Indian Journal of Novel Drug Delivery 14(3), Jul-Sep, 2022, 159-163

Indian Journal of Novel Drug Delivery

IJNDD
An Official Publication of
Karnataka Education and
Scientific Society

Research Article
Antioxidant Activity of Mentha Suaveolens Ehrh. Extracts
MOUNIRA MERGHEM*, WAFA NOUIOUA
Laboratory of Phytotherapy Applied to Chronic Diseases, Faculty of Natur and Life Sciences, Ferhat Abbas University, Sétif-1, Algeria.
ARTICLE DETAILS ABSTRACT
Article history: Mentha is a genus belonging to the family of Lamiaceae, whose plants are among
Received on 22 April 2022 the most aromatic and spread in diverse environments worldwide. The aim of this
Modified on 13 June 2022 study is to evaluate the total phenolics and flavonoids contents and the in vitro
Accepted on 18 June 2022
antioxidant activities of aqueous and ethanolic extract of Mentha suaveolens. The
Keywords: Folin–Ciocalteu method was used to determine the total phenols content while
Mentha Suaveolens, flavonoids were estimated according to the aluminum chloride colorimetric
Antioxidant Activity, method. Also the antioxidant activities were determined by two methods: 2,2-
Polyphenols, diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and β-carotene linoleic
DPPH Scavenging, acid. The total polyphenols content of the aqueous extract was 240.66±30.42µg
Β-Carotene. GAE/ mgE and flavonoids were 18.35±0.57µg QE/mgE. The total polyphenols
content of the ethanolic extract was 219.217±20.63µg GAE/mgE and flavonoids
were 20.59±0.6µg QE/ mgE. In the DPPH assay the aqueous extract showed the
higher scavenging capacity (IC50 = 0.035 ± 0.001 mg/ml), followed by ethanolic
extract with IC50 of 0.049 ± 0.000 mg/ml. In the test of β-carotene /linoleic acid,
the percentage of inhibition was 66.954 ± 2.64% of aqueous extract and 74.329 ±
2.03% of ethanolic extract.

© KESS All rights reserved

INTRODUCTION Herbal polyphenolic compounds are secondary

Reactive oxygen species (ROS) will act as metabolites with a characteristic aromatic
damaging or signaling molecule depends on the structure, which can be classified into fifteen
delicate equilibrium between ROS production groups according to the basic part of their
and scavenging. Because of the multifunctional molecule, as for example phenols, phenolic
roles of ROS, it is necessary for the cells to acid, flavonoids, anthocyanins, qui-nones,
control the level of ROS tightly to avoid any catechins and tannins, just to name a few [3].
oxidative injury and not to eliminate them Since ancient times, the human has been
completely. Scavenging or detoxification of exploring the potential of plants to improve their
excess ROS is achieved by an efficient health, often attributed to the presence of
antioxidative system comprising of the phenolic compounds with strong antioxidant
nonenzymic as well as enzymic antioxidants. ROS properties. The antioxidant ability of phenolic
are liberated by virtue of stress, and thus, an components occurs mainly through a redox
imbalance is developed in the body that damages mechanism and allows the components to act as
cells in it and causes health problems [1]. reducing agents, hydrogen donors, singlet
oxygen quenchers, and metal chelators [4].
Natural antioxidant compounds exhibit their Polyphenolic compounds are commonly found in
antioxidant activity by various mechanisms both edible and inedible plants, and they have
including chain breaking by donation of been reported to have multiple biological effects,
hydrogen atoms or electrons that convert free including anti oxidant activity [5].
radicals into more stable species and
decomposing lipid peroxides into stable final The Lamiaceae family includes about 220 genera
products [2]. and 3300 species which are widely used for
various purposes worldwide [6]. Plants belonging
to the Labiatae family are rich in polyphenolic
compounds and a large number of them are well
*Author for Correspondence: known for their antioxidant properties [7, 8]. The
Email: [email protected] genus Mentha is an important member of the

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family. It includes eighteen species and eleven reaction mixture was incubated at room
hybrids, among which several species have temperature for 1h 30 min and the absorbance of
economic importance due to their high-valued oil the mixture was measured at 760 nm, Gallic acid
and good taste [9, 10]. Mentha (20-140 mg/L) was used as standard for the
Suaveolens (Lamiaceae family) locally is calibration curve. The total polyphenols content
traditionally used to treat cold, digestive system was expressed as micrograms of gallic acid
ailment, hemorrhoid, fever, intestinal swelling, equivalents (GAE) per milligram of extract. All
flu against chill, and as carminative and samples were analyzed in three replications.
conception and fertility booster [11 - 14]. This study
aims to determine the phenol and flavonoid 5. Determination of Total Flavonoids
contents of Mentha Suaveolens extracts and Contents
evaluate the antioxidant activity. The total flavonoids in plant extracts were
determined using the aluminum trichloride
MATERIALS AND METHODS (AlCl3) method [16]. Briefly, 1 mL of 2% AlCl3 in
1. Chemicals methanol was mixed with 1 mL of the extract.
Folin-Ciocalteu, aluminum chloride (AlCl3), gallic After incubation in dark at room temperature for
acid, quercetin, 2,2-diphenyl-1-picrylhydrazyl 10 min, the absorbance of the reaction mixture
hydrate (DPPH), gallic acid and tween 40 were was measured at 430 nm. Quercetin (1-40 mg/L)
purchased from Sigma Chemical Co. (St. Louis, were used as standards for calibration curve and
MO). Linoleic acid, β-carotene and butylated the total flavonoids content was expressed as
hydroxytoluene (BHT) were obtained from Fluka micrograms quercetin equivalents (QE) per
Chemical Co. (Buchs, Switzerland). milligram of extract.

2. Plant Material 6. Evaluation of Antioxidant Activity

Mentha suaveolens plant material was collected 6.1. DPPH Free Radical-Scavenging Assay
from Bougaa région, Wilaya of Sétif Northeast of The free radical scavenging activity of the
Algeria. extracts was measured by 2,2- diphenyl-1-
picrylhydrazyl (DPPH) assay [17]. After dissolving
3. Preparation of Plant Extract the extracts, the solution of DPPH in methanol
3.1. Aqueous Extract (0.04mg/ mL) was prepared and 1250 µL of this
The aerial parts of plant material were cleaned solution was added to 50µL of extracts solution
with tap water, dried in the shade at room at different concentration and kept in the dark
temperature for 2 weeks and ground into for 30 minutes at room temperature. Then, the
powder using an electric grinder. The aqueous absorbance of this solution was measured at
extract was prepared by boiling 100g of Mentha 517nm. All tests were performed in triplicate.
suaveolens powder in distilled water for 15 Radical-scavenging activity was calculated using
minutes, The resulting mixture was filtered using the following equation:
Whattman filter paper and then evaporated in
rotary vacuum evaporator at 45°C. A −A
RSA (%) = × 100
A
3.2. Ethanolic Extract
The ethanolic extract was obtained by Where,
maceration in water/ethanol mixture (20:80) for RSA: Radical Scavenging Activity
24 h. The resultant extract was filtered through A blank: Absorbance of the control.
Whattman paper and the solvent was removed A sample: Absorbance of extract.
by rotary evaporator under reduced pressure at
45°C. 6.2. β-Carotene/Linoleic Acid Assay
In this test, the antioxidant capacity of the
4. Determination of Total Polyphenol Content extracts was determined according to the
Total phenolic content was determined using method described by Kartal et al. [18]. The β-
Folin-Ciocalteu method, according to Li and al, carotene solution was prepared by dissolving 0.5
[15] with slight modifications. A volume of 100 µL
mg β-carotene in 1 mL of chloroform, one
of the extract was mixed with 500 µl of Folin– milliliter of this solution was pipetted to a flask
Ciocalteau (diluted 10% in distilled water). After covered with aluminum foil. Then 25 µL of
4 min, 400 µL of sodium carbonate solution linoleic acid and 200 mg of tween 40 were added
Na2CO3 (75 g/L) was added to the mixture, the in the foil and the chloroform was evaporated

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using evaporator. After evaporation, 100 mL of Natural antioxidant compounds exhibit their
distilled water saturated with oxygen was added. antioxidant activity by various mechanisms
Then 2.5 mL of this mixture were transferred to including chain breaking by donation of
test tubes, and 350 µL of the extracts (2mg/mL hydrogen atoms or electrons that convert free
methanol) were added and the absorbance was radicals into more stable species and
reading at 490 nm after 1h, 2h, 6h, 24h and 48h decomposing lipid peroxides into stable final
respectively. The same procedure was repeated products [20].
with butylated hydroxyl toluene (BHT) as a
positive control and with distilled water and The antioxidant activities of Mentha suaveolens
methanol as a negative control. The antioxidant extracts were determined by DPPH scavenging
activity of extracts was calculated using the activity assay and the β-carotene bleaching
following equation: inhibition. DPPH radical scavenging assay is the
most popular and frequently used for the
𝐴 determination of antioxidant activity of plant
𝐴𝐴% = × 100
𝐴 extracts. Bleaching ability of 𝛽-carotene in
linoleic acid system is another simple,
Where,
reproducible, and time efficient method for rapid
A sample: Absorbance of the Extract.
evaluation of antioxidant properties [21].
A BHT: Absorbance of positive control BHT.
β-carotene undergoes rapid decolorization in the
Statistical Analyses
absence of an antioxidant. The presence of
The results are expressed as the mean ±
different antioxidants can hinder the extent of β-
standard deviation. One-way analysis of variance
carotene bleaching by neutralizing linoleate free
(ANOVA) was performed to assess differences
radicals and other radicals formed in the system
between groups. [22].

RESULTS AND DISCUSSION

2.1. DPPH Radical Scavenging Activity
1. Total Polyphenols and Flavonoids Contents
In DPPH, the sample was able to reduce the
Total polyphenols compounds, total flavonoids of
stable violet DPPH radical to the yellow DPPH-H,
Mentha suaveolens extracts were estimated
the results were expressed as IC50 values (Table
spectrophotometrically. The results are shown in
2). Aquous extract of Mentha suaveolens revealed
Table 1, and are expressed as μg of gallic acid or
higher antioxidant capacities with the value of
quercetin equivalent per miligram extract,
(0.035 ± 0.001 mg/mL) than ethanolic extract
respectively, the aqueous extract is richer on
(0.049 ± 0.000 mg/mL).
polyphenols (240.66±30.42 μg GAE/mg extract)
compared to ethanolic extract (219.217±20.63
Table 2: DPPH scavenging activity of Mentha
μg GAE/mg extract). However, the ethanolic
suaveolens extracts and standards
extract showed a high total flavonoids with value
of (20.59±0.6 μg QE/mg extract), followed by the Extracts IC50(mg/mL)
aqueous extract (18.35±0.57 μg QE/mg extract). AqE 0.035 ± 0.001
EE 0.049 ± 0.000
Table 1: Total polyphenols and flavonoids
Gallic acid 0.001 ± 0.000#
content of Mentha suaveolens extracts.
BHT 0.043 ± 0.003#
Extract Polyphenols Flavonoids #: μg/ml. Each value represents the mean ± SD (n = 3).
μg GAE/mg extract μg QE/mg extract
2.2. β-carotene/Linoleic Acid Bleaching Assay
AqE 240.66±30.42 18.35±0.57
The highest antioxidant capacity was observed in
EE 219.217±20.63 20.59±0.6 the ethanolic extract (74.329 ± 2.03%) folewed
AqE : aqueous extract, EE : ethanolic extract, GAE: gallic by aquous extract (66.954 ± 2.64%) (Table 3).
acid equivalent, QE: quercetin equivalent. Each value
represents the mean ± SD (n = 3).
The DPPH radical-scavenging capacity of the
2. Antioxidant Activity Evaluation extracts could be explained by the presence of
Antioxidants have the ability of protecting phenolic components [23]. The high correlation
organisms from damage caused by free radical- between the values of phenols concentration in
induced oxidative stress [19]. plant extracts and antioxidant activity is well
documented [24]. Sugihara et al (1999), discussed

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