Antioxidant activity of selected medicinal plants used in folk medicine in

Libya
Salmin Alshalmani1*, Ghazalla M Benhusein2, Ebtisam A Absomaha3, Marwa I Meshri3, Hamdoon A.
Mohammed1 and Jamal S. Mezogi2

1
Faculty of Pharmacy, University of Benghazi, Libya
2
Faculty of Pharmacy, University of Tripoli, Libya
3
Medical Research Centre, Zawia, Libya

*Salmin K Alshalmani
E mail: salalshalmani2002@yahoo.com
Tel: 00218(0)917281665
Abstract
Eight wild medicinal plants used by Libyan and growing in Al-Jebel Al-Akhdar, Libya, were
suspected to estimate the antioxidant activity using 2,2-Diphenyl-1-Picrylhydrazyl stable free
radical (DPPH). Incidences of purple colour reduction of the DPPH by testing extracts in
addition to quercetin and vitamin C as positive controls reflect its ability to scavenge free
radicals. All testing plants extract showed noticeable strength as antioxidant regarding its
abilities to scavenge DPPH with an especial regards to Sarcopoterium spinosum.

Abbreviations: Antioxidant; scavenging activity; Folk medicine; Methanol extracts.

 1. Introduction
Since ancient times, the medicinal properties of plants have been investigated in the recent
scientific developments throughout the world, due to their potent antioxidant activities. As
antioxidants have been reported to prevent oxidative damage caused by free radical, it can
interfere with the oxidation process by reacting with free radicals, chelating, and catalytic
metals and also by acting as oxygen scavengers (1, 2). The potentially reactive derivatives of
oxygen, attributed as reactive oxygen species (ROS), are continuously generated inside the
human body. The generated ROS are detoxified by the antioxidants present in the body.
However, overproduction of ROS and/or inadequate antioxidant defence can easily affect and
persuade oxidative damage to various bio-molecules including proteins, lipids, lipoproteins
and DNA (3). This oxidative damage is a critical etiological factor implicated in several
chronic human diseases such as diabetes mellitus, cancer, atherosclerosis, arthritis and
neurodegenerative diseases and also in the ageing process(4). Antioxidant defence
mechanisms in biological systems mainly consist of both enzymatic and non-enzymatic
reactions. The non-enzymatic antioxidant include nutrient antioxidants like water and fat
soluble vitamins, carotinoids, α-tocopherol, ascorbic acid, glutathione, flavonoids, uric acid
and plasma proteins such as albumin, transferrin, ceruloplasmin, metal othionein etc.(5)
The use of synthetic antioxidants has been decreased due to their suspected activity as
promoters of carcinogenesis (6,7), as well as general consumer’s rejection of synthetic food
additions (8). There is a current worldwide interest in finding new and safe antioxidants from
natural sources e.g. plant material to prevent oxidative deterioration of food and to minimize
oxidative damage to living cells (9,10). There are the natural antioxidants present in grapes,
berry crops, tea, herbs nutmeg, apple etc. All the herbs and plants contain natural antioxidants
compounds including flavonoids, isoflavones, flavones, anthcyanins, coumarins, lignans,
catechin, isocatechin, gallic acid, etc. (11). Recently there has been an upsurge of interest in
the therapeutic potentials of plants, as antioxidants in reducing free radical induced tissue
injury.
Although several synthetic antioxidants, such as butylated hydroxyl anisole (BHA) and
butylated hydroxyl toluene (BHT), are commercially available, but are quite unsafe and their
toxicity is a problem of concern. Hence, strong restrictions have been placed on their
application and there is a trend to substitute them with naturally occurring antioxidants.
Natural antioxidants especially phenolics and flavonoids from tea, fruits, vegetables and
spices are already exploited commercially either as antioxidant additives or as nutritional
supplements (12). Also many other plant species have been investigated in the search for
novel antioxidants (13-15), but generally there is still a demand to find more information
concerning the antioxidant potential of plant species as they are safe and also bioactive.
Therefore, in recent years, considerable attention has been directed towards the identification
of plants with antioxidant ability.
The present work exerts the antioxidant potency for eight of the medicinal plants which are
widely distributed in Libya. Since these plants used by the indigenous people and consumed
by animal in Al-Jabel Al-Akhdar, Libya. The longer life span of these people might be
reflecting the effect of the environments and how natural sources can save the life.
2. Materials and Methods:
2.1. Plant collection and preparation of extracts:

Fresh herbs of plants under investigation were collected from the Green Mountain (Al-Jabal
Al-Akhdar) area, in Libya and identified by comparison with standard samples in the
herbarium of the faculty of science, University of Tripoli, Libya. The plants were washed
with tap water and left for drying in the open air place. 100 gram of the plants powders were
extracted by soxhlet apparatus with methanol (500 ml). The methanol extracts were
evaporated to dryness using rota vapour (IKA-WERKE, GMBH & Co. Kg, D-79219 Staufen,
Germany), and methanol extracts were reconstituted in their extraction solvent to give the
required concentration needed in this study.

2.2. Scavenging activity of 2, 2-Diphenyl-1-Picrylhydrazyl (DPPH)

radical:
Scavenging activity of different plants methanol extracts was measured by exerting the effect
of extracts on DPPH stable free radical according to method described by Sami G. A. et al.
(16), Briefly, stock solutions of all extracts of 1 mg/ml were prepared in pure methanol.
Aliquot of 400μl of 0.1μM of DPPH solution was added to 1 mL cuvett. Extract solution at
different concentration (1-50 µg) were added. Then, 600 µl of pure ethanol was added and
and the mixture was shaken vigorously and allowed to stand in dark place at room
temperature for 5 min. The absorbance of the mixture was measured spectrophotometrically
at 517 nm. Quercetin and ascorbic acid (vitamin C) were used as standard antioxidants. The
percentage of free radical scavenging was calculated according to the following equation:
% scavenging = (1-Sample absorbance at 517nm/blank absorbance at 517nm)X100.
Results showed in table 1 as a mean of three independent tests ± SD.
 3. Results
Antioxidant activity of the plants ethanol extracts are expressed as the ability of these extracts
to inhibit the DPPH stable free radical purple colour. IC50 of the plants extracts in addition to
quercetin and butaylated hydroxy anisole (positive controls), are expressed in the table 1.
Antioxidant potency comparison between different extracts at 10 and 30 µg/ml are showed in
table 1 and Figure 1.

Table 1: scavenging activity of the methanol extracts of different plants as shown by the
IC50 in µg / ml in addition to inhibition percentage at 10 and 20µg/ml.

DPPH inhibition in μg/ml

Plants and IC50 Inhibition % at 10 Inhibition % at 20
controls µg / ml µg/ml µg/ml
Helichrysum stoochs 20.1±1.27 24.9±1.50 49.8±2.95
Sarcopoterium spinosum 14.26±0.18 48.1±2.76 68.3±3.11
Cistus parrifloras 46.3±0.21 12.4±0.87 23.9±2.32
Euphorbia cnavasis 17.8±0.56 27.2±1.25 56.5±2.59
Quercus coccifera 19.1±0.35 26.5±0.10 54.1±3.02
Myrtus communis 26.0±0.14 20.1±1.73 37.1±1.39
Phlomis flocosa 30.0±0.70 17.1±1.01 33.0±1.99
Juniperus phoenicea 78.5±0.28 7.2±0.07 13.7±0.91
Ascorbic acid 1.40±0.14 100 % inhibition of DPPH at both
Quercetin 1.36±0.028 concentration (10 and 20 µg/ml)
Results were expressed as mean ± slander deviation of three independent measures.
Inhibition % Fig 1: DPPH scavenginge activity of different
of DPPH
80
plant extraxts at 10 and 20µg/ml
Inhibition % at 10 µg/ml
70
60 Inhibition % at 20 µg/ml

50
40
30
20
10
0
Helichrysum Sarcopoterium Cistus Euphorbia Quercus Myrtus Phlomis Juniperus
stoochs spinosum parrifloras cnavasis coccifera communis flocosa phoenicea

Ethanol extracts of plants at 10 and 20 µg/ml

Fig1: DPPH scavenging activity of different plant extracts at 10 and 20 µgl ml

4. Discussion
Oxidative stress, considered as one of the modern healthy problems around the world. ROS
is very damaging, since they attack lipids in cell membranes, carbohydrates, DNA. Enzymes
and proteins in tissues, to induce oxidative modifications, which cause membrane damage,
DNA damage and loss of protein function (17). This oxidative damage playing a causative
role in ageing and several degenerative diseases associated withit, such as heart disease,
congestive dysfunction and cancer (18). Humans and animals have evolved antioxidant
systems to protect against free radicals. These systems include some antioxidants produced in
the body (endogenous antioxidants) and others obtained from the diet (exogenous
antioxidants) (19).

The investigated plants shows a noticeable scavenging activity for the DPPH free radical
compared to the reference controls quercetin and vitamin C. Sarcopoteriums pinosumthat,
widely used as an antidiabetic drug by Bedouin healers (20) showed the strongest effect
among all investigated plants for the scavenging effect with IC50 equal to 14.2µg/ml. On the
other hand, 20 µg/ml of Sarcopoterium able to inhibit 68% of the DPPH free radical.
According to antioxidant effect, Euphorbia cnavasis, Quercus coccifera and Helichrysum
stoochs showed ability to scavenging the free radical with IC50 equal to 17.8, 19.1 and 20.1
µg/ml, respectively. Moderate activity appears in the result obtained from Myrtus communis,
Phlomis flocosa, and Cistus parrifloras with IC50 equal to 26, 30 and 47 µg/ml, respectively.

The present results gave clear impact for the abundance of these plants with phytochemicals
that, responsible for the noticeable antioxidant activity and ability to scavenge free radical.
Further studies for the identification of these phytochemicals may be a worthwhile research
point in our future.

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