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Document heading
Analgesic
doi: 10.1016/S1995-7645(14)60275-3
3*
Farhana Tasleem , Iqbal Azhar , Syed Nawazish Ali , Shaista Perveen , Zafar Alam Mahmood
Department of Pharmacognosy, Faculty of Pharmacy, University of Karachi, Karachi, Pakistan
Department of Chemistry, University of Karachi, Karachi, Pakistan
3
Colorcon Limited, Crossways, Victory Way, Kent, Dartford, England
1
2
ARTICLE INFO
ABSTRACT
Article history:
Received 4 Jul 2014
Received in revised form 14 Jul 2014
Accepted 3 Aug 2014
Available online 17 Aug 2014
Objective: To evaluate and compare the analgesic and anti-inflammatory activity of pure
compound, piperine along with hexane and ethanol extracts of Piper nigrum L. fruit in mice and
rats.
Methods: The analgesic activity was determined by tail immersion method, analgesy-meter, hot
plate and acetic acid induced writhing test. While the anti-inflammatory activity was evaluated
by carrageenan-induced paw inflammation in rats.
Results: Piperine at a dose of 5 mg/kg and ethanol extract at a dose of 15 mg/kg after 120 min
and hexane extract at a dose of 10 mg/kg after 60 min exhibited significant (P<0.05) analgesic
activity by tail immersion method, in comparison to ethanol extract at a dose of 10 mg/kg using
analgesy-meter in rats. However, with hotplate method, piperine produced significant (P<0.05)
analgesic activity at lower doses (5 and 10 mg/kg) after 120 min. A similar analgesic activity was
noted with hexane extract at 15 mg/kg. However, in writhing test, ethanol extract significantly
(P<0.05) stopped the number of writhes at a dose of 15 mg/kg, while piperine at a dose of 10
mg/kg completely terminated the writhes in mice. In the evaluation of anti-inflammatory effect
using plethysmometer, piperine at doses of 10 and 15 mg/kg started producing anti-inflammatory
effect after 30 min, which lasted till 60 min, whereas hexane and ethanol extracts also produced a
similar activity at a slightly low dose (10 mg/kg) but lasted for 120 min.
Conclusions: It is concluded from the present study that Piper nigrum L possesses potent
analgesic and anti-inflammatory activities.
Keywords:
1. Introduction
Secondary metabolites are utilized by plants as shielding
molecules and accountable for their biological efficacy
and are distinctly effective for human beings because of
their great influence in healthcare system[1]. Raw materials
produced by plants are the absolute onset of curative
radical and their chemical structures can be used for
the exploration of new compounds[2]. Piper nigrum L. (P.
nigrum) is one of the best known species of Piperaceae
family. It is cultivated in Pakistan, the hills of southwestern India as well as in South America and Africa. It
is robust-woody climbing perennial plant. Piperine is a
major alkaloid constituent of P. nigrum L. The pungency of
*Corresponding author: Dr. Zafar Alam Mahmood, Colorcon Limited, Crossways,
Victory Way, Kent, Dartford, England.
E-mail: zmahmood@colorcon.com
Foundation Project: Supported by the Research & Development Fund of the Dean,
Faculty of Pharmacy, University of Karachi-Pakistan (Grant No. 0852469-15/4/13).
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Farhana Tasleem et al./Asian Pac J Trop Med 2014; 7(Suppl 1): S461-S468
(1%),
Farhana Tasleem et al./Asian Pac J Trop Med 2014; 7(Suppl 1): S461-S468
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2.6. Equipments
Eddys hot plate (Life Science No. 8 Model 39), Ugo Basile
(GP-1)
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3. Results
3.1. Acute toxicity
The pure compound piperine, hexane and ethanol extracts
of P. nigrum L. were evaluated for acute toxicity in mice and
rats by intraperitoneal and oral administration of extracts
at 15, 25, 50 and 75 mg/kg and 15, 50, 100 and 200 mg/kg,
respectively. Results are presented in Tables 1 and 2.
Table 1
Toxicity assessment of P. nigrum L. in mice.
Extracts and
Compound
Piperine
(mg/kg)
15
25
50
P. nigrum
L. (hexane
extract)
P. nigrum
L. (ethanol
extract)
Control
75
15
25
50
75
15
25
50
75
of symptoms
0/6
6/6
6/6
6/6
0/6
6/6
6/6
6/6
0/6
6/6
6/6
6/6
0/6
of mortality
0/6
6/6
6/6
6/6
0/6
6/6
6/6
6/6
0/6
6/6
6/6
6/6
0/6
of mortality
0
100
100
100
0
100
100
100
0
100
100
100
0
(mg/kg)
15
50
100
P. nigrum
L. (hexane
extract)
P. nigrum
L. (ethanol
extract)
Control
200
15
50
100
200
15
50
100
200
of symptoms
6/6
6/6
6/6
6/6
6/6
6/6
6/6
6/6
6/6
6/6
6/6
6/6
6/6
of mortality
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0
0
0
0
0
0
0
0
0
0
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Dose (mg/kg)
0 min
30 min
120 min
Mean
3.6200.170
6.1800.070
7.1400.060
7.7000.130
6.1600.107
2.6120.023
7.1680.248
3.9900.011
11.6580.129
6.3570.102
N/R
1.6220.001
1.7080.008
1.7720.001
**
10
3.5660.003
6.5560.008
0.8540.079
2.0600.083
2.0540.083
2.0740.175
2.4400.123
15
0.2320.294
10
1.3220.103
1.0400.117
15
10
15
0.9900.113
3.0800.018
1.1700.013
**
3.6000.004
**
5.2840.111
0.2860.129
3.7800.030
**
3.0000.095
5.5680.032
2.0600.083
1.7570.082
2.0370.201
4.4000.113
4.8220.133
2.9960.115
4.5960.172
5.1660.003
1.7250.003
4.0300.379
8.2840.207
3.3760.144
2.0200.007
9.1520.024
1.7980.004
2.1270.131
5.0640.070
3.2060.011
**
**
3.9400.045
9.6020.041
3.5190.125
3.8080.015
4.1830.026
n=5. *: P<0.05 significant, **: P<0.01 most significant as compared to control. Reaction time is the time taken by mice to withdraw the tail,
recorded by stop watch in seconds.
Table 4
Analgesic activity of P. nigrum L. tested by analgesy-meter method in rats.
Extracts and Compounds
Dose (mg/kg)
0 min
30 min
60 min
120 min
Mean
10
2.4800.001
8.5720.002
8.6600.002
9.2940.001
7.2500.001
4.6800.023
5.4400.033
8.2800.001
4.6000.050
5.7500.026
6.7000.001
**
9.4000.004
N/R
2.2400.005
3.8000.003
5.0800.006
**
10
2.8000.002
4.5000.008
2.6000.008
4.9000.003
13.0000.006
6.2000.001
8.9000.045
15
10
5.2000.002
2.6000.035
1.8000.006
13.5000.002
1.6000.005
8.4000.068
4.2000.013
10
2.3000.002
15
**
1.5000.003
15
5
**
4.4000.020
**
**
3.4000.005
**
8.7000.011
6.4000.001
**
3.2000.013
12.0000.003
**
**
*
14.3000.007
20.9000.023
9.2000.021
**
8.1000.021
**
4.8000.005
**
3.6300.004
5.6000.005
7.3500.007
**
5.9250.007
**
5.2250.011
6.2000.001
6.3750.015
10.4000.074
9.5000.029
**
2.4000.008
**
2.6000.020
**
**
8.0000.005
**
5.4500.028
n=5,*: P<0.05 significant, **: P<0.01 most significant as compared to control. a: Sensitivity of animals to pain by reaction time recorded in second.
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Table 5
Analgesic activity of P. nigrum L. tested by hot plate method in rats.
Extracts and Compounds
Dose (mg/kg)
0 min
30 min
20
4.3300.030
7.8900.050
3.5000.001
15
2.8780.001
N/R
4.3300.030
3.3900.003
4.4500.001
9.9900.005
5.3320.002
8.7560.006
6.5360.002
6.4120.013
**
1.9380.006
1.5080.004
15
1.9540.006
10
15
1.6760.002
1.1020.020
1.9260.008
1.2440.007
Mean
6.6220.033
12.8700.003
10
120 min
7.0200.015
3.4740.001
10
60 min
7.2500.040
**
**
**
2.0180.016
5.1300.011
**
10.2480.016
2.3820.025
2.6600.053
8.9080.009
**
**
5.6430.003
1.4520.094
1.70800.022
1.7130.033
2.4860.023
2.3560.030
1.3280.132
1.7080.002
1.6460.009
**
4.4040.003
**
1.9740.051
**
5.1780.016
**
9.0400.017
1.3700.145
2.0220.390
4.8400.009
*
*
**
1.7680.057
**
2.1220.022
2.7380.035
2.0090.059
1.9910.115
0.7960.019
1.4390.040
0.7220.037
1.3450.027
n=5,*: P<0.05 significant, **: P<0.01 most significant as compared to control. Reaction time is a licking time in seconds of animals for fore paw.
acid)
Control (acetic acid)
Piperine
Dose
No. of writhes
23.0002.480
0.1
70.6000.000
10
0.0000.000
100.00
0.2000.374
99.71
(mg/kg)
0.2000.374
15
7.0000.205
10
0.2000.374
15
5
10
15
***
0.8000.242
0.4000.178
1.2000.109
0.0000.000
***
% Protection
54.90
99.71
90.08
99.71
98.86
99.43
98.30
100.00
**
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Table 7
Anti-inflammatory activity of P. nigrum L in rats. mL.
Extracts and compounds
Dose (mg/kg)
0 min
Reaction time
30 min
60 min
120 min
Mean
10
0.9700.030
1.2200.060
1.3300.060
1.2900.060
1.2020.052
0.0440.022
0.2540.006
0.4540.004
0.2580.003
0.2520.008
0.3620.001
0.4840.001
0.5480.001
0.5880.001
0.4950.001
N/R
0.1280.045
10
0.0720.007
0.2340.002
15
10
0.3620.007
0.1100.001
15
10
15
0.0400.005
**
0.2820.002
**
**
0.3380.001
**
0.2480.004
**
0.4220.001
0.2280.001
**
0.1040.009
0.3620.001
**
**
0.3500.001
**
0.4700.005
**
0.0520.024
0.1360.001
0.2940.013
0.2840.006
0.3800.001
**
**
**
0.1180.001
**
0.1040.002
0.0560.001
**
0.4840.001
**
**
0.0600.016
**
0.2340.004
**
**
0.4160.004
**
0.4420.003
**
0.0500.170
0.0710.016
**
0.2370.002
**
**
0.3340.002
**
0.3800.004
**
0.1880.049
0.2320.007
0.1490.002
0.2840.006
0.2410.008
**
0.4320.001
**
**
**
0.3500.001
**
**
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