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Studies on diuretic and laxative activity of bark extracts of Neolamarckia cadamba (Roxb.)Bosser

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Abstract

The diuretic and laxative activity of various extracts of the barks of Neolamarckia cadamba (Roxb.) Bosser (Family: Rubiaceae) were studied in Wistar albino rats. Furosemide (10 mg/kg, p.o.) and agar-agar (300 mg/kg, p.o.) were used as reference standards respectively for activity comparison. The methanol extract significantly increased the urinary out put as well as urinary electrolyte concentration at the tested dosage regimen that is comparable with the reference standard except in increasing the urinary out put. The chloroform extract produced significant laxative activity. Presence of different phytoconstituents in various extracts of N. cadamba may be responsible for the specific activities
Drug Invention Today Vol.1.Issue 1.November 2009
Mondal S et al. / Drug Invention Today 2009, 1(1),78-80
78-80
Research Article
Available online through
www.ditonline.info
*Corresponding author.
Mondal S
Matushree V. B. Manvar College of Pharmacy,
Dumiyani, Rajkot district,
Gujarat -360 440.India
Tel.: + 91-9375484757
E-mail:logonchemistry@yahoo.co.in
INTRODUCTION
Neolamarckia cadamba (Roxb.) Bosser, syn.
Anthocephalus cadamba var A. chinensis (Family: Rubiaceae)
commonly known as Kadam is a large tree up to 45 m high, fre-
quently found in moist deciduous evergreen forests and widely
distributed through out the greater part of India. The bark is gray,
smooth in young trees, rough and longitudinally fissured in old
trees1. The dried stem bark is used as folk medicine in the treat-
ment of anemia, uterine complaints and for improvement of semen
quality2. The stem bark is also reported to possess astringent,
febrifuge and antiseptic properties and is given in cough3-5. Chlo-
rogenic acid isolated from the leaves has been reported to pos-
sess hepatoprotective activity in vitro and lipid peroxidation in
liver microsomes in vivo6. In our present study, we report diuretic
and laxative activity of different extracts from the bark.
MATERIALS AND METHODS
Plant material
The plant material was collected from the herbal garden
of Regional Plant Research Centre, Bhubaneswar in July 2007 and
Studies on diuretic and laxative activity of bark extracts of Neolamarckia cadamba (Roxb.)
Bosser
Mondal S1*, Dash G. K1, Acharyya A1 , Acharyya S 1 and Sharma H.P2
1Matushree V. B. Manvar College of Pharmacy, Dumiyani, Rajkot district, Gujarat -360 440.
2Sri Balaji College of Pharmacy, Jaipur, Rajasthan – 302 013.
Received on: 22-07-2009; Revised on:01-08- 2009; Accepted on:15-10-2009
ABSTRACT
The diuretic and laxative activity of various extracts of the barks of Neolamarckia cadamba (Roxb.) Bosser (Family: Rubiaceae) were studied
in Wistar albino rats. Furosemide (10 mg/kg, p.o.) and agar-agar (300 mg/kg, p.o.) were used as reference standards respectively for activity
comparison. The methanol extract significantly increased the urinary out put as well as urinary electrolyte concentration at the tested dosage
regimen that is comparable with the reference standard except in increasing the urinary out put. The chloroform extract produced significant
laxative activity. Presence of different phytoconstituents in various extracts of N. cadamba may be responsible for the specific activities
Keywords: Acute toxicity, diuretic, laxative, Neolamarckia cadamba.
identified by the taxonomists of the Botanical Survey of India,
Shibpur, Howrah. A voucher specimen [Sp. No: CNH/ I-I / (255)/
2008Tech.II] has been kept in our research laboratory for further
reference. After authentication, fresh bark material was collected
in bulk, washed, shade dried and pulverized in a mechanical
grinder to obtain coarse powder.
Preparation of extract
The powdered barks (500 g) was extracted successively
with 2 lit each of petroleum ether (40-600 C), chloroform, methanol
and water for 48 h in a soxhlet extractor. Following extraction, the
liquid extracts were concentrated under vacuum to yield dry ex-
tracts. Standard methods were used for preliminary phytochemi-
cal screening of the different extracts to know the nature of
phytoconstituents present within them 7,8. The results are shown
in Table-I.
Animals
Swiss albino mice (20–25 g) of either sex were used for
acute toxicity study and adult Wistar albino rats (150-200 g) of
either sex were used for evaluation of pharmacological studies.
The animals were kept in standard polypropylene cages at room
temperature of 34 ± 2 0C and at 60-65 % relative humidity during
the experimental work. The institutional Animal ethics committee
approved all the experimental protocols (Regd. No. 1212/ac/08/
CPCSEA).
Drug Invention Today Vol.1.Issue 1.November 2009
Mondal S et al. / Drug Invention Today 2009, 1(1),78-80
78-80
Acute toxicity study
The test was carried out as suggested by Ganapaty et
al.,9. Selected animals were divided into different groups of six in
each. The control group received 1% Tween-80 in normal saline
(2 ml/kg, p.o.). The other groups separately received 100, 200,
300, 600, 800, 1000, 2000 and 3000 mg/kg of the test extracts re-
spectively in a similar manner. Immediately after dosing, the ani-
mals were observed continuously for the first 4 hours for any
behavioral changes. They were then kept under observation up
to 14 days after drug administration to find out the mortality if
any.
Diuretic activity
The method of Lipschitz et al., 1943 was employed for
the assessment of diuretic activity 10, 11. In this method, male al-
bino rats weighing between 150-200g deprived of food and water
for 18 hours prior to the experiment, were divided into five groups
of six rats in each. The animal groups were administered orally
either with vehicle (1% Tween-80 in normal saline, 25 ml/kg) The
first group of animals serving as control, received normal saline
(25 ml/kg, p.o.), the second group received furosemide (10 mg/kg,
p.o) in saline12; Group-III, IV, V and VI received different extracts
separately at doses of 300 mg/kg in a similar manner. Immediately
after administration, the animals were placed in metabolic cages
(2 per cage), specially designed to separate urine and faces, kept
at 200 ± 0.50C. The volume of urine collected was measured at the
end of 5 h. During this period, no food and water was made avail-
able to animals. The parameters taken were the body weight be-
fore and after test period, total urine volume, concentration of
Na+, K+ and Cl- in the urine. Na+ and K+ concentrations were de-
termined by flame photometer and Cl- concentration was estimated
by titration with silver nitrate solution (N/50) using 3 drops of 5%
potassium chromate solution as indicator 13-15. The results are
depicted in Table II.
Laxative activity
The test was performed according to method of Bose et
al 16 on rats of either sex, fasted for 12 h before the experiment, but
with water provided ad libitum. The animals were divided into
five groups of six in each. The animal groups were administered
orally with vehicle (1% Tween-80 in normal saline, 25 ml/kg), ref-
erence standard agar-agar (300 mg/kg, p.o.) in saline 9 and doses
of extracts (300 mg/kg) in a similar manner. Immediately after dos-
ing, the animals were separately placed in cages suitable for col-
lection of faces. After 8 h drug administration, the faces were
collected and weighed. Thereafter, food and water were given to
all rats and faecal outputs were again weighed after a period of 16
h (Table III).
Statistical analysis
The data obtained in the studies were subjected to one
way of analysis of variance (ANOVA) for determining the signifi-
cant difference. The inter group significance was analyzed using
Dunnet’s t- test. A p-value less than <0.05 were considered to be
significant. All the values were expressed as Mean ± SEM.
RESULTS
The results of the preliminary phytochemical screening
of different extracts revealed presence of steroids, terpenoids,
flavonoids, tannins, alkaloids, saponins and sugars in the test
extracts (Table-I). In acute toxicity study, it was found that the
chloroform and methanol extract induced sedation, diuresis, pur-
gation and temporary postural defect at all tested doses. How-
ever, there was no mortality in any of the extracts at tested doses
till the end of 14 days of observation.
Diuretic activity
Data represented in Table-II reveal that the methanol ex-
tracts were produced significant increase in excretion of sodium,
potassium and chloride ions at the tested dose level (300 mg/kg,
p.o.), when compared to other extracts. The order of increased
urinary output was found to be methanol extract > aqueous ex-
tract > chloroform extract > petroleum ether extract. However, the
order of increased urinary electrolytes excretion was found to be
higher in the methanol extract.
Laxative activity
Results of the evaluation of laxative activity in Table-III
revealed that the chloroform extract produced significant activity
at the tested dose level (300 mg/kg, p.o.). The order of activity for
the other extracts was methanol extract > aqueous extract > petro-
leum ether extract.
DISCUSSION
Diuretics are useful in reducing the syndrome of volume
overload, including orthopnea and paroxysmal nocturnal dysp-
noea by decreasing plasma volume and subsequently venous re-
turn to the heart (preload). This decreases cardiac workload, oxy-
gen demand and blood pressure 17. Thus, diuretics play an impor-
tant role in hypertensive patients. In present study, the methanol
extract of N. cadamba significantly increased the urinary out put
as well as urinary electrolyte concentration at the tested dosage
regimen that is comparable with the reference standard except in
increasing the urinary out put. Further, the chloroform and metha-
nol extracts were found to be more effective in enhancing urinary
electrolyte concentration for all the three ions tested (Na+, K+, Cl-
). Petroleum ether extract on the other hand did not increase uri-
nary electrolyte concentration. The increase in the ratio of con-
centration of excreted sodium and potassium ions indicates that
the extracts increase sodium ion excretion to a greater extent than
Drug Invention Today Vol.1.Issue 1.November 2009
Mondal S et al. / Drug Invention Today 2009, 1(1),78-80
78-80
potassium, which is a very essential requirement of an ideal di-
uretic with lesser hyperkalaemic side effect.
The laxative activity study revealed significant activity of the
chloroform extract up to 8 h of observation. Simultaneously, the
methanol and aqueous extracts were found to be the less potent
and petroleum ether extract was found to be least active.
Presence of phytoconstituents like flavonoids, terpe-
noids, saponins, have been previously found to be responsible
for diuretic and laxative activities in plants 18-20. The presence of
the said constituents in different extracts of N. cadamba may be
responsible for the observed activities. The exact mechanism ex-
hibited by the extracts can only be established after further in-
vestigation.
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Extract Phytoconstituents present
Pet ether extract Lipids, Steroids, Terpenoids.
Chloroform extract Lipids, Steroids, Terpenoids, Flavonoids, Tannins, Saponins, Alkaloids.
Methanol extract Flavonoids, Tannins, Saponins, Sugars
Aqueous Extract Flavonoids, Tannins, Saponins, Sugars
Table I: Phytochemical screening of different extract of the bark of Neolamarckia cadamba (Roxb.) Bosser
Table II: Diuretic Activity of Different Extracts of Neolamarckia cadamba Barks.
Group Treatment Dose Urine Concentration of ions (mEq/l) Na+/ K+ ratio
Volume(ml) Na+K+Cl-
IControl 25 ml/kg 3.05 ± 079 52.12 ± 2.86 141.72 ± 2.68 87.85 ± 3.88 0.36
II Furosemide 10mg/kg 12.1± 2.28** 108.13 ± 3.71** 187.55 ± 1.98** 164.83 ± 6.89** 0.58
III Pet-Ether extract 300 mg/kg 4.06±0.57 54.30 ± 2.09 143.32 ± 1.83 85.70 ± 2.59 0.37
IV Chloroform extract 300 mg/kg 5.26± 0.92 72.13 ± 1.47** 151.07 ± 3.35 108.87 ± 2.96** 0.48
V Methanol extract 300 mg/kg 10.45± 1.55** 101.05 ± 2.42** 182.55 ± 3.05** 130.61 ± 3.69** 0.55
VI Aqueous extract 300 mg/kg 8.3± 1.61** 67.85 ± 4.22 147.50 ± 2.49 104.95 ± 1.69*0.46
Values are expressed as mean ± S.E. (n = 6). All columns are significant using ANOVA.. * P<0.05, * * P<0.01 when compared to control; Dunnet’s t-test.
Table III: Laxative activity of Different Extracts of Neolamarckia cadamba Barks.
Group Treatment Dose Faecal Output (g)
8h 8-16h
I Control ___ 0.652±0.097 0.385 ± 0.055
II Agar-agar 300mg/kg 1.053±0.046** 0.360 ± 0.041
III Pet-Ether extract 300 mg/kg 0.747±0.075 0.321±0.028
IV Chloroform extract 300 mg/kg 1.035±0.072** 0.336±0.034
VMethanol extract 300 mg/kg 0.909±0.105 0.232±0.031*
VI Aqueous extract 300 mg/kg 0.859±0.029 0.189±0.021**
Values are expressed as mean ± S.E. (n = 6). All columns are significant using ANOVA..* P<0.05, * * P<0.01 when compared to control; Dunnet’s t-test.
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