Indian J Physiol Pharmacol 2013; 57(1) : 51–62 Pharmacological Activities of Ginger Oil 51

ANTIOXIDANT, ANTI-INFLAMMATORY AND ANTINOCICEPTIVE

ACTIVITIES OF ESSENTIAL OIL FROM GINGER

KOTTARAPAT JEENA*, VIJAYASTELTER B. LIJU*

AND RAMADASAN KUTTAN**

Amala Cancer Research Centre,

Amala Nagar, Thrissur – 680 555, Kerala, India
( Received on June 8, 2011 )

Abstract : Chemical compositions of ginger oil as well as its antioxidant,

anti-inflammatory and antinociceptive potential were evaluated in the
present study. The main constituents as detected by GC/MS analysis was
V-zingiberene which constituted 31% of the total area, ar-curcumene (15.4%)
and a -sesquiphellandrene (14.02%). Ginger oil scavenged superoxide, DPPH,
hydroxyl radicals and inhibited tissue lipid peroxidation in vitro .
Intraperitoneal administration of ginger oil was found to inhibit phorbol-
12-myristate-13-acetate induced superoxide radicals elicited by macrophages.
Oral administration of ginger oil for one month, significantly increased
superoxide dismutase, glutathione and glutathione reductase enzymes level
(P<0.001) in blood of mice and glutathione-S-transferase, glutathione
peroxidase and superoxide dismutase enzymes in liver. Ginger oil produced
significant reduction in acute inflammation produced by carrageenan and
dextran and formalin induced chronic inflammation (P<0.001). It also
exhibited significant reduction in acetic acid induced writhing movements
(P<0.001). The present report revealed that ginger oil possesses antioxidant
activity as well as significant anti-inflammatory and antinociceptive property.
Key words : antioxidant activity anti-inflammatory
antinociceptive essential oil GC/MS

INTRODUCTION pungency. The essential oil of ginger has

been found to possess antibacterial, antiviral
Ginger, the rhizome of Zingiber officinale and antifungal properties (1, 2). The present
Roscoe, is one of the most widely used spices study reports a detailed pharmacological
and a traditional remedy in Indian, Chinese screening of ginger oil evaluating its
and Oriental medicine against pain, antioxidant, anti-inflammatory and
inflammation and gastrointestinal disorders. antinociceptive properties. It is anticipated
Ginger oil is produced from fresh rhizomes that this study will throw further light on
of Zingiber officinale. It possesses the the pharmacological and medicinal properties
aroma and flavor of the spice but lacks the of ginger oil.

*Corresponding author : Dr. Ramadasan Kuttan, Ph.D, Research Director, Amala Cancer Research Centre, Amala
Nagar, Thrissur, Kerala – 680 555, India. Tel: 91-487-2307968; Fax: 91-487-2307968;
Email: [email protected], [email protected]
52 Jeena et al Indian J Physiol Pharmacol 2013; 57(1)

MATERIALS AND METHODS Research Laboratories Pvt. Ltd., (Mumbai,

India), 2,2-Diphenyl-1-picryl hydrazyl (DPPH)
Drugs and 2,2-azobis-3-ethylbenzthiozoline-6-
sulphonic acid (ABTS) were purchased from
The essential oil isolated by steam Sigma Aldrich (St.Louis, USA) and phorbol-
distillation from the fresh rhizomes of ginger 12-myristate-13-acetate (PMA) was a gift from
was supplied by Kancore Ingredients Limited, Dr. Allen Conney, USA. All other chemicals
Angamali, Kerala. In order to get a uniform and reagents used were of analytical reagent
suspension of ginger oil for in vitro studies, grade. Carrageenan was purchased from
the oil was dissolved in hexane (100 mg/10 Sigma Aldrich, USA.
ml) and 10 µl of Triton X 100 (Sigma-Aldrich)
was added and further evaporated to dryness Gas chromatography/mass spectrometry analysis o f
and finally made up to 10 ml with distilled ginger oil
water. The oil was dissolved in paraffin oil
for all in vivo studies. Ginger oil was subjected to gas
chromatography/mass spectrometry (GC/MS)
Animals analysis using a Hewlett–Packard gas
chromatograph (Model 6890) coupled with a
Balb/c mice (20–25 g) were used in the quadruple mass spectrometer (Model HP
study. They were purchased from Small 5973) and a HP – 5MS capillary column
Animal Breeding Station, Mannuthy, Kerala, (5% phenylmethylsiloxane; 30 m × 320U m ×
India and were housed in well ventilated 0.25U m). The interphase, ion source and
cages under controlled conditions of light and selective mass detector temperatures were
humidity and provided with normal mouse maintained at 243°C, 230°C and 150°C,
chow (Sai Durga Food and Feeds, Bangalore, respectively. Helium was used as a carrier
India) and water ad libitum . All the animal gas at a flow rate of 1.4 ml/min. For the
experiments were done as per the instructions essential oil, the oven temperature was
prescribed by the Committee for the programmed linearly at 60°C; then increased
Purpose of Control and Supervision of from 60°C to 243°C at the rate of 3°C/min.
Experiments on Animals (CPCSEA), Ministry
of Environment and Forest, Government of Determination of antioxidant activity of ginger
oil by in vitro method
India, and implemented through the
Institutional Animal Ethical Committee of Superoxide radical scavenging activity
the Research Centre.
Superoxide radical scavenging activity
Chemicals and reagents was determined by the NBT reduction
method (3). Different concentrations of the
Nitroblue tetrazolium (NBT), glutathione, oil (10-200 µg) were added to the reaction
glutathione oxidized (GSSG), nicotinamide medium containing 3 µg NaCN in 6 µM
adenine dinucleotide phosphosphate reduced EDTA, riboflavin (2 µM), NBT (50 µM) and
(NADPH) and 5-5’dithiobis 2-niotrobenzoic phosphate buffer (67 mM, pH 7.8) in a final
acid (DTNB) were purchased from Sisco volume of 3 ml. The tubes containing
Indian J Physiol Pharmacol 2013; 57(1) Pharmacological Activities of Ginger Oil 53

reaction mixture were uniformly illuminated ml), thiobarbituric acid (TBA-0.8%, 1.5 ml),
with an incandescent lamp for 15 minutes. and acetic acid 20%, pH-3.5, 1.5 ml). The
The optical density was measured at 560 nm total volume was then made up to 4 ml with
before and after illumination. The percentage distilled water and incubated for 1 hr at 100
inhibition of superoxide generation was 4°C water bath. After cooling, 1 ml of distilled
evaluated by comparing the absorbance value water and 5 ml of mixture of n-butanol and
of the control and experimental tubes. pyridine (15:1, v/v) were added and vortexed
and after centrifugation the absorbance of
Hydroxyl radical scavenging activity the organic layer was measured at 532 nm.
The percentage inhibition of lipid peroxidation
The hydroxyl radical scavenging activity was determined by comparing the results of
was measured by studying the competition the test compound with those of the control
between deoxyribose and the extract for (4).
hydroxyl radicals generated from the
Fe 3+ /ascorbate/EDTA/H 2O 2 system (Fenton Determination of DPPH radical scavenging activity
reaction). The reaction mixture (1.0 ml final
volume) contained deoxyribose (2.8 mM), Ginger oil at different concentrations
FeCl3 (0.1 mM), EDTA (0.1 mM), H2O (1 mM), (0.1-1 mg) were mixed with 0.375 ml
ascorbate (0.1 mM), KH 2PO 4 – KOH buffer methanolic solution of DPPH (1, 1-diphenyl-
(20 mM, pH 7.4) and different concentrations 2-picrylhydrazyl-0.25 g/L). Total volume
of the oil (10-200 µg). The reaction mixture was made up to 2 ml with methanol.
was incubated for 1 hr at 37°C. Deoxyribose The disappearance of DPPH was read
degradation was measured as TBARS and spectrophotometrically at 515 nm after 20
percentage inhibition calculated. (4). The min of incubation at room temperature in
percentage inhibition was calculated dark (5).
according to the formula: Inhibition
percentage (Ip) = [(A B –A A )/A B ]· 100 where A Determination of ABTS radical scavenging activity
B and A are the absorbance values of the
blank sample and of essential oil solution, ABTS radical scavenging activity of
respectively. the extract was determined using ferryl
myoglobin/ABTS protocol (6). The stock
Determination of inhibition of lipid peroxidation solutions and 500 µM ABTS diammonium
salt, 400 µM myoglobin (MbIII), 740 µM
Different concentrations of the oil (10- potassium ferricyanide, and 450 µM H 2O2 was
200 µg) were added to rat liver homogenate prepared in phosphate-buffered saline (PBS;
(0.1 ml, 25% w/v), ascorbic acid (0.06 mM), pH 7.4). Metmyoglobin was prepared by
30 mM KCl, 0.16 mM FeSO 4 solution and mixing equal amounts of myoglobin and
final volume made up with Tris buffer (40 potassium ferricyanide solutions. The test
mM, pH-7) to 0.5 ml. Tubes were incubated tubes contained ABTS (150 µM), MbIII (2.25
for one hour at 37°C. Aliquots of the µM), varying concentrations of essential oil
incubation mixture (0.4 ml) were treated (10-200 µg) and PBS (pH-7.4) in 2 ml. The
with sodium dodecyl suphate (SDS-8%, 0.2 reaction was initiated by adding 75 µM H 2 O 2
54 Jeena et al Indian J Physiol Pharmacol 2013; 57(1)

and lag time in seconds was recorded before oil on the inhibition of superoxide generation
absorbance of ABTS at 734 nm began to in the macrophages were measured by
increase. inhibition in the reduction of NBT to
formazan (8). The percentage inhibition was
Ferric reducing antioxidant power assay (FRAP determined by comparing the absorbance
assay) values of untreated and treated animals.

The ferric reducing ability was measured

Determination of effect of ginger oil on in vivo
at low pH. FRAP reagent contained 25 ml antioxidant enzyme levels
0.3 M acetate buffer, 2.5 ml 4,6-Tris-2-pyridyl-
(s)-Triazine (TPTZ) and 2.5 ml ferric Balb/c mice (4-6 weeks) weighing 20-25 g
chloride. Different concentration of the oil were divided into 4 groups of five animals
(10-200 µg) was made up to one ml with and they were treated orally with ginger oil
freshly prepared FRAP reagent. The mixture dissolved in paraffin oil at different doses
was incubated at 37°C for 15 minutes and for 30 days. Group I was kept as normal.
read against distilled water at 595 nm. Group II was kept as control treated with
Standard graphs were constructed using paraffin oil only. Group III and Group IV
known concentrations of ferrous salt in were treated with 100 mg ginger oil/kg b.wt
water/methanol to replace FeCl3 and and 500 mg ginger oil/kg b.wt respectively.
expressed as EC 1 values. EC 1 is defined as
concentration of an antioxidant having a At the end of the experiment, animals
ferric reducing ability equivalent to that of were sacrificed by ether anesthesia, and
1 mM ferrous sample (7). blood was collected by heart puncture, liver
was excised and washed in ice-cold Tris Hcl
Determination of the effect of extract on PMA-
buffer (0.1 M, pH-7.4), and cytosolic samples
induced superoxide radical generation in
peritoneal macrophages of liver homogenate were prepared by
centrifuging at 10,000 rpm for 30 mins at
Balb/c mice (4-6 weeks) weighing 20-25 g 4°C.
were used for the study. Animals were
divided into 5 groups (3 animals/group). All The total protein was estimated by the
the animals were injected (i.p) with sodium method of Lowry, Rosenbroug, Farr and
caseinate (5%) to elicit macrophages. Group Randall, 1951 (9). Hemoglobin was estimated
I was kept as normal. Group II was kept as by the cyanmethemoglobin solution using
control. Group III, IV and V were treated Drabkin’s method (10). Whole blood was used
with single dose of ginger oil at different for determination of superoxide dismutase,
concentration (10, 50 and 250 mg/kg body catalase and glutathione levels. Serum was
weight, respectively). On the fifth day 1 hr used for glutathione reductase estimation.
after oil administration, peritoneal Liver tissue homogenate was used for super
macrophages were activated in vivo by oxide dismutase, catalase, glutathione peroxidase,
injecting phorbol-12-myristate-13-acetate glutathione reductase, glutathione –S-transferase
(PMA-100 ng/animal). After 3h, peritoneal and glutathione assessment as given
macrophages were harvested. The effects of below.
Indian J Physiol Pharmacol 2013; 57(1) Pharmacological Activities of Ginger Oil 55

Superoxide dismutase activity was In the dextran model, edema was induced
measured by the NBT reduction method (3). by injection of dextran. Group VI was kept
Catalase activity was estimated by the as control goup, dextran (1% w/v). Group VII
method of Aebi, 1974 (11) by measuring the was treated with standard drug, diclofenac,
rate of decomposition of hydrogen peroxide (10 mg/kg.i.p). Group VIII, IX and X were
at 240 nm. Glutathione activity was assayed treated with different concentrations
by the method of Moron, Depierre and of ginger oil (100, 500, 1000 mg/kg),
Manner, 1979 (12) based on the reaction with intraperitoneally for 4 consecutive days. One
DTNB. The assay of glutathione peroxidase hour, after the fourth dose of ginger oil
was done by the method of Hafeman, Sundae administration, acute inflammation was
and Houestra, 1974 (13) based on the induced by sub-plantar injection of 0.02 ml
degradation of H 2O 2 in the presence of GSH. dextran (1% w/v in 0.1% carboxy methyl
Glutathione reductase activity was measured cellulose) in the right hind paw (17). The
by the method of Racker, 1955 (14). The thickness of paw was measured using Vernier
method of Habig, Pabst and Jakoby, 1974 calipers at 0, 1, 2, 3, 4 and 6 hours and
(15) was followed to assay the activity of finally after 24 hrs. The percentages of
glutathione -S-transferase (GST) which is inhibition were calculated according to the
based on the rate of increase in conjugate
following formula :
formation between GSH and 1-chloro-2, 4-
nitrobenzene (CDNB).
% Inhibition = [(VT _ V0)control _ (VT _ V0)treated group]
Determination of anti-inflammatory activity of x 100/(VT _ V0)]control
ginger oil
Where V T is paw thickness 3 hr after
Acute inflammation models using carrageenan and
injection V 0 is the initial paw thickness.
dextran induced paw edema in mice

The experimental mice were divided into Chronic inflammation model using formalin
10 groups (6 animals/group). Group I was induced paw edema in mice

kept as control group -carrageenan (1% w/

v). Group II was treated with standard drug, Balb/c mice were divided into 5 groups
Diclofenac (10 mg/kg, i.p). Group III, IV and (6 animals/group). Group I was kept as
V were treated with different concentrations control and group II was treated with
of ginger oil (100, 500, 1000 mg/kg), standard drug diclofenac (10 mg/kg, i.p).
intraperitoneally for 4 consecutive days. One Group III, IV and V were treated with
hour, after the fourth dose of ginger oil different concentrations of ginger oil (100,
administration, acute inflammation was 500, 1000 mg/kg, i.p). Chronic inflammation
induced by sub-plantar injection of 0.02 ml was induced by sub-plantar injection of
carrageenan (1% w/v, in 0.1% carboxy methyl freshly prepared 0.02 ml of 2% formalin on
cellulose) in the right hind paw (16). The the right hind paw in all groups (18). Drug
thickness of paw was measured at 0, 1, 2, 3, treatment was started 1 hour prior to
4, 6 and 24 hr using Vernier calipers after formalin injection and continued for 6
carrageenan injection to determine the consecutive days. The inflammation was
formation of edema. measured using Vernier calipers before and
56 Jeena et al Indian J Physiol Pharmacol 2013; 57(1)

after injection of formalin and for 6 RESULTS

consecutive days. The percentage inhibition
was calculated using the formula given Composition of ginger oil
above.
A number of compounds were identified
Determination of antinociceptive activity of ginger by GC/MS analysis of ginger essential oil
oil and the most prominent components are
given in Table I. The principal constituent
Acetic acid-induced writhing in mice of ginger oil was zingiberene (31.08%), a
sesquiterpene hydrocarbon, followed by ar-
Balb/c mice were divided into 5 groups curcumene (15.4%) and α-sesquiphellandrene
(5 animals/group). Acetic acid (0.6% v/v, 10 (14.02%). Other compounds include bisabolene
ml/kg) was injected into the peritoneal (13.80%) and sabinene (8.27%). α-Zingiberene
cavities of mice, which were placed in a large has been reported to be the major constituent
observation boxes, and the intensity of by Singh, Kapoor, Singh, Heluani and
nociceptive behavior was quantified by Lampasona, 2008 (20) while Agarwal, Walia,
counting the total number of writhings Dhingra and Khambay, 2001 (21) has
occurring between 0 and 20 mins after reported curcumene as the major constituent.
stimulus injection (19). The vehicle, paraffin
oil and ginger oil (100, 500 and 1000 mg/kg, Antioxidant activities of ginger oil in vitro
i.p) were given 30 mins prior to acetic acid
injection. Aspirin (100 mg/kg, i.p) was used The essential oil of ginger was found to
as the standard reference drug. The writhing scavenge superoxide, hydroxyl radicals and
response consisted of a contraction of the inhibit tissue lipid peroxidation (Fig. 1).
abdominal muscle together with a stretching Ginger oil gave an IC 50 of 36 µg/ml for
of the hind limbs. The antinociceptive
activity was expressed as the writhing scores TABLE I : Chemical composition of ginger oil.
over a period of 20 min. Antinociceptive
effect was expressed as reduction of number No. Compound (% of total area)
of writhes between control and treated
1. Camphene 5.14
groups, using the formula: [(C-D)/C] x 100 2. Sabinene 8.28
where C is the average number of writhings 3. Isoborneol 1.14
4. α-Cubebene 0.75
for control group of mice and D is the 5. α-Elemene 1.57
average writhings of the drug treated mice. 6. γ-Elemene 0.58
7. Aromadendrene 0.34
8. Germacrene 2.40
9. Ar-Curcumene 15.35
Statistical analysis
10. beta-Panasinsene 1.47
11. Zingiberene 31.08
12. Beta Bisabolene 13.81
The values were expressed as mean± 13. α-Sesquiphellandrene 14.02
standard deviation (SD). Statistical evaluation 14. Verbenone 0.29
15. Elemol 0.45
of the data was done by one way ANOVA 16. Thujopsene 1.04
followed by Dunnet’s test (post-hoc) using 17. Silane 1.61
18. Farnesol 0.70
Instat 3 software package.
Indian J Physiol Pharmacol 2013; 57(1) Pharmacological Activities of Ginger Oil 57

Fig. 1 : In vitro free radical scavenging activity of ginger oil. (A) Superoxide radical scavenging
activity. (B) Hydroxyl radical scavenging activity. (C) Inhibition of lipid peroxidation.
(D) DPPH radical scavenging activity.(E) ABTS radical scavenging activity.

scavenging superoxide radicals but IC 50 for showed only a mild capacity to scavenge
inhibition of hydroxyl radicals and lipid ABTS radical.
peroxidation was found to be greater than
200 µg/ml and 400 µg/ml respectively and so Effect of ginger oil on PMA-induced superoxide
could not be calculated. The ferric reducing radical generation
activity for 50 µg of ginger oil was found
to be 1.8 mM for ginger oil. Ginger oil Superoxide radicals generated during the
possessed only mild DPPH radical scavenging activation of PMA in sodium caesinate
ability as IC 50 was more than 1000 µg/ml. It induced macrophages in vivo were found to
58 Jeena et al Indian J Physiol Pharmacol 2013; 57(1)

be scavenged by ginger oil. The percentage groups at 100 mg/kg bw (P<0.01) and 500
inhibition was 18.25% for ginger oil at 250 mg/kg bw (P<0.001). Glutathione reductase
mg/kg bw respectively. was elevated by the administration of ginger
oil (P<0.05) in all animals treated with 100
Effect of administration of ginger oil on mg/kg bw and significant activity was shown
antioxidant enzymes and glutathione at 500 mg/kg bw (P<0.001).

Antioxidant enzymes in blood and serum Ginger oil also showed a significant effect
of mice were increased after administration on some of the antioxidant enzymes in liver
of ginger oil for a period of 30 days (Table tissue of mice after treatment for 30 days
II). Catalase was found to be increased in all (Table III). Catalase did not change
animals treated with 100 and 500 mg/kg bw singnificantly in any of the treated groups.
of ginger oil (P<0.05). The essential oil Superoxide dismutase activity was elevated
administration significantly elevated in the group treated with 500 mg/kg.bw
superoxide dismutase (P<0.001) at both (P<0.01) ginger oil. The level of glutathione
concentrations. Glutathione level was also peroxidase enzyme was significantly
found to be significantly increased in treated increased by ginger oil at 500 mg/kg bw

TABLE II : Effect of oral administration of ginger oil for one month on antioxidant systems in blood.

Catalase Superoxide Glutathione Glutathione

Treatment (k/g Hb) dismutase reductase (nmol/ml)
(U/g Hb) (U/g Hb)

Normal 34.74±8.18 478±43.97 2.88±0.56 46.5±4.9

Control 32.52±7.38 445±39.62 2.76±0.25 44.8±7.3
100 mg/kg bw 57.91±8.30* 610±31.07*** 6.58±.82* 68.6±6.4**
(↑ 27.12%) (↑ 58%) (↑ 34.69%)
500 mg/kg bw 51.22±18.19* 872±24.77*** 8.15±1.62*** 78.4±5.7***
(↑ 48.95%) (↑ 66.14%) (↑ 42.85%)

Each value represents the mean±S.D (n=5). *P<0.05 compared with control, **P<0.01 compared with control,
***P<0.001 compared with control. ↑ indicates % increase.

TABLE III : Effect of oral administration of ginger oil for one month on antioxidant systems in liver.

Catalase Superoxide Glutathione Glutathione Glutathione Glutathione

Treatment (U/mg protein) dismutase peroxidase -S-transferase reductasea (nmol/ml)
(U/mg protein) (U/mg protein) (nmol/mg
protein)

Normal 5.29±0.66 0.84± 0.12 8.13±0.30 41.57±1.66 88.83±12.53 12.7±1.6

Control 4.97±0.76 0.83± 0.03 8.82±1.51 39.46±3.43 79.00±20.16 11.4±1.7
100 mg/kg bw 5.48±0.98 0.90±0.16 12.98±6.09 32.00±11.61 79.79±17.49 12.2±2.3
500 mg/kg bw 7.36±0.45 1.10± 0.13* 18.89±7.34 103.09±12.6*** 69.78±9.53 13.8±3.4
(↑ 24.54%) (↑ 53.30%) (↑ 61.72%)

*P<0.05 compared with control, **P<0.01 compared with control, ***P<0.001 compared with control. ↑ indicates
% increase. Each value represents the mean±S.D (n=5). a(nmol of NADPH consumed/min/mg Protein).
Indian J Physiol Pharmacol 2013; 57(1) Pharmacological Activities of Ginger Oil 59

(P<0.01). Even though glutathione was found

to be increased, it was not found significant.
The level of glutathione reductase was found
to be unaltered in both the treated groups.
Glutathione-S-transferase (GST) level was
found to be elevated in animals and
significant activity was shown at 500 mg/k
bw (P<0.01) treated group.

Effect of administration of ginger oil in acute

inflammation models

Anti-inflammatory effect of ginger oil was

evaluated in acute edema models. As
observed in Fig. 2a, intraperitonieal
treatment with ginger oil in mice at 100,
500 and 1000 mg/kg was capable of reducing
the odema formation by carrageenan in a
dose dependent manner by 27.8, 44.4 and
61.1% (P<0.001) when compared to control.
Diclofenac (10 mg/kg) gave a percentage
inhibition of 55.6%.

The inhibitory values of edema at 3 hr

post dextran treatment were 35, 41.2 and
52.9% (P<0.001 for 100, 500 and 1000 mg/kg
of ginger oil respectively (Fig. 2b) while
diclofenac gave similar activity at 10 mg/kg
body weight.
Fig. 2 : Anti-inflammatory activity of ginger oil. (A)
Effect of administration of ginger oil in chronic Carrageenan model. (B) Dextran model. (C)
inflammation models Formalin model.

Ginger oil showed significant suppression Effect of ginger oil on acetic acid induced writhing
of inflammation in chronic model of model
inflammation induced by formalin in a
concentration dependent manner. Percentage Ginger oil showed marked and significant
inhibition of 54.17, 62.5 and 70.8% (P<0.001) (P<0.001) reduction in the number of
were shown at doses of 100, 500 and writhings induced by acetic acid. As shown
1000 mg/kg respectively (Fig. 2c) while in Fig. 3, the analgesic activity at all tested
diclofenac (10 mg/kg) showed an inhibition doses (100, 500 and 1000 mg/kg) indicated a
of 54.8%. dose dependent relationship in the inhibition
60 Jeena et al Indian J Physiol Pharmacol 2013; 57(1)

(GPx) are thought to be the most important.

The current study indicates that ginger

oil could inhibit the oxygen radicals as seen
from the inhibition of lipid peroxidation,
scavenging of superoxide and hydroxyl radical
in vitro . However they showed only a mild
scavenging activity against stable free
radicals such as DPPH and ABTS. Ginger
Fig. 3 : Effect of ginger oil in nociceptive behaviour oil also scavenged the superoxide radicals
of mice evaluated in acetic acid induced generated in vivo in mice peritoneal
abdominal writhing model. Groups of
animals were pretreated with aspirin (10 macrophages. The antioxidative potential of
mg/kg bw), ginger oil (1000, 500 and 100 mg/ ginger oil may be due to the mixture of
kg bw, i.p) 3 0 m i n s b e f o r e a c e t i c a c i d
injection. Nociception was measured by the dozens of compounds of different functional
number of writhes the animal showed 20 mins groups, polarity and chemical behavior which
following intraperitonieal 0.6% acetic acid
injection. Values are reported as mean±S.D. produces either synergistic or antagonistic
**P<0.01, ***P<0.001.
effect on antioxidant activity. Many reports
emphasis that phenolic groups play an
of writhing reflux by 13.08, 70.64, and important role in antioxidant activity (22).
92.15%. The antinociceptive activity of In the present study, percentage of phenolic
ginger oil was compared with that of the compounds was found to be low compared to
standard drug at aspirin (10 mg/kg). previous reports (20). Administration of
ginger oil was capable of protecting the
DISCUSSION cells from intracellular oxidative damage
invivo by increasing the serum antioxidant
Reactive oxygen species (ROS) are formed status as clearly observed from the data.
in both physiological and pathological
conditions in mammalian tissues. The Carrageenan and dextran are widely used
uncontrolled production of free radicals is as noxious agents to induce experimental
considered to be an important factor in tissue inflammation to screen anti-inflammatory
damage which can induce pathophysiological agents. The irritant effect induced by
changes. Free radicals also play an important carrageenan is the result of activation of
role in inflammation that can mediate tissue kinin and complement cascades and the
destruction. Cells have several antioxidant release of anti-inflammatory mediators such
enzymes and other antioxidant mechanisms as vasoactive amines (histamine, serotonin,
to protect themselves from the harmful 5-hydroxytryptamine and bradykinins) during
effects of oxidants. The latter include the early phase of inflammation and kinin
glutathione (GSH) and numerous GSH like substances and eicasanoids (prostaglandins,
dependent enzymes, metal binding proteins, proteases and lysosomes) during the later
and vitamins. Major antioxidant enzymes are phase of inflammation (23, 24). Dextran
the superoxide dismutase (SOD), catalase and induces paw edema mainly by releasing
peroxidases, of which glutathione peroxidases histamine and 5-hydroxytryptamine to the
Indian J Physiol Pharmacol 2013; 57(1) Pharmacological Activities of Ginger Oil 61

site of inflammation from the mast cells (25). for peripheral analgesic activity. In the
In the present study, treatment with present study, ginger oil showed dose
essential oil of ginger oil was effective in dependent and significant inhibition of acetic
reducing the oedamatogenic response evoked acid induced writhes in mice suggesting
by carrageenan and dextran in a dose that ginger oil has strong antinociceptive
dependant manner. In chronic inflammatory activity and the mode of action might
models, inhibition of formalin induced involve the inhibition of synthesis of
oedema is considered as one of the most arachidonic acid metabolite mediated by COX
suitable test procedures to screen anti inhibition.
inflammatory agents as it closely resembles
human arthritis (26). Inflammation induced Thus, in addition to imparting flavor to
by formalin comprises an early neurogenic food, ginger oil possesses potential health
response mediated by substance P and benefits by scavenging the free radicals
bradykinin followed by a tissue mediated formed in the body. The study also indicates
response where histamine, 5-hydroxytryptamine, the efficacy of ginger oil as an efficient
prostaglandins and bradykinin are involved therapeutic agent in acute and chronic
(27). Ginger oil displayed strong anti-inflammatory inflammatory conditions.
activity in chronic inflammation model and
the mode of action may be due to the ACKNOWLEDGEMENTS
inhibition of prostaglandin release.
Authors are thankful to Spice Board,
Acetic acid induces pain by enhancing India, for providing financial assistance to
the levels of endogenous substances like carry out this work We assure you that this
PGE 2 and PGF 2 (28) in the peritoneal cavity. manuscript has not been published already
This indicates that acetic acid acts indirectly in part or whole in any journal or magazine
in the stimulation of nociceptive neurons by for private or public circulation. Kottarapat
the release of endogenous mediators. Thus, Jeena Vijayastelter B. Liju Ramadasan
acetic acid is used to screen compounds Kuttan.

REFERENCES

1. Singh G, Maurya S, Catalan, C, Lampasona MP. peroxide in animal tissue by thiobarbituric acid
Studies on essential oils, Part 42: chemical, reaction. Anal Biochem 1979; 95: 351–358.
antifungal, antimicrobial and sprout suppressant
5. Aquino R, Morellis S, Lauro MR, Abdo S, Saija
studies on ginger essential oil and its oleoresin.
Flavour Frag J 2005; 20: 1–6. A, Tomaino A. Phenolic constituents and
antioxidant activity of an extract of Anthurium
2. Koch C, Reichling J, Schneele J. Inhibitory effect vesicular leaves. J Nat Prod 2001; 64: 1019–
of essential oils against herpes simplex virus 1023.
type-2. Phytomedicine 2008; 15(1-2): 71–80.
6. Alzoreky N, Nakahara N. Antioxidant activity of
3. Mc Cord JM, Fridovich I. Superoxide dismutase some edible Yemeni plants evaluated by Ferryl
enzyme function for erythrocaprein. J Biochem myoglobin/ABTS assay. J Food Sci Technol Res
1969; 244: 6049–6056. 2001; 7: 144–146.
4. Ohkawa H, Oshishi N, Yagi K. Assay for lipid 7. Benzie IFF, Strain JJ. The ferric reducing ability
62 Jeena et al Indian J Physiol Pharmacol 2013; 57(1)

of plasma (FRAP) as a measure of “antioxidant 18. Chau TT. Analgesic testing in animals models.
power” - The FRAP assay. Anal Biochem 1996; Alan R Liss Inc. In: Pharmacological methods in
239: 70–76. the control of inflammations. New York, 1989:
448–452.
8. Dwivedi PP, Verna AS, Ray P. Induction of
immune rejection of tumours by protein A in 19. Koster R, Anderson M, De Beer J. Acetic acid
mice bearing transplantable solid tissue Dalton’s for analgesic screening. Fed Proc 1959; 18: 412–417.
lymphoma tumors. Immunopharmacol Immunotoxicol
1992; 14 (1-2): 105–128. 20. Gurdip S, Kapoor IPS, Pratibha S, Carola S de
Heluani, Marina P de Lampasona and Cesar AN
9. Lowry OH, Rosenbrough NJ, Farr AL, Randall Catalan. Chemistry, antioxidant and antimicrobial
RJ. Protein measurement with Folin Wu investigations on essential oil and oleoresins of
reagent. J Biol Chem 1951; 193: 265–275. Zingiber officinale. Food Chem Toxicol 2008; 46:
3295–3302.
10. Drabkin DL, Austin M. Spectrometric studies;
spectrometric constants for common haemoglobin 21. Agrawal M, Walia S, Dhingra S, Khambay BPS.
derivatives in human, dog and rabbit blood. J Insect growth inhibition, antifeedant and
Biol Chem 1932; 98: 719–733. antifungal activity of compounds isolated/derived
from Zingiber officinale Roscoe (ginger) rhizomes.
11. Aebi M. Catalase estimation. In: HV Berg Meyer, Pest Manag Sci 2001; 57: 289–300.
eds. Methods of enzymatic analysis New York,
Verlag, Chemie 1974: 673–684. 22. Ruberto G and Baratta MT. Antioxidant activity
of selected essential oil components in two lipid
12. Moron MA, Depierre JW, Manner Vick B. Levels model systems. Food Chem 2000; 69: 167–174.
of glutathione, glutathione reductase and
glutathione-S-transferase activities in rat liver. 23. Larsen GL, Henson PM. Mediators of inflammation.
Biochim Biophys Acta 1979; 582: 67–68. Annu Rev Immunol 1983; 1: 335–359.
13. Hafeman DG, Sundae RA, Houestra WG. Effect 24. Vinegar R, Traux JF, Selph JL, Jonhston PR,
of dietary selenium on erythrocyte and liver Venable AL, McKenzie RK. Pathway to
glutathione peroxidase in the rat. J Nutr 1974; carrageenan induced inflammation in the hind
104: 580–587. limb of the rat. Fed Proc 1957; 46: 118–126.
14. R a c k e r E . G l u t a t h i o n e r e d u c t a s e ( L i v e r a n d 25. Lo TN, Almeida AP, Beaven MA. Dextran and
yeast). In: Sidney PC, Nathen OK, eds. Methods carrageenan evoke diferent inflammatory
of Enzymology. New York, Academic Press 1955: responses in rat with respect to composition of
722–725. infiltrates and effect of indomethacin. J
Pharmacol Exp Ther 1982: 221: 261–267.
15. Habig WH, Pabst MJ, Jakoby WR. Glutathione-
S-transferase, the first enzymatic step in 26. Greenwald RA. Animal models for evaluation of
mercapturic formation. J Biol Chem 1974; 247: arthritic drugs. Meth Find Clin Pharmacol 1991;
7130–7139. 13: 75–83.
16. Winter CA, Risly EA, Nuss CW. Carrageenan 27. W h e e l e r - A c e t o H , C o w a n A . N e u r o g e n i c a n d
induced edema in hind paw of the rats as assay tissue mediated components of formalin- induced
for anti inflammatory drugs. Proc Soc Exp Bio oedema. Agents Actions 1991; 34: 264–269.
Med 1962; 111: 549–554.
28. T a e s o t i k u l T , P a n t h o n g A , K a n j a n a p o t h i D ,
17. Maity TK, Mandal SC, Mukherjee PK. Studies Verpoorte R, Scheffer JJC. Antiiflammatory,
on anti inflammatory effect of Cassia tora leaf antipyretic and antinociceptive activities of
extract. (fam. Leguminosae). Phytother Res Tabernaemontana pandacaqui Poir. J Ethnopharmacol
1998; 12: 221–224. 2003; 84: 31–35.