Analgesic and antipyretic activity of methanolic extract of Acacia leucophloea bark

1. 390
_____________________________________
* Corresponding author: R.Thirumurthy
E-mail address: priyajoshikumar@gmail.com
Available online atwww.ijrpp.com
Print ISSN: 2278 – 2648
Online ISSN: 2278 - 2656 IJRPP | Volume 2 | Issue 2 | 2013 Research article
Analgesic and antipyretic activity of methanolic extract of Acacia leucophloea bark
*1
R.Thirumurthy, 2
J.Priya
1
Department of pharmaceutical chemistry S.S.M.College Of Pharmacy, Jambai, Erode, Tamilnadu.
2
Department of pharmaceutical chemistry SSRCP College Of Pharmacy, Mahabubnagar, Andrapradesh.
Abstract
The pharmacological and chemical constituents of plant from Acacia leucophloea which is widely used in folk medicine.
The aim of present study is to investigate the analgesic and anti pyretic activity of Methanolic extract of Acacia leucophloea
bark. The analgesic activity was found out by Eddys hot plate by using the standard drug Pentazocine. The anti pyretic
activity was found out by yeast induced pyrexia method by standard drug paracetamol. The preliminary phytochemical
screening showed the presence of glycosides, alkaloids, phytosterols, saponins, fixed oils and fats, flavonoids
and coumarins are present in extracts.
Keywords: Acacia leucophloea, antipyretic, analgesic activity.
Introduction
Acacia leucophloea, synonyms alba wild family
(Mimosaceae). It finds throughout the dry region area
especially in Punjab, Madhya Pradesh and
Tamilnadu. It is a moderate sized tree upto 30m in
height with spreading branches, crooked and gnarled
stems, white spines and pale yellowish grey to nearly
white bark with pale red inside (Orient Longman,
1995) the leaves are Bipinnate, 2.5-5 cm long, main
rachis pubescent with a cup shaped gland between
each pair of pinnae, 5-15 pairs of pinnae of 12- 30
pairs linear – oblong, obtuse (Orient Longman ,1995)
Flowers in large terminal tomentose panicles, heads
numerous, globose. The fruits are sessile, thin, flat,
slightly covered with pale brown tomentum (Orient
Longman,1995). The seeds are 10-20 per pod.
(Orient Longman,1995). The medicinal uses of the
bark is astringent, bitter, thermogenic, styptic,
alexeteric, anthelmintic, vulnerarys demulcent,
constipating, expectorant and antipyretic. It is useful
in vitiated conditions of kapha, cough, inflammation,
wounds, skin diseases, leukoderma, leprosy, pruritus,
vomiting, diarrhoea, dysentery, internal and external
hemorrhages, dentalcaries,oral ulcer, stomatitis and
fever (Orient Longman, 1995). Various parts are used
in ayurveda medicine.
Materials and methods
The plant Acacia leucophloe Willd. belonging to
family Mimosaceae are grown widely throughout
India. The plants were collected from Bhavani, Erode
district, Tamilnadu, India in the month of July 2009
and were authenticated by Mr. G.V.S. Murthy, Join
Director, comparing it with authentic specimen at
botanical survey of India, Ministry of Environment
and Forests, Government of India, Coimbatore (No.
BSI / SC /5 / 23 /09- 10 Tech. 438).voucher
specimens have been kept in our laboratory.
International Journal of Research in
Pharmacology & Pharmacotherapeutics

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Extraction
The bark of Acacia leucophloea was collected and
dried under shade and then made into a coarse
powder with a mechanical grinder. The powder was
passed through sieve No .60 stored in air tight
container for further use. About 650gm of coarsely
powdered bark was extracted with petroleum ether at
40-60
o
C, by continuous hot percolation, using
soxhlet apparatus. The extraction was carried out by
using solvents of increasing polarity starting from
petroleum ether and chloroform respectively. The
extraction was continued for 72 hrs. The petroleum
ether extract was filtered and concentrated to a dry
mass by using vacuum distillation. A dark green
residue was obtained (3.5gm). The mark left, after
petroleum ether extraction was taken out, and
subsequently extracted with chloroform for 72 hrs.
The chloroform extract was then filtered and
concentrated to the dry mass. A dark brownish green
residue was obtained (3.8 gm).The mark left after the
chloroform extract was taken out, and subsequently
extracted with methanol upto 72 hrs. The methanol
extract was then filtered and concentrated to a dry
mass. A dark green colored residue was obtained
(20.2 g).
Preliminary phyto chemical investigation
The preliminary phyto chemical studies were done in
the extracts of the bark of acacia leucophloea for the
presence of carbohydrates, glycosides, proteins,
fixed oils & fats, flavanoids, gums & mucilages,
phytosterol, saponins and alkaloids were present. The
phytochemical constituent present in the plant was
shown in the table.no. 1
Table no. 1. Qualitative phytochemical analysis of extracts of A. leucophloea
S.No TESTS
PET.ETHER
EXTRACT
CHLOROFORM
EXTRACT
METHANOL
EXTRACT
1 Carbohydrates _ + +
2 Glycosides _ _ +
3 Proteins _ _ +
4 Fixed oils & fats _ _ _
5 Gums & mucilages _ _ _
6 Alkaloids _ + +
7 Phytosterols + + +
8 Flavonoids _ + +
9 Phenolic compounds _ + +
10 Saponins _ _ +
( + ) = indicates presence. (−) = indicates absence.

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Pharmacological studies
Acute oral toxicity
The procedure was followed by using OECD
guidelines 423 (Acute toxic class method). The acute
toxic class method is a step wise procedure with 3
animals of single sex per step. The method uses
defined doses (5, 50, 300, 2000 mg / kg body weight)
and the results allow a substance to be ranked and
classified according to the Globally Harmonised
System (GHS) for the classification of chemical
which cause acute toxicity.
Procedure
Twelve animals (Albino mice, 20-25gm) were
selected for studies. The starting dose level of
methanolic extracts of bark of A. leucophloea was
2000 mg/kg body weight p.o. Most of the crude
extracts possess LD50 value more than 2000 mg/kg of
the body weight of animal used. Death was observed
at the dose level of 2000 mg /kg body weight. The
observed toxic signs at the dose level of 2000 mg / kg
was Increase in the lachrymal secretion, miosis,
tremor, change in eye colour from pink to dark red,
sedation, decrease in loco motor activity, loss in
touch sensation, increase in heart rate and respiratory
rate. So as per down procedure in OECD we
administered 1000 mg /kg to observe the toxic sign
once again. The dose level of 1000 mg /kg body
weight does not produce any toxic signs as earlier.
Hence, the biological dose was fixed 100 & 200 mg/
kg body weight for the extract.
Analgesic activity
Analgesic activity was performed using eddy’s hot
plate (Inco,India) maintained at a temperature of 55±
1˚C. (Ghosh MN. Fundamentals of experimental
pharmacology 3rd edition). Healthy Wistar rats of
both sexes weighing 150-200g were used for the
experiment. The animals were housed at 25±2˚C
under a 12 hour light/12 h dark cycle and has free
access to standard pellet diet (Purina chow) and tap
water. Each animal was used once in an experiment.
The basal reaction time of all animals toward thermal
heat was recorded. The animals which showed for
paw licking and jumping response with in 6 to 8
seconds were selected for the study. Paw licking
response was recorded. Two doses of methanolic
extract of A. leucophloea (100 and 200 mg/kg, p.o.)
significantly increased the latency time in a dose-
dependent manner. At 200mg/kg the methanolic
extract of A. leucophloea markedly increased the
response latency, the highest inhibition of response
latency was exhibited at a higher dose of 200 mg/kg
of and the maximum effect of A. leucophloea was
observed at 60 min after drug administration.
Pentazocine (5 mg/kg, p.o.) also significantly
increased the response latency time with a maximum
protective effect.
Table no. 2. Effect of methanol extract of A. leucophloea on eddy’s hot plate
Group
Reaction time in Seconds
0 min 30 min 60 min 120 min 180 min
Control 4.74±0.04 4.74±0.08 4.59±0.10 4.82±0.04 4.67±0.06
Standard
(pentazocine)
4.65±0.06 4.95±0.11 5.73±0.03c
7.41±0.13 c
5.4±0.02 c
Sample-I
(100 mg)
4.69±0.06 5.61±0.02 c
7.37±0.03 c
9.46±0.11 c
7.58±0.03 c
Sample-II
(200 mg)
4.60±0.04 5.65±0.04 c
8.74±0.05 c
11.62±0.08 c
8.6±0.11c
Comparision between O min and other time value. a
= P<0.05, b
= P< 0.01, c
= P<0.001 N=6
*P<0.05 vs control, Data were expressed as mean ± SEM

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Antipyretic activity
The antipyretic activity of the extracts was screened
by using yeast induced hyperpyrexia method. Wistar
rats of either sex weighing between 150-200 gm were
selected and divided into seven groups each having
six animals. They were maintained at a constant
temperature of 24-25
0
for 24 hr. before pyrexia was
induced by S.C injection of 2 ml of 15% brewer’s
yeast suspension in saline solution. After 18 hrs of
yeast injection, the extracts were suspended with
1%CMC and administered orally. Paracetamol (200
mg/kg) was used as a standard drug for comparison
of antipyretic activity .Rectal temperatures were
noted at 30 min intervals. The methanol extract of
A.leucophloea was suspended in 2 % CMC. This
suspension was administered orally to the animals, at
the dose of 100 and 200 mg/kg body weight doses,
respectively. Paracetamol (100 mg) was administered
orally to the animals with the help of an intra gastric
catheter, at the dose of 100 mg/kg body weight. The
rats were kept fasting for a period of 48 h prior to the
experiment and were divided into four groups of six
animals each. The antipyretic activity of the extract
was evaluated using Brewer’s yeast induced pyrexia
(Vimala, et al., 1997). Fever was induced by
injecting subcutaneously, 2 ml/kg body weight of
20% w/v aqueous suspension of Brewer’s yeast in
normal saline and rectal temperature was recorded by
clinical thermometer. Prior to the experiment, the rats
were maintained in separate cages for 7 days and the
animals with approximately constant rectal
temperature (38.0 -38.5 C) were selected for the
study. The experimental pyrexia rats showed a mean
increase of 2 C in rectal temperature at 20 h, after
Brewer’s yeast injection. Group I animals were
served as control and received 2% sodium CMC.
Group II received the standard paracetamol at 100
mg/kg body weight. Groups III and IV received the
methanol extract of A. leucophloea bark at 100 and
200 mg/kg body weight doses, respectively. The fall
in rectal temperature was recorded after 1 h interval
for 4 h, using a digital thermometer. Rectal
temperature was observed using Tele- thermometer at
30 min interval upto 180 min (Kang et.al. 2008).
Results and discussion
The analgesic property of A. leucophloea can also
probably be due to the blockade of the effect or the
synthesis and/or release of PGs and/or other
endogenous substances that excite pain nerve endings
(Juan, 1979), Although the mechanism of action of
this effect was not proved in this study. This effect is
very useful in painful hemorrhoid. The present study
proved the traditional use of A. leucophloea as an
analgesic drug in folk medicine.
Analgesic activity of
Acacia leucophloea bark extract
Normal 100 mg 200 mg Pentazocaine
0
1
2
3
4
5
6
7
8
9
10
11
12
0 min
30 min
60 min
120 min
180 min
c
c
c c
c
c
c
c
c
c
c
Groups
Values

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ANTIPYRETIC ACTIVITY
Brewer’s Yeast Induced Pyrexia Method
The fever was evaluated using Tele thermometer
method. Administration of 20 % of 2 ml /kg of
Brewer’s Yeast increases the body temperature to 0.3
to 0.5 ˚C. The methanolic extract of A. leucophloea
decreases the body temperature in a dose dependent
manner. The extract at the dose of 200 mg / kg
reduces the body temperature significantly in the
fever induced group compared with the 100 mg / kg
of MEAL. The standard drug paracetamol also
decreases the body temperature significantly. The
comparison of the antipyretic activity was shown in
the table – 3
Table no. 3. Effect of methanol extract of Bark of A. leucophloea on brewer’s yeast induced
pyrexia in rats.
Group
Tim e in seconds
Basal body
Tempareture
0 hrs 1 hrs 2 hrs 3 hrs 4 hrs
Control 37.38 ±0.10
39.41
±0.22
39.33
±0.19
39.20
±0.22
39.10
±0.13
39.03
±0.14
Standard
100mg/kg 37.51 ±0.10
39.68
±0.15
38.76
±0.11a
38.21
±0.10b
37.58
±0.13c
37.53
±0.04c
MEAL
(100 mg ) 37.42 ±0.05
39.36
±0.09
39.06
±0.06
38.38
±0.11a
38.15
±0.09c
37.85
±0.02c
MEAL
(200 mg) 37.46 ±0.08
39.26
±0.26
38.48
±0.13b
38.30
±0.11b
37.96
±0.06c
37.71
±0.07c
*Values are expressed as mean ± SE, (n = 6), a
P<0.05, b
P<0.01 and c
P<0.001 between control and treated groups
In addition, all the extracts derived from the bark
of A. leucophloea showed anti-pyretic activity in
mice made hyperthermic by dried yeast injection.
The response of methanol extracts was almost
comparable to that of paracetamol.
Antipyretic activity of MEAL
BBT 0 min 1 hr 2 hr 3hr 4 hr
0
10
20
30
40
c o n tro l
S td 1 0 0 m g
M E A L 1 0 0 m g
M E A L 2 0 0 m g
a ab b b c c c
c
c c
H r s
Temperature

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