EXCLI Journal 2011;10:230-239 – ISSN 1611-2156

Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

Original article:

IMMORTELLE (XERANTHEMUM ANNUUM L.) AS A NATURAL

SOURCE OF BIOLOGICALLY ACTIVE SUBSTANCES
Milan S. Stanković*, Ivana D. Radojević, Olgica D. Stefanović, Marina D. Topuzović,
Ljiljana R. Čomić, Snežana R. Branković

Department of Biology and Ecology, Faculty of Science, University of Kragujevac, Radoja

Domanovića 12, 34000 Kragujevac, Republic of Serbia
∗
corresponding author: Tel.: +381 34 336 223; Fax: +381 34 335 040
e-mail: [email protected]

ABSTRACT

Antioxidant and antimicrobial effects, total phenolic content and flavonoid concentrations of
methanolic, acetone and ethyl acetate extracts from Xeranthemum annuum L. were investi-
gated in this study. The total phenolic content was determined using Folin-Ciocalteu reagent
and ranged between 101.33 to 159.48 mg GA/g. The concentration of flavonoids in various X.
annuum extracts was determined using spectrophotometric method with aluminum chloride
and the results varied from 22.25 to 62.42 mg RU/g. Antioxidant activity was monitored spec-
trophotometrically using DPPH reagent and expressed in terms of IC50 (µg/ml), and it ranged
from 59.25 to 956.81 µg/ml. The highest phenolic content and capacity to neutralize DPPH
radicals were found in the acetone extract. In vitro antimicrobial activity was determined by
microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal
concentration (MMC) have been determined. Testing was conducted against 24 microorgan-
isms, including 15 strains of bacteria (standard and clinical strains) and 9 species of fungi.
Statistically significant difference in activity between the extracts of X. annuum L. was ob-
served and the acetone extract was found most active. The activity of acetone extract was in
accordance with total phenol content and flavonoid concentration measured in this extract.
The tested extracts showed significant antibacterial activity against G+ bacteria and weak to
moderate activity against other microorganisms. Based on the obtained results, X. annuum can
be considered as a rich natural source of polyphenolic compounds with very good antioxidant
and antimicrobial activity.

Keywords: immortelle, Xeranthemum annuum, antimicrobial capacity, antioxidant, phenols,

flavonoids

INTRODUCTION white-tomentose beneath and about 30 mm

long and 2-7 mm wide. Flowers are light-
Immortelle, Xeranthemum annuum L.,
purple or pink forming individual capitula,
belongs to the family Asteraceae (Compo-
up to 5 cm wide, having a hemispherical
sitae), subfamily Cichorioideae, tribe
involucre. It is a thermophilic species and
Cardueae, subtribe Carlininae. Four species
inhabits arid rocky meadows and dry,
belong to the genus Xeranthemum in the
rocky, sunny and thermophilic habitats
flora of Europe. X. annuum is an annual
along roadsides, fields and vineyards in
herb, growing to 50 cm, with a thin and
Southern Europe and Anatolia (Xeran-
spindle-shaped root and a tree with a small
themum, 1975).
number of linear to oblong leaves that are

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

A large number of known medicinal gated on bacterial and fungal strains by mi-
species belonging to the family Asteraceae crodilution method.
are used in phytomedicine and pharmacy.
For very long time medicinal plants of this MATERIAL AND METHODS
family have been used in the treatment of
Plant material
many diseases of the digestive, respiratory,
In August 2009 aerial parts of X. annu-
and cardiovascular systems and skin dis-
um were collected from natural populations
eases, as well as for the preparation of bev-
on Plackovica hill in the region of Vranje
erages, and as culinary spices. The species
sity in south Serbia: (position:
of the family Asteraceae are very rich in
42°34′51.94′′N, 21°53′53.90′′E, altitude:
phenolic compounds with very strong bio-
1075.00 m, exposition: E, habitat: arid
logical activity and they exhibit strong anti-
thermophilic rocky meadows). Plants
oxidant, antibacterial, antifungal, antiviral
identified were confirmed and voucher
and antiproliferative effects (Özgen et al.,
specimens deposited at the Herbarium of
2004; Boussaada et al., 2008; Jayaraman et
the Department of Biology and Ecology,
al., 2008; Muley et al., 2009; Kasim et al.,
Faculty of Science, University of Kragu-
2011).
jevac. The collected plant material was air-
Literature data indicate that the X.
dried in darkness at ambient temperature
annuum is a medicinal plant in traditional
(20 °C). The dried plant material was cut up
medicine and it is applied as a source of
and stored in tightly sealed dark containers.
active substances (Vogl-Lukasser and Vogl,
2004; Watson and Preedy, 2008). However,
Chemicals
there is very little data on laboratory
Organic solvents and sodium hydrogen
phytochemical studies and biological
carbonate were purchased from „Zorka
activity of extracts and isolated components
pharma“ Šabac, Serbia. Gallic acid, rutin
from X. annuum, which points to the fact
hydrate, chlorogenic acid and 2,2-diphenyl-
that the plant has not been explored
1-picrylhydrazyl (DPPH) were obtained
completely. The existing data often provide
from Sigma Chemicals Co., St Louis, MO,
chemical properties of secondary
USA. Folin-Ciocalteu phenol reagent, and
metabolites from X. annuum and the
aluminium chloride hexahydrate (AlCl3)
description of the most common
were purchased from Fluka Chemie AG,
metabolites (Zemtsova and Molchanova,
Buchs, Switzerland. Nutrient liquid
1979; Skaltsa et al., 2000).
medium, a Mueller–Hinton broth was
Bearing in mind that the family
purchased from Liofilchem, Italy, while a
Asteraceae comprises species that are well
Sabouraud dextrose broth was obtained
known in phytomedicine and pharmaceu-
from Torlak, Belgrade. An antibiotic, doxy-
tical industry, it is obvious that the
cycline, was purchased from Galenika
evaluation of X. annuum as a new source of
A.D., Belgrade, and antimycotic, flucona-
natural medicinal substances may contri-
zole was from Pfizer Inc., USA. All other
bute to the knowledge about the use and
solvents and chemicals were of analytical
importance of the family.
grade.
Therefore, the purpose of this study was
to explore X. annuum as a new potential
natural source of effective antioxidant and Preparation of plant extracts
antimicrobial agents, as well as its total Prepared plant material (10 g) was
phenolic content and flavonoid concentra- transferred to dark-coloured flasks with
tions. Antioxidant activity, phenolic content 200 ml of solvent (methanol, ethyl acetate,
and flavonoid concentrations are deter- acetone) and stored at room temperature.
mined by spectrophotometric methods and After 24 h, infusions were filtered through
in vitro antimicrobial activity was investi- Whatman No. 1 filter paper and residue was
re-extracted with equal volume of solvents.

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

After 48 h, the process was repeated. ing the method described by Tekao et al.
Combined supernatants were evaporated to (1994), adopted with suitable modifications
dryness under vacuum at 40 °C using rotary from Kumarasamy et al. (2007). The stock
evaporator. The obtained extracts were kept solution of the plant extract was prepared in
in sterile sample tubes and stored in a methanol to achieve the concentration of
refrigerator at 4 °C. 1 mg/ml. Dilutions were made to obtain
concentrations of 500, 250, 125, 62.5,
Determination of total phenolic contents 31.25, 15.62, 7.81, 3.90, 1.99, 0.97 µg/ml.
in the plant extracts Diluted solutions (1 ml each) were mixed
The total phenolic content was deter- with 1 ml of DPPH methanolic solution
mined using spectrophotometric method (80 µg/ml). After 30 min in darkness at
(Singleton et al., 1999). The reaction mix- room temperature (23 °C) the absorbance
ture was prepared by mixing 0.5 ml of was recorded at 517 nm. The control sam-
methanolic solution (1 mg/ml) of extract, ples contained all the reagents except the
2.5 ml of 10 % Folin-Ciocalteu’s reagent extract. The percentage inhibition was cal-
dissolved in water and 2.5 ml 7.5 % Na- culated using the equation: % inhibition =
HCO3. The samples were incubated at 100 x (A control – A sample)/A control),
45 °C for 15 min. The absorbance was de- whilst IC50 values were estimated from the
termined at λmax = 765 nm. The samples % inhibition versus concentration sigmoidal
were prepared in triplicate and the mean curve, using a non-linear regression analy-
value of absorbance was obtained. Blank sis. The data were presented as mean values
was concomitantly prepared with methanol ± standard deviation (n = 3).
instead of extract solution. The same proce-
dure was repeated for the gallic acid and the Test microorganisms
calibration line was construed. The total Antimicrobial activity of acetone, ethyl
phenolic content was expressed in terms of acetate and methanolic extract was tested
gallic acid equivalent (mg of GA/g of ex- against 24 microorganisms including fifteen
tract). strains of bacteria (standard strains:
Escherichia coli ATCC 25922,
Determination of flavonoid concentrations Staphylococcus aureus ATCC 25923,
in the plant extracts Enterococcus faecalis ATCC 29212,
The concentrations of flavonoids was Pseudomonas aeruginosa ATCC 27853,
determined using spectrophotometric Bacillus subtilis ATCC 6633, Bacillus
method (Quettier et al., 2000). The sample pumilus NCTC 8241 and clinical strains:
contained 1 ml of methanolic solution of Escherichia coli, Staphylococcus aureus,
the extract in the concentration of 1 mg/ml Enterococcus faecalis, Pseudomonas
and 1 ml of 2 % AlCl3 solution dissolved in aeruginosa, Proteus mirabilis, Sarcina
methanol. The samples were incubated for lutea, Salmonella enterica, Bacillus subtilis
an hour at room temperature. The absorb- and Bacillus cereus) and nine species of
ance was determined at λmax = 415 nm. The fungi: Penicillium italicum PMFKG-F29,
samples were prepared in triplicate and the Penicillium digitatum PMFKG-F30,
mean value of absorbance was obtained. Penicillium chrysogenum PMFKG-F31,
The same procedure was repeated for the Trichothecium roseum PMFKG-F32,
rutin and the calibration line was construed. Botrytis cinerea PMFKG-F33; Aspergillus
The concentration of flavonoids in extracts niger ATCC 16404; Candida albicans
was expressed in terms of rutin equivalent (clinical isolate); Rhodotorula sp. PMFKG-
(mg of RU/g of extract). F27 and Saccharomyces boulardii
PMFKG-P34. All clinical isolates were a
Evaluation of DPPH scavenging activity generous gift from the Institute of Public
The ability of the plant extract to scav- Health, Kragujevac. The other microorga-
enge DPPH free radicals was assessed us- nisms were provided from a collection held

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

by the Microbiology Laboratory, Faculty of DMSO on the growth of microorganism. It

Science, University of Kragujevac. was observed that 10 % DMSO did not in-
hibit the growth of microorganism. Also, in
Suspension preparation the experiment, the concentration of DMSO
Bacterial and yeast suspensions were was additionally decreased because of the
prepared by the direct colony method. The twofold serial dilution assay (the working
turbidity of initial suspension was adjusted concentration was 5 % and lower). Each
by comparing with 0.5 McFarland’s stand- test included growth control and sterility
ard (Andrews, 2005). Initial bacterial sus- control. All tests were performed in dupli-
pension contains about 108 colony forming cate and MICs were constant.
units (CFU)/mL and yeast suspension con-
tains 106 CFU/mL. 1:100 dilutions of initial Statistical analysis
suspension were additionally prepared into Data are presented as means ± standard
sterile 0.85 % saline. The suspensions of deviations where appropriate. All statistical
fungal spores were prepared by gentle analyses were performed using SPSS pack-
stripping of spores from slopes with grow- age. Mean differences were established by
ing aspergilli. The resulting suspensions Student’s t-test. Data were analyzed using
were 1:1000 diluted in sterile 0.85 % saline. one-way analysis of variance (ANOVA). In
all cases P values <0.05 were considered
Microdilution method statistically significant.
Antimicrobial activity was tested by de-
termining the minimum inhibitory concen- RESULTS AND DISCUSSION
tration (MIC) and minimum microbicidal
Total phenol content and flavonoid con-
concentration (MMC) using microdilution
centrations
method with resazurin, an indicator of mi- Three extracts from X. annuum were
crobial growth (Sarker et al., 2007). The prepared using different solvents (methanol,
96-well plates were prepared by dispensing acetone, ethyl acetate) in order to examine
100 µL of nutrient broth, Mueller–Hinton the total phenolic content, flavonoid con-
broth for bacteria and Sabouraud dextrose centrations and free radical scavenging ac-
broth for fungi, into each well. A 100 µL tivity. Different solvents were used to ex-
from the stock solution of tested extracts tract active ingredients of diverse polarities.
(concentration of 80 mg/mL) was added The choice of solvents proved to be very
into the first row of the plate. Then, two- effective in earlier studies (Stanković et al.,
fold, serial dilutions were performed by us- 2010).
ing a multichannel pipette. The obtained The results of the total phenolic content
concentration range was from 40 to determination of the examined plant ex-
0.156 mg/mL. MIC was defined as the low- tracts, using Folin-Ciocalteu method, are
est concentration of tested extracts that pre- presented in Table 1. The total phenolic
vented resazurin color change from blue to content in extracts was expressed as gallic
pink (Vasić et al., 2010). acid equivalents ranged between 101.33 to
Minimum microbicidal concentration 159.48 mg GA/g. Total phenolic content
(MMC) was determined by plating 10 µL of was the highest in all extracts from X. an-
samples from wells, where no indicator nuum, among which the acetone extract
color change was recorded, on nutrient agar (159.48 mg GA/g) contained the highest
medium. At the end of the incubation pe- concentracion of phenolic compounds.
riod the lowest concentration with no Analyzing the results of total phenolic con-
growth (no colony) was defined as mini- tent in all extracts, it was noticed that the
mum microbicidal concentration. highest concentration of phenolic com-
Doxycycline and fluconazole were used pounds in the extracts were obtained using
as a positive control. Solvent control test the solvents of moderate polarity. Other au-
was performed to study the effects of 10 %

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

thors reported that high dissolubility of picrylhydrazyl (DPPH) reagent. DPPH

phenols in polar solvents provides high method has also been used to quantify anti-
concentration of these compounds in the oxidants in complex biological systems in
extracts obtained using polar or moderately recent years and it’s based on the reduction
polar solvents for the extraction (Zhou and of methanolic solution of colored free radi-
Yu, 2004). cal DPPH by free radical scavenger. The
scavenging activity was measured as the
1
Table 1: Total phenolic contents and concen- decrease in absorbance of the samples ver-
trations of flavonoids in X. annuum extracts sus DPPH standard solution.
The antioxidant activity of three differ-
Total ent extracts from X. annuum is expressed in
Flavonoid
phenolic
Type of
content
concentration terms of IC50 (µg/ml) values. Along with
extract (mg RU/g of the examination of the antioxidant activity
(mg GA/g of
extract)
extract) of the plant extracts, the values for chloro-
methanolic 101.33 ± 0.99 36.52 ± 0.98 genic acid as reference substance were ob-
acetone 159.48 ± 1.12 62.42 ± 1.01 tained and compared to the values of the
ethyl acetate 105.32 ± 1.24 22.25 ± 0.85 antioxidant activity of X. annuum. IC50 val-
1
Each value in the table was obtained by calculating ues for antioxidant activity of the extracts
the average of three analyses ± standard deviation are given in Table 2. The antioxidant activ-
ity values examined by DPPH radical scav-
The results of the analysis of different enging activity ranged from 59.25 to
extracts indicate that acetone is the most 956.81 µg/ml. The largest capacity to neu-
effective solvent for extraction of phenolic tralize DPPH radicals was measured in the
compounds from X. annuum, and that mod- acetone extract from X. annuum, which
erately polar solvents should be used for the neutralized 50 % of free radicals at the very
purpose. small concentrations of 59.25 µg/ml. In the
The concentration of flavonoids in vari- measuring of total phenolic content and fla-
ous extracts of X. annuum was determined vonoid concentrations acetone extract
using spectrophotometric method with showed the highest activity from the tested
AlCl3. The content of flavonoids was ex- extracts.
pressed in terms of rutin equivalents. The
summary of quantities of flavonoids identi- Table 2: Antioxidant (DPPH scavenging) activ-
fied in the tested extracts is shown in Table ity1 of investigated plant extracts and standard
substance presented as IC50 values (µg/ml)
1. The concentrations of flavonoids in plant
extracts ranged from 22.25 to 62.42 mg
RU/g. High concentrations of flavonoids Type of extract IC50 values (µg/ml )
were measured in acetone extracts. The methanolic 91.31 ± 1.32
concentration of flavonoids in plant extracts acetone 59.25 ± 0.93
depends on the polarity of solvents used in ethyl acetate 956.81 ± 1.84
the extract preparation (Min and Chun- chlorogenic acid 11.65 ± 0.52
Zhao, 2005). Based on the obtained values 1
Each value in the table was obtained by calculating
of the concentration of flavonoids in the the average of three analyses ± standard deviation
examined extracts of X. annuum, it was
Based on these results, each extract of
found that the highest concentration of
X. annuum showed a phenol concentration-
these compounds was in the extracts ob-
dependent scavenging effect. Numerous
tained using solvents of moderate polarity.
investigations of the antioxidant activity of
plant extracts have confirmed significant
Antioxidant activity
linear correlation between total phenolic
The antioxidant activity of different
content and antioxidant activity (Katalinić
plant extracts of X. annuum was determined
et al., 2004).
using methanol solution of 2,2-diphenyl-1-

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

In previous studies of quantitative and In addition, the phenolic contents of ex-

qualitative composition of the main bio- tracts depend on the solvent used for extrac-
logical active ingredients of X. annuum, tion, and not only the phenolic content but
luteolin (3’,4’,5,7-tetrahydroxyflavone), 1, also the properties of these compounds do
and quercetin (3,3’,4’,5,7-pentahydroxy- contribute to the activites of different ex-
flavone), 2, were identifed as the predomi- tracts.
nant flavonoids and ursolic acid - penta-
cyclic triterpene acid, 3, as dominant triter- Antimicrobial activity
penoid compound (Zemtsova and Molcha- The results of in vitro antibacterial and
nova, 1979). A number of pharmacological antifungal activities of acetone, ethyl ace-
studies showed that luteolin and quercetin tate and methanolic extracts of X. annuum
exhibit antioxidant, antibacterial, antifun- are shown in Table 3 and Table 4. For
gal, anti-inflammatory and anticancer ac- comparison, MIC and MMC values for
tivities (López-Lázaro, 2009; Gusdinar et doxycycline and fluconazole are also listed
al., 2011). Ursolic acid exist widely in most in Table 3 and Table 4. The solvent (10 %
of the active components of medicinal DMSO) did not inhibit the growth of the
herbs, spices, fruits, and vegetables. Several tested microorganisms.
stydies demonstrated the strong antioxidant, Antimicrobial activity of tested extracts
antimicrobial and antitumor activity of this was evaluated by determining MICs and
important compound (Assimopoulou et al., MMCs in relation to the 24 species of mi-
2005; Feng et al., 2009). croorganisms. MICs and MMCs values
were in range from 0.625 mg/ml to
40 mg/ml. The tested extracts showed dif-
ferent degree of antimicrobial activity in
relation to the tested species. The intensity
of antimicrobial action varied depending on
the groups of microorganisms and on the
type of the extracts.
Figure 1: Structure of luteolin (1) In general, the tested extracts demon-
strated selective and moderate antimicrobial
activity, while showing more potent inhibi-
tory effects on the growth of G+ bacteria
than to the other tested microorganisms
(p < 0.05). Statistically significant differ-
ence in activity between the extracts of X.
annuum was observed (p < 0.05). The ace-
tone extract showed the strongest activity,
and ethyl acetate appeared the weakest. At
Figure 2: Structure of quercetin (2) the acetone extract statistically significant
difference can be seen in the activity on G+
bacteria compared to other microorganisms.
In relation to the tested standard and clini-
cal strains of bacteria, all tested extracts
demonstrated approximately similar activi-
ty. An exception was observed in S. aureus,
where the clear difference can be seen at
acetone and methanolic extract.
Figure 3: Structure of ursolic acid (3)

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
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Table 3: Antibacterial activities of acetone, ethyl acetate and methanolic extracts of X. annuum
against tested microorganisms based on microdilution method

Acetone Ethyl acetate Methanolic Doxycycline

Species 1 2
MIC MMC MIC MMC MIC MMC MIC MMC
Escherichia coli ATCC 25922 10 10 5 20 20 20 15.625 31.25
Escherichia coli 20 40 40 40 40 40 7.81 15.625
Pseud. aeruginosa ATCC
5 5 10 20 5 10 62.5 125
27853
Pseud. aeruginosa 2.5 10 10 20 5 10 250 > 250
Salmonella enterica 10 10 40 40 20 20 15.625 31.25
Proteus mirabilis 1.25 2.5 20 40 2.5 5 250 > 250
Staph. aureus ATCC 25923 10 10 20 20 20 20 0.224 3.75
Staph. aureus 0.625 1.25 5 10 2.5 2.5 0.448 7.81
Enter. faecalis ATCC 29212 10 10 20 40 20 20 7.81 62.5
Enter. faecalis 20 40 20 40 20 40 7.81 62.5
Bacillus subtilis ATCC 6633 1.25 2.5 5 5 2.5 5 1.953 31.25
Bacillus subtilis 1.25 2.5 10 10 1.25 5 0.112 1.953
Bacillus cereus 0.625 1.25 1.25 5 1.25 2.5 0.977 7.81
Bacillus pumilus NCTC 8241 0.625 1.25 1.25 5 1.25 2.5 0.112 7.81
Sarcina lutea 2.5 2.5 10 20 5 5 < 0.448 3.75
1
Minimum inhibitory concentration (MIC) and 2minimum microbicidal concentration (MMC) values are given as
mg/ml for plant extract and µg/ml for antibiotics. Antibiotic: doxycycline

Table 4: Antifungal activities of acetone, ethyl acetate and methanolic extracts of X. annuum against
tested microorganisms based on microdilution method

Acetone Ethyl acetate Methanolic Fluconazole

Species
1 2
MIC MMC MIC MMC MIC MMC MIC MMC
Candida albicans 20 40 20 40 20 20 62.5 1000
Rhodotorula sp. 5 10 5 10 5 10 62.5 1000
Saccharomyces boulardii 20 20 20 20 20 40 31.25 1000
Aspergillus niger ATCC 16404 10 40 1.25 5 0.625 0.625 62.5 62.5
Penicillium italicum 2.5 2.5 20 40 1.25 20 1000 1000
Penicillium digitatum 40 40 20 40 20 20 31.25 31.25
Penicillium chrysogenum 40 40 20 40 10 20 62.5 500
Trichothecium roseum 10 20 20 40 20 20 500 500
Botrytis cinerea 10 40 20 40 10 20 31.25 500
1
Minimum inhibitory concentration (MIC) and 2minimum microbicidal concentration (MMC) values are given as
mg/ml for plant extract and µg/ml for antibiotics. Antibiotic: fluconazole

The tested extracts showed high anti- 20 mg/mL. The acetone extract showed a
bacterial activity against G+ bacteria, significant effect against B. pumilus NCTC
especially for species of the genus Bacillus 8241 and B. cereus. Clinical isolate of S.
(clinical isolates and standard strains). aureus also showed a promising sensitivity
MICs values were in range from to acetone extract, MIC was at
0.625 mg/mL to 10 mg/mL, and MMCs 0.625 mg/mL and MMC at 1.25 mg/mL.
values were from 1.25 mg/mL to

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

The tested extracts did not affect the REFERENCES

growth of clinical isolates and standard
strains of G- bacteria or their activities were Andews JM. BSAC standardized disc sus-
very low (MIC and MMC ranged from ceptibility testing method (version 4). J An-
2.5 mg/mL to 40 mg/mL). The exception is timicrob Chemother 2005;56:60-76.
in the activity of the acetone extract against
P. mirabilis, where MIC was at Assimopoulou AN, Zlatanos SN, Papa-
1.25 mg/mL and MMC at 2.5 mg/mL. georgiou VP. Antioxidant activity of natu-
The tested extracts showed low to mod- ral resins and bioactive triterpenes in oil
erate antifungal activity. Methanol extract substrates. Food Chem 2005;92:721-7.
showed a significant effect on A. niger
ATCC 16404 with MIC and MMC at Boussaada O, Ben Halima Kamel M, Am-
0.625 mg/mL. mar S, Haouas D, Mighri Z, Helal NA. In-
Stamatis et al. (2003) have investigated secticidal activity of some Asteraceae plant
the activity of different plants of Greek extracts against Tribolium confusum. B In-
herbal medicines on clinical isolates of He- sectol 2008;61:283-9.
licobacter pylori and have determined that
X. annuum has no effect on these bacteria. Dogan Y, Nedelcheva AM, Yarci C. Plant
Although it is mentioned in the papers as a taxa used as brooms in several southeast
wild medicinal herb of these areas (Jaric et European and west Asian countries. Nat
al., 2007; Dogan et al., 2008; Radanova, Croat 2008;17:193-206.
2009), with the exception from Stamatis et
al. (2003), there are no other data about Feng J, Chen W, Zhao Y, Ju X. Anti-tumor
antimicrobial activity of X. annuum. In this activity of oleanolic, ursolic and glycyr-
study antimicrobial activity of acetone, rhetinic acid. The Open Nat Prod J
ethyl acetate and methanolic extracts of X. 2009;2:48-52.
annuum against wide range of G+, G-
bacteria and fungi was investigated for the Gusdinar T, Herowati R, Kartasasmita RE,
first time. The results of our research Adnyana IK. Anti-inflammatory and anti-
indicate good antimicrobial potential of X. oxidant activity of quercetin-3,3’,4’-tri-
annuum. Thus, X. annuum can be consider- acetate. J Pharm Toxicol 2011;6:182-8.
ed as a source of potential antimicrobial
substances. Jaric S, Popovic Z, Macukanovic-Jocic M,
Djurdjevic L, Mijatovic M, Karadzic B et
CONCLUSION al. An ethnobotanical study on the usage of
wild medicinal herbs from Kopaonik
The results of our study suggest the Mountain (Central Serbia). J Ethnopharma-
great value of X. annuum in phytotherapy, col 2007;111:160-75.
pharmacy and food industry. Therefore, the
aerial parts of this plant are potential natural Jayaraman S, Manoharan MS, Illanchezian
source of biologically active substances S. In vitro antimicrobial and antitumor ac-
with strong antioxidant and antimicrobial tivities of Stevia rebaudiana (Asteraceae)
effects. For the extraction of active leaf extracts. Trop J Pharm Res 2008;7:
components from X. annuum acetone is an 1143-9.
effective solvent.
Kasim LS, Ferro VA, Odukoya OA, Ukpo
ACKNOWLEDGEMENTS GE, Seidel V, Gray AI et al. Evaluation of
This investigation was supported by the cytotoxic and antimicrobial activities of
Ministry of Education and Science of the Struchium sparganophora (Linn.) Ktze As-
Republic of Serbia, grants No. III41010 and teraceae. J Med Plant Res 2011;5:862-7.
OI173032.

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 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

Katalinić V, Miloš M, Kulišić T, Jukić M. Singleton VL, Orthofer R, Lamuela RRM.

Screening of 70 medicinal plant extracts for Analysis of total phenols and other oxida-
antioxidant capacity and total phenols. tion substrates and antioxidants by means
Food Chem 2004;94:550-7. of Folin-Ciocalteu reagent. Meth Enzymol
1999;299:152–78.
Kumarasamy Y, Byres M, Cox PJ, Jasapars
M, Nahar L, Sarker SD. Screening seeds of Skaltsa HD, Lazari DM, Constantinidis T.
some Scottish plants for free-radical scav- Composition of the essential oil of Xeran-
enging activity. Phytother Res 2007;21: themum annuum L. from Greece. J Essent
615-21. Oil Res 2000;12:742-4.

López-Lázaro M. Distribution and biologi- Stamatis G, Kyriazopoulos P, Golegou S,

cal activities of the flavonoid luteolin. Min- Basayiannis A, Skaltsas S, Skaltsa H. In
Rev in Med Chem 2009;9:31-59. vitro anti-Helicobacter pylori activity of
Greek herbal medicines. J Ethnopharmacol
Min G, Chun-Zhao L. Comparison of tech- 2003;88:175-9.
niques for the extraction of flavonoids from
cultured cells of Saussurea medusa Maxim. Stanković MS, Topuzović M, Solujić S,
World J Microb Biot 2005;21:1461-3. Mihailović V. Antioxidant activity and con-
centration of phenols and flavonoids in the
Muley BP, Khadabadi SS, Banarase NB. whole plant and plant parts of Teucrium
Phytochemical constituents and pharmacol- chamaedrys L. var. glanduliferum Haussk.
ogical activities of Calendula officinalis L. J Med Plant Res 2010;4:2092-8.
(Asteraceae): A review. Trop J Pharm Res
2009;8:455-65. Tekao T, Watanabe N, Yagi I, Sakata K. A
simple screening method for antioxidant
Özgen U, Mavi A, Terzi Z, Coşkun M, Yil- and isolation of several antioxidants pro-
dirim A. Antioxidant activities and total duced by marine bacteria from fish and
phenolic compounds of some Asteraceae shellfish. Biosci Biotechnol Biochem
species. Turkish J Pharm Sci 2004;3:203- 1994;58:1780-3.
16.
Vasić GP, Glodjović VV, Radojević ID,
Quettier DC, Gressier B, Vasseur J, Dine T, Stefanović OD, Ćomić LjR, Djinović VM
Brunet C, Luyckx C et al. Phenolic com- et al. Stereospecific ligands and their com-
pounds and antioxidant activities of buck- plexes. V. Synthesis, characterization and
wheat (F. esculentum Moench) hulls and antimicrobial activity of palladium(II) com-
flour. J Ethnopharmacol 2000;72:35–42. plexes with some alkyl esters of (S,S)-
ethylenediamine-N,N0-di-2-propanoic acid.
Radanova S. Variety of medicinal plants in Inorg Chim Acta 2010;363:3606-10.
a cultigenic ecosystem. XI Anniversary
Scientific Conference. Biotechnol & Bio- Vogl-Lukasser B, Vogl CR. Ethnobotanical
technol Eq 2009;23:389-92. research in homegardens of small farmers
in the alpine region of Osttirol (Austria):
Sarker SD, Nahar L, Kumarasamy Y. Mi- An example for bridges built and building
crotitre plate-based antibacterial assay in- bridges. Ethnobot Res Appl 2004;2:111-37.
corporating resazurin as an indicator of cell
growth, and its application in the in vitro Watson R, Preedy VR. Botanical medicine
antibacterial screening of phytochemicals. in clinical practice (p 421). Trowbridge,
Methods 2007;42:321-4. UK: CAB International, 2008.

238
 EXCLI Journal 2011;10:230-239 – ISSN 1611-2156
Received: September 14, 2011, accepted: November 20, 2011, published: November 24, 2011

Xeranthemum. In: Flore de la Republique

Socialiste de Serbie VII, Acad. Serb. Sci. &
Arts. Gajić M, Belgrade (in Serbian), 1975,
(p 176).

Zemtsova GN, Molchanova LP. Flavonoids

and triterpenoids of Xeranthemum annuum
L. Chem Nat Comp 1979;15:762.

Zhou K, Yu L. Effects of extraction solvent

on wheat bran antioxidant activity estima-
tion. LWT-Food Sci Technol 2004;37:717-
21.

239