 109
Acta Biologica Hungarica 66, 2015
Table 5
The quantitative determination of phenolic compounds (mg/100 g) in different extracts of Cynara cornigera and Cichorium indivia
Phenolic compound
Cynara cornigera Cichorium indivia
Ethyl acetate Butanol Water Ethyl acetate Water Ethanol
Flower head Leaf Root Root Root
Gallic acid 1.65 ND ND 1.64 5.84 ND 6.05 20.35
Pyrogallic ND ND ND 9.66 ND ND 2.94 86.26
Chlorogenic acid 135.63 ND ND ND ND ND 2.26 44.03
Resorcinol ND ND ND 2.43 7.49 ND 4.27 ND
Catechin 16.99 ND 15.65 ND 5.84 ND 38.03 25.14
p-Hydroxy benzoic acid 121.17 3.24 ND ND 0.49 41.01 0.50 54.45
Luteolin-3-methoxy-7-
rutinoside ND 6.92 1.42 0.68 ND ND 0.19 ND
Caffeic acid 28.22 6.93 10.56 53.14 0.47 ND 0.27 2.87
Vanillin 2.67 79.79 27.06 69.31 0.20 5.76 1.93 ND
Ferulic acid 22.76 18.32 ND 84.89 ND 2.64 ND 1.55
Rutin ND 2.22 17.58 ND ND ND 0.62 7.71
Phenol 12.43 48.01 5.14 3.50 0.19 16.41 ND ND
p-Coumaric acid 56.66 8.87 ND ND ND 6.92 0.01 4.81
Luteoline ND 0.63 0.47 0.32 ND ND 0.03 0.23
Quercetin 5.09 11.57 2.77 ND 0.06 ND ND 8.27
Salicylic acid ND 8.27 ND ND ND ND ND ND
Apigenin ND ND ND ND ND ND 0.01 ND
Euganol ND ND ND 0.45 ND 5.96 ND ND
5,7-Dihydroy 4-methoxy
isoavone 1.53 0.79 0.50 ND ND 1.19 0.01 ND
Chrysin ND ND ND ND 0.72 ND ND 2.25
Sum of compounds 404.84 195.61 81.20 255.34 21.33 79.91 57.38 264.76
ND = Not detected.

110 AhmAd K. hegAzy et al.
Acta Biologica Hungarica 66, 2015
organs while gallic acid, luteoline, luteolin-3-methoxy-7-rutinoside, p-hydroxyl ben-
zoic acid, 5-7-dihydroxy-4-methisoavone and ferulic acid detected in three out of
ve extracts of C. cornigera (Table 5).
The avonoid composition including catechin rutin and quercetin was the most
abundant in C. endivia, while chrycin, 5,7-dihydroy 4-methoxy isoavone, luteolin,
luteolin-3-methoxy-7-rutinoside and apigenin detected as traces. On the other hand,
the total avonoids content found in C. endivia was 43.61 mg/100 g and 38.90
mg/100 g in ethanol and water extracts, respectively, followed by ethyl acetate in
C. cornigera root (38.43 mg/100 g).
The order of yield in the most abundant constituents of different C. cornigera
extracts was catechin > luteolin-3-methoxy-7-rutinoside > rutin > quercetin >
5.7-dihydroy 4-methoxy isoavone, followed by, luteolin-3-methoxy-7-rutinoside,
luteoline and chrycin. Flavonoid content ranged from traces to 38.039 mg/100 g
(Table 5). It was abundant in ethyl acetate root extract except for quercetin in leaf
extract and 5,7-dihydroy 4-methoxy isoavone in ower head extract. Total avo-
noids of plant organs are in the order of root (54.59 mg/100 g) > ower head (23.62
mg/100 g) > leaf (22.15 mg/100 g). Luteolin was extremely low in all organs of
C. cornigera and also showed an almost zero level of apigenin. Phenolic compounds
were abundant in ethanolic extract of C. endivia with about 19 out of 20 compounds.
In general, C. cornigera extracts had higher phenol concentration than C. endivia.
The most abundant compounds were pyrogallic acid in the ethanolic extract
(86.26 mg/100 g) and catechin in the aqueous extract of C. endivia (38.03 mg/100 g).
Lipids and triterpenes
The plant extracts contained 22 n-alkane hydrocarbons (Tables 6 and 7). The identi-
ed compounds of methylene chloride formed 75.2–84.7%, where eicosane, octade-
cane, tridecane and heneicosane were found to be the major constituents in C. corni-
gera (Table 6). The petroleum ether fractions of the leaf and root were present in quite
similar amounts, like the methylene chloride fractions. The amounts of octadecane
(C17, C18) and tridecane were signicantly higher in the roots (13.97, 12.18 and
11.85%, respectively) than ower heads (4.41, 5.77% and 4.54%, respectively), fol-
lowed by leaves. The amount of eicosane was higher in leaves (18.99%) than in the
ower heads (10.27%). Petroleum ether extract of the ower heads contained eight
compounds higher than the leaves & roots which were characterized by high content
of eicosane, C18 and docosane (18.02, 16.6 and 11.7%), while the roots contained
high amounts of tridecane and tetradecane (16.2–11.2%). The identied compounds
in petroleum ether extract constituted 83.4–95.1% of the total content. Octadecane,
eicosane, docosane, tridecane and tetracosane were the major constituents in C. endi-
via (Table 7). The petroleum ether extract contains more hydrocarbons (93.43%) than
the crude ethanol extract (62.60%).
Two sterol compounds were identied as stigmasterol and β-sitosterol. The lowest
contents were measured in the methylene chloride leaf extract. The major sterol

Phytochemical components in Cichorium and Cynara 111
Acta Biologica Hungarica 66, 2015
components in C. cornigera were 4.33% in petroleum ether ower head extract, fol-
lowed by root extract (4.08%) then leaf extract (3.99%). The main component of
methylene chloride extract is α-amyrin found in the ower head (1.36%), followed by
β-sitosterol and stigmasterol (1.35% and 1.30%, respectively). α-Amyrin was not
found in petroleum ether ower head extract, while β-sitosterol was detected in
ower head and root extracts with values1.84% and 1.52%, respectively. Stigmasterol
Table 6
Percentage composition of hydrocarbon and sterol components in Cynara cornigera obtained by GC
Hydrocarbons
Relative %
Root Leaf Flower head
Petroleum
ether Methylene
chloride Petroleum
ether Methylene
chloride Petroleum
ether Methylene
chloride
n-Nonane (C9) ND ND ND 4.99 ND 3.26
n-Decane (C10) ND 2.64 1.32 4.95 ND 1.28
n-Henedecane (C11) 1.15 2.22 1.26 2.25 ND 4.67
n-Dodecane (C12) 2.40 2.71 2.07 3.11 ND 6.70
n-Tridecane (C13) 16.19 11.85 4.55 4.23 ND 4.54
n-Tetradecane (C14) 11.17 1.92 7.53 2.06 2.87 2.56
n-Pentadecane (C15) 4.25 1.02 9.34 7.67 7.71 6.72
n-Hexadecane (C16) 1.66 1.32 3.47 3.75 10.43 4.45
n-Heptadecane (C17) 0.73 13.97 1.51 2.88 ND 4.41
n-Octadecane (C18) 12.95 12.18 14.49 4.52 16.63 5.76
n-Nonadecane (C19) 0.65 2.11 0.53 4.23 3.72 3.16
n-Eicosane (C20) 12.92 3.72 12.48 18.99 18.02 10.26
n-Heneicosane (C21) ND 9.50 0.30 4.98 1.21 4.07
n-Docosane (C22) 8.06 2.54 10.10 4.33 11.71 3.17
n-Tricosane (C23) 1.98 ND 0.34 3.37 ND 4.75
n-Tetracosane (C24) 6.96 6.07 7.09 2.91 9.55 2.66
n-Pentacosane (C25) 0.48 1.95 0.52 2.14 ND 2.44
n-Hexacosane (C26) 4.30 3.67 4.30 1.32 5.91 1.77
n-Heptacosane (C27) 0.21 1.24 0.55 1.51 ND 1.75
n-Octacosane (C28) 3.03 2.37 3.48 1.20 3.70 1.29
n-Triacontane (C30) 0.32 ND 2.55 ND ND 1.90
n-Hentriacontane (C31) ND ND 1.54 ND 1.23 ND
Total 89.50 83.09 89.42 85.50 92.75 81.66
Sterols and triterpene
α-Amyrin 0.36 0.90 0.49 0.71 ND 1.32
β-Setosterol 1.52 0.68 0.45 0.96 1.84 1.35
Stigmasterol 2.19 1.20 3.05 0.19 2.49 1.30
Values rounded to the nearest two decimals. ND = Not detected.

112 AhmAd K. hegAzy et al.
Acta Biologica Hungarica 66, 2015
was high in petroleum ether shoot extract of C. endivia (3.05%), followed by that of
the ower head extract (2.49%) and root extract (2.19%) of C. cornigera.
Two types of triterpenes were detected in both C. cornigera and C. endivia
extracts. Petroleum ether extract of C. cornigera ower heads showed the highest
amount (54.7%) of oleanolic acid (Table 8), while C. endivia had the highest amount
of ursolic acid (88.5%) (Table 9).
Table 7
Percentage composition of hydrocarbon and sterol components
in Cichorium endivia obtained by GC
Hydrocarbons Relative %
Ethanol Petroleum ether
n-Octane (C8) ND 2.59
n-Nonane (C9) ND 7.44
n-Decane (C10) ND 9.24
n-Henedecane (C11) 10.72 11.12
n-Dodecane (C12) 5.20 8.71
n-Tridecane (C13) 1.70 7.38
n-Tetradecane (C14) 2.29 5.85
n-Pentadecane (C15) 4.01 ND
n-Hexadecane (C16) 2.15 7.38
n-Heptadecane (C17) 2.88 0.37
n-Octadecane (C18) 3.80 7.51
n-Nonadecane (C19) 2.11 ND
n-Eicosane (C20) 4.19 5.77
n-Heneicosane (C21) 2.75 1.67
n-Docosane (C22) 1.41 4.62
n-Tricosane (C23) 1.93 ND
n-Tetracosane (C24) 5.68 4.20
n-Pentacosane (C25) 1.45 3.40
n-Hexacosane (C26) 6.03 1.30
n-Heptacosane (C27) 2.51 2.48
n-Octacosane (C28) 3.65 1.69
n-Triacontane (C30) ND 0.63
n-Hentriacontane (C31) ND ND
Total 93.43 62.60
Sterols and triterpene
α-Amyrin 11.33 1.17
β-Setosterol 6.76 0.97
Stigmasterol 3.27 1.61
Values rounded to the nearest two decimals. ND = Not detected.

Phytochemical components in Cichorium and Cynara 113
Acta Biologica Hungarica 66, 2015
DISCUSSION
The results of phenolic compounds and avonoids showed that apigenin was not
detected in the extracts of C. cornigera, while traces of chrysin was found in water
extract of the root. These two compounds are reported here for the rst time in the
two species studied. Apigenin and luteolin were identied in C. cardunculus [31].
The chlorogenic acid was identied in the aqueous and alcoholic extracts of Aster
ageratoides ower buds [11].
The results indicate that C. cornigera organs, especially the ower heads could
represent an important source of polyphenols with therapeutic activity [34].
According to Gupta et al. [22], avonoids are generally found in substantial concen-
trations in plant leaves or owers. Brás et al. [8] reported the presence of quercetin
and rutin in the leaves of Eupatorium littorale (Asteraceae). Twelve biologically
active avonoids, mainly avonol and avone derivatives, including two glycosides
were detected in the glandular trichomes of Chromolaena species [25], while a total
of 135 different compounds were reported from the Cichorieae (Lactuceae) tribe of
the Asteraceae [44]. In C. cornigera and C. endivia, the high and variable active
compound concentrations are related to the arid climate conditions, such as, hot tem-
perature, high solar exposure, drought and salinity which stimulate the biosynthesis
of secondary metabolites as polyphenols [13].
The presence of biologically active secondary metabolites such as tannins (gallic
acid and catechin), sterols and terpenes contributes to its medicinal value [47].
Tannins have been reported to have several pharmacological activities such as spas-
Table 8
Retention time (min) and percentage (%) of triterpene compounds in Cynara cornigera extracts
of petroleum ether and ethanol
Compound
Cynara cornigera
Root Leaf Flower head
Retention time
(min) %Retention time
(min) %Retention time
(min) %
Oleanolic acid 2.21 1.70 2.60 2.20 0.98 54.70
Ursolic acid 0.98 95.5 0.96 96.3 0.26 32.00
Table 9
Retention time (min) and percentage (%) of triterpene compounds
in Cichorium endivia extracts of petroleum ether and ethanol
Compound
Cichorium endivia
Petroleum ether Ethanol
Retention time (min) %Retention time (min) %
Oleanolic acid 2.58 3.40 2.26 4.40
Ursolic acid 0.93 96.60 0.93 88.50

114 AhmAd K. hegAzy et al.
Acta Biologica Hungarica 66, 2015
molytic activity in smooth muscle cells [50]. Gallic acid has antineoplastic and bac-
teriostatic activities and anticancer properties in prostate carcinoma cells [28], and
anti-inammatory and antibacterial activity [46]. The present study revealed that
gallic acid is in range from 1.64 mg/100 g in C. cornigera parts to 20.35 mg/100 g in
C. endivia. There were variations in total phenolic content of different aromatic and
medicinal plants ranging between 6.80–32.10 mg gallic acid/g dry weight bases [4,
21]. p-Coumaric and ferulic acids are also present in combination with betanidin
monoglucoside in fruits of Basella rubra [20]. Hydroxycinnamic acid derivatives
were found in C. endivia var. crispum and var. latifolium; three derivatives
(5-O-caffeoylquinic acid, 3,4-di-O-caffeoylquinic acid, and 5-O-feruloylquinic acid)
were found in both chicories, while 3,5-di-O-caffeoylquinic acid was typical of var.
crispum and cis-caftaric acid of var. latifolium [40].
The presence of kaempferol 3-O-(6-O-malonyl) glucoside in endive is reported
here for the rst time. Methanol extract of Lactuca sativa L. varieties (Romaine,
Iceberg and Baby) and C. endivia L. variety “escarole” have yielded a high phenolic
content, where chicory extracts being the richest with 50 mg phenolics/g of freeze-
dried extract [14, 35]. Tannins, sesquiterpene lactones, cinnamic acid, caffeic and
chlorogenic acids were reported in chicory [27]. Monocaffeoyl tartaric acid, chloro-
genic acid, and chicoric acid were detected in different varieties of Cichorium inty-
bus, while cyanidin 3-O-glucoside, delphinidin 3-O-glucoside, and cyanidin
3-O-glucoside were the main phenolic compounds in the red varieties [26]. Also,
apigenin and apigenin-7-0-α-1-arabinoside were isolated from Cichorium intybus
aerial parts [55].
The p-hydroxybenzoic acid (4-hydroxybenzoic acid) found in C. cornigera
showed a broad range of pharmacological activities, for instance, antifungal, anti-
mutagenic, antisickling, estrogenic and antimicrobial effects [10, 43]. Chlorogonic
acid has been reported to have a number of biological properties, such as antibacte-
rial, antioxidant, hepatocyte protective and antimutagenic activity [51]. Artichoke
capitula was characterized by signicant amounts of chlorogenic acid and various
dicaffeoylquinic esters, especially 1,3-dicaffeoylquinic acid, known as cynarin [7].
Eight phenolic compounds were isolated from n-butanol leaf fraction of Cynara
scolymus which is known for its antimicrobial activity [56, 57]. Also, Orlovskaya et
al. [39] have previously reported qualitative and quantitative results on phenolic com-
pounds on raw material and extracts of C. scolymus leaves. The phenolic contents in
leaf and seed of Cynara cardunculus were similar and two times higher than those in
ower heads [16]. The organic subfraction of artichoke is rich in polyphenolic com-
pounds, with caffeoylquinic acids and avonoids as the major chemical constituent
[32]. In leaf extracts of commercially available artichoke (C. scolymus) a number of
compounds were identied, including 8-deoxy-11-hydroxy-13-chlorogrosheimin,
cryptochlorogenic acid, chlorogenic acid, neochlorogenic acid, cynarin, cynaratriol
(tentatively), grosheimin, 8-deoxy-11,13-dihydroxygrosheimin, luteolin-7-O-rutino-
side, luteolin-7-O-glucoside, and cynaropicrin [18]. Phenolic acids, up to 2%, caffeic
acid, mono- and dicaffeoylquinic acid derivatives, e.g., cynarin, and chlorogenic acid

Phytochemical components in Cichorium and Cynara 115
Acta Biologica Hungarica 66, 2015
were reported to be present in artichoke [24]. The avonoids, triterpenoids and sapo-
nines were reported to possess hepatoprotective activity in animals [3].
As for lipid and triterpene content, twenty-two compounds of the hydrocarbons
were identied in petroleum ether of C. cornigera parts. The major components con-
stitute up to 85% of the plant chemical composition, while the minor components
reported in trace amounts [37]. A high content of derivatives was also characteristic
in C. endivia extracts, including henedecane, decane, dodecane, octane and heptade-
cane. The presence of terpenoids in C. cornigera and C. endivia has also been
reported to show several biological and pharmacological activities such as antioxida-
tive [48], anti-inammatory [38], hepato-protective effects [33]. The mechanism of
action of terpenes is not fully understood but it is speculated to involve membrane
disruption by the lipophilic compounds [23].
In conclusion, our ndings indicate that C. cornigera and C. endivia are rich
sources of eight important biologically active avonoids, twelve phenolic acids, tow
triterpenes and three steroids, which were detected for the rst time in the two species
studied. Considering that chlorogenic acid and rutin are highly valuable natural poly-
phenol compounds used as medical and industrial materials, the identication and
quantication of these main polyphenols in the present study can be important for
potential use at commercial level in the pharmaceutical industry.
ACKNOWLEDGEMENT
We thank the Deanship of Scientic Research, College of Science Research Center, King Saud University,
Riyadh, Saudi Arabia for supporting this publication.
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