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com
International Journal of Current Pharmaceutical Review and Research; 7(2); 110-116

ISSN: 0976 822X

Research Article

Antioxidant and Antimicrobial Activity of Flavonoids Fraction Extract

from Arnebia Decumbens (Vent) Growing in South East Algeria
Tamma N. Eddine1, Gherraf N. Eddine2, Laoini Salah Eddine1, Kefi Serra3, Selmi Sowsen3,
Limam Ferid3
1
Vaolization and technology of resource Saharian laboratory, Faculty of Technology, El Oued University El Oued
39000, Algeria.
2
Laboratory of Biomolecules and Plant, Breeding, Larbi Ben Mhidi University, Oum El Bouaghi 04000, Algeria.
3
Laboratory of Bioactive Substances, Center of Biotechnogy, Borj Cedria (CBBC), BP 901- Hammam Lif - 2050,
Tunisia.

Available Online:27th February, 2016

ABSTRACT
This study intends to investigate plants that grow in southeast Algeria namely Arnebia Decumbens (Vent.) which are
commonly used by medical science for a treatment. More especially this study shed light on the antioxidant effect and
biological activity of the plant whereby extraction of the active ingredients flavonoids is taken into account. The active
ingredients were identified through the scanning device using High-performance liquid chromatographic (HPLC). In-
order to get deep insight into the body of knowledge towards extraction process; current study utilized both qualitative
and quantitative approach. The data was analyzed separately on the antibacterial activity and oxidation of the extracts
flavonoids. The obtained results revealed a significant effect on the proliferation of some bacterial strains and free
radical. In addition, extracts of flavonoids have shown an active effect on bacteria; Staphylococcus aureus ATCC 6816
and Staphylococcus aureus methicillin resistant and a greater efficacy than the antibiotic chosen "polymyxin B". Hence,
based on the empirical evidences it can be stated that from various concentrations approach; the sensitivity of each type
of bacteria against each extract can be determined.

Keywords: Arnebia Decumbens (Vent.), flavonoids, DPPH, antioxidant activity, antimicrobial.

INTRODUCTION towards liver damage and carcinogenesis or toxic4,5,6.

Arnebia Decumbens (Vent.) it called in the region of Thus, in order to avoid certain unhealthy circumstances, it
south east Algeria in the name homaire , it is a small is necessary to focus on others natural antioxidant
red pigment which is usually found in the crust of the extracts from plants. Several chemical compounds
roots of an herbaceous plant in the region of south extracted from plant leaves, however, the most important
Algeria. The size of the homaire does not exceed 25 cm. is the flavonoids. Flavonoids are secondary metabolites
It is covered with stiff bristles of the latter turn into a ubiquitously distributed in all higher plants7. Flavonoids
semblance of thorns thin when they reach the plant and (or bioflavonoids) (from the Latin word flavus meaning
the beginning of drying Arnebia Decumbens (Vent.) yellow, their color in nature). Chemically, they have the
elongated leaves and do not have a clear neck. The general structure of a 15-carbon skeleton, which consists
flowers are yellow in color and collects in nurat apical of two phenyl rings (A and B) and heterocyclic ring (C).
dense1. Herbal treatment for curing certain medical This carbon structure can be abbreviated C6-C3-C68,9.
diseases is a common practice in Africa, statistically it is Contemporary studies confirmed the antimicrobial
estimated that over 80% of the total population produced activity of flavonoids occurring in vegetable foods and
a wide array of phytochemical most of which are used medicinal plants. These antimicrobial activities facilitate
from the plant. The main reason for preferring herbal in diverse ways such as anti-allergic, antimicrobial, anti-
treatment is to avoid the undesirable secondary effects inflammatory, vasoprotector and anti-tumour agents
that are commonly known as unwanted side effects of respectively10. The fruits date is rich with phytochimicals
some synthetic chemical drugs2. Moreover, research has like phenolic acids, sterols, procyanidins, flavanoids,
indicated that there is an inverse relationship between the carotenoids and anthocyanidin. Research has also
dietary intake of antioxidant-rich foods and the incidence revealed that the Arnebia decumbens (Vent.) Coss et Kral
of human disease3. Two synthetic antioxidants namely are beneficial in a biological and pharmacological
butylated hydroxytoluene (BHT) and butylated viewpoint. Because Arnebia decumbens (Vent.) Coss et
hydroxyanisole (BHA) which are more used in the food Kral contains antiviral, antibacterial, anti-inflammatory,
industry and also considered as a major contributor antitumor, these activities strength the immunity system,

*Author for Correspondence

 Tamma et al. / Antioxidant and Antimicrobial

Table 1: Gradient elution After drying in a dry, ventilated area, away from sunlight,
Temps Solvent Solvent the plant is crushed and then be weighted (100 g). Plant
(min) A (%) B (%) material obtained is then maceration in a hydro alcoholic
0 10 90 mixture (methanol /water; 80/20; V/ V). This maceration
5 20 80 process repeated in three times with solvent renewed
10 30 70 every after forty-eight hours. After went through the
process of filtration and concentration in vacuum, the
30 50 50
methanolic extract is then diluted with water distilled at
40 60 40
50 ml per 100 g of dry matter, the rest is left in solution
45 70 30 overnight then filtered. After filtration, the solution has
50 90 10 undergone successive liquid-liquid extractions type using
55 50 50 solvents of increasing polarity starting with chloroform
60 10 90 and ethyl acetate and finally with n-butanol16-19.
Analyses using High performance liquid chromatography
also it contains anti-oxidant activity11-15. Although all (HPLC)
studies conducted in appointments, in our knowledge, Flavonoids compounds have been separated and
there is no scientific information and empirical evidence identified by liquid chromatography system high
on the study of the plant, the ability of anti-oxidation of performance reverse phase mark (Agilent Technologies
bacterial activity on the Arnebia decumbens (Vent.) Coss 1260, Germany) equipped with a UV diode array detector
et Kral plant. There the current the study was conducted (DAD) and equipped with a chromatographic column
to estimate the chemical composition, the effect of an filled with a grafted silica gel, octadecyl type
anti-microbial and anti-oxidant from the extraction of ZorbaxEclipse XDB- C18 (4.6 x 100 mm, 3.5 microns).
natural products effective "flavonoid" of Arnebia For the various extracts, a conventional chromatographic
decumbens (Vent.) Coss et Kral plant that grows in the condition is usually adapted. Indeed, the detector (DAD)
southeast of Algeria. It is expected that the obtained is adjusted to a scan of scanning from 200 to 400 nm,
results can be taken as a guideline and might considered whereby the column temperature was maintained at 25
as a new source of agent antioxidant and antimicrobial. C. The volume injected is 20 l and the mobile phase
used is made up of two solvents A and B: Solvent A
MATERIAL AND METHODS (Methanol), Solvent B (water containing 0.1% formic
Chemicals and reagents acid). The speed of this phase is set at 0.4 ml / min. The
Methanol, ethanol absolute, chloroform (CHCl3), n- separation method adopted is the gradient elution in
butanol, ethyl acetate and ultra pure water were which the program is shown in the Table 1. Identification
purchased from VWR Merk (France), Diphenyl-1 of flavonoids compounds was performed by comparing
picrylhydrazyl (DPPH), BHT and chlorogenic acid were the retention times of peaks obtained for those flavonoids
procured from SigmaAldrich Inc (Paris, France). All standards injected in the same chromatographic
other chemicals and reagents were analytical-reagent, conditions.
sodium carbonate (Na2CO3), gallic acid, sodium nitrate Determination of total flavonoids
(NaNO2), aluminium chloride (AlCl3), sodium hydroxide Two reagents were used namely sodium nitrite colorless
(NaOH), catechin, hydrochloric acid (HCl), quercetin, solutions (NaNO2, 5%) and aluminum chloride (AlCl3,
linoleic acid, sodium phosphate, ferric chloride (FeCl3), 10%). The principle of the method is based on the
sulfuric acid (H2SO4) and ammonium molybdate. The oxidation of the flavonoids by these reagents; it leads to
following reagents were used for the microbial activity the formation of a brownish complex, which absorbs at
namely Nutrient agar and sabouraud dextrose agar. Plant 510 nm. Comparing the OD observed to that obtained by
materialThe aerial parts of Arnebia decumbens (Vent.) in a known concentration of catechin standard used to
March 2014 from Douilatte located in Wilaya of El-Oued evaluate the total content of flavonoids. The total
south east Algeria (33 07" 00" N 7 11' 00" E). This flavonoids are measured calorimetrically in a flask of 10
species was identified by Pr. Gerraf Noureddine; ml were introduced successively 250 l of extract of
Laboratory of Biomolecules and Plant, Breeding, Larbi known concentration in leaves and 75 l of a solution of
Ben Mhidi University, Oum El Bouaghi. Prior using the NaNO2 (5%). After 6 minutes was added 150 l of AlCl3
extraction of the plant for medical purpose, the leaves (10%) and 500 l of NaOH (1N) and in 1525 l of
were dried in well-ventilated spaces at room temperature. distilled water was added to the mixture successively. A
After, powdered and sifted in a sieve (0.75 m). calibration curve is prepared at different concentrations
Extraction of flavonoids with standard solutions of catechin. The absorbance of

Table 2: Mass yield and flavonoids content EC mg /g DM) of leaves extract obtained by methanol 80% of Arnebia
Decumbens (Vent.). Results are expressed as the mean and standard deviation of three independent experiments.
Total antioxidant activity mg GAE/g
Yield (%) w/w Flavonoids content
DW
Arnebia Decumbens (Vent.) 10,25 0,07 129.45 3.43 9.181 0.23

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 Tamma et al. / Antioxidant and Antimicrobial

mAU DAD1 A, Sig=254,4

7.271
600

500

400

300 10.326

200

28.053
8.597

17.892

30.701
24.015
24.999
25.848
14.918
100
5.9806.199

26.490
10.047

21.494
15.977

31.252
26.812
22.200
15.101

18.254

27.633
22.898
22.752

31.918
24.337
17.369
16.963

27.396

28.983
13.588

24.547

29.640
18.601

22.429

29.462
19.990

28.571

30.204
11.153

38.032
23.622

34.532
14.602

21.189

36.708
23.277
20.691
16.760

32.972
32.350
32.783
33.450
21.920

34.004

35.560
12.164

14.440

19.216
8.967

35.119
14.255

37.241
13.037

15.792
5.201

36.075
13.963
12.789

38.473
16.410
12.489
11.381
7.474
6.501

8.251
7.803
5.736

9.838
6.787

9.348
6.951

0 5 10 15 20 25 30 35 min
Figure 1: HPLC profile of leaves extract from Arnebia Decumbens (Vent.) recorded in UV at 254 nm

Table 3: Quantification of flavonoids compounds identified in methanolic 80% leaves extract from Arnebia
Decumbens (Vent.).
Retention time (min) Pic surface Identification Quantification g/ml
8.597 690.4 Gallic acid 63.676
7.271 3992.9 No identified -
10.326 1493.3 Catechin 122.584
24.015 625.6 p-comaric acid 101.451
25.848 715.8 Coumarine 82.478
30.701 783.5 Isoquercetrin 557.084
28.053 1502.3 No identified -
12.622 255 Querecetin 20.30

the mixture obtained is directly measured by UV-visible free radicals is therefore to measure its ability to scavenge
spectrophotometer at 510 nm and the results are free radicals and therefore slow or inhibit the creation of
expressed in mg catechin equivalent /g of dry matter (EC free radicals. In the case of the evaluation of the
/g DM). The data was analyzed with three separate antioxidant activity according to the equity standard
experiments. The obtained correlation coefficient of the polyphenols, the method comprises of comparing the
calibration curve was R2 = 0.988 while the result of the absorbance of targeted samples to that of a calibration
obtained result is presented as mean ( SEM)20-24. straight line, which connects the absorbance to the
Determination of the antioxidant activity concentration in standard. The types of radicals that are
The measurement of antioxidant potential was carried out used to evaluate the antioxidant activity of extracts of the
by determining the products resulting from the oxidation leaves are reducing power, the radical ABTS test, the
or by assessing the ability to trap reaction models radical OH, and the DPPH radical25-28.
radicals. The first mode requires prior knowledge of the Total antioxidant activity
compounds from oxidation. Indeed these methods seek During this test, hydrogen and electron are transferred
some functional groups (aldehydes, ketones, dicarbonyl) from the reducing compound (extract-antioxidant) to the
in the derivatives of the original components. The second oxidizing complex. This transfer phenomenon depends on
mode links the amount of trapped radicals of used the redox potential of the medium pH and on the structure
antioxidant. These dual modes and forms of expression of the antioxidant compound. The test is based on the
are preferred to use the percent inhibition (IC) and / or reduction of the molybdenum of the oxidation stage (VI)
equivalence standards polyphenols obtained by UV- the oxidation state (V). This reduction is materialized by
Visible spectroscopy. The percent inhibition for assessing forming a greenish complex (phosphate / Mo (V)) at an
antioxidant activity of a sample is determined using the acidic pH that measures the decrease in the coloration of
following formula: IC (%) = [(A-B) / (A)] 100 molybdenum (VI) complex in the presence of antioxidant.
Where a = absorbance of the oxidized solution in the Unlike other tests, this test makes it possible not only to
absence of antioxidant agents, b = absorbance of the quantify the contribution of the antioxidant activity of the
oxidized solution in the presence of antioxidant agents. polyphenols but also other antioxidants such as vitamins.
Evaluation of the ability of the compound (extract) to trap The method comprises introducing into an Eppendorff

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Figure 2: DPPH scavenging radical activity of leaves extract from Arnebia Decumbens (Vent.).

Table 2: Antibactrial activity of methanolic extracts calculated using the formula below: % I C = [(At0 -
leaves of Arnebia Decumbens (Vent.) At 30) / At0 100)]
Microorganisms Diameter of zone inhibition Where At 0: absorbance of control (containing no
Bacteria extract polymyxine B antioxidant) 30 minutes At 30: excerpts absorbance
Staphylococcus aureus measured after 30 minutes. The activity of radical
13 0.6 12,5 0,5 scavenging is usually expressed in IC50 (g/ml), the dose
ATCC 6816
Staphyloccocus aureus 14 0.5 11,0 0,4 radical scavenging required to cause 50% inhibition. All
Bacillus cereus ATCC results presented are averages ( SEM) and analyzed with
8 0,2 8,5 0,1 three repetitions. By varying the concentration of the
14579
extracts and calculating the percentage concentration for
tube 100 l of the extract of the leaves mixed with 1 ml each corresponding IC, a linear regression was
of a reactive compound of H2SO4 (0.6 M), NaH2PO4 (28 determined between the different concentrations and
mM) and ammonium molybdate (4 mM), the tube percentage of inhibition. Prediction also leads towards
incubated at 95 C for 90 min. After cooled, the deduction of the corresponding IC 50 value31,32.
absorbance is measured at 695 nm, the control consisted Antimicrobial activity assays
of 100 l of methanol mixed solvent with 1 ml of the Microorganisms
reagent mentioned above29,30. For the calibration curve In total, three bacterial cells were used in a study. The
and controls are incubated under the same conditions. bacterial cells assayed: Staphylococcus aureus ATCC
The results are expressed in mg of gallic acid equivalents 6816, Staphyloccocus aureus, Bacillus cereus ATCC
per gram of dry matter (EAG mg / g DM). The calibration 14579. All strains were obtained from the Laboratory of
curve is established with a correlation coefficient R2 = Bioactive Substances, Center of Biotechnogy, Borj
0.998. Cedria (CBBC), and BP 901- Hammam Lif - 2050,
DPPH assay Tunisia33-35.
DPPH test (2, 2-diphenyl-1-picrylhydrazyl) is a method Incubation conditions
widely used in the analysis of an antioxidant activity. Nutrient agar was used culture medium for bacteria which
Indeed, the DPPH is characterized by its ability to was incubated for 24 h at the temperature of 37 C and
generate stable free radicals. This stability is due to the yeasts were cultured in sabouraud dextrose agar (SDA;
relocation of the free electrons in the molecule. The 4% dextrose, 2% neopeptone and 1,7% agar) for 24-48
presence of DPPH radicals resulted in a dark purple color hours at the temperature of 30C36,37.
of the solution, which absorbs at about 517 nm. The Disc diffusion assay
reduction of DPPH radicals by an antioxidant agent Methanol water extracts Arnebia Decumbens (Vent.) were
results in a discoloration of the examined solution. dissolved in methanol-water 50% for a final
Evaluation of the antioxidant capacity is performed as concentration 10 mg/ml and filter-sterilized through a
follows, in 250 l of DPPH solution (7.8 mg DPPH in 0.45 membrane filter. The antimicrobial activity was
100 ml of methanol) was mixed with 1 ml of leaves estimated by method of disc diffusion, 100 l of
extract. The resulting mixture was then kept sheltered suspension for each microorganism 108 colony-forming
from the light at room temperature for 30 minutes. The units (CFU)/ml containing 20 ml of nutrient agar for
absorbance is measured at 517 nm. Sample preparation bacteria, after were placed in the petri sterilized filter
and the control is carried out under the same operating paper disc (6mm in diameter). Later it was soaked with
conditions. The decrease in the absorbance is measured 15 l of the 50 mg/ml of each methanolic extracts (150
spectrophotometrically and IC% (percent inhibition) is g/disc). The methanol 50% was used as a negative

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control whereas polymyxine B was taken as the positive extracts of the leaves of Arnebia Decumbens (Vent.).
control. However, both positive and negative control was These results indicate that the extract from the leaves of
prepared with the same procedure. The detailed described Arnebia Decumbens (Vent.), the extract is rich in
on the procedure was mentioned-earlier except for the flavonoids; the results are shown in table 2. The reference
methanol extract which was substituted by 15 l of compound used in the preparation of this curve is
positive control at 50 mg/ml. The diameter of the catechin. The curve is calculated with a correlation
inhibition zone around each disc was measured for three coefficient R2 = 0.988.
replicates. Total antioxidant activity
Statistical analysis The total antioxidant activity of methanolic 80% leaves
Data was analysed using statistical tests whereby the extracts from Arnebia Decumbens (Vent.) is 129.45
obtained results were presented in mean values, and 3.43 mg GAE/g DW. These results exhibit strong values
standard deviations (SD). Since, all measurements were and confirmed the high antioxidant activity of leaves
carried out in three experiments therefore, the all the extract of Arnebia Decumbens (Vent.) founded in DPPH
analyses in the present study were analyzed three times reducing power. Study estimated the total antioxidant
determinations). Statistical calculations were carried out capacity of Korkobb, Tunisian date fruit. The authors
by OriginPro version 8 software (Prolab), correlations suppose that the highest level of flavanoid in this variety
were obtained by Pearson correlation coefficient using is responsible for the higher total antioxidant capacity36,
bivariate correlations test. P value were set at 0.05. the result are presented in Table 2.
Therefore, the obtained value less P value (0.05) was DPPH radical scavenging activity
regarded as a statistically significant and P values < 0.01 The DPPH radical scavenging activity of methanolic
was regarded extremely statistically significant. water extract of Arnebia Decumbens (Vent.) is
represented in Figure 2. The crude extract of leaves
RESULTS AND DISCUSSION displayed the high value (IC50=12.050.08 g/mL). The
Extract yield antioxidant capacity of leaves extract is higher than the
The methanol is a solvent extract significant amount of positive control BHT (IC50= 14.46 0.06 g/mL). The
flavonoids compounds and recently used in several phenolic compounds were considered to be the most
studies. It is considered as the best solvent of active antioxidant derivatives in plants and are well
antimicrobial substances compared with the other known as antioxidant and scavenger agent against free
solvents and given an elevated antioxidant activity. The radicals111. In living system, the free radicals are
results of extract yield of Arnebia Decumbens (Vent.) are constantly generated and their associated with oxidative
mentioned in Table 3, which shows the extraction yield extensive damage to tissues. Different therapeutic
(g/100 g dry weight), the mass yield obtained for approaches can be used to decrease the oxidative stress
methanolic water extract of leaves from Arnebia including scavenging of free radicals. Inhibition of these
Decumbens (Vent.) is found 10,25%. radicals produces enzymes and enchains antioxidant
HPLC analysis system by targeting the signaling routes. Many synthetic
The identification of compounds flavonoids Extract the drugs protect against oxidative damage but they have
majority of plant extract Arnebia Decumbens (Vent.). adverse side effects37. The present study showed that the
HPLC was carried out based on the comparison of their leaves methanolic water extracts of Arnebia Decumbens
retention times with those obtained for the same standard (Vent.) have good antioxidant as well as free radicals
compounds. This comparison allowed us to confirm the scavenging properties.
presence of two major flavonoids isoquercitrin with a Antimicrobial activity
retention time of 30 701 min, and catechin with a The diameter inhibition of leaves extract from Arnebia
retention time 10 326 min. Show in Figure 2 and table 4 Decumbens (Vent.) is 130.6 mm. Then we can say that
with rates respectively 557 084 mg / 100 g of plant the bacteria Staphylococcus aureus ATCC 6816 sorting
extract and 122 584 g / 100 g of plant extract, and in very sensitive to flavonoids. The results showed that the
small, we find a Querecetin compound with a retention diameter inhibition 12.5 0.5 mm for Polymyxin B are
time 12.622 min. Minor peaks were also recorded with against the bacteria Staphylococcus aureus ATCC
retentions times ranging from 8597 min probably are 6816. Through reading the results, we conclude that the
phenolic compounds gallic acid with a rate of 63 676 g / most effective flavonoids to polymyxin B because the
100 g and compounds coumarine p-coumaric acid 24.015 diameter of inhibition is greater. The diameter of
min a time of retention, and coumarine at retention time inhibition of leaves extract from Arnebia Decumbens
of 25 848 min. with rates respectively 101.451g / 100g (Vent.) is 14 0. 5 mm, then we can say that the
and 82.478g / 100 g of plant extract. Flavonoids rates methicillin-resistant Staphylococcus aureus bacteria
are determined in plant extracts according to the sensitive to the extracts flavonoids. The results give
calibration curve (peak areas as a function of the inhibition diameter 11.0 0.4 Polymyxin B for against
concentration of the standards). bacteria Staphylococcus Aureus Methicillin resistant.
Flavonoids content Through reading the results, we conclude that flavonoids
The results obtained are expressed as mg catechin are more effective than polymyxin B against bacteria
equivalents per gram of dry mass (EC mg / g DM). The Staphylococcus aureus methicillin resistant. The diameter
results of the quantitative analyzes of flavonoids in the of inhibition of leaves extract from Arnebia Decumbens

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 Tamma et al. / Antioxidant and Antimicrobial

(Vent.) is 8 0, 2 and from these results we can say that 4. Pekkarinen S.S., Heinonen I.M., Hopia A.I. (1999):
the bacteria Bacillus cereus ATCC 14579 Medium Flavonoid quercetin, myrcetin, kaemferol and (+)-
Sensitivity against flavonoid. Through reading the results, catechin as antioxidants in methyl linoleate. Journal of
we conclude that flavonoids are less effective than the Science of Food and Agriculture, 79: 499506.
polymyxin B against to bacteria Bacillus cereus ATCC 5. Whysner, J., Wang, C. X., Zang, E., Iatropoulos, M.
14579. J., Williams, G. M., 1994. Dose response of
promotion of butylated hydroxyanisole in chemically
CONCLUSION initiated tumors of the rat fore stomach. Food and
We think that the present study is the first investigation Chemical Toxicology. 32, 215-222.
and comparing the phytochemical composition, 6. Moure, A., Cruz, J. M., Franco, D., Dominguez, J. M.,
antioxidant and antimicrobial activity of extracts of three Sineiro, J., Dominguez, H., et al., 2001. Natural
varieties of Arnebia decumbens (Vent.) growth in antioxidants from residual sources. Food Chemistry,
Southeast Algeria. This study showed that considerable 72(2), 145-171.
variance exists between the three extracts of leaves for 7. Maria Daglia., 2011. Polyphenols as antimicrobial
plant Arnebia decumbens (Vent.) Cosset Kral" agents. Current Opinion in Biotechnology. 23, 174-
flavonoids". We found highest amount of flavonoids. On 181.Mi-Yae, S., Tae-Hun, K., Nak-Ju, S., 2003.
the other hand, the results of antioxidant activity tests Antioxidants and free radical scavenging activity of
present the strong capacity of extracts of leaves for plant Phellinus baumii (Phellinus of Hymenochaetaceae)
Arnebia decumbens (Vent.), higher than the standards extracts. Food Chemistry. 82, 593-597.
antioxidants (BHT). Finally, all extracts Appears the high 8. Naught MC, Alan D; Wilkinson, Andrew; IUPAC
antimicrobial activity for the Microorganisms tests (1997), "IUPAC Compendium of Chemical
(bacteria) exceeded most of the time the positive control. Terminology", Flavonoids (isoflavonoids and
A strong correlation was found between activity and neoflavonoids) (2 ed.), Oxford: Blackwell
phytochemical contents indicates that the effects observed Scientific, doi:10.1351/goldbook.F02424
could be attributed to flavonoids compounds. This data 9. Jump up "The Gold Book". doi :10.1351/gold book.
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ACKNOWLEDGEMENTS 13. Tanaka S, Kuwai Y, Staba M (1986) A comparative
The authors want to thank gratefully Dr. Touhami Lanez study on anti-inflammatory activities of enantiomers,
Director of Valorization and Technology of resource shikonin and alkannin. J Nat Prod 49: 466469)
Saharian laboratory (El-Oued University, Algria) for or 14. Sankawa U, Ebizuka Y, Miyazaki T, Isomura Y,
her continuous supported during the work and the use of Otsuka H, Shibata S, Inomata M, Fukuoka F (1977)
all laboratory materials. Thanks are also to Dr Farid Antitumor activity of shikonin and its derivatives.
Limam Director of Laboratory of Bioactive Substances, Chem Pharm Bull 25:23922395
Center of Biotechnogy, Borj Cedria (CBBC), Tunisia and 15. Kashiwada Y, Nishizawa M, Yamagishi T, Tanaka T,
Dr Chedly Abdelly Director of Biotechnogy Center, Nonaka G-I, Cosentino LM, Snider JV, Lee KH
Ecopark of Borj Cedria, Tunisia for their help during the (1995) Anti-AIDS agents 18. Sodium and potassium
experimental process and for the explanation of all salts of caffeic acid tetramers from Arnebia euchroma
techniques used in this study. as anti-HIV agents. J Nat Prod 58: 392400
16. Seshadri TR. (1962) In the chemistry of flavonoid
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