Algérian Mélissa Antioxydant

Arabian Journal for Science and Engineering (2018) 43:3349–3357
https://doi.org/10.1007/s13369-017-3000-6
RESEARCH ARTICLE - BIOLOGICAL SCIENCES
Estimation of Total Phenolic Contents and In Vitro Antioxidant

and Antimicrobial Activities of Various Solvent Extracts of Melissa
officinalis L.
H. Mabrouki1 · C. M. M. Duarte2 · D. E. Akretche1
Received: 7 August 2017 / Accepted: 19 November 2017 / Published online: 27 November 2017
© King Fahd University of Petroleum & Minerals 2017
Abstract
This study aimed to evaluate the antioxidant and antimicrobial activities of Melissa officinalis L. extracts. In order to estimate
the best conditions to extract polyphenols from the plant studied, different solvents such as hexane, acetone, and ethanol
were used. The polyphenols content was estimated by the Folin–Ciocalteu method. The antioxidant activity was assessed by
different methods such as ORAC test and the scavenging of free radical DPPH, while ascorbic acid was used as a control.
The antimicrobial activity of M. officinalis L. was assessed by the method of disk diffusion. The results obtained showed that
ethanol was the most efficient solvent for extracting polyphenols from the M. officinalis L. The ethanol extract exhibited strong
antioxidant and antimicrobial activities with an EC 50 of 18.16 ± 0.64 µg/mL and a strong inhibition against Staphylococcus
aureus. The results obtained suggest that Algerian M. officinalis L. can be used as an important source of natural antioxidant
and antimicrobial agent for food and pharmaceutical industries.
Keywords Melissa officinalis L. · Polyphenols · ORAC test · Antioxidant activity · Antimicrobial activity
1 Introduction propyl gallate (PG) which are being restricted due to their
toxicity [4].
Medicinal and aromatic plants constitute a rich source of For this reason, several studies were focused on the study
biologically active compounds. Among the different kinds of natural resources and their phenolics content, antioxi-
of phytochemicals compounds, polyphenols (phenolic acids, dant and antimicrobial activities [5–7]. Melissa officinalis
flavonoids) are qualified to possess an important antioxidant L., commonly known as “Trendjane” in Algeria, is an aro-
potential. Several studies revealed that antioxidants can scav- matic perennial plant of the family of Lamiaceae that is native
enge the free radicals, delay or inhibit the oxidative process to the Mediterranean region (including Algeria), Southern
and reduce the risk of developing chronic diseases such as Europe, Asia and Southern parts of North America [8–10].
cancer, arteriosclerosis and Parkinson disease [1,2]. Melissa officinalis L. is traditionally used as a spasmolytic,
In recent years, there has been an emerging interest in sedative and antimicrobial agent. In addition, M. officinalis L.
finding natural antioxidants [3] as an alternative to synthetic possesses antiseptic, antiviral, gastric, antitumor and antiox-
antioxidants such as butylate hydroxyl toluene (BHT) and idative properties [9].
Several investigations showed that M. officinalis L. pos-
sesses a very pronounced antioxidant potential which is
related to the richness in phenolic compounds [11,12].
B H. Mabrouki
[email protected] According to literature data, M. officinalis L. contains
0.02–0.3% essential oil, comprising more than forty com-
1 Laboratory of Hydrometallurgy and Inorganic Molecular pounds of which more than 60 % are monoterpenes, such as
Chemistry, Faculty of Chemistry, University of Sciences and
citronellal, neral, geranial, etc. and more than 35% sesquiter-
Technology Houari Boumediene USTHB, BP 32, 16111
El-Alia, Bab-Ezzouar, Algiers, Algeria penes, such as β-caryophyllene and β-caryophyllene oxide;
2 monoterpene alcohols: nerol, geraniol and citronellol.
Instituto de Biologia Experimental e Tecnológica, Avenida da
República, Quinta-do-Marquês, Estação Agronómica Actually, essential oil of M. officinalis L. and their compo-
Nacional, Apartado 12, 2781-901 Oeiras, Portugal nents are gaining increasing interest because of their potential
123
3350 Arabian Journal for Science and Engineering (2018) 43:3349–3357
use as antimicrobial agents in the field of pharmacology, and 2.3 Preparation of Extracts
food preservation.
The study of the antimicrobial activity of the M. officinalis The dried and grounded leaves of M. officinalis L. (10 g)
L. extracts revealed a great bactericidal effect, a significant were extracted for 6 h by Soxhlet apparatus with 300 mL of
anti-inflammatory activity and a protective effect on ethanol- hexane, acetone, and ethanol respectively. The extracts were
induced gastric ulcer [12]. evaporated to dryness under vacuum, at 40 ◦ C in a Heidolph
The first main objective of this research was to extract rotary evaporator (Schwabach, Germany). The residues were
natural bioactive components from M. officinalis L. using weighted for the determination of the yield of extraction and
different solvents (such as ethanol, acetone, and hexane) to stored at 4 ◦ C in darkness for further use.
optimize the extraction yield and to compare the antioxidative
activities of the different extracts. 2.4 Extraction Yield
The phenolic and flavonoid contents were evaluated using
colorimetric assays [13]. The antioxidative abilities of leaf The method of Zhang et al. was used to determine the extrac-
extracts of M. officinalis L. were assessed by different tests tion yield. It was calculated and expressed as the percentage
such as DPPH method [14], β-carotene bleaching test, ferric- of the crude extract weight to the raw material (10 g) [17].
reducing antioxidant power (FRAP) [15].
However, this work assessed for the first time the antiox- Mass of extract
Extraction yield (%) = × 100 (1)
idant activity of Algerian M. officinalis L. using ORAC and Mass of raw material
H-ORAC tests. These tests are claimed to be more rele-
vant because they use a biologically relevant radical source 2.5 Total Phenolic Content
[16].
The second aim of this study is the valorization of M. Gallic acid was used as a standard to express the concentra-
officinalis L. which can be used as a new source of natural tions of TPC:
and safe antioxidants for pharmaceutical and food industries.
Abs = 0.00116 × C + 0.00245 (2)
where Abs was the absorbance of the sample; C was the

2 Materials and Methods concentration of gallic acid (mg/mL).
The amount of TPC was expressed as milligrams of Gallic
2.1 Reagents acid equivalent (GAE) per g of dry extract (mg GAE/g of dry
extract) according to the following equation:
DPPH (2,2 -diphenyl-1-picrylhydrazyl), aluminum chloride
hexahydrate, quercetin, ethanol, acetone, hexane, gallic acid, C×V
TPC (mg/g) = (3)
Folin–Ciocalteu reagent, Tween 40, β-carotene, linoleic M
acid, rosmarinic acid and potassium ferricyanide were from
Sigma-Aldrich. Ferric chloride, trichloroacetic acid, ascorbic where C, V, and M are the concentration of gallic acid estab-
acid, and sodium carbonate were purchased from VWR Pro- lished from the calibration curve (mg/mL), the volume of the
labo. Sodium acetate was from Riedel de Haen. Chloroform extract solution (mL) and the weight of extract (g), respec-
was from Fisher Scientific. tively [18,19].
An Optizen 3220 UV Spectrophotometer was used for
absorbance measurements, and an FL800 microplate fluo- 2.6 Total Flavonoid Content
rescence reader (Bio-Tek Instruments, Winooski, VT, USA)
was used for ORAC and H-ORAC tests A colorimetric method using AlCl3 reagent was used to esti-
mate the total flavonoid content (TFC) [20–22]. 2 mL of
aluminum trichloride (2%, w/v) was mixed with 2 mL of
2.2 Plant Materials extracts solution. After 10 min of incubation at room tem-
perature, the absorbance was read at 415 nm (Optizen 3220
The aerials parts (leafs) of M. officinalis L. were collected in UV/Visible spectrophotometer) against a blank (a mixture of
April 2015 from the northern part of Algeria (Rouiba area, 2 mL of extract solution and 2 mL ethanol). The quercetin
located in the eastern suburbs at 22 km from Algiers). The calibration curve (10–100 µg/mL) was used to determine the
plant was authenticated at the National Institute of Agron- TFC of each extract.
omy in Algiers (INA) by Professor H. Abdelkrim. The herb
material was dried, pulverized and stored until use. Abs = 17,374 × C + 0.02 (4)
123

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Arabian Journal for Science and Engineering (2018) 43:3349–3357

RESEARCH ARTICLE - BIOLOGICAL SCIENCES

Estimation of Total Phenolic Contents and In Vitro Antioxidant

where Abs was the absorbance of the sample; C was the

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