Preliminary Phytochemical
and Anti-pyretic Screening of Crude extract of the leaf of Clerodendrum colebrookianum
K.G.Mahajan1*,
S.Tamilvanan2, H.S.Sawarkar1, R.R.Thenge1,
V.S.Adhao1 and P.S.Gangane1,
1College of
Pharmacy, Malkapur, Dist. – Buldana,
443101
2S.B.College
of Pharmacy, Sivakasi, Dist – Virudunagar,
626130
ABSTRACT
The
current study was focused to evaluate the antipyretic potential of the hexane
extract (HECC) and methanol extract (MECC) of the whole plant Clerodendrum colebrookianum (Family: Verbenaceae).
HECC and MECC on normal body temperature and yeast - induced pyrexia in albino
rats. The whole plant Clerodendrum colebrookianum
was collected from Arvind Herbal lab, Rajapalayam and authenticated. The pulverized plant
material (1000 gm) was extracted successively with hexane and methanol solvents
in a soxhlet apparatus. Adult albino rats of either
sex weighing 180 - 200g were taken for the experiment. Yeast suspension (10 ml
/kg body wt.) increased rectal temperature after 17 hours of subcutaneous
injection. The HECC and MECC at doses of 100mg, 300mg and 500mg / kg body temperature
and yeast - provoked elevated temperature in a dose dependent manner. The
effect also extended up to 5 hours after the drug administration. The
anti-pyretic effect of HECC and MECC was comparable to that of a standard
antipyretic agent paracetamol (150 mg / kg body wt, p.o).
KEYWORDS: Antipyretic
activity, HECC, MECC, Clerodendrum colebrookianum
INTRODUCTION
Clerodendrum colebrookianum
is a shrub, which has foetid smell. It is a common
medicinal plant used for rheumatic pains. Roots with bark are helpful in cough,
scrofulous affections and asthma. Clerodendrum colebrookianum belonging to the family Verbenaceae.1
It is widely distributed throughout
MATERIALS
AND METHODS:
Collection
of Plant Material:
The
plant was collected from Arvind Herbal Lab, Rajapalayam, Tamilnadu. The
authentication of the plant was established by Dr. Stefen,
Dept. of Botany, The American College,
PHYTOCHEMICAL ANALYSIS AND SCREENING OF CLERODENDRUM COLEBROOKIANUM
Table 1: Effect of extract of Clerodendrum colebrookianum
on normal body temperature
|
Treatment mg/Kg
body Wt. |
Rectal
Temperature (°C) before and after treatment |
|||||
|
0
hr |
1
hr |
2
hr |
3
hr |
4
hr |
5
hr |
|
|
Control
( |
37.0 ± 0.2 |
37.3 ± 0.1 |
37.3 ± 0.2 |
37.4 ± 0.3 |
37.2 ± 0.1 |
37.4 ± 0.2 |
|
HECC
100 |
37.2 ± 0.1 |
36.8 ± 0.3 |
36.7 ± 0.2 |
36.6 ± 0.3 |
36.9 ± 0.2 |
37.0 ± 0.2 |
|
HECC
300 |
37.3 ± 0.1 |
36.6 ± 0.2 |
36.4 ± 0.3 |
36.3 ± 0.2 |
36.1 ± 0.1 |
36.7 ± 0.1 |
|
HECC
500 |
37.3 ± 0.2 |
36.2 ± 0.3 |
35.9 ± 0.2 |
35.7 ± 0.2 |
35.6 ± 0.1 |
36.0 ± 0.1 |
|
MECC
100 |
37.4 ± 0.1 |
37.0 ± 0.1 |
35.7 ± 0.2 |
35.5 ± 0.1 |
36.7 ± 0.3 |
36.7 ± 0.2 |
|
MECC
300 |
37.2 ± 0.1 |
36.4 ± 0.2 |
36.3 ± 0.1 |
36.2 ± 0.2 |
36.1 ± 0.1 |
36.6 ± 0.1 |
|
MECC
500 |
37.1 ± 0.1 |
36.9 ± 0.2 |
36.7 ± 0.3 |
36.6 ± 0.1 |
36.6 ± 0.2 |
37.0 ± 0.1 |
Values expressed as
mean ± SEM, n = 6 in each group.
Table 2: Anti-pyretic activity of Clerodendrum colebrookianum
|
Treatment mg/Kg body Wt. |
Rectal Temperature
(°C) after Yeast injection at |
|||||
|
0 hr |
17 hr |
18 hr |
19 hr |
20 hr |
21 hr |
|
|
Control ( |
37.5
± 0.02 |
39.5
± 0.01 |
39.4
± 0.02 |
39.2
± 0.03 |
39.0
± 0.01 |
39.4
± 0.02 |
|
Paracetamol 150 |
37.6
± 0.03 |
39.5
± 0.02 |
38.4
± 0.01 |
38.1
± 0.01 |
37.7
± 0.02 |
37.6
± 0.01 |
|
HECC 100 |
37.3
± 0.01 |
39.5
± 0.01 |
39.3
± 0.03 |
38.7
± 0.02 |
38.4
± 0.01 |
37.8
± 0.01 |
|
HECC 300 |
37.5
± 0.03 |
39.6
± 0.02 |
38.7
± 0.02 |
38.3
± 0.01 |
37.8
± 0.02 |
37.0
± 0.03 |
|
HECC 500 |
37.7
± 0.02 |
39.6
± 0.01 |
38.8
± 0.03 |
37.8
± 0.02 |
37.4
± 0.03 |
37.4
± 0.01 |
|
MECC 100 |
37.6
± 0.01 |
39.6
± 0.02 |
39.3
± 0.03 |
38.5
± 0.02 |
37.9
± 0.03 |
34.4
± 0.02 |
|
MECC 300 |
37.5
± 0.02 |
39.6
± 0.01 |
37.7
± 0.02 |
37.4
± 0.01 |
36.9
± 0.02 |
33.9
± 0.01 |
|
MECC 500 |
37.4
± 0.01 |
39.7
± 0.03 |
37.5
± 0.01 |
36.8
± 0.01 |
35.9
± 0.02 |
33.9
± 0.01 |
Values expressed as
mean ± SEM, n = 6 in each group
Preparation
of Extracts:
The dried
and pulverized plant material (1000gm) was subjected to hot continuous
extraction with hexane (63 – 700C) and methanol (950C) as
solvents respectively in a soxhlet extractor. After
each extraction, the solvent was removed in vacuo in
a rotary evaporator to provide dry extracts. The dried mass was kept in a
refrigerator and was used, as and when required for the experiment. The
percentage extractive values of hexane and methanolic
extracts was found to be 8.58% and 5.2% respectively. The extracts were
subjected to preliminary qualitative tests in order to identify the various phytoconstituents present in plants.6 From
preliminary phytochemical screening, both the
extracts showed the presence of steroids, flavanoids,
triterpenoids, phenols and coumarins
which were further confirmed by thin-layer chromatography study.
Animals:
Adult
albino rats (wistar strain) of either sex weighing
180 - 200 g were used. The animals were maintained under suitable nutritional
and environmental conditions through out the experiment. The animals were
maintained under standard laboratory condition for an acclimatization period of
seven days prior to performing the experiment.
Study on
normal body temperature:
Rats of either
sex were divided into seven groups comprising six in each group for this
experiment. The body temperature of each rat was measured rectally at
predetermined intervals before and for 5 hours after administration of either
2% aqueous tragacanth solution (control) or HECC and
MECC at doses of 100, 300 and 500 mg/kg body weight orally.
Induction
of yeast induced pyrexia:
Rats were
divided into eight groups of six rats each. The normal body temperature of each
rat was measured rectally at predetermined intervals and recorded.7
Fever was induced as per the method described.8 The rats were
trained to remain quiet in a restraint cage. A thermister
probe was inserted 3-4cm deep into the rectum and fastened to the tail by
adhesive tape. The temperature was measured on a thermometer. After measuring
the basal rectal temperature, animals were given a subcutaneous injection of
10ml/kg body wt of 15% w/v yeast suspended in 0.5% w/v methyl cellulose
solution. Rats were then returned to their housing cages. After 17 hours of
yeast injection, the animals were again restrained in individual cages for the
recording of their rectal temperatures as described previously.
Drug
administration:
After 17
hours of yeast injection, the HECC and MECC were administered orally at doses
of 100, 300 and 500 mg / kg body wt. to six groups of animals respectively. A
similar volume (5ml/kg body wt) of 2% aqueous tragacanth
solution was administered orally to the control group of animals. The eighth
group of animals received the standard drug paracetamol (150mg/kg body wt.)
orally. Rats were restrained for recording of their rectal temperatures at the
seventeenth hour, immediately before HECC or MECC or paracetamol administration
and again at one hour's interval up to twenty - first hour after yeast
injection.
Statistical
Analysis:
Data was
expressed as mean ± standard error means. Statistical analysis was made by
using Kruskal Wallis (non - parametric) Anova test at different time intervals.9-11
RESULTS:
Effect of
the HECC and MECC on normal body temperature in rats was presented in Table 1. It was found that the HECC and
MECC at doses of 100 mg / kg body wt. caused significant lowering of body
temperature up to 4 hours following its administration. This effect was maximal
at doses of 300 and 500 mg/kg body wt. in a dose dependent manner and it caused
significant lowering of body temperature upto 5 hours
after its administration. The subcutaneous injection of yeast suspension
markedly elevated the rectal temperature after 17 hours of administration.
Treatment with HECC and MECC at doses of 100, 300 and 500 mg/kg body wt
decreased the rectal temperature of the rats in a dose dependent manner. The
antipyretic effect started as early as 1 hour, and the effect was maintained
for 4 hours, after its administration. The standard drug paracetamol at 150
mg/kg body wt. dose significantly reduced the yeast provoked elevation of body
temperature. The results obtained from the standard drug treated and HECC, MECC
treated rats were compared with the control (2% aqueous tragacanth
solution) group and we observed a significant reduction in the yeast - elevated
rectal temperature (Table 2).
DISCUSSION:
Fever may
be a result of infection or one of the sequelae of
tissue damage, inflammation, graft rejection or other disease states.
Antipyretics are drugs which reduce elevated body temperature. Regulation of
body temperature requires a delicate balance between the production and loss of
heat, and the hypothalamus regulates the set point at which body temperature is
maintained. In fever this set point is elevated and drugs like paracetamol do
not influence body temperature when it is elevated by factors such as exercise
or increases in ambient temperature.12 The present results shows
that the HECC and MECC possesses a significant antipyretic effect in yeast
provoked elevation of body temperature in rats, and its effect is comparable to
that of standard drug paracetamol. Furthermore, the HECC and MECC also
significantly reduced the normal body temperature, and this is to be studied
further for the exact mechanism of action.
CONCLUSION:
The
antipyretic activity of Clerodendrum colebrookianum
supports its use in the traditional medicine to reduce fever but further
studies are needed to elucidate the exact mechanism by which Clerodendrum colebrookianum
plant extract exerts the antipyretic effect.
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T.Pullaiah, Encyclopaedia of World Medicinal Plants; P.593.
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Gupta RK, Chandra S, Mahadevan
V. Indian J Pharmacy 1967; 29: 102.
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Subramanian SS, Nair AGR. J Indian Chem Soc 1972; 49: 1069.
5.
Harborne JB. Phytochemical methods: A guide to modern techniques of
plant analysis.
6.
Trease GE, Evans
WC. Text Book of Pharmacognosy. 12th Edn,
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Murugesan T, Mandal SC, Bhakta T, Das T, Pal
M, Saha B.P., Phytomedicine
2000; 7(3) : 231.
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Kulkarni SK. Hand
Book of Experimental Pharmacology. 2nd Edn,
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Sundar Rao PSS, Richard J. An Introduction to Biostatistics. 3rd Edn.
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Goodman, Gilman. The Pharmacological Basis of
Therapeutics. 9th Edn,
Received on
01.09.2009
Accepted on
10.10.2009
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Publication all right reserved
Research
Journal of Pharmacognosy and
Phytochemistry. 1(3): Nov. – Dec. 2009, 191-193