Asma Saeed et al., J.Chem.Soc.Pak., Vol. 38, No.

02, 2016 333

Evaluation of Antioxidant Effects and Inhibitory Activity of Medicinal Plants against

Lipid Peroxidation Induced by Iron and Sodium Nitroprusside in the Mouse Brain
1
Asma Saeed, 1Saif Ur Rehman, 2Muhammad Akram, 3Muhammad Zeeshan Bhatti, 4Rubina Naz, 5Asif Latif,
3
Amjad Ali, 6Ayaz Ahmad and 7Ahmad Saeed
1
Department of Biological sciences, Gomal University, Dera Ismail Khan 29050, Pakistan.
2
Department of Eastern Mdeicine and Surgery, Faculty of Medical and Health Sciences, University of Poonch,
Rawalakot, Azad Kashmir, Pakistan.
3
Institute of Biomedical Sciences, School of Life Science, East China Normal University,
Dongchuan Road, Shanghai, China.
4
Department of Chemistry, Gomal University, Dera Ismail Khan 29050, Pakistan.
5
Department of Horticulture, Faculty of Agriculture, Gomal University, Dera Ismail Khan 29050, Pakistan.
6
Department of Biotechnology, Abdul Wali Khan University, Mardan 23200,Pakistan.
7
Department of Chemistry, Qurtuba University of Science and Information Technology,
Dera Ismail Khan 29050, Pakistan.
asaeedti@gmail.com*

(Received on 14th July 2015, accepted in revised form 20th November 2015)

Summary: The present study compares the protective properties of aqueous extracts of five
medicinal plants, Myristica fragrans, Illicium verum, Curculigo orchioides, Glycyrrhiza glabra
and Embelia ribes against lipid peroxidation in mice brain. The antioxidant activities were
analyzed by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical assay, lipid peroxidation assay and
total antioxidant activity. The plant extracts exhibited inhibitions against thiobarbituric acid
reactive species (TBARS) induced by pro-oxidant (10μM sodium nitroprusside or FeSO4) in mice
brain. Antioxidant activity of the extracts was investigated by the scavenging of DPPH radical. All
the extracts had shown high antioxidant activity and the order of their antioxidant activity was G.
glabra>I. verum>C. orchioeides> E. ribes> M. fragrans. The inhibitory effect and antioxidant
activity of under study medicinal plants may be due to presence of higher phenolic contents, free
radical scavenging activity and reducing ability. These plants may be used to prevent oxidative
stress in brain.

Key Words: Antioxidant activity, Medicinal plants, Phenolics, Oxidative stress, Mice brain.

Introduction

Free radicals that arise either from various
environmental chemicals or metabolic processes,
cause many diseases ranging from shocks, ischemia,
skin irritation, inflammation, to tumors and cancer,
due to oxidation of fundamental macromolecules
resulting in the cell disfunction [1]. The antioxidants
occur in the normal cells, act as interceptors of free
radicals which defend the biological systems from
oxidative damage [2]. Several kinds of natural
secondary metabolites are produced by different parts
of the plants such as flavonoids, phenols, phenolic,
glycosides and saponins which act as natural
antioxidants and have specific pharmacological
activities [3]. Phenolic compounds have got much
importance for their significance antioxidant
properties, prevent oxidative damage and for the
perfection of health toward various diseases [4].
butylated hydroxyanisole (BHA) and butylated
hydroxytoluene (BHT) are in use but these are
prohibited due to their in-vivo carcinogenic effects
[6]. Nowadays, the trend has been changing to
replace synthetic antioxidants with natural
antioxidants of plant origin. From the past few years,
researchers are trying to discover the natural
antioxidants from medicinal plants [7] as well as their
role in detoxifying free radicals because the isolated
natural products are nontoxic, available at
economical prices and have no or fewer side effects
[8]. Chalcas koenigii for example, belongs to family
Rutaceae (common name is ‘curry patta’) is popular
as a spice and condiment. Their antioxidants serve as
free radical scavengers, inhibit lipid peroxidation and
protect food from oxidation by free radicals during
food processing [5].

Synthetic antioxidants are also widely used Myristica fragrans (local name Jaiphal)
as preservatives of food stuff that are specifically belongs to family Myristicaceae. It has antioxidant
retard deterioration, rancidity or discoloration due to property [9]. Its main constituents are terpenes,
oxidation [5]. Synthetic antioxidants such as geraniol, terpineol, volatile oil and myristicin. It is

*
To whom all correspondence should be addressed.
Asma Saeed et al.,
prescribed for the treatment of impotency, cardiac
weakness and premature ejaculation. Its oil is applied
externally for the treatment of headache, arthritis and
paralysis. Illicium verum (Common name Badian-i-
khatai) belongs to family Illiciaceae. Seed part is
used. The fruit contain higher bitter principle, tannins
and essential oil (9-10%), consisting of anethole (85-
90%), α-pinene, limone, β-phellandrene, α-terpineol,
farnesol and safrol. Its pharmacological actions
include insecticidal, antifungal, antibacterial,
antioxidant, carminative, stomachic, stimulant,
expectorant, diuretic, digestive, expectorant,
deodorant. The Illicium verum herb is reported to be
antioxidant, antibacterial and antifungal [10, 11]. It
can increase production of milk of new mother and
its seed oil is used in rheumatism. Curculigo
orchioides (Common name Musli siah) belongs to
family Amaryllidaceae. Syringic acid, phenolic
glycosides and antioxidant phenols have been
isolated from roots of C. orchioides [12] which have
various pharmacological effects such as anticancer,
antimicrobial, hypoglycemic properties, wound
healer and antioxidant [13]. It is used as general
tonic, aphrodisiac and for skin diseases. It is also
prescribed in urinary infections, jaundice, impotence,
skin disorders, gonorrhea, diarrhea, piles, asthma,
cough, deafness and stress [14]. Glycyrrhiza glabra
(Common name Mulethi), is under shrub or a hardy
herb with multifoliate leaves. Liquorice has been
isolated from it, which is anti-inflammatory,
hepatoprotective, expectorant, anti-anxiety and
antioxidant [15, 16]. Other constituents found, are
coloring matter, volatile oil, resins, bitter principles,
asparagines, starch, mannite, sucrose and glucose.
Embelia ribes Brum (Common name Baberang),
belongs to family myrsinaceae. It contains vilagin,
tannins, quercitol, embelin, christembine and embolic
acid. E. ribes is used as anthelmintic drug as an
effective remedy for tapeworm. It is useful in
flatulence, anorexia and abdominal pain. This plant is
also used for fever, skin diseases, diarrhea, cough,
chest ailments, abdominal and cystic tumors.
The present work deals with the study of the
antioxidant and inhibitory effect of above mentioned
plants with important medicinal properties on Fe (II)
and sodium nitroprusside (SNP) induced lipid
peroxidation in mice brain in vitro.
Experimental
Chemicals
Thiobarbituric acids (TBA), 1, 10-
phenanthroline, gallic acid, quercetin, 2, 2-diphenyl-
1-picrylhydrazyl (DPPH) and malonaldehyde-bis-
J.Chem.Soc.Pak., Vol. 38, No. 02, 2016 334
dimethyl acetal (MDA) were purchased from Sigma
Aldrich (St. Louis, MO, USA). Ferrous sulphate was
obtained from Biocehmicals (Lahore) and sodium
nitroprusside (SNP) sodium dodecyl sulphate (SDS)
were from Merck (Darmstadt, Germany).
Preparation of the Plant Extracts
M. fragrans, I. verum, C. orchioides, G.
glabra and E. ribes Brum were collected and
authenticated by a botanist. The voucher specimens:
M. fragrans (UPR/FMHS/EMS/S01070), I. verum
(UPR/FMHS/EMS/S01071), C. orchioides
(UPR/FMHS/EMS/S01072), G. glabra
(UPR/FMHS/EMS/S01073) and E. ribes Brum
(UPR/FMHS/EMS/S01074) were deposited in
Herbarium of Faculty of Medical and Health
Sciences, University of Poonch, Rawalakot Azad
Kashmir, Pakistan. Dried and finely ground plant
sample was suspended in deionized water at the rate
of 5g /250 mL and kept for 48 hours at room
temperature. It was then filtered and the residue was
further extracted twice. The combined filtrate was
concentrated by vacuum distillation apparatus using
Rotavapor R-20 and dried at 40–500C in an oven to
obtain a powder form to achieve overall yield of 17-
19.2 % of the starting plant material. Dilutions were
made to obtain desired concentrations of extract for
experiment. The aqueous extracts of the plants were
used as these are non toxic and suitable for their use
in traditional medicine. Animals used in this study
were handled under NIH Guidelines provided. Male
balb c mice (2.0-2.5 months and 24-30g), were
purchased from National Institute of Health,
Islamabad and were kept in cages at temperature of
22o C with supply of food and water.
Production of Thio-Barbituric Acid Reactive Species
(TBARS) from Animal Tissues.
The production thio-barbituric acid reactive
species (TBARS) and its assay were determined by
applying a modified method of Ohkawa et al [17].
The mice were given anesthesia of ether and
decapitated. The brain tissues were excised and
placed in Petri-dish pre-cooled on ice. One gram
brain was homogenized in a homogenizer and was
centrifuged at 1400 rpm for 10 min. The pellet was
discarded and supernatant was collected for assays.
To 0.1 mL of supernatant, 0.05 mL of 10 µM
nitroprusside or iron was added, followed by the
additions of various concentrations of plant extract
and water to adjust the volume up to 0.3 mL. The
mixture was incubated at 37o C for 1 h. Similarly
control was prepared in which oxidant was omitted.
The color was developed by adding 0.2 mL of 8.1%
Asma Saeed et al.,
SDS, 0.5 mL of acetic acetate buffer, pH 3.4 and 0.5
mL of 0.6% TBA. The reaction mixtures were
incubated at 97o C for 1 hour with dilution of 0.03
mM MDA and the absorbance of each tube was
measured at 532 nm.
Antioxidant Activity by DPPH Radical Scavenging
DPPH is a stable free radical with red color.
If free radicals have been scavenged, DPPH will
generate its yellow color. It is used to study the
radical scavenging effects of some plant extracts
containing natural products, accompanied by the
disappearance of initial red color [18].
Antioxidant activity of the extract was
performed as described by Hatano et al [19]. The
aqueous extract(s) (25-200 µg/mL) were added to 0.5
mL methnolic solution of 0.25 mM DPPH. The
mixtures were vortexed and incubated at room
temperature for 30 min. The absorbance of reaction
mixture was measured at 517 nm. Ascorbic acid was
used as positive control. The DPPH scavenging
activity was calculated as below.
Inhibition of DPPH (%) = (Absorbance control -
Absorbance sample / Absorbance control) x 100
Total Antioxidant Assay
Total antioxidant assay
(phosphomolybdenum assay) was determined by
the method of Prieto et al [20]. The aqueous
extract of different concentrations (50µg/mL to
250µg/mL) was added to 3 mL of the reagent
solution, consisting of 28 mM sodium phosphate,
0.6 M H2 SO4 and 4 mM ammonium molybdate.
The content was mixed by vortex and incubated
at 95 oC for 90 minutes. After cooling to room
temperature the absorbance was measured at 695
nm. Ascorbic acid was used as control and the total
antioxidant activity was calculated as ascorbic acid
equivalent (µg/mL).
Determination of Phenolics Content
For determination of total phenolics content,
2.5 mL of 10% Folin-Ciocalteu reagent was mixed
with 0.5 mL of the aqueous extract and 2.0 mL of 7.5
% sodium carbonate. The mixture was incubated at
45o C for 40 minutes and the absorbance was
measured at 765 nm. The calibration curve for gallic
acid was constructed as standard phenol and the total
phenolics content was expressed as micrograms of
gallic acid equivalents/ mg of extract.
J.Chem.Soc.Pak., Vol. 38, No. 02, 2016 335
Statistical Analysis
The results were expressed as means ±
standard deviation. The data was analyzed by one
way ANOVA and different group means were
compared by Duncan multiple ranges (DMR) test
where necessary. P < 0.05 was considered significant
in all cases. The software Package statistic was used
for analysis of data.
Results and Discussion
Mice brain homogenates were induced with
iron and sodium nitroprusside to cause lipid
peroxidation and the effect of M. fragrans, I. verum,
C. orchioeides, G. glabra and E. ribes were
determined. These plants were chosen as they are
frequently used as antihypercholesterolemic agents in
a number of herbal formulations.
Lipid Peroxidation in Mice Brain Induced with
Sodium Nitroprusside and Iron
Fig. 1 (a) shows the antioxidant effect of M.
fragrans, I. verum and C. orchioides, in mice brain.
The results revealed that treatment with 5 µM sodium
nitroprusside caused a significant (P <0.05) increase
in thiobarbituric acid reactive substances (TBARS)
compared to the basal. Three separate controls were
used for M. fragrans, I. verum and C. orchioides,
However, treatment with different concentrations of
extracts (50-250µg/mL) caused a significant decrease
in lipid peroxidation. I. verum showed a higher
decrease in lipid peroxidation compared to the other
plant species. M. fragrans showed a pro-oxidant
effect at 50 µg/mL by causing increase in lipid
peroxidation but from concentration 100 µg/mL to
250 µg/mL, it showed a regular decrease in lipid
peroxidation. I. verum showed a gradual decrease in
lipid peroxidation at all concentrations (50-
250µg/mL). C. orchioeides also showed antioxidant
effect by a gradual decrease in lipid peroxidation
from 50-250µg/ml extract concentrations. The order
of decreasing antioxidant activity was I. verum>C.
orchioeides>M. fragrans. Similarly, Fig. 1 (b) shows
the antioxidant effect of G. glabra and E. ribes
extracts in mice brain. The results revealed that
treatment with 5 µM sodium nitroprusside caused a
significant (P <0.05) increase in thiobarbituric acid
reactive substances (TBARS) compared to the basal.
Two separate controls were used for G. glabra and
E.ribes. Treatment with different concentrations of G.
glabra and E. ribes extracts caused a significant
decrease in lipid peroxidation. G. glabra showed
tremendous antioxidant activity at all concentrations
whereas E. ribes showed pro-oxidant effect at
 Asma Saeed et al., J.Chem.Soc.Pak., Vol. 38, No. 02, 2016 336

50µg/mL extract concentration but lipid peroxidation
was significantly reduced from concentration of
100µg/mL upto 250µg/mL. The order of decreasing
antioxidant activity was G. glabra > E. ribes. In an
early report, acetone extract has comparatively higher
antioxidant activity than aqueous extract. The
antioxidant activity could be due to the presence of β-
pinene, α-pinene, 1, 8-cineole, myrcene, carvacrol,
terpinen-4-ol, isoeugenol and eugenol. At or less than
50µg/mL of aqueous extract concentrations, E. ribes
showed pro-oxidant effect by causing increase in
lipid peroxidation which might be due to lesser
amount of the presence of β-pinene, α-pinene, 1, 8-
cineole, myrcene, carvacrol, terpinen-4-ol,
isoeugenol and eugenol. At 250µg/mL of aqueous
extract concentration, these compounds are quite
sufficient to cause antioxidant activity which results
in the smallest TBARS value.
M. F
Fig. 2 (a) shows the antioxidant effect of M.
fragrans, I. verum and C. orchioeides in mice brain.
Here, the TBARS was induced with 10 µM iron. The
results revealed that treatment with Fe (II) caused a
significant (P <0.05) increase in thiobarbituric acid
reactive substances (TBARS) compared to the basal.
Three separate controls were used for M. fragrans, I.
verum and C. orchioeides. Treatment with different
concentrations of M. fragrans, I. verum and C.
orchioeides extracts caused a marked decrease in
lipid peroxidation. I. verum showed a higher
percentage decrease in lipid peroxidation compared
to M. fragrans and C. orchioeides. At lower
concentrations, M. fragrans and C. orchioeide
extracts showed pro-oxidant effect. The order of
decreasing antioxidant activity was > I. verum > M.
fragrans > C. orchioeides.
M.F I.V C.O
I.V C. O
600

500
TBAR S(n mol/g.tissue)

T BARS (n mo l /g .ti ssu e )

500

400
400

300
300

200
200

100
100

0 0
Basal Cont r ol 50 100 150 200 250 Basal Cont r ol 50 100 150 200 250

Extract concentrations (microgram/ml)

Extract concentration(microgram/ml) (a)
(a) G.G E.R
G.G E.R
500

500
T BARS (n mo l /g .t i ssu e )

400
T BARS (n mo l /g .ti ssu e )

400

300
300

200 200

100
100

0
Basal Cont r ol 50 100 150 200 250 0
Basal Cont r ol 50 100 150 200 250
Extract concentrations (microgram/ml)
Extract concentrations (microgram/ml)
(b) (b)
Fig. 1: (a) Effect of aqueous extracts of Myristica
fragrans (M.F), Illicuim verum (I.V) and Fig. 2: (a) The effect of aqueous extracts of
Curculigo orchioeides (C.O) against lipid Myristica fragrans (M.F), Illicuim verum
peroxidation in mice brain induced by (I.V), Curculigo orchioeides (C.O) against
sodium nitroprusside. (b) Effect of lipidperoxidation in mice brain induced by
Glycyrrhiza glabra and Embelia ribes iron. (b) The effect of aqueous extracts of
against lipid peroxidation in mice brain Glycyrrhiza glabra (G.G) and Embelia ribes
induced by sodium nitroprusside. (E.R) against lipid peroxidation in mice
brain induced by iron.
 Asma Saeed et al., J.Chem.Soc.Pak., Vol. 38, No. 02, 2016 337

Fig. 2 (b) shows the antioxidant effect of G.
glabra and E. ribes in mice brain lipid peroxidation
induced with iron. Treatment with Fe (II) stimulated
the TBARS production. However, the lower
concentrations of G. glabra and E. ribes extracts
/mg of extract and 263 ± 0.65 µg /mg of extract of G.
glabra, I. verum, M. fragrans, C. orchioeides and E.
ribes, respectively.

Ascorbic Acid equivalent(microgram/ml)

were found to be pro-oxidant but at high
M. F I.V C.O GG E.R
concentrations they showed decrease in lipid
peroxidation. The order of decreasing antioxidant 250
activity was G. glabra > E. ribes.
DPPH Radical Scavenging Activity 200

Fig. (3) shows the DPPH radical scavenging 150

activity of studied plants. All the extracts had shown

100
high antioxidant activity which was evident by their
ability to scavenge DPPH radical to more than 50%. 50
However, the order of their antioxidant activity was
G. glabra > I. verum > C. orchioeides > E. ribes > 0
50 100 150 200 250
M. fragrans.
Extract concentration(microgram/ml)
M. F I.V C.O GG E .R
% scavenging of DPPH radical

100
Fig. 4: Total antioxidant activity measured by
80 phosphomolybdenum reduction method,
60
Values are mean ± SD (n=3).

40 Table-1: Total phenolic content (µg of gallic acid

20
equivalents / mg of extract) of extract of plants.
Plants Total phenolic content
0 Myristica fragrans 293± 2.33
50 100 150 200 250 Illicuim verum 319.5±0.2
Curculigo orchioeides 287±4.1
Extract concentration(microgram/ml) Glycyrrhiza glabra 339±1.25
Embelia ribes 263±0.65
Fig. 3: Antioxidant activity of aqueous extract of The results are expressed as means ± SD of three replicate analyses.

medicinal plants. DPPH radical scavenging

activity of different plants. Values are means
± SD (n=3).
G. glabra, I. verum and C. orchioeides
showed higher % scavenging of DPPH radical even
at lower tested concentrations.
Total Antioxidant Activity
Fig. 4 shows the total antioxidant activity of
different plants extract expressed as ascorbic acid
equivalent. All the extracts showed their reducing
activity. However, the order of their reactivity was G.
glabra >I. verum >C. orchioeides > M. fragrans > E.
ribes.
Total Phenolics Content
Mean values for total phenolic content are
shown in Table-1. The phenolic contents were found
339± 1.25µg / mg of extract, 319.5± 0.2 µg/mg of
extract, 293 ± 2.33 µg/mg of extract, 287 ± 4.1 µg
Oxidative stress is associated with
pathologies and normal aging process [21]. There are
various plants having antioxidants, have therapeutic,
biological and pharmacological activities against
oxidative damage caused by free radicals [22]. The
protections against the free radicals offered by the
aqueous extracts of M. fragrans, I. verum, C.
orchioeides, G. glabra and E. ribes suggest that they
will be beneficial to treat brain toxicities. Aqueous
extracts of M. fragrans, I. verum, C. orchioeides, G.
glabra and E. ribes have provided protection in
tissues against lipid peroxidation induced by sodium
nitroprusside. These plants may prevent the diseases
occurring from sodium nitroprusside overload. These
findings also indicate that the extracts of these plants
show defense against various neurotoxins (Iron and
SNP) at very low concentrations and in many cases,
has the capability to decrease the formation of
TBARS which is smaller than the amount of TBARS
production in basal case. Various research studies
indicate that medicinal plants, grains, fruits and
vegetables are major source of bioactive compounds,
polyphenols that are considered to be good
Asma Saeed et al.,
antioxidant with DPPH scavenging activity. Free
radicals produced in cells during the metabolic
processes, are involved in various diseases such as
tumors, cancer and cardiovascular diseases [23]. The
high DPPH scavenging activity of these plants
suggests their use in these and some other diseases
which are arising from oxidative damage to
macromolecules such as DNA, proteins and etc., by
free radicals. The total antioxidant assay is used to
evaluate fat- and water-soluble antioxidant activity
(phosphomolybdenum assay), the plant extracts
showed electron-donating ability and act as chain
breakers, converting the reactive free radicals into
non-reactive species [24]. Phenolic compounds are
good antioxidants [25]. Flavonoid compounds have
scientific and therapeutic value. Flavonoids reduce
reactive oxygen species and scavenge dangerous free
radicals [26]. Polyphenolic flavonoids exhibit their
antioxidant activity through different mechanisms
such as chelation of metal ions, scavenging of
reactive oxygen radicals and prevention of lipid
peroxidation [27]. In the present study following
traditional herbals like M. fragrans, I. verum, C.
orchioeides, G. glabra and E. ribes were tested with
respect to their anti-oxidant activity against lipid
peroxidation in mice brain, total phenolic content,
DPPH radical-scavenging test and total anti-oxidant
assay. Among all the herbals, G. glabra and I. verum
showed highest antioxidant activity against lipid
peroxidation in mice brain induced by sodium
nitroprusside and iron. These results were also
proved by total phenolic contents because G. glabra
and I. verum have highest phenolic contents among
all five herbals. DPPH radical scavenging and total
antioxidant assay also proved that G. glabra and I.
verum have highest anti-oxidant activity. Whereas
other three herbals, M. fragrans, C. orchioeides, E.
ribes also showed anti -oxidant activity against lipid
per oxidation in mice brain induced by sodium
nitroprusside and iron but was low as compared to G.
glabra and I. verum due to lesser phenolic contents
compared to them. Their lesser anti-oxidant activity
was also proved by DPPH radical scavenging test and
total phenolic assay. Dinesha et al [28] conducted a
study to determine the antioxidant activity of the
aqueous extract of I. verum against H2O2 induced
DNA damage and human peripheral lymphocyte cell
death. The antioxidant activities were evaluated by
lipid peroxidation, hydroxyl radical scavenging
activity, superoxide radical scavenging activity and
DPPH radical scavenging activity. It was concluded
in this study that I. verum possesses effective
prevention ability against H2O2 induced cell death
and DNA protection. Cheng et al [29] conducted a
study in which results showed that the ethyl acetate
fractions of I. verum have free radical scavenging
J.Chem.Soc.Pak., Vol. 38, No. 02, 2016 338
effects and reducing power effect. The strong
correlation between its DPPH and TEAC values with
those obtained from the reducing power assay
implied that the antioxidants in the extracts were
capable of scavenging free radicals and reducing
oxidants. The antioxidant components of I. verum
were also characterized by thin-layer chromatography
and GC-MS in this study. G. glabra is
a legume native to India, Southern Europe and other
countries of Asia. Many components have been
isolated form G. glabra including water soluble,
biologically active complex. This complex is
composed of flavonoids, triterpene polysaccharides,
saponins, pectins, amino acids, simple sugars,
mineral salts and various other substances [30].
Visavadiya and Narasimhacharya [30] reported the
antioxidant effects and hypocholesterolemic effects
of G. glabra root powder in hypercholesterolemic
male albino rats in a previous report. A four week
administration of G. glabra root powder (5 and 10
gm% in diet) to hypercholesterolaemic rats led to
significant reduction in plasma, cholesterol,
triglycerides, hepatic total lipids, low-density
lipoprotein and VLDL-cholesterol as well as
significant increases in HDL-cholesterol levels. In
another study conducted by Visavadiya et al [31] to
examine antioxidant activity in the extracts obtained
from G. glabra root. The aqueous and ethanolic
extracts exhibited scavenging activity against nitric
oxide with 50% inhibition. M. fragrans (nutmeg) is
an aromatic evergreen tree and is used as medicinal
plant, possesses phenolic compounds reported to
have DPPH free radical scavenging activity and
ability to chelate with metallic elements to form
complexes [32, 33]. Ashish et al [34] reported the
antioxidant and antimicrobial activities of nutmeg
seed extracts. Acetone, ethanol, methanol, butanol
and water were used for seeds extraction. All the
extracts have shown significant antioxidant potential.
Acetone extract has shown the highest antioxidant
potential. This high antioxidant activity could be due
to presence of β-pinene, α-pinene, 1, 8-cineole,
myrcene, carvacrol, terpinen-4-ol, isoeugenol and
eugenol. Suchandra et al [35] also reported the
phenolic compounds in M. fragrans which have great
potential to scavenge DPPH free radicals and inhibit
lipid peroxidation.
In another study conducted by Su et al [33]
in which M. fragrans was extracted with 80%
methanol and 50% acetone, free radical-scavenging
activities against cation radicals (ABTS+.), hydroxyl
(HO-1.) radicals, DPPH. and peroxyl (ORAC) radicals
were evaluated. The 80% methanol extract of M.
fragrans had greater ORAC, ABTS+ and TPC values
compared to the 50% acetone extract. The data
Asma Saeed et al.,
indicated that M. fragrans may serve as source of
natural antioxidants for improving health. Devyani et
al [36] evaluated the in vitro antioxidant activity of
C. orchioides, and revealed that C. orchioides
possesses potential effects in lipid peroxidation,
DPPH antiradical, nitric oxide scavenging,
superoxide scavenging and protection against
superoxide induced damage to erythrocytes
supporting the results of our study. Various
pharmacological activities of E. ribes have been
reported such as cell lines like antitumour, anti-
inflammatory and analgesic activities [37]
chemopreventive [38], anticancer activity [39] and
antioxidant activity by DPPH method [40].
Conclusion
These plants can be used by the people to
treat various diseases, including infectious ones and
serve as dietary source of natural antioxidants.
Therefore, food manufacturers use these sources for
the perfection of health food. The phytochemical
screening also revealed the presence of alkaloids,
flavonoids, saponins, steroids and tannins. Finally,
the results in this study indicate that antioxidant
activity against lipid peroxidation in mice brain is
due to the certain amounts of polyphenols. Hence it is
proving them to be perfect sources of antioxidants.
Aknowledgements
This research work was carried out under
M.Phil program in the Department of Biological
Sciences, Gomal University, D.I.Khan, Pakistan.
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