19 - Pharmacological evaluation of Anti inflammatory, Analgesic and Antipyretic activity of Bauhinia variegate stem bark

1. 336
_______________________________________________
* Corresponding author: Krishnaveni Janapareddi
E-mail address: krishnavenij153@gmail.com
Available online at www.ijrpp.com
Print ISSN: 2278 – 2648
Online ISSN: 2278 - 2656 IJRPP | Volume 2 | Issue 1 | 2013 Research article
Pharmacological evaluation of Anti inflammatory, Analgesic and Antipyretic
activity of Bauhinia variegate stem bark
*1
Krishnaveni Janapareddi, 2
Swetha Jannapureddy, 2
Manjula Pulluru, 2
Sudheer K
Dundigalla
1
Krishnaveni Janapa reddi, Asst. Professor, University College of Pharmaceutical Sciences,
Kakatiya University, Warangal, AP-506009, India.
2
Care College of Pharmacy, Oglapur (V), Atmakur (M), Warangal Dist. 506 006 A.P.
ABSTRACT
NSAIDs are one of the most widely used categories of drugs presently and there are many clinical situations where
one needs to use them for fairly long periods. As a result of prolonged use, side effects especially gastric ulceration
may worsen the clinical manifestations in the patient. Therefore there is a need to search for analgesics without the
accompanying problems even after a prolonged use. A literature survey of herbs with analgesic and anti-
inflammatory effects led to Bauhinia variegate which was quoted in the Ayurvedic literature as having many
therapeutic activities some of them being analgesic and anti-inflammatory. Lot of research work is reported on this
plant but the stem bark was not evaluated for analgesic and anti-inflammatory activities. A hydro ethanolic extract
was prepared and evaluated for analgesic, anti-inflammatory and antipyretic activities using conventional
pharmacological methods and animal models. The results indicated that the bark extract at a dose of 200mg/kg
showed significant analgesic, anti-inflammatory and antipyretic activities compared to the control group.
Key words: NSAIDs, Bauhinia variegata, analgesic, anti-inflammatory, antipyretic, stem bark
INTRODUCTION
Herbs are widely used in disease therapy as well as
in cosmetics, teas, foods. About 60% of the
pharmaceutical preparations that are in use at
present are plant based. (Rao K and Nigam 1976,
Singh PB and Aswal BS 1992). This indicates the
importance of herbal formulations in disease
therapy and since NSAIDs are one of the widely
used drug categories, a herb with analgesic, anti-
inflammatory and antipyretic activities was
selected for pharmacological evaluation. Bauhinia
variegata (Linn) contains sterols, flavonoids,
saponins and tannins in various parts of the plant
like roots, bark, leaves and flowers (Gupta et al
1980, Gupta AK and Chauhan J 1984) and is
known to possess many pharmacological
properties (Bakhru HC 1997, Balajirao N.S et al
1995) and anti inflammatory, analgesic and
antipyretic properties are among them The non
steroidal anti inflammatory drugs (NSAIDs)
presently available are associated with a number of
side effects like gastric irritation, effect on renal
blood flow in the compromised kidney and a
tendency to prolong bleeding through inhibition of
platelet function (Hardman JG et al 2002). Hence
we have made an attempt to evaluate
International Journal of Research in
Pharmacology & Pharmacotherapeutics

2. 337
Krishnaveni Janapareddi et al / Int. J. of Res. in Pharmacology and Pharmacotherapeutics Vol-2(1) 2013 [336-343]
www.ijrpp.com
pharmacologically these properties of stem bark of
the plant Bauhinia variegate.
MATERIALS & METHODS
COLLECTION & EXTRACTION
The stem bark of Bauhinia variegate was collected
from the campus of Care College of Pharmacy,
Warangal, A.P, India. The plant was taxonomically
identified by Dr. Jaganmohan Reddy, Botanist,
Kakatiya University, Warangal, A.P, India. The
stem bark of Bauhinia variegata was shade dried
and powdered in a mechanical grinder and passed
through sieve No#40 .The coarse powder (1000g)
was extracted with 1 liter of 60% alcohol by
continuous hot percolation using soxhlet apparatus
at 60°c for 12 hr. After completion of extraction,
solvent was removed under reduced pressure and
washed with petroleum ether and dried in
desiccator.
Acute Toxicity studies
The hydro ethanolic extract of the stem bark of
Bauhinia variegatawas subjected to chemical tests
for the identification of chemical constituents and
acute toxicity and gross behavioral studies were
carried out in mice in accordance with OECD
guidelines to determine the test doses. All animal
studies were carried out after the approval of
Institutional Animal Ethical committee. Acute
toxicity test was performed in mice as per OECD
guidelines 2002 in mice.
PHARMACOLOGICAL EVALUATIONS
The extract of Bauhinia variegate stem bark was
studied at doses of 100mg/kg and 200mg/kg for the
analgesic, anti-inflammatory and antipyretic
activities. The extracts and the standard drugs were
given as a suspension by oral route.
Anti-inflammatory activity
Carrageenan- induced paw oedema in rats
(Parmar N.S, Shiv Prakash, 2006)
Wister rats (150-200g) of either sex were
randomized into 4 groups of six each and food was
witheld 10 hours prior to experimentation but free
access to water was allowed. All animals were
administered the preparations by oral route as a
suspension in 2% gum acacia, at doses 100mg/kg
and 200mg/kg. Control group were treated with 2%
gum acacia. Diclofenac sodium at a dose of
30mg/kg was used as standard. Inflammation in the
hind paw was induced in each animal by injecting
0.1ml of freshly prepared carrageenin suspension
(1% w/v) in normal saline into the sub plantar
surface of the rat hind paw. Paw oedema was
measured by wrapping a piece of cotton thread
round the paw and measuring the circumference
with a meter rule (Hess and Miloning, 1972,
Bamgbose and Noamesi, 1981). Measurement was
carried out immediately before and 5 hours
following carrageenan injection. The inhibitory
activity was calculated by the following formula.
Percentage inhibition= (Ct-C0)Control-(Ct-Co) treated X `100
(Ct-C0) Control
(Ct-Co) Average increase in paw circumference at time t
Analgesic activity
Analgesic activity was studied by hot plate
method and Writhing test.
Hot plate method (Hendershot LC., et al.,1959)
Albino mice were placed individually on the hot-
plate, maintained at constant temperature (55°c ±
1°c) for a period of 30 seconds. The reaction time
i.e., time taken by the animal to lick its hind paw or
to leap (turner) out after placing it on the hot plate
(reaction) was recorded. Analgesics increase the
reaction time. Diclofenac sodium 10mg/kg was
given for standard group. Mice were treated with
standard and extract, 60 min prior to placing the
animals on the hot plate. Animals were placed on
the heated surface and the time between placing and
shaking or licking of the paw or jumping was
recorded. An automatic 30s cut-off was used to
prevent tissue damage.The reaction time for mouse
was recorded at 0, 30, 60, 120, and 180 min.
Writhing test (Gerhard Vogel H, 2002)
Male albino mice weighing 20-25g showing
writhing response for acetic acid injection were
selected for this study and divided into four groups
containing six animals in each group; these animals

3. 338
Krishnaveni Janapareddi et al / Int. J. of Res. in Pharmacology and Pharmacotherapeutics Vol-2(1) 2013 [336-343]
www.ijrpp.com
were fasted for 10-12 hours prior to experiment.
The treatment to different groups was as in the
earlier groups, the standard drug being diclofenac
sodium 30 mg/kg.
Mice were injected intra peritonially 0.1ml of 0.6%
acetic acid after 30 min of the above treatment. The
mice were then placed individually into glass
beakers and observed for ten minutes and the
number of writhes were recorded in each animal.
The percentage protection from writhing is
calculated as follows,
% Protection= (Avg writhesin control group – Avg writhes instreated group) X 100
Avg Writhesin control group
Antipyretic activity
2,4 DNP was used in our study to induce pyrexia.
Hyperthermia developed within 30 min of DNP
administration. Male Wister rats weighing 150-
170g were divided into four groups containing six
animals each were treated with 2,4 DNP 10mg/kg
i.p and 30 min later the groups were treated as in
earlier study , the standard drug being Paracetamol
100 mg/kg. The rectal temperatures in the animals
were recorded using Tele- thermometer for a
period of four hours at hourly intervals.
RESULTS
The bark extract at dose of 2000mg/kg was
studied in acute toxicity and no signs of toxicity
and behavioral changes were observed.
Anti-inflammatory activity
The anti inflammatory activity by carrageenan
induced paw odema is shown in table1andfigure1.
The anti-inflammatory effect was found to be
significantly high at 100mg/kg and 200mg/kg
doses of the extract compared to the control group
and the effect at 200mg/kg dose was comparable
with diclofenac sodium 30mg/kg.
Analgesic activity
Hot plate method
Theanalgesic activity by Hot plate method is
shown in table 2 and figure2. The reaction timeis
significantly (P<0.001) increased and dose
dependently when compared to the control group
and the effect was less when compared with
diclofenac sodium 10mg/kg.
Writhing test
The result of analgesic activity by acetic acid
induced writhing test is shown in table 3&4 and
figure 3. The percent protection to writhing is
significantly (P<0.001) high when compared to the
control group and the effect was less when
compared with diclofenac sodium 10mg/kg.
Antipyretic activity
The rectal temperatures in the animals were
recorded using Tele- thermometer for a period of
four hours at hourly intervals were shown in table5
and fig 4.The extract at doses 100&200mg/kg
significantly (P<0.001)reduced the temperature
when compared with standard drug paracetamol
100mg/kg.
Table 1.Anti inflammatory activity
Treatment dose
(mg/kg)
Difference in paw circumference in cm at time points
1hr 2hr 3hr 4hr 5hr
Control 0.93±0.28 1.23±0.16 1.46±0.15 1.01±0.21 0.73±0.15
Diclofenac sodium 30
0.5±0.10***a
0.11±0.07***a
0±0.01***a
0±0.07***a
0.02±0.08**a
Extract 100 0.38± 0.08***a 0.29±0.05***a
0.21±.05***a*b
0.23±0.05***a
0.2±0 .05***a
Extract 200 0.12±0.05***ab
0.1±0.04***a
0.04± 0.10***a
0.06±0.08***a
0.08±0.04***a
n=6, Values are expressed as Mean±SEM.
a: when compared with control, b: when compared with reference standard. (**P<0.01, ***P<0.001)

4. 339
Krishnaveni Janapareddi et al / Int. J. of Res. in Pharmacology and Pharmacotherapeutics Vol-2(1) 2013 [336-343]
www.ijrpp.com
Table 2: Analgesic Effect of Bauhinia variegata bark extract (Eddy’s Hot plate test)
Group Dose mg/kg
Reaction time (sec)
(Mean± SEM)
Control(vehicle) 1ml/100g 3.08± 0.16
Diclofenac sodium 10 14.52± 0.30***a
Extract 100 4.56± 0.37**b
Extract 200 9.61± 0.17***a, b
n=6, Values are expressed as mean±SEM.
a: when compared with control, b: when compared with reference standard. (**P<0.01,***P<0.001)
Table 3: Effect on acetic acid induced writhing in mice.
Treatment dose (mg/kg)
Average number of writhes
30 min 60 min 120 min 180 min
Control 13± 1.67 14± 1.87 14.8± 1.36 12.5± 1.87
Diclofenac sod. 10mg 1.66±1.96***a
3.83±1.72***a
6.0±1.41***a
10.16±1.16***a
Extract 100mg 2.83±2.31***a 8.16±2.48***a
9.66±1.36***a
11.33±3.06***a
Extract 200mg 2.33±2.06***a
6.66±3.32***a
8.0± 2.09***a
10.83±2.31***a
n=6, Values are expressed as mean±SEM, a (***P<0.001) when compared with control
Table 4: Percent Protection of writhing effect
Group
% Protection
30 min 60 min 120 min 180 min
Diclofenac sodium10
mg/kg 87 72 59 23
Extract 100mg/kg
78 41 34 9
Extract 200mg/kg
82 52 45 13

5. 340
Krishnaveni Janapareddi et al / Int. J. of Res. in Pharmacology and Pharmacotherapeutics Vol-2(1) 2013 [336-343]
www.ijrpp.com
Table 5: Effect of hydro-alcoholic extract of bark of Bauhinia variegataon 2,4 DNP induced pyrexia in rats.
Temperature in °C
Treatment dose (mg/kg)
0hr 1hr 2hr 3hr 4hr
Positive Control
35.66±0.81 36.16±0.75 36.83±0.75 37.48±0.51 36.91±0.80
PCM
100mg
37.43±0.68***a
36.11±0.49**a 35.58±0.49**a
35.36±0.81***a
34.53±0.60***a
Extract
100
33.45±0.54***ab
33.0±0.83***ab
31.53±0.47***ab
32.0± 0.34***ab
33.0±0.74***ab*
Extract 200 34.58±0.80***b
33.36±0.58***ab
31.65±0.63***ab
32.4± 0.45***ab
34.55±0.63***a
n=6, Values are expressed as mean±SEM, a: when compared with control, b: when compared with
reference standard. (*P<0.05, **P<0.01, ***p<0.001)
PCM- Paracetamol.
Figure 1: Anti inflammatory activity by paw oedema method
0
20
40
60
80
100
120
1hr 2hr 3hr 4hr 5hr
%inhibitionofpawedema
Time in hr
Diclofenac sodium
Extract 100mg
Extract 200mg

6. 341
Krishnaveni Janapareddi et al / Int. J. of Res.
Figure 2: Analgesic activity by
Figure 3:Analgesic activity of
Figure 4: Percent Protection Vs time plot of writhing effect
0
5
10
15
20
control
Reactiontimeinsec
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
30 min
%Protection
et al / Int. J. of Res. in Pharmacology and Pharmacotherapeutics Vol-2(1) 201
www.ijrpp.com
Figure 2: Analgesic activity by hot plate method
Analgesic activity of Bauhinia variegataby Acetic acid induced writhing method.
Figure 4: Percent Protection Vs time plot of writhing effect
control Diclofenac sod.Extract 100 Extract 200
Eddy's Hot-plate test
30 min 60 min 120 min 180 min
Diclofenac sod.
Extract 100mg
Extract 200mg
Time intervals
2013 [336-343]
by Acetic acid induced writhing method.
Diclofenac sod.
Extract 100mg
Extract 200mg

7. 342
Krishnaveni Janapareddi et al / Int. J. of Res. in Pharmacology and Pharmacotherapeutics Vol-2(1) 2013 [336-343]
www.ijrpp.com
Figure 5 : Anti pyretic activity of Bauhinia variegataby 2, 4 DNP induced pyrexia.
Discussion
A review of the plant Bauhinia variegata
indicates that the various parts of the plant
contain many chemical constituents which may
be responsible for the various medicinal
properties attributed to it.(Pandit CK and Suresh
1992, Jain et al 2004, Kirthikar and Basu 1999).
(Rao et al 2008, Yadava RN and
ReddyVM.2001 Yadava RN and Reddy VM
2003). Present study is conducted to evaluate
bark extract with the activities which were
reported earlier for extract of leaves. The results
proved that the bark of Bauhinia variegate also
contains similar active principles present in
other parts. Anti pyretic activity was
significantly high and isolation of active
principles will potentiate the effect.
Conclusions
The extract of Bauhinia variegate stem bark was
studied at doses of 100mg/kg and 200mg/kg for the
analgesic, anti-inflammatory and antipyretic
activities. The extract exhibited Anti inflammatory
activity reduced inflammation in Carrageenan
induced rat paw oedema model and the effect was
comparable with Diclofenac sodium. The extract
also showed analgesic activity when tested by hot
plate method and acetic acid induced writhing test.
The extract reduced pyrexia induced by 2,4 DNP
and the Anti pyretic activity was significantly
higher than that of Paracetamol.
REFERENCES
Bakhru HC. (1997) Herbs that heal I Orient Longman Ltd, New Delhi, 17
Balajirao N.S, Rajasekhar D, Narayana R and D. Raju.(1995) Ethno-medical therapyamong the
Chenchus of Nallamalai forest of Andhra Pradesh.Biosci. Res. Bull. 11:81-5.
Gerhard vogal H, Drug discovery and Evaluation, pharmacological assays 2nd
edition. Pg 716
Gupta A.K and J.S.Chauhan.( 1984) Constituents from the stem of Bauhinia variegata. NatlAcadSci
Lett.7:174-76.
Gupta A.K, T.J. Vidyapati and J.S. Chauhan.(1980) Chemical examination of the stem of Bauhinia
variegata.Planta Med. 38: 174-76.
Hardman JG, Limbird LE and Gillman GA, (2001) Goodman andGilman’s the Pharmacological Basis of
Therapeutics, Mc-Graw Hill Medical Publishing Division USA, 10th
Edn, pp. 687-707.
28
29
30
31
32
33
34
35
36
37
38
0hr 1hr 2hr 3hr 4hr
positive control
Paracetamol 100mg
Extract 100mg
Extract 200mg
Temperaturein°c

8. 343
Krishnaveni Janapareddi et al / Int. J. of Res. in Pharmacology and Pharmacotherapeutics Vol-2(1) 2013 [336-343]
www.ijrpp.com
Hendershot LC,Forsaith J (1959). Antagonism of the frequency of phenylquinone- induced writhing in the
mouse by weak analgesics and non analgesics.J Pharmacol.Exp Therap.125: 237-240.
Jain R,Nagpal S, Jain S, (2004) Chemical and biological evaluation of bauhinia species.J.Med.Arom.Plant
sci.26.
Kirtikar K.R,B.D.Basu, Indian Medicinal Plants, (International Book Distributor,Dehradun, 1999) pp. 892-901.
Koster R, Anderson M, de Beer EJ (1959).Acetic acid for analgesic screening.Fed proc. 18: 412.
OECD/OECD Guidelines for the testing of chemicals, Revised Draft Guidelines 423; acute oral toxicity- acute
toxic class method revised document, October 2000.
Pandit RK, Suresh K. (1992) Kanchnarguggulu- A critical review.Journal of research and Education in
Indian Medicine11(3): 39-42.
Parmar N.S, Shiv Prakash, 2006 screening methods in Pharmacology, Pg: 213-33.
Rao K., Nigam MC. (1976) Indigenous herbs of potential value having curative properties. Indian drugs, 14(3):
59.
Rao YK, Fang SH, Tzeng YM. (2008) Anti-inflammatory activities of flavonoids and triterpenecaffeate
isolated from Bauhinia variegata. Phytother Res. 22: 957-62.
Singh PB and Aswal. B.S. (1992) Medicinal plants of Himachal Pradesh used in Indianpharmaceutical
industry. Bull. Med. Ethnobot. Res. 13: 172-208.
YadavaRN and V.M. Reddy.(2003) Anti-inflammatory activity of a novel flavonol glycoside from
the Bauhinia variegata Linn. J.Nat.Prod.Res.17 (3): 165-69.
Yadava RN, Reddy VMS, (2001) a new flavone glycoside 5-hydroxyl 7,3‫-׳5,׳4,׳‬tetra-methoxyflavone 5-o-β-D-
xylopyronosyl (1-›2) a L- rhamnopyroanoside from Bauhinia variegata Linn. Journal of Asian Natural Product
Research3: 341-46.
*******************************