https://doi.org/10.52973/rcfcv-e34431

Received: 30/03/2024 Accepted: 20/05/2024 Published: 25/07/2024

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Revista Científica, FCV-LUZ / Vol. XXXIV, rcfcv-e34431

ABSTRACT

Mentha rotundifolia is a member of the Lamiaceae family and

is distributed mainly in traditional medicine in Algeria and the

Mediterranean locality. This study aimed to demonstrate how the

extraction technique can determine the type of active ingredients

obtained, as well as their potential pharmacological and therapeutic

effects. The methanolic extracts were obtained by maceration and

Soxhlet apparatus. The study of the chemical composition was

determined by colourimetric methods and liquid chromatography–

mass spectrometry (LC–MS/MS); the antioxidant activity was

evaluated invitro by three tests: DPPH test, ferrous ion chelating

test and reducing power test. The antimicrobial effect of extracts was

evaluated by agar diffusion assay against six microbial strains. The

results demonstrate that the best yield of extraction, Total phenolic

and avonoid contents, and antioxidant activity were found to be

higher in the extract obtained by Soxhlet than those obtained by

maceration. The phytochemical screening tests identied polyphenols,

avonoids, tannins, terpenoids, and quinones, and anthraquinones and

saponins were absent in both extracts. The LC–MS/MS revealed the

presence of several phenolic compounds, the predominant in both

extracts was rosmarinic acid. The antimicrobial activity shows that

both extracts have no effect. In conclusion, this study reveals that

extraction by Soxhlet is more suitable than extraction by maceration

for this plant.

Key words: Antioxidant; extraction; LC/MS/MS; maceration; Mentha

rotundifolia; Soxhlet

RESUMEN

Mentha rotundifolia es miembro de la familia Lamiaceae y se

distribuye ampliamente en la medicina tradicional en Argelia y la

localidad mediterránea. El objetivo de este estudio fue demostrar

cómo la técnica de extracción puede determinar el tipo de

principios activos obtenidos, así como sus potenciales efectos

farmacológicos y terapéuticos. Los extractos metanólicos se

obtuvieron mediante maceración y aparato Soxhlet. Se determinó el

estudio de la composición química mediante métodos colorimétricos

y espectrometría de masas por cromatografía líquida (LC–MS/MS).

La actividad antioxidante se evaluó in vitro mediante tres pruebas:

prueba de DPPH, prueba de quelación de iones ferrosos y prueba

de poder reductor. El efecto antimicrobiano de los extractos se

evaluó mediante un ensayo de difusión en agar contra seis cepas

microbianas. Los resultados demuestran que el mejor rendimiento

de extracción, el contenido de fenólicos y avonoides totales, la

actividad antioxidante fue mayor en los extractos obtenidos por

Soxhlet que los obtenidos por maceración. Las pruebas de detección

toquímica permitieron identicar polifenoles, avonoides, taninos,

terpenoides, quinonas y la ausencia de antraquininas y saponinas

en ambos extractos. La LC–MS/MS reveló la presencia de varios

compuestos fenólicos, el predominante en ambos extractos fue el

ácido rosmarínico. La actividad antimicrobiana muestra que ambos

extractos no tienen ningún efecto. En conclusión, este estudio revela

que la extracción mediante Soxhlet es más adecuada que la extracción

por maceración para esta planta.

Palabras clave: Antioxidante; extracción; LC/EM/EM; maceración;

Mentha rotundifolia; Soxhlet

Comparative study of chemical composition Antioxidant and Antimicrobial

activity of Methanolic Extracts of Mentha rotundifolia

Estudio comparativo de la composición química, actividad antioxidante y

antimicrobiana del extracto metanólicos de Mentha rotundifolia

Siham Ferdjioui* , Rachid Belhattab

Ferhat Abbas University, Department of Biochemistry, Laboratory of Applied Microbiology. Setif, Algeria.

*Corresponding author: ferdjioui_89@yahoo.fr

Comparative study of Mentha rotundifolia methanolic extracts / Ferdjioui and Belhattab ___________________________________________

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INTRODUCTION

Due to its richness in secondary metabolites, plants can be considered

a rich source of therapeutic compounds. Variation in extraction methods

usually depends on the duration of the extraction period, the solvent

used, the pH of the solvent, temperature, the particle size of the plant,

and the solvent–to–sample ratio. The majority of extraction methods

involve the separation of medicinally active compounds of plants from

the inactive components by using selective solvents [1, 2].

The solubility of compounds in the solvent depends on their

chemical natures, which vary for compounds of others; this structural

diversity is responsible for diversity in molecules’ physicochemical

properties. For this reason, it is dicult to develop a universal method

for extracting all effective compounds from plants [3].

Oxidative stress can be dened as the state in which the free

radicals in our body outnumber our antioxidant defenses. An

antioxidant is a molecule capable of inhibiting the oxidation of another

molecule. Antioxidants break the free radical chain of reactions by

sacricing their own electrons to feed free radicals without becoming

free radicals themselves [4]. Thus, it is essential to develop effective

and natural antioxidants that can protect the body from free radicals

and retard the progress of many chronic diseases [5].

Mentha rotundifolia is one of the Lamiaceae species that is widely

distributed in Algeria. It is used as a condiment, and it has been applied

in traditional medicine for a wide range of actions: stimulative, tonic,

stomachic, carminative, analgesic, antispasmodic, anti–inammatory,

sedative, hypotensive and insecticidal [6]. This plant was also the subject

of several scientic studies, which made it possible to determine its

therapeutic effect as an antioxidant [7], anti–inammatory [8], and

antimicrobial [9]. The objective of this study was to investigate the

effect of temperature in the extraction of phenolic compounds from M.

rotundifolia and its antioxidant and antimicrobial activity.

MATERIAL AND METHODS

Plant materials

The plant M. rotundifolia was collected in the region of Djemila

Wilaya of Setif (Algeria) during the period of plain Florissant. Professor

Laouer H, a botanist in the laboratory of Botanical Sciences, Ferhat

ABBES Setif–1 University, Algeria, carried out the botanical identity

of the plant (family, genus, and species). The aerial parts were air–

dried in the shade at room temperature away from humidity and then

powdered by an electric grinder.

Extraction

Extraction by maceration

Twenty g of plant powder was put into a maceration bottle and

lled with 96% methanol for 48 hours (h), with the solvent renewed

after 24 h. The ratio between plant powder and ethanol was 1:10.

Afterwards, the ltrate separated from the residue using Watman

lter paper was collected and evaporated using a rotary evaporator

(Rotavator, Büchi; Swiss) at 45°C until a dry extract was obtained.

Soxhlet extraction

Twenty g of aerial parts powder was packed in a Watman lter paper

and placed in the Soxhlet extractor (Büchi; Swiss). Then, 200ml of

methanol was poured into the roundbottom ask. The solvent was

heated using the dismantle, which began to evaporate, moving through

the apparatus to the condenser. The condensate then dripped into the

reservoir containing the plant extract. The process was made to run

for a total of 6 h. Finally, the extract was collected, and the methanol

was evaporated using a rotary evaporator (BUCHI rotavap Suiss) at

45°C. The extract was stored at room temperature for further use.

Phytochemical screening

Phytochemical tests were performed on methanolic extracts to verify

the presence of some compounds (polyphenols, avonoids, tannins,

terpenoids, quinines, and saponins). Their detection is achieved

using the methods described by Bagre et al.[10], Khaldi etal.[11],

Vayalakshmi et al. [12].

Determination of total phenolic compounds

The polyphenols in extracts was quantied using Folin–Ciocalteu

reagent according to the method describedby Li et al.[13]. Briey,

an aliquot of 200 μL of the extract was mixed with 1 ml of Folin–

Ciocalteu reagent for 4 min, followed by the addition of 800 μL of

aqueous solution (7,5%). The absorbance was measured

(SECOMAN Spectrophotometer, French) at 765 nm after two hours

of incubation. The polyphenol content was expressed as mg gallic

acid equivalent (GAE)·g extract

-1

Determination of total avonoids

The total avonoid content of each extract was determined by a

colourimetric method as described by Kosalec et al. [14]. In brief, 1 ml of

each extract was added to 1 ml of aluminum chloride (AlCl

) methanolic

solution (2%) and allowed to stand for 30 min, the absorbance of the

mixture was measured at 430 nm. The total avonoid content was

reported as mg of quercetin equivalent (QE)·g extract

-1

Identication and quantication of phenolic compounds in extracts

using liquid Chromatography/ Mass Spectrometry (LC/MS/MS)

Methanol was used to dissolve the extracts and standard at a rate

of 1.0 mg·mL

-1

. For every analysis, a volume of 20 μL was continuously

injected at a ow rate of 1.0 mL·min

-1

onto an Agilent Zorbax 150 mm

× 4.6 mm C18 column. A gradient solvent system was used for the

study, with aqueous–formic acid (0.10%) serving as solvent (A) and

acetonitrile (100%) serving as solvent (B). A 35–min run time was

allocated to a ve–step linear gradient elution, wherein solvent A

was reduced to 10% and solvent B was increased to 90%.

A triple–quadrupole mass spectrometer (API 3200; MDS Sciex,

Concord, ON, Canada) received the full flow from the high–

performance liquid chromatography (HPLC). The mass spectrum

information was collected in negative ion mode with a capillary voltage

of 4500 V, an Electrospray Ionization (ESI) ion source, a cone voltage

of 70 V, a collision energy of 35 eV, a drying temperature of 650°C, N

as the drying gas at a ow rate of 4.0 L·min

-1

, and Analyst software

version 6. The eluted samples and standards were found at 280 nm [7].

Antioxidant activity

Diphenyl–1–picrylhydrazyl (DPPH•) radical scavenging assay

All substances that can donate a hydrogen atom or an electron

to DPPH can be considered antioxidants and, therefore, radical

TABLE I

Amounts of total phenolic compounds and

avonoids in Mentha rotundifolia extracts

Extract Polyphenols

(a)

Flavonoids

(b)

Methanolic extract obtained by maceration

(MEM)

141.571 ± 0.143 15.636 ± 0.030

Methanolic extract obtained by Soxhlet

(MES)

168.642 ± 1.642 33.045 ± 0.76

(a)

: Expressed as mg galic acid equivalent (GAE) per gram of extract,

(b)

: Expressed as

mg quercitin equivalent (QE) per gram of extract

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scavengers. The degree of discolouration of the violet colour of DPPH

as it gets reduced indicates the radical scavenging potential of the

antioxidant [15]. The capacity of extracts to scavenge DPPH was

determined following the method of Sarikurkcu et al. [16]. Briey: 0.1

mL of extracts of different concentrations was mixed with 1 ml DPPH

solution (0.008% in ethanol). The mixture was shaken and left to stand

for 2 h at room temperature. The absorbance was measured at 515

nm. The scavenging activity of extracts was compared with that of

Butylated hydroxytoluene (BHA) as standard. The capability to scavenge

the DPPH• radical was calculated using the following equation:

Where A

is the absorbance of the control, and A

the absorbance

of the sample.

Metal ion chelating assay

Transition metal species such as ferrous iron (Fe

) can facilitate

the production of ROS within animal and human systems; the capacity

of compounds to chelate iron can provide a valuable antioxidant

capability [17]. The Metal ion chelating effect of the extract was

estimated using the method reported by Le et al. [17]. Briey, one

ml of each extract at a different concentration was mixed with 1 ml

of FeCl

(0.6 mM). After 5 min, the reaction was initiated by adding 1

ml ferrozine (5 mM). The tubes were vortexed and allowed to stand

for 10 min at room temperature. Absorbance was measured at 562

nm. The ratio of ferrozine–Fe

complex formation inhibition was

calculated as follows:

100

control

controlsample

The EDTA was used as a positive control.

Reducing power assay

The plant extract components’ reducing power might be a signicant

indicator of its potential antioxidant activity [18]. The presence of

reductants such as antioxidant substances in the samples causes

the reduction of the Fe

/ferricyanide complex to the ferrous form.

Therefore, Fe

can be monitored by measuring the formation of Perl’s

Prussian blue at 700 nm [17]. The reducing power of extracts was

assessed by the method described by Beyhan et al. [17], 1 ml of extracts

was mixed with 2.5 ml phosphate buffer (pH=6.6) and 2.5 ml of 1%

potassium ferricyanide [K

Fe(CN)

] solution. The mixture was incubated

at 50°C for 20 min, and then 2.5 mL of 10% Trichloroacetic acid was

added. After vigorous agitation, 2.5 mL of this solution was mixed

with 2.5 mL of distilled water and 0.5 ml ferric chloride (FeCl

) (0.1%).

The absorbance was measured at 700 nm. The reducing power of the

extracts was compared with that of Butylated Hydroxyanisole (BHA) as

a positive control. Higher absorbance indicates higher reducing power.

Antimicrobial activity

The study of antimicrobial activity was performed by the method

of agar diffusion against two Gram–negative bacteria: Pseudomonas

aeruginosa ATCC 27853 and Escherichia coli ATCC 25922, one gram–

positive bacteria Staphylococcus aureus ATCC 25923 and three fungi

(Aspergillus niger 2CA936, Aspergillus avus NRRL391, Candida albicans

ATCC1024). Sterilized Whatman paper discs of 6 mm diameter were

impregnated with 20 µl of solution of extracts (200–500 mg·ml

-1

and the discs were then placed in Petri dishes previously inoculated

by Muller–Hinton agar for bacteria, Potato Dextrose Agar (PDA) for

fungus, and Sabouraud + Chloramphenicol for Candida albicans.

Different standard antibiotics (according to the type of bacteria)

(Gentamicin (GEN), Imipenem (IMP), Cefoxitin (CX), Pristinamycin (RP),

Vancomycin (VA), Piperacillin (PRL), Ciprooxacin (CIP), Clindamycin

(CD)) and antifungal (Amphotericin B (AM), Chloramphenicol (CTR),

Nystatin (NY)) were used as positive control.

The Petri dishes are incubated (Memmert ovens, Germany) at

37°C·24h

-1

for bacteria, at 37°C for 48 h for yeast, and at 27°C for 72 h

for fungi. The negatif control was an impregnated disk DMSO (Dimethyl

sulfoxide) alone. Different antibiotic and antifungal disks were used as

positive control. Three repetitions were performed for each test [19].

Statistical analysis

The results were presented as the mean ± standard deviation (SD).

Statistical data analysis was performed using the GraphPad Prism 5

program with the Dunnett test; the level of signicance was set at P<0.05.

RESULTS AND DISCUSSION

Phytochemical analyses

In the present study, methanolic extracts of M. rotundifolia were

evaluated for their phytochemical screening, polyphenolic contents,

antioxidant, and antimicrobial activity.The best extract yield was

recorded by the Soxhlet apparatus with 16.05 % against 9.2 % for the

maceration. The phytochemical screening revealed the presence of

polyphenols, avonoids, tannins, quinones, and terpenoids. However,

saponins and anthraquinones were absent in both extracts.

Amounts of total polyphenols and avonoids

The contents of total polyphenols, and total flavonoids were

determined in plant extracts by using Folin–Ciocalteu and AlCl

reagents respectively, the results are shown in TABLE I. As can be

seen, the Soxhlet apparatus seems to be signicantly (P<0.05) better

than the maceration to extract total polyphenols and avonoids.

Identication and quantication of phenolic compounds using

LC/MS/MS

The results of this study demonstrate that among the 24 standards

used, 17 phenolic compounds were identied in MEM and 15 in MES.

Among the phenolic compounds identied: phenolic acids (Vanilic

acid, Rosmarinic acid, Ellagic acid, Syringic acid…), avonols (Myrtillin,

Quercetin, Rutin, Isoquercitin), flavone (Apigenin, Luteolin) and

anthocyanins (Cyanin chloride).The predominant compound in both

TABLE II

Phenolic prole of methanolic extracts obtained by maceration and Soxhlet, identied by LC– MS/MS

Compounds C

(ng·ml

-1

) MEM C

(ng·ml

-1

) MES RT Q1 Q3 MWT

Apigenin 213.00 2.99 8.40 269.000 151.0 270.120

Isoquercitrin 57.60 nt 7.00 464.900 300.0 464.400

Catechol nt nt 5.50 109.000 109.0 110.110

Epicatechin nt nt 6.57 289.100 108.8 290.300

Gallic Acid 152.00 nt 1.53 169.000 124.6 170.120

Procyanidin B

nt nt 6.40 577.100 407.0 / 289.0 578.520

Quercetin 3–O–Galactoside 328.00 60.00 7.00 463.000 301.0 464.379

Luteolin 275.00 118.00 8.00 285.000 217.0 286.240

Chlorogenic Acid 357.00 96.50 5.94 353.155 190.4 /84.8 / 93.1 354.310

Epigallocatechin Gallate nt nt 6.66 456.579 168.2 / 168.5 / 124.8 458.372

Cyanin Chlorid 304.00 203.00 8.00 286.198 133.4 /132.8 / 150.6 287.100

Myrtillin 65.40 8.62 7.01 462.178 299.8 / 270.8 / 254.7 500.800

Quercetin nt nt 8.01 300.604 150.4 302.200

Rutin 337.00 168.00 6.84 609.419 299.0 / 299.9 / 270.9 610.520

Caec Acid 156.00 95.10 6.50 178.465 134.2 / 106.4 / 89.1 18.160

Ellagic Acid 7.98 nt 8.01 300.703 149.7 / 150.6 / 149.9 302.197

Ferulic Acid 109.00 56.00 7.20 192.807 133.9 / 133.4 / 177.8 194.180

Hydroxybenzoic Acid 198.00 84.70 6.48 134.779 88.4 / 106.8 / 89.1 135.120

P–Cumaric Acid 178.00 153.00 7.06 162.756 118.8 / 118.1 / 92.7 164.160

Rosmaric Acid 18900.00 7690.00 7.38 358.319 160.7 / 161.0 / 132.7 360.320

Syringic Acid 116.00 126.00 6.48 196.718 120.4 / 120.7 / 152.3 198.170

Transcinamaldehyde Acid nt 975.00 7.95 131.797 103.7 / 102.8 / 101.9 132.160

Vanilic Acid 206.00 221.00 6.38 166.660 107.8 / 151.2 / 151.7 168.150

Hypericin nt nt 10.53 503.000 405.0 504.450

Q1: compound molecular weight, Q3: fragment molecular weight, MWT: molecular weight, nt: not found

Comparative study of Mentha rotundifolia methanolic extracts / Ferdjioui and Belhattab ___________________________________________

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extracts was Rosmaric Acid (18900 and 7690 ng·mL

-1

for MEM and

MES respectively) (TABLEII). Furthermore, the study showed that the

amounts of the majority of phenolic compounds identied are higher

in MEM than in MES. However, the quantities of syringic acid, vanillic

acid, and trans–cinnamaldehyde were higher in MES.

The medicinal value of the plant is due to the phytochemical

constituents they produce, which exhibit certain physiological actions

on human body [20].The result of this study shows the presence of

polyphenols, avonoids, tannins, quinones, and terpenoidsM. rotundifolia

extracts. This diversity of compounds can lead to a diversity of plant

therapeutic properties. The determination of the amount of total

polyphenols and avonoids demonstrate the richness of both extracts

on this compounds with a predominance in the extract obtained by

Soxhlet. In a study carried out by Boussouf etal. [8] on the leaves of the

same plant from the northern Algeria, the methanolic extract obtained by

maceration gave respective contents of polyphenols and avonoids of

350.10 ± 0.96 mg galic acid equivalent (GAE·g

-1

) of extract and 79.44 ± 0.76

mg quercitin equivalent (EQ·g

-1

) of extract. These differences with our

study may be due to several factors that can inuence the contents

and the nature of phenolic compounds in extracts, such as the region

and period of the harvest of the plants, the part used, the time and

the extraction temperature, the polarity of the solvent [1]. In order to

identify the phenolic compounds of extracts, LC–MS/MS was carried

out. Based on comparing their chromatographic proles and retention

times with those of the standards used, several phenolic compounds

with therapeutic interest are identied. Rosmarinic acid was the major

compound and has been the subject of numerous scientic studies

demonstrating its therapeutic effects, such as antioxidant properties

[21], anticancer [22], anti–inammatory [23].

Antioxidant activity

DPPH radical scavenging assay

The IC50 for DPPH radical–scavenging activity reported in TABLEIII

demonstrates that the radical–scavenging activity of extracts

increased with increasing concentration of extract. The methanolic

extract obtained by the Soxhlet apparatus had shown better (P<0.05)

scavenging activities than those obtained by maceration. However,

the activity of BHA was much more marked (P<0.001) than extracts.

Determination of metal chelating activity

The results of this test also demonstrate that the extract

obtained by maceration has a significantly smaller chelating

TABLE III

Values of Mentha rotundifolia Extracts and standard for DPPH Scavenging

Activity, Metal chelating activity and EC

for reducing power

Test MES (µg·ml

-1

) MEM (µg·ml

-1

) BHA (µg·ml

-1

) EDTA (µg·ml

-1

)

DPPH

scavenging

133.160 ± 11.346 265.491 ± 2.221 20.701± 0.065 NT

Metal chelating

activity

2194.000 ± 0.038 3417.000 ± 0.011 NT 11.100 ± 0.060

Reducing

power

468.000 ± 1.000 550.325 ± 3.613 89.000 ± 0.285 NT

NT: not tested

TABLE IV

Antimicrobial activities of Mentha rotundifolia extracts against the bacterial and fungal strains tested

Escherichia coli

ATCC 25922

Pseudomonas aeruginosa

ATCC 27853

Staphylococcus aureus

ATCC 25923

Aspergillus niger

2CA936

Aspergillus avus

NRRL391

Candida albicans

ATCC1024

MEM 200 and 500 (µg·ml

-1

) NI NI NI NI NI NI

MES 200 and 500 (µg·ml

-1

) NI NI NI NI NI NI

Antibiotics

27 (CX) 34 (PRL) 30 (RP) 9 (AM) 8 (AM) 19 (AM)

28 (GEN) 39 (CIP) 30.5 (CD) 11 (NY) 11 (NY) 21 (NY)

37 (IMP) 38 (IMP) 16 (VA) 20(CTR) 24 (CTR) 33 (CTR)

NI: No inhibition zone observed around the discs (6 mm) impregnated with 20 µl of methanolic extracts.

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power capacity (IC50=3417 ± 0.011 µg·ml

-1

) than the Soxhlet extract

(IC50=2194 ± 0.038µg·ml

-1

) (TABLE III).

Reducing power

In this assay, The EC50 values indicated that the BHA showed a good

reducing power (EC50= 89 ± 0.285 µg·ml

-1

); the EC50 values of extracts

demonstrate that the activity of the extract obtained by maceration

is signicantly lower (P<0.001) than of Soxhlet extract (TABLE III).

No correlation was noted between levels of some phenolic

compounds determined by HPLC/ MS–MS and antioxidant activity; this

may be, explained by the presence of other compounds not identied

in this study in soxhlet extract with a good antioxidant effect.

Antimicrobial activity

The antimicrobial activity of methanolic extracts was evaluated

against two Gram–negative bacteria: Pseudomonas aeruginosa ATCC

27853 and Escherichia coli ATCC 25922, one Gram–positive bacteria

Staphylococcus aureus ATCC 25923 and three fungus (Aspergillus niger

2CA936, Aspergillus avus NRRL391, Candida albicans ATCC1024).

The results indicated that M. rotundifolia extracts have no signicant

antimicrobial activity against all microbial strains tested (TABLE IV).

Infectious diseases caused by pathogenic microorganisms

affect millions of people worldwide [33]. This study did not report a

signicant antimicrobial effect.

According to Gulluce et al. [34], the methanolic extract of

Mentha longifolia L. ssp. longifolia was inactive against 38 different

microorganisms (bacteria, fungus, and yeast). Moreover, the essential oil

showed a high activity against all microorganisms. Adiguzel et al. (2009)

[35] studied the antimicrobial activity of other Lamiaceae species (Nepta

quataria) against 40 different microorganisms (24 bacteria, 15 fungi, and

yeast); the methanolic extract of this species was inactive against 28

microorganisms (Pseudomonas aeroginosa ATCC–9027, Staphylococcus

aureus ATCC–29213, Streptococcus pyogenes ATCC–176…).

The inactivity of our extracts may be due to the absence of phenolic

oligomers. According to Karou et al. (2005) [36], the mechanism of

toxicity of polyphenols towards microorganisms is either through the

deprivation of metal ions such as iron or by non–specic interactions

such as the establishment of hydrogen bonds with cell wall proteins or

enzymes. However, an important factor that governs the antimicrobial

activity of polyphenols is their molecular weight; monomers are too

small to establish enough hydrogen bonds, while high molecular

weight polymers are too large to cross the bacterial cell wall.

Therefore, the ideal molecular weight would be that of oligomers.

CONCLUSION

The results of this study reveal the riches of M. rotundifolia in

polyphenols and avonoids. The extract obtained by Soxhlet had

the highest amount of total phenolic compounds and the best

antioxidant activity compared to the extract obtained by maceration.

The results of the antimicrobial assay show that both extracts have

no antimicrobial activity. Further studies are needed to determine

the toxicity and other biological properties of this plant.

Phenolic and flavonoid compounds are responsible for the

antioxidant activity of plant materials [24]. Polyphenols’ antioxidant

activity was attributed to their redox properties, which allow them

to act as reducing agents, hydrogen donators, and singlet oxygen

quenchers, some of which show metal chelation properties [25, 26].

However, some plant polyphenols may generate reactive secondary

radicals during the cycling process. Furthermore, phenoxyl radicals

produced in the course of radical scavenging by some phenolic

compounds are capable of oxidizing both proteins and lipids [2].

Due to the varied characteristics of phytochemicals, the antioxidant

activity of plants could not be properly assessed using either one

antioxidant assay method [27]. In the present study, three different

assays were employed to determine the mode of action and compare

the antioxidant properties of M. rotundifolia extracts.

The antioxidant results demonstrate a correlation between

the amount of total polyphenols and avonoids in extract and the

antioxidant activity; a high antioxidant activity in all tests was

observed in Soxhlet extract in comparison with those obtained by

maceration. This correlation was observed by other researchers [28,

29, 30].But, for others, no correlation existed [31, 32].

Comparative study of Mentha rotundifolia methanolic extracts / Ferdjioui and Belhattab ___________________________________________

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ACKNOWLEDGMENT

The authors are grateful for the nancial support received from

theMinistry of higher education and scientic research of Algeria.

Conict of interest

The authors declare no conict of interest.

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