Phenolic characterization and antioxidant activity of two ...

Moroccan Journal of Chemistry

ISSN: 2351-812X

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Amine & al./ Mor. J. Chem. 6N°1 (2018) 01-13

Mor. J. Chem. 6N°1 (2018) 01-13

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Phenolic characterization and antioxidant activity of two endemic

wormwood species of Morocco: Artemisia ifranensis J. Didier and Artemisia

mesatlantica

S. AMINE

a,b* , H. EL AZZOUZI

a,b, F. RADI

a,b, Z. KHIYA

a,b, S.AMALICH

a,b, CH.

SEKKATEa,b

, M. MAHJOUBIa,b

, M. BOURAKHOUADARa,b

, T. ZAIRa,b

aResearch team of Chemistry Bioactive Molecules and the Environment, University Moulay Ismail, Faculty of Sciences, BP

11201, Zitoune, Meknes, Morocco

b Laboratory of Materials Chemistry and Biotechnology of Natural products, University Moulay Ismail Faculty of Sciences,

B.P. 11201 Zitoune, Meknès, Morocco

* Corresponding author:

[email protected].

Received 25 Jan 2017,

Revised 02 Oct 2017,

Accepted 26 Oct 2017

Abstract

Artemisia ifranensis and Artemisia mesatlantica are two endemic species in

Morocco belonging to the Asteraceae family. Located in the region of

Moroccan Atlas, they are very much sought after by the local population for

their therapeutic properties. However, given the limited studies conducted on

these species and in order to contribute to their valorization, we selected them in

full bloom in the region of Timahdite for a phytochemical study and an

evaluation of their antioxidant properties. The results of phytochemical

screening revealed their richness in polyphenols, sterols, triterpenes and

mucilages. The extraction yields of the total polyphenols by maceration in a

methanol-water mixture (80/20) were 10.10% for A. mesatlantica and 16.66%

for A. ifranensis. The determination of the total phenols by the Folin-Ciocalteu

method after fractionation of the polyphenolic extracts showed the abundance of

these compounds in the butanolic fraction of A. Mesatlantica and the

methanolic extract of A. Ifranensis. For the total flavonoids dosed with

aluminum trichloride, the ethyl acetate fractions of A. Mesatlantica and

butanolic acid. Ifranensis were the richest. In addition, the crude extract of A.

Mesatlantica was particularly rich in condensed tannins. Evaluation of the

antioxidant activity by reduction of iron (FRAP) and trapping of the free radical

DPPH * showed that all the extracts have a moderate antioxidant power. These

early studies on A. mesatlantica and A. ifranensis of Timahdite reveal their

abundance of important chemical compounds. Also, due to the ability of these

plants to trap radicals, they can be considered as sources of natural antioxidants.

Keywords: Artemisia mesatlantica, Artemisia ifranensis, Antioxidant activity, Flavonoids, Tannins.

Mor. J. Chem. 6N°1 (2018) 01-13

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1. Introduction The Asteraceae family, constituted from 13 000 plant species, is by far the most important in the flowering

trees [1]. It accounts for 10% of the world's flora and contains the largest number of endemic plant species

and sub-species [2]. Of the 1500 genera registered in this family, the genus Artemisia is distinguished by its

specific richness ranging from 200 to 400 species of herbaceous plants generally found in arid and semi-arid

zones [3]. In Morocco, which is the African country that is the "richest in Artemisia species", these play a

very important economic role. Indeed, the country alone provides 90% of the essential oils of wormwood

that feed the world market and which particularly integrate the cosmetics and perfumery industries [4].

Moreover, thanks to the experimental and ethnobotanical studies carried out on certain species of the genus,

their menstrual cycle regulating properties and their use as a remedy for several diseases such as diabetes,

bronchitis, abscesses, diarrhea, Respiratory and digestive disorders [5]. However, of the twelve species in

Morocco, only the species Artemisia herba-alba has been studied extensively in Morocco, Spain, Tunisia [6]

and Algeria [7]. This is particularly the case for endemic species such as A. ifranensis, A. mesatlantica, A.

atlantica var maroccana, A. flahauti and A. arborescens, which according to our knowledge have been only

subject to a limited number of studies relating to the chemical composition of their essential oil and their

biological activity [8-10]. The aim of this work is therefore to contribute to a better knowledge of these

species with a view to the possible identification of new natural molecules of economical interest. In this

study, we were interested in the valorisation of two endemic species: Artemisia mesatlantica (Artemisia

blue) and Artemisia ifranensis J. Didier which are very widespread between the two regions of Boulmane

and Ifrane located in the Middle Atlas of Morocco [11]. Thus, we carried out a phytochemical study and

evaluated the antioxidant power of polyphenols and flavonoids extracted from the two species: Artemisia

mesatlantica and Artemisia ifranensis J. Didier

2. Materials and Methods

2.1. Phytochemical tests

2.1.1. Plant material

The species selected for this study are Artemisia ifranensis J. Didier and Artemisia mesatlantica harvested

respectively in April 2015 and March 2015 at the time of their flowering in the region of Timahdite. They

were then dried in the shade for about ten days. The botanical identification of the two species was carried

out in the laboratory of Floristics of the Scientific Institute in Rabat.

2.1.2. Phytochemical Screening

This is a qualitative study aimed at the research of the main chemical groups in the extracts of the studied

plants. The analytical techniques undertaken are those described in the work of Bekro [12] and Bruneton

[13]. The search for alkaloids was possible thanks to the reagent of Dragendorff (reagent to the iodo-

bismuthate of potassium). The alcoholic solution of ferric chloride allowed the characterization of the

polyphenols. Acetic anhydride and concentrated sulfuric acid were required for sterols and polyterpenes. For

the characterization of gallic tannins, we used the Stiasny reagent, sodium acetate and ferric chloride. And

for the catechic tanins we used the reagent of Stiasny and the acetate of sodium. Chloroform, ammonia and

hydrochloric acid permitted the investigation of quinone substances. Hydrochloric alcohol, magnesium chips

and isoamyl alcohol were used to search for flavonoids. 2.1.2. Reactions of characterization

Mor. J. Chem. 6N°1 (2018) 01-13

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2. 1.3. Preparation of the extracts of A. Mesatlantica and A. ifranensis J. Didier

The extraction of the polyphenols was carried out at ambient temperature by maceration of 30 g of

vegetable powder (leaves and flowered tops) in a 300 ml hydro-alcoholic mixture (methanol / water, 80/20,

V / V) for 48 hours with stirring. The filtrates obtained were evaporated under vacuum at 50 ° C to remove

the solvent. The aqueous phase was successively extracted (fractionation) of liquid-liquid type using organic

solvents of increasing polarity (ethyl acetate and n-butanol). After evaporation of the various extraction

solvents, the series of extractions made it possible to obtain four fractions: the hydrometanol fraction (F0),

the ethyl acetate fraction (F1), the n-butanol fraction (F2), and the aqueous fraction (F3). The extracts are

kept cold in the absence of light until they are used.

2. 1.4. Determination of total polyphenols of A. Mesatlantica and A. ifranensis J. Didier

The total phenolic compound contents were evaluated using the Folin-Ciocalteu colorimetric method [14].

Of each plant extract, 20 μl are mixed with 1.5 ml of a sodium carbonate solution (75 g / l). Then 1.5 ml of

the 10% Folin Ciocalteu reagent (V / V) are added and the tubes are incubated for two hours at ambient

temperature and protected from light. The reading is carried out at 765 nm against a blank using a

spectrophotometer of the Shimadzu UV-MINI 1240 type. A calibration curve is carried out in parallel under

the same operating conditions using gallic acid as a positive control at Different known concentrations. The

results are expressed in milligrams of gallic acid equivalent per gram of dry matter (mg EAG / g of plant).

The total phenol content is calculated according to the following formula:

C: Concentration measured by calibration curve.

V: Volume of the overall sample.

D: Dilution factor.

2.1.5. Determination of flavonoids A. Mesatlantica and A. ifranensis J. Didier

The total flavonoid content of A. Mesatlantica and A. ifranensis J. Didier are estimated by the method of

aluminum chloride (AlCl3), according to the modified protocol of Kumaran [15]. 0.1 ml of the extract from

A. Mesatlantica and A. ifranensis J. Didier are mixed with 0.1 ml of 10% aluminum chloride followed by 20

ml of distilled water and supplemented with absolute methanol to 50 ml and then left for two hours in the

dark. The absorbance of each solution is determined at 433 nm. Under the same conditions, a concentration

range (5 to 35 μg / ml) was prepared from the quercetin stock solution with a concentration of 0.1 mg / ml

(25 mg / 250 ml). Thus, a curve of regression of the calibration range was established with quercetin (y =

0.073x-0.081). The flavonoid concentrations of each extract were calculated from this equation and

expressed as (mg equivalent quercetin / g dry plant material). The flavonoid contents are calculated by the

following formula:

T: Total flavonoid content (mg quercetin equivalent / g dry plant material)

C0: Concentration of gallic acid calculated from the equation of regression of the calibration range (μg / ml).

V0: Volume of solubilization of product obtained (ml).

Vf: Volume of the volumetric flask (50ml).

Vi: Volume of the extract (ml). and M: Weight of dry plant material (g).

Mor. J. Chem. 6N°1 (2018) 01-13

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2.2. Determination of the condensed tannins of A. Mesatlantica and A. Ifranensis J. Didier

The amounts of condensed tannins are estimated using the vanillin method (Julkunen-Titto, 1985) [16].

20 mg of catechin are mixed with 10 ml of distilled water and from this solution we have prepared the

various concentrations of (0.2 to 1.8). From each concentration we removed 50 μl and then added 3 ml of the

vanillin / methanol solution (4%, m / v). Then 1.5 ml of concentrated hydrochloric acid (HCl) were added

and allowed to react at the temperature Ambient for 20 min. The absorbance at 499 nm is measured against

white. The results of the plants studied are expressed in milligrams (mg) equivalent of the catechin per gram

of the dry plant material (mg EC / g) from the calibration curve.

2.3. Evaluation of the Antioxidant Activity of the Extracts of A. Mesatlantica and A. ifranensis J. Didier

by trapping the free radical: 2, 2-diphenyl-1-picrylhydrazyl (DPPH *)

The antiradical power of the initially prepared fractions was tested using the method dictated by Brand-

Williams et al [17]. The different fractions obtained from the crude extracts of each plant are subjected to

pre-tests in order to identify the most active fractions which will be used for the rest of the study. To carry

out the pre-tests and formal tests for the anti-radical activity, a solution of DPPH * was prepared by

solubilizing 2.4 mg of DPPH * in 100 ml of absolute ethanol. 200 μl of each ethanolic solution of the n-

butanol fraction and ethyl acetate are added to 2.8 ml of the ethanolic solution of DPPH * (0.024 g / l).

Solutions of ascorbic acid (reference antioxidant) are also prepared under the same conditions. The negative

control (white) consists only of 200 μl of ethanol with 2.8 ml of the ethanolic solution of DPPH *. The

reading of the absorbance is made against a white at 515 nm after 30 min incubation in the dark and at

ambient temperature. The positive control is represented by a solution of a standard antioxidant, ascorbic

acid, the absorbance of which was measured under the same conditions as the samples and for each

concentration; the test is repeated three times. The results were expressed as percent reduction of DPPH *

(PI %) according to Shariffar et al. [18].

-A control: Absorbance of the solution containing only radical DPPH solution

-A sample: Absorbance of the sample solution to be tested in the presence of DPPH*

Concentration values to inhibit or reduce 50% of the initial concentration of DPPH* (IC50) were determined

graphically by linear regression.

2.4. Evaluation of the Antioxidant Activity of the Extracts of A. Mesatlantica and A. ifranensis J. Didier

by the Ferric reducing antioxidant power (FRAP)

The iron reduction activity of our extracts is determined according to the method described by Oyaizu [19]

based on the reduction of Fe3 +

present in the K3Fe (CN)

6 complex to Fe

2 +. 400μl of the extract at different

concentrations are mixed with 2.5 ml of a 0.2 M phosphate buffer solution (pH 6.6) and 2.5 ml of a 1% K3Fe

(CN) 6 potassium ferricyanide solution. The whole is incubated in a water bath at 50

° C. for 20 minutes and

then 2.5 ml of 10% trichloroacetic acid are added to stop the reaction. The tubes are centrifuged for 10

minutes 2.5 ml of the supernatant are mixed with 2.5 ml of distilled water and 0.5 ml of a freshly prepared

0.1% ferric chloride solution. The reading of the absorbance is done at 700 nm against a prepared white,

replacing the extract with distilled water which makes it possible to calibrate the apparatus (UV-VIS

spectrophotometer). The positive control is represented by a solution of a standard antioxidant. Ascorbic

Mor. J. Chem. 6N°1 (2018) 01-13

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acid whose absorbance was measured under the same conditions as the samples. An increase in absorbance

corresponds to an increase in the reducing power of the extracts tested [20].

3. Results and discussion

3.1. Phytochemical tests

3.1. 1. Phytochemical Screening

This study has been carried out to identify the different families of secondary metabolites; it is based on

color and precipitation reactions. The phytochemical screening results are reported in the table 1.

Table 1: The results of the flowering phytochemical screening of A. mesatlantica and A. ifranensisJ. Didier

Tests Artemisia mesatlantica Artemisia ifranensisJ. Didier

Tannins +++ ++

Gallic tannins ++ --

Tannins catechics + +

Alkaloids - -

Flavones - +

Flavonole ++ -

Flavonone - ++

Sterols and triterpenes + +

Coumarin ++ -

Mucilage ++ ++

Anthocyanin - -

Saponiside - -

(+ ): Weak presence (++): Moderate presence (+++): Strong presence ( - ): Absence

Based on the results in the table 1, the Artemisia ifranensis and Artemisia mesatlantica contain important

secondary metabolites. A strongly positive response was observed in both species of Artemisia showing the

presence of tannins, Sterols, triterpenes and the mucilages.Instead for alkaloids, power and anthocyanins, a

negative reaction is noted for both species. The coumarins are presented in A. mesatlantica while they are

absent in A.ifranensis. On the other hand, the presence of flavones and flavonones are observed only in A.

ifranensis and flavonols are a characteristic of A. mesatlantica. By comparison with various work on the

species of the genus Artemisia, our results are consistent with the later reported work by Aljaiyash et al.,

(2014) [20]; Makhloufi, (2014) [21]; Elazzouzi et al.., (2014) [22]; Majdouli, (2015) [23]. Indeed, the

Artemisia species studied Artemisia herba alba, Artemisia herba alba Asoo, Artemisia ifranensis J.

Didier,Artemisia herba alba hugueii are all rich in polyphenols such as tannins, flavonoids and sterols.

However, the presence of alkaloids, saponins and mucilages varies from one species to another. Thus the

richness and variability of the compounds of these species are a real asset that assigns them various

important therapeutic properties and facilitates their integration in the areas of medical and cosmetic [2].

Similarly, in medicine, these species are very used against several diseases like diabetes, the

Mor. J. Chem. 6N°1 (2018) 01-13

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bronchitisabscesses, diarrhea, diseases of digestive and respiratory devices, the ulcer, etc. [5] and the

highlighting of therapeutic compounds via the phytochemical tests supports their use.

3.1. 2.Yields of extracts

Yields gross of A. mesatlantica and A. ifranensis J. Didier hydromethanolic extracts are the order of 10.01%

and 16.66%, respectively (table 2). For splitting of raw extracts of these two species, the results show that,

for A.mesatlantica the ethyl acetate fraction represents the highest yield (37.51%) followed by the aqueous

fraction (30,86%) then the butanolic (17.78%) fraction. However for A. ifranensis J. Didier highest

performance is obtained with the butanolic fraction (37, 66%), followed by the fraction in ethyl acetate

(31.16%) and Finally the aqueous phase containing a low yield compared to other fractions(12.33%). Yields

are calculated compared to 1 g of dry matter.

Table 2: Yields Extracts and fractions of A .mesatlantica and A .ifranensisJ.Didier

Yields of extracts and fractions in (%)

Species A .mesatlantica A. ifranensisJ. Didier

Brut extract 10 16.66

Ethyl acetate fraction 37.51 31.16

butanolic Fraction 17.78 37,66

Aqueous phase 30,86 12.33

Figure 1: The yields of extracts and fractions for A. mesatlantica and A.ifranensis J. Didier

Mor. J. Chem. 6N°1 (2018) 01-13

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3.1. 3. Determination of polyphenols, flavonoids and tannins total

In order to assess the levels of polyphenols of the different extracts and fractions of the species studied,

curves to calibration from Gallic acid, quercetin and of the vanillin were established (Figure 2). The

concentrations of phenols, flavonoids and tannins total in the extracts and fractions were then respectively

estimated equivalent per g of dry matter of Gallic acid, equivalent per g of dry matter of quercetin and

equivalent per g of dry matter of vanillin.

Figure 2: Calibration curves of Quercetin (a) acid Gallic (b) and vanillin(c)

So the highest levels of total phenols were detected in the butanolic fraction of A. mesatlantica (1.51 mg

EAG / 30g of dry matter) and the brut extract for A. ifranensis J. Didier (2.10 mg EAG / 30 g of dry matter)

while low levels have been found in the ethyl acetate fraction for A. mesatlantica (0, 17 mg EAG / 30 g of

dry matter) and the aqueous phase for a. ifranensis J. Didier (0.13 mg EAG / 30 g of dry matter). According

to our knowledge, there is very few works on the compounds phenolic A. mesatlantica and A. ifranensis J.

Didier. We chose to compare our results with other species of the genus Artemisia. We cite as an example,

the total phenolic compounds of Artemisia arvensis, Artemisia campestris and Artemisia herba alba

Djeridane and al., [24] .These authors showed that the rate in total phenols of these plants is respectively of

the order of 3.3 mg, 2,038 mg and 1.6 mg equivalent of Gallic acid / g of dry matter. These results are closer

to our work. The quantitative determination of total flavonoids by the method of the aluminum reveals that

the fractions of ethyl acetate and butanolic are the rich in flavonoids with a values of (1.49 µg EQ/mg of

Mor. J. Chem. 6N°1 (2018) 01-13

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extract) and (4.22µg EQ/mg of extract) respectively for A. mesatlantica and A. ifranensis J. Didier. Then

comes the aqueous extract (0.30 µg EQ/mg of extract) and (2.76 µg EQ/mg of extract) as for A.ifranensis J.

Didier ethyl acetate fractions and the crude extract for A. mesatlantica, their levels remain low (0, 22µg

EQ/mg of extract) and (0, 13µg EQ/mg of extract) respectively. The dosage of tannins condensed by the

method vanillin shows that the raw extract of A. mesatlantica gives the highest value: 424.8 mg EC / g of the

plant compared to A. ifranensis j. Didier: 342.1 EC mg / g of the plant. However, it is difficult to compare

these results with those of the bibliography because the use of different methods of extraction reduces the

reliability of a comparison between studies. Several factors can influence the content of phenolic

compounds, recent studies have present that extrinsic factors (such as geographical and climatic factors),

genetic factors, but also the degree of maturation of the plant and the duration of storage have a strong

influence on the content in polyphenols [25-26].

Figure 3: Polyphenols contents of fractions for A. mesatlantica and A. ifranensis

Figure 4: Flavonoid contents of fractions for A. mesatlantica and A. ifranensis

Mor. J. Chem. 6N°1 (2018) 01-13

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Figure 5: Tannin contents of extracts for A.mesatlantica and.A.ifranensis

3.2. Antioxidant activity

The antioxidant activity of different extracts from the two plants studied was evaluated in vitro by two

different methods, DPPH radical scavenging activity and Ferric reducing-antioxidant power.

3.2.1. Free radical DPPH * scavenging

DPPH* is a stable radical, which presents a characteristic absorption between 515 nm solution giving it

violet color. This color disappears when the DPPH * is reduced by a sensor of free radicals and becomes

pale yellow which translates into a change in optical absorption. The Figure 6 shows the results of

measurement of percentage of inhibition the DPPH*radical depending on the concentration of extracts ethyl

acetate for A. mesatlantica butanolic for A. ifranensis J. Didier and Ascorbic acid.

0,0 0,5 1,0 1,5 2,0

0

20

40

60

80

100

I n

h i

b i

t i

o n

p

e r

c e

n t

a g

e (

%)

Conc (mg/ml)

butanolic extract of A.mesatlantica

Ethyl acetate of A.ifranensis

Ascorbic acid

Figure 6: The percentage of inhibition of the DPPH radical of the different concentrations of extracts of A.

mesatlantica ethyl acetate, butanolic of A. ifranensis J. Didier and Ascorbic acid.

Mor. J. Chem. 6N°1 (2018) 01-13

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IC50

0,0

0,2

0,4

0,6

0,8

1,0

1,2

IC

50

(m

g/m

l)

A

Ascorbic acid

Butanolic extract of A.mesatlantica

Ethyl acetate of A.ifranensis

Figure 7: The Value IC50extractsof ethyl acetate for A. mesatlantica, and butanolic for A. ifranensis J. Didier

and Ascorbic acid

These results show that, of ethyl acetate fraction for A. mesatlantica and butanolic fraction for A. ifranensis

J. Didier have significant antioxidant activity, with an inhibitory concentration 50%: IC50 of (125 µg/ml and

110 /ml) respectively. However, low the antioxidant activity of the other fractions. These activities are twice

lower than of Ascorbic acid that remains the most effective antioxidant with an IC50 of the order of 65

µg/ml. A low value of IC50 indicates strong antioxidant activity. The obtained results of the activity

antioxidant are in agreement with those of Lopes-Lutz et al, [27] which are confirmed in a study done in a

few species of Artemisia that the antioxidant activity of these plants is moderate. Kang and al., [28] have

suggested that the extracts from the plants containing polar molecules show high anti-radical activity. The

extraction method and the used solvents for the extraction could be the origin of these results [29]. The

phenolic compounds are known as powerful antioxidants [30]. They are very important components in the

extracts and their ability to scan of free radicals is due to their [31] hydroxyl groups.

3.2.2. Reduction of iron: FRAP (Ferric reducing-antioxidant power):

The reducing power of an extract is associated with its antioxidant power. The reducer of the iron of our

extracts activity is determined according to the method described by Oyaizu (1986), based on the chemical

reaction of reduction of the Fe3 +

to Fe2 +

.The absorbance of the reaction medium is determined at 700 nm.

An increase in absorbance corresponds to an increase of the reducing power of the tested excerpts [32].

The curve of Ascorbic acid that we measure the reducing power of the two extracts according to their

absorbance. The values presented in the figure 7 shows the reducing power of the two extracts at different

concentrations.

Mor. J. Chem. 6N°1 (2018) 01-13

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Figure 8: The reduction of iron in the presence of ethyl acetate, butanolic extracts and Ascorbic acid.

Figure 9: power reducing the two extracts and Ascorbic acid expressing the effective concentration 50% in

mg/ml.

The results presented in the figure 9 show that the capacity of reduction is proportional to the decrease of the

concentration of the samples. The two extracts of plants present antioxidant activities lower than the

reference product (Ascorbic acid). The reductive power was estimated at 0.796 mg/ml for the butanolic of A.

ifranensis J. Didier and 1.102 mg/ml for the acetate ethyl fraction of A. mesatlantica. Then that of the

positive control (Ascorbic acid) is 0.04 mg/ml. The activities of our extracts are medium-sized. The reducing

power of the two extracts of Artemisia is due to the presence of hydroxyl group in phenolic compounds that

can be used as electron donor. As a result, antioxidants can be considered inactivates of oxidants and

0,0 0,5 1,0 1,5 2,0

0,00

0,56

1,12

1,68

Ascorbic acid

Ab

sorb

an

ce i

n 7

00

nm

Conc (mg/ml)

0 2 4 6 8 10

0,0

0,5

1,0

1,5

2,0 Butanolic extract of A.ifranensis

Ab

so

rba

nce

in

70

0 n

m

Conc (mg/ml)

0 1 2 3 4 5 6

0,0

0,5

1,0

1,5

2,0 Ethyle acetate of A.mesatlantica

Ab

sorb

an

ce i

n 7

00

nm

Conc (mg/ml)

0,044

1,102

0,796

IC50

0,0

0,2

0,4

0,6

0,8

1,0

IC5

0 (

mg

/ml)

Ascorbic acid

Butanolic extract of A.ifranensis

Ethyl acetate extract of A. mesatlantica

Mor. J. Chem. 6N°1 (2018) 01-13

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reductants [33].Some previous studies have also shown that the reducing power of a compound can be used

as significant of his potential antioxidant activity indicator [34].

4. Conclusion

During this work, we have tried to quantify the contents of polyphenols in the two species, A. mesatlantica

and A. ifranensis J. Didier. We found that the two plants are rich in secondary metabolites (tannins,

flavonoids, sterols triterpenes, and mucilages).The dosage Results of phenols, flavonoids totals and tannins

condensed show that butanolic fraction of A. mesatlantica and hydromethanolic extract of A. ifranensis J.

Didier are rich in polyphenols, then for total flavonoids of A. mesatlantica ethyl acetate fraction and the

fraction butanolic of A. ifranensis J. Didier present the highest levels. However, the tannins condensed from

brut extract of A. mesatlantica gives the highest value compared to A. ifranensis. The study in vitro

antioxidant activity by the method of reduction of iron and that of the DPPH free radical trapping showed

that the fraction butanolic and ethyl acetate have moderate antioxidant activity. These excerpts could be an

alternative to synthetic additives. This activity remains significantly lower than Ascorbic acid.

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