Pharmacology & Pharmacy, 2013, 4, 473-477
http://dx.doi.org/10.4236/pp.2013.46068 Published Online September 2013 (http://www.scirp.org/journal/pp)
473
Evaluation of Some Biological Activities of Albizia lebbeck
Flowers
Mohamed Farag1*, Ali El Gamal1,2, Ashraf Kalil1,2, Adnan Al-Rehaily1, Omar El Mirghany3,
Kamal El Tahir3
1Pharmacognosy Department, Faculty of Pharmacy, King Saud University, Riyadh, KSA; 2Pharmacognosy Department, Faculty of
Pharmacy, Mansoura University, Mansoura, Egypt; 3Pharmacology Department, Faculty of Pharmacy, King Saud University, Riyadh,
KSA.
Email: *dr.farag@gmail.com
Received May 17th, 2013; revised June 21st, 2013; accepted July 5th, 2013
Copyright © 2013 Mohamed Farag et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
ABSTRACT
Reviewing the current literature for the importance of the plant Albizia lebbeck L. growing worldwide revealed
many biological interests. However, the species growing in Saudi Arabia has not received due attention. The pre-
sent study was undertaken to study antipyretic, analgesic, estrogenic and anti-inflammatory activities of five different
fractions from successive extraction of Albizia lebbeck flowers: n hexane, dichloromethane, ethyl acetate, n-butanol as
well as the 70% total alcohol. The flowers showed reasonable antipyretic, analgesic, estrogenic and anti-inflammatory
activities.
Keywords: Herbal Medicine; Albizia lebbeck; Anti-Inflammatory Effect; Antipyretic Effect; Analgesic Effect;
Estrogenic Effect
1. Introduction
The genus Albizia (Fabaceae) comprises approximately
150 species, mostly trees and shrubs native to tropical
and subtropical regions of Asia and Africa [1]. Albizia
lebbeck was imported many years ago from India and
well adapted to the harsh environmental conditions of the
central part of Saudi Arabia. The current literature re-
vealed that some plants of the genus Albizia have great
medicinal values. A. lebbeck is a tree well known in the
Indian subcontinent for its range of uses. A. lebbeck is
used in Indian folk medicine to treat several inflamma-
tory pathologies such as asthma, arthritis and burns [2]. A.
lebbeck inhibited the passive cutaneous anaphylaxis and
mast cell degranulation in rat. In addition, it could protect
the sensitized guinea pig from antigen induced anoxic
convulsion [3]. Recently, it was found that the alcoholic
extract of A. lebbeck has antihistaminic property, by neu-
tralizing the histamine directly or due to corticotrophic
action as evidenced by raising cortisol levels in plasma
[4]. Moreover, Saponins of A. lebbeck have been claimed
to be useful in treatment of Alzheimer’s and Parkinson’s
diseases [5]. Leaves have been claimed to have anticon-
vulsant activity [6] and nootropic effect [7] which may
be due to the presence of certain important compounds
like alkaloids and flavanoids. Moreover, the aqueous
extract of A. lebbeck leaves showed antioxidant activity
in diabetic rats [8]. The saponins of the seeds of A. leb-
beck exhibited antiovulatory properties. The seeds had
anti-fertility effect on male rats [9] and antidiarhoeal
activity studied on conventional rodents models of diar-
rhea [10]. In addition, the flowers are being commonly
used to treat anxiety, depression and insomnia in tradi-
tional Chinese medicine [11]. In this study, we report
evaluation of antipyretic, analgesic, estrogenic and anti-
inflammatory activities of different extracts of A. Leb-
beck flowers.
2. Materials and Methods
2.1. Plant Material
The air-dried flowers of A. lebbeck were collected from
Riyadh district, Saudi Arabia in Spring 2008. The plant
was identified and kindly authenticated by Professor Dr.
Ahmad Alfarhan, Department of Botany, College of
Science, King Saud University. A voucher specimen was
deposited at the Pharmacognosy Department, College of
*Corresponding author.
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Evaluation of Some Biological Activities of Albizia lebbeck Flowers
474
pharmacy, King Saud University.
2.2. Extraction and Isolation
The air-dried flowers were ground to a coarse powder. A
sample of 700 gm was soaked in 70% ethanol for 3 days
with occasional shaking. This process was repeated four
times until complete exhaustion. The alcoholic extract
was then concentrated to dryness under reduced pressure
at 40˚C using a rotary vacuum evaporator. The crude
dried alcoholic extract (95 gm) was then liquefied in wa-
ter-alcohol mixture (20:80) and successive extracts were
prepared using shaking in separating funnel with n-hex-
ane, dichloromethane, ethyl acetate and n-butanol, re-
spectively.
2.3. Animals
Healthy male adult Swiss albino mice, weighing between
20 - 25 g and albino Wistar rats were obtained from the
Experimental Animal Care Centre, College of Pharmacy,
King Saud University, Riyadh. The conduct of experi-
ments and the procedure of sacrifice (using ether) were
approved by the Ethics Committee of the Experimental
Animal Care Society, College of Pharmacy, King Saud
University, Riyadh, Saudi Arabia.
2.4. Screening for Antipyretic Activity
Six male albino mice, weighing 20 - 25 g, were fasted
overnight before the experiments. Pyrexia was induced
by a subcutaneous injection of 20% w/v brewer’s yeast
suspension (10 ml/kg) into the animal’s dorsum region.
Seventeen hours after the injection, the rectal tempera-
ture of each mouse was measured using a digital ther-
mometer. Only mice that showed an increase in tem-
perature of at least 0.7˚C were used for experiments. A
dose of 1 g/kg of each extract, except n-butanol extract
which was administered in dose of 0.25 g/kg, was ad-
ministered intraperitoneally, and the temperature meas-
ured at 15, 30, 60, 120 and 180 minutes after injection.
Water ad libitum was used as a negative control and as-
pirin (200 mg/kg) was used as a positive control [12].
2.5. Screening for Analgesic Activity—Hot Plate
Method
Male albino white mice, weighing 20 - 25 g, were used.
They were gently placed on a hot plate thermostatically
maintained at 55˚C [13]. The time in sec at which the
animals displayed nociceptive responses exhibited as
licking of the front paws or fanning (blowing) the hind
paws was recorded and the animals were removed from
the plate. A cut-off time of 40 sec was used to avoid
damage of the paws. Crude extracts (500 mg/kg) and a
reference analgesic drug, aspirin (200 mg/kg) were ad-
ministered (i.p) [14].
2.6. Screening for Estrogenic Activity
Two groups each of three immature female albino rats,
weighing 100 g, were housed under a temperature and
light controlled room. They were maintained in a well
ventilated animal quarter. They had free access to water
and commercially available food. To determine estro-
genic activity, a dose of 500 mg of each crude extract/
kg/day was administered intraperitoneally in 1 ml saline
for 3 consecutive days and on the 4th day animals were
sacrificed and uteri were removed and weighed. Normal
saline was used as a negative control and 17-ß-estradiol
(0.32 μg/animal/day) was used as a positive control [15].
2.7. Screening for Anti-Inflammatory
Activity—Carrageenan-Induced
Inflammation
Carrageenan-induced rat hind paw oedema was induced
following the method of [16]. Initially the volumes of the
hind paws of male Wistar rats, weighing 150 - 200 g,
were measurd using hydroplethysmometer (Model 7150,
Ugo Basile, Caemerio, Italy). For this purpose each paw
was marked at the level of the lateral malleoulus and then
dipped gently into the 0.45% NaCl fluid in the chamber
of plethysmometer. This instrument measures the volume
of the paw in ml. Then 0.1 ml of carregeenin (2% w/v in
sterile saline) was injected in one paw under the planter
aponeurosis. The paw volume was then measured hourly
for 4 - 5 hours. Inflammation (or oedema) was expressed
as volume (in ml) increase above the original volume of
the paw or as a percentage increase in the paw volume.
Animals were then injected (i.p) with various doses of
the crude extracts 60 min. before injection of carrageenan.
Then carrageenan was injected as described above and
the paw volume determined hourly for 4 - 5 hours there-
after. Diclofenac sodium was used as standard drug at
concentration of 20 mg/kg of body weight. The influence
of the treatment on the induced inflammation was evalu-
ated [14].
2.8. Statistical Analysis
Values are given as arithmetic means ± standard devia-
tion of the mean (S. D. M.). Data was statistically ana-
lyzed by using the Student’s t-test or ANOVA as appro-
priate. Significance with respect to control group.
3. Results
3.1. Antipyretic Activity
The effects of the different extracts administered at doses
of 1 g/kg except the n-butanol extract which was admin-
istered at a dose 0.25 g/kg (i.p) are shown in Table 1. All
of the treatments decreased the body temperature sig-
nificantly. The maximum decrease of 8˚C was shown by
the dichloromethane extract.
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Evaluation of Some Biological Activities of Albizia lebbeck Flowers 475
Table 1. Results of antipyretic study.
A. lebbeck Extract Basal body
temperature
Temperature
after 90 min
Decrease in
temperature
(˚C)
n-butanol (0.25 g/kg) 37.1 34.8 2.3 ± 0.2*
Dichloromethane (1 g/kg) 36.6 28.6 8.0 ± 0.4*
Ethyl acetate (1 g/kg) 36.6 31.6 5 ± 0.9*
Total alcohol (1 g/kg) 36.8 32.1 4.7 ± 0.2*
Aqueous (1 g/kg) 36.6 33.9 2.7 ± 0.9*
n-hexane (1 g/kg) 37.2 35.5 1.7 ± 0.2*
Aspirin (200 mg/kg) 37.2 34.1 3.1 ± 0.7*
The values marked with asterisks differ significantly from the control group,
(*p < 0.05), Number of animals = 5.
3.2. Analgesic Activity
The effect of the different extracts of A. Lebbeck on pain
sensation was tested using hot plate method [13]. Ad-
ministration of the different extracts at doses of 1 g/kg I.p,
except n-butanol extract which was administered in dose
of 0.25 g/kg, induced variable increases in the pain
threshold in the hot plate test. The percentage increases
of pain threshold are shown in Table 2. A reference an-
algesic drug, aspirin (200 mg/kg) was administered as a
positive control [14].
3.3. Estrogenic Activity
Table 3 shows the effects of different extracts of A. leb-
beck on the uteri of immature rats. Normal saline was
used as a negative control and 17-ß-estradiol (0.32 μg/
animal/day) was used as a positive control [15].
3.4. Anti-Inflammatory Activity
Administration of the different A. lebbeck inflorescences
extracts to rats in doses of 1 g/kg I.P (except the n-bu-
tanol extract 0.25 g/kg) with experimental carrageenan-
induced inflammation suppressed inflammation to vari-
ous degrees. The best anti-inflammatory activity was
observed 2 hours after administration of carrageenan. All
extracts were administered 1 hour before carrageenan.
The results are shown in Table 4.
4. Discussion
4.1. Antipyretic Activity
Screening of the antipyretic activity of ethanolic extract
of A. lebbeck seeds was already reported [17], but nothing
was reported concerning flowers growing in Saudi Ara-
bia. By screening of the flowers, it was found that di-
chloromethane and ethyl acetate fractions have signifi-
cant decreases in fever dropped by 8˚C & 5˚C, respec-
tively. Moreover, total alcohol, n-butanol, aqueous and
n-hexane extracts showed less activity than those men-
Table 2. Results of analgesic activity screening.
Reaction time on hot plate
A. lebbeck Extract
Zero time 90 min. after
administration
% increase in
pain threshold
n-butanol (0.25 g/kg) 6.4 ± 0.1 7.4 ± 1.6 14.2 ± 7.6
Dichloromethane (1 g/kg)7 ± 0.2 8.8 ± 0.7 25.7 ± 2.9*
Ethyl acetate (1 g/kg) 7 ± 0.7 8 ± 0.9 14.2 ± 2.2
Total alcohol (1 g/kg) 6.6 ± 0.4 7.6 ± 0.2 15.1 ± 1.1
Aqueous (1 g/kg) 6.8 ± 0.2 6.6 ± 0.7 Not active
n-hexane (1 g/kg) 6.8 ± 0.4 7.6 ± 0.2 8.8 ± 0.9
Aspirin (0.2 g/kg) 5.7 ± 0.2 9.6 ± 0.9 68.4 ± 8
The values marked with asterisks differ significantly from the control group,
(*p < 0.05), Number of animals = 5.
Table 3. Results of estrogenic activity screening.
A. lebbeck Extract
Mean ratio of
uterine horns to
body weight
% change in uterine
weight/total body ratio
Control 0.00547 ± 0.0018 -
n-butanol
(500 mg/kg/day)- Very toxic (Not tested)
Dichloromethane
(500 mg/kg/day)0.00504 7.8% decrease
Ethyl acetate
(200 mg/kg I.P) 0.00409 25.2% ± 5.8% decrease*
Total alcohol
(500 mg/kg/day)0.01144 ± 0.0002 109.141% ± 2.2% increase**
Aqueous
(500 mg/kg/day)0.00574 4.9% increase
n-hexane
(500 mg/kg/day)0.00520 4.93% decrease
The values marked with asterisks differ significantly from the control group,
(*p < 0.05), (**p < 0.001), Number of animals = 4.
Table 4. Results of of anti-inflammatory activity screening.
A. lebbeck Extract
% suppression of inflammation
2 hours after carrageenan
administration
n-butanol (0.25 g/kg) Zero
Dichloromethane (1 g/kg) 71.6 ± 8.5%*
Ethyl acetate (1 g/kg) 60.3 ± 7.3%*
Total alcohol (1 g/kg) 33.9 ± 6.4%*
Aqueous (1 g/kg) 37.7 ± 10.2%*
n-hexane (1 g/kg) 50.9 ± 3.1%*
Diclofenac sodium (20 mg/kg) 67.4 ± 5.4%
The values marked with asterisks differ significantly from the control group,
(*p < 0.05), Number of animals = 3; N. B. Maximum paw edema 2 hours
after carrageenan intra-plantar injection was 0.53 ± 0.05 ml.
tioned above by dropping the temperature by 2.3˚C,
4.7˚C, 2.7˚C and 1.7˚C, respectively as shown in Table 1.
4.2. Analgesic Activity
It was reported that the bark and seeds extracts of A. leb-
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Evaluation of Some Biological Activities of Albizia lebbeck Flowers
476
beck have analgesic activity but flowers extracts didn’t
take due attention [18]. For flowers extracts as shown in
the Table 2, a maximum of 25.7% increase in the reac-
tion time was shown by the dichloromethane extract. The
aqueous extract was inactive. Maximum increases in the
pain threshold were observed 90 minutes after admini-
stration of each extract.
4.3. Estrogenic Activity
Estrogens are steroid hormones with important functions
in the regulation of specific sexual processes in the fe-
male. It was reported that methanolic pod extract of A.
lebbeck has antifertility effect but nothing was reported
about estrogenic effect [19]. At a dose of 500 mg/kg I.P.
to immature rats, only total alcohol extract caused sig-
nificant increase in the ratio of weight of the two uterine
horns to the total body weight by 109.14%. This is a very
high percentage and indicates the potent estrogenic ac-
tivity of the total alcohol extract that can point to further
researches. Whereas, the ethyl acetate extract exerted
significant depression of uterine weight/body weight
ratio.
4.4. Anti-Inflammatory Activity
Screening of the anti-inflammatory activity of seeds and
bark of A. lebbeck was reported but nothing about flow-
ers of species growing in Saudi Arabia was reported
[17,18]. The best extract which showed anti-inflamma-
tory activity was the dichloromethane extract with 71.6%
followed by the ethyl acetate extract with 60.3% inhibi-
tors.
In conclusion, these findings clearly suggest that flow-
ers of Albizia lebbeck growing in Saudi Arabia have in-
teresting biological activities and we recommend further
research for isolation of the components responsible for
the estrogenic and anti-inflammatory activities.
5. Acknowledgements
This research project was supported by a grant from the
research Center of the Center for Male Scientific and
Medical Colleges in the King Saud University.
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