Hypotensive activity and toxicology of constituents from Bombax ceiba stem bark

Article abstract of DOI:10.1248/bpb.26.41

A novel constituent, shamimicin, 1?,1??′-bis-2- (3,4-dihydroxyphenyl)-3,4-dihydro-3,7-dihydroxy-5-O-xylopyranosyloxy-2H-1- benzopyran alongwith lupeol, which possesses potent hypotensive activity has been isolated from Bombax ceiba stem bark. BCBMM - one of the most active hypotensive fractions has revealed its adverse effects on heart, liver and kidneys of mice at the dose of 1000 mg/kg/d.

Full text of DOI:10.1248/bpb.26.41

January 2003
Biol. Pharm. Bull. 26(1) 41—46 (2003)
41
Hypotensive Activity and Toxicology of Constituents from Bombax ceiba
Stem Bark
,
a
a
a
b
Rubeena SALEEM,* Syed Iqbal AHMAD, Mohammad AHMED, Zareen FAIZI,
c
c
c
Sadia ZIKR-UR-REHMAN, Muhammad ALI, and Shaheen FAIZI
a
b
Dr. HMI Institute of Pharmacology and Herbal Sciences, Hamdard University; Karachi-74600, Pakistan: Faizi Clinic,
c
Sharah-e-Faisal Colony No. 3; Karachi, Pakistan: and HEJ Research Institute of Chemistry, University of Karachi;
Karachi-75270, Pakistan. Received July 22, 2002; accepted October 16, 2002
A novel constituent, shamimicin, 1ꢀ,1ꢀꢁꢁ-bis-2-(3,4-dihydroxyphenyl)-3,4-dihydro-3,7-dihydroxy-5-O-xylopy-
ranosyloxy-2H-1-benzopyran alongwith lupeol, which possesses potent hypotensive activity has been isolated
from Bombax ceiba stem bark. BCBMM—one of the most active hypotensive fractions has revealed its adverse
effects on heart, liver and kidneys of mice at the dose of 1000 mg/kg/d.
Key words novel benzopyran; Bombax ceiba; hypotensive activity; histopathology
1
Hypertension is a disease, which if left untreated affects 500 spectrometer operating at 500 MHz for H and 125 MHz
1
3
all important organs of human body. It is known as silent for C nuclei, with spectra referenced to residual solvent sig-
killer as without showing significant symptoms, it may qui- nals. The chemical shifts are in ppm (d) and coupling con-
etly lead to stroke, brain hemorrhage, cardiac disorders, renal stants (J) are in Hz. For flash column chromatography (FCC)
failure and vision loss. Hypertension has affected 10—15% and vacuum liquid chromatography (VLC) silica gel 9385
of global population and killed a large number of human race (E. Merck) and silica gel 60 GF254 (Merck) were used, re-
in every region of world. Drugs used for its treatment on one spectively. Purity of compounds was checked on silica gel 60
hand are very expensive and beyond the reach of a common GF254
man while on the other hand, side effects associated with
Plant Material Stem bark was collected in February
.
these drugs restricted the people to use them. Hence develop- 2000 from Karachi University Campus. The plant was au-
ment of a cost effective antihypertensive drug with minimum thenticated at the Department of Botany, University of
or no side effects from commonly available indigenous medi- Karachi and a voucher specimen (No. 66854KU) was de-
cinal plants is highly required. Keeping this in view as well posited in the same department.
as blood pressure reducing property of Bombax ceiba present
work describes the hypotensive evolution of its stem bark
alongwith toxicology and histopathology of active fraction.
Bombax ceiba (syn. Bombax malabaricum DC) commonly
Isolation of Lupeol Fresh, uncrushed stem bark of B.
1,2)
known as silk cotton tree is an important medicinal plant.
Earlier investigations led to the identification of a sesquiter-
3)
pene lactone, a potent growth inhibitor in fungi and a
flavonol glycoside possessing significant hypotensive and hy-
4)
poglycaemic activities. Both of these compounds were iso-
lated from roots and leaves of B. ceiba respectively. This is
the first report of hypotensive evaluation of it’s stem bark
which has resulted in the isolation of a novel benzopyran
5)
dimer, shamimicin, lupeol and fractions possessing hy-
potensive activity. In addition to this, methanolic extracts of
different parts of plant, including flowers, pulp, stem and
stem bark have also been studied for their effect on mean ar-
terial blood pressure (MABP) of rats.
MATERIALS AND METHODS
Instrumentation Ultraviolet spectra were recorded in
MeOH on Hitachi-U-3200 and infrared spectra were mea-
sured in CHCl on JASCO-A-302 spectrophotometers. The
3
electron impact (EI) mass spectra were recorded on a Finni-
gan MAT-112 instrument. Field desorption (FD-MS), fast
atom bombardment (FAB-MS positive and negative) and
exact mass measurements were carried out on a Finnigan
MAT-312 instrument. High-resolution mass spectra were
1
13
recorded on a JMS HX-110 spectrometer. The H- and C-
NMR spectra were run in CDCl on a Bruker Aspect AM-
3
∗
To whom correspondence should be addressed. e-mail: rs127pk@yahoo.com
© 2003 Pharmaceutical Society of Japan

42
Vol. 26, No. 1
ceiba (2.55 kg) was extracted twice with petroleum ether. from E. Merck, tween 80 from BDH chemicals, atropine sul-
®
The extracts were combined and evaporated on rotavapour fate from Boehringer Ingelheim, and Pentothal sodium from
under reduced pressure to give a residue (BCBP, 5 g). A Abbott Karachi. Acetylcholine (1 mg/kg) was used as a posi-
white matter separated out when BCBP was treated with tive control while saline (0.9% NaCl) and 10% tween-80 (in
methanol. It was filtered and washed thrice with methanol to the case of BCBP, lupeol and its acetate) as negative controls.
6)
give white shinning needles of lupeol (mp 214.7 °C, lit.
Hypotensive Activity Normotensive Sprague–Dawley
2
15—216 °C, 1.5 g, 0.06%, Rf, 0.40, PE : EA, 6 : 4). Spectral rats (either sex, 200—250 g) were anaesthetized with pen-
7,8)
®
data of lupeol is in full agreement with literature values.
tothal sodium (50 mg/kg i.p.). The trachea was exposed and
Acetylation of Lupeol A solution of lupeol (50 mg) in cannulated to facilitate spontaneous respiration. Drugs were
pyridine (0.5 ml) was treated with acetic anhydride (0.5 ml) injected (vol. 0.2—0.25 ml) through a polyethylene cannula
and left over night at room temperature. Reaction mixture inserted into the external jugular vein followed by a saline
when evaporated under hood gave white needles. Needles ob- flush (0.2 ml). The arterial blood pressure was recorded from
tained from bottom of flask were recrystallized with CHCl3– the carotid artery via polyethylene arterial cannula connected
MeOH, which gave pure white bunch of needles [35 mg, mp to a Research Grade Blood Pressure Transducer (Harvard,
6)
2
14 °C, lit. mp 215—216 °C ].
60-3003) coupled with four channel Harvard Universal Os-
Isolation of Novel Shamimicin Marc left after the ex- cillograph (Curvilinear, 50-9307). The temperature of the an-
traction of B. ceiba stem bark with petroleum ether was ex- imals was maintained at 37 °C by use of over head lamp. An-
tracted successively with methanol (thrice) and 50% aqueous imals were allowed to equilibrate for at least 15 min before
methanol. Combined methanolic extracts and 50% methano- administration of any drug. Mean arterial blood pressure
lic extract were evaporated under reduced pressure to give (MABP) was calculated as sum of the diastolic blood pres-
concentrated mass BCBM (41.3 g) and BCBM-50 (31.2 g). A sure plus one-third pulse width. Changes in blood pressure
solid matter settled at the bottom of flask containing BCBM were expressed as the percent of control values, obtained im-
4)
when kept in cold for three days. It was filtered to give solu- mediately before the administration of test substance.
ble (BCBMM, 36.5 g) and insoluble fraction (BCBMI, 4.5 g). Hypotensive Activity of BCBMM through Oral Route
Vacuum liquid chromatography (VLC) of BCBMM [silica Two groups of three male rats in each group were taken.
gel 60 GF254 260 g, using petroleum ether (PE, 2.7 l), ethyl Group-I treated (p.o.) with BCBMM at the dose of 200
acetate (EA, 5.5 l), methanol (6.8 l) and water (1.9 l) with mg/kg/d for five consecutive days. Group-II serving as con-
gradually increasing polarity by 10%, volume eluted 500 ml] trol was treated with saline. On fifth day after two hours of
®
gave 35 fractions. Fractions 1—13 (PE 100%–EA 100%), last dose, all animals were anaesthetized with pentothal
1
4—17 (MeOH, EA, 1,9–MeOH, EA, 4,6), 18—21 (MeOH, sodium (25 mg/kg, i.p.) and cannulated as described earlier.
EA, 1,1–MeOH, EA, 8,2), 22—26 (MeOH, EA, 9,1–MeOH Toxicology Acute toxicity of BCBMM was measured
00%) and 27—35 (MeOH, H O, 9,1–MeOH, H O, 2,8) in mice (20—30 g) by oral route of administration. Three
1
2
2
were combined on the basis of thin layer chromatography sets of each containing 10 mice (males and female) were
and evaporated under reduced pressure to give residues BA taken. Group-1 and II were treated with 100 mg/kg/g and
(
2.1 g), BB (2.4 g), BC (7.0 g), BD (0.53 g) and BE (12.7 g) 1000 mg/kg/d (vol. of each dose was 0.7—0.8 ml) of
4)
respectively. First three fractions were further chromato- BCBMM respectively for seven consecutive days. Group-
graphed through FCC and work is still under progress. III served as control, and was given saline. All the animals
FCC of BB [silica gel 9385 (73 g), solvent used PE (1.5 l), were kept under constant observation for two hours to ob-
EA (2 l), MeOH (2 l) and water (1.2 l) in order of increasing serve any change in general behavior or other physical activi-
polarity by 5%, volume eluted 100 ml] resulted in 64 frac- ties. Numbers of survivors were noted after seven days. Sur-
tions, of which fraction No. 26 gave a colourless gelly like vived mice were killed by hitting on the head for tissue
substance when kept at room temperature. It was decanted to analysis. Heart, liver and kidneys of killed as well as freshly
give a pure new compound shamimicin (140 mg) showing dead animals were removed, blotted and weighed immedi-
single pink spot on TLC (Rf, 0.5, EA, MeOH, 8.5, 1.5). ately on electronic balance.
Structure of shamimicin was elucidated through UV, IR, MS,
H-NMR, C-NMR and 2D-NMR (including NOESY, formalin. After usual processes of dehydration, clearing and
Histology Heart, liver and kidneys were fixed in 10%
1
13
COSY, HMQC, HMBC) as 1ꢀ,1ꢀꢁꢁ-bis-2-(3,4-dihydroxy- infiltration, tissues were embedded in paraffin wax and sec-
phenyl)-3,4-dihydro-3,7-dihydroxy-5-O-xylopyranosyloxy- tioned into 5-mm slices through Leica RM 2145-Rotation
2
H-1-benzopyran.
Microtom. The tissues were stained with haematoxylin and
Methanolic Extracts of Different Parts of B. ceiba eosin. The slides were studied and photographed through
Fresh, undried flowers (390 g), pulp of fruit (228 g), stem Nikon Advance Trinocular Research Microscope OP-
310 g) and stem bark (255 g) of B. ceiba were extracted di- TIPHOT Model X2T-21E equipped with Nikon Micropho-
(
rectly with MeOH. Extract of each part evaporated separately tography system; Model UFX-DX-35 and phase contrast N
on rotavapour to give residues BCF (5.68 g), BCP (4.0 g), plan.
BCS (4.76 g) and BCB (4.23 g) respectively.
Statistical Analysis Changes in blood pressure and
Animals and Drugs Animals used in this study were weights of vital organs were compared using analysis of vari-
Sprague–Dawley rats (200—250 g) and NMRI mice (20— ance followed by Student’s t-test. Values of pꢂ0.01, pꢂ0.001
30 g). They were housed at the Animal House of Dr. HMI In- were considered to be significant.
stitute of Pharmacology and Herbal Sciences, Hamdard Uni-
versity and were given a standard diet and tap water ad-libi-
tum. Drugs used were acetylcholine and sodium chloride

January 2003
43
RESULTS AND DISCUSSION
potension but the effect of BC lasted for more than 2 min.
Duration of activity in rest of the fractions was mostly less
Petroleum ether extract of B. ceiba stem bark (BCBP) than one minute (Table 1).
caused 58% fall in MABP of rats at the dose of 10 mg/kg. In
Shamimicin, a new rarely occurring glycoside caused no
some cases third administration of the same dose (after 20— significant fall in blood pressure at the dose of 15 mg/kg.
30 min of last administration) caused death of rat. Lupeol the Shamimicin is a dimeric ent-epicatechin glycoside (vide
main component (30%) of non polar extract and an anti-in- structure). Since ent-epicatechins are known to possess car-
9
)
14)
flammatory agent exhibited 44% and 52% reduction in diac stimulating, hyperglycaemic, hypoglycaemic
hair
15)
16)
blood pressure for 1.7ꢃ0.6 min and 2.8ꢃ1.8 min at the dose growth promoting and antioxidant activities, it is possi-
of 5 mg/kg and 15 mg/kg respectively. This is the second re- ble that shamimicin might possess activity at higher doses.
port of hypotensive activity of lupeol. Earlier lupeol from The detailed spectroscopy and elucidation of structure for
Acosmium dasycarpum was reported to decrease blood pres- shamimicin is submitted for publication elsewhere.
sure at 20—100 mg/kg. Lupeol acetate, an anti-inflamma-
tory triterpene exhibited 43% fall in MABP at 3 mg/kg for fect when examined in rats pretreated with atropine sulfate
10)
BCBM and BCBM-50 did not exhibit their hypotensive ef-
9)
1.8ꢃ1.1 min. Derivative did not show any significant effect at (69.5 mg/kg). This behavior matches exactly to that of acetyl
1 mg/kg. BCBP and lupeol were not completely soluble in choline (1 mg/kg), which did not produce, any fall in blood
10% tween-80 and some residue remained settled at the bot- pressure in the presence of atropine sulfate. It may therefore
tom of vial. Since lupane type of triterpenes have reported to be concluded that muscarinic M receptors on cardiac mus-
2
11)
12)
posses hepatoprotective, anti-HIV and tumor growth in- cles as well as vascular dilation by endothelium derived re-
hibitory properties, lupeol which constitutes 0.06% of laxing factor may contribute to the cholinergic hypotensive
plant’s stem bark can also be studied for similar type of activ- effect of BCBM and BCBM-50. Conversely, hypotensive
13)
ities.
property of BCBMM remained unaltered in atropinized rats.
Methanolic extract of defatted stem bark (BCBM) and its Rest of the constituents could not examined due to paucity of
fraction BCBMI produced 30% and 51—57% hypotension at compound or shortage of experimental animals.
10 mg/kg and 30 mg/kg respectively (Table 1). The 50%
Methanolic extract of flowers (BCF) and pulp (BCP) were
methanolic extract (BCBM-50) at both doses of 10 mg/kg found as most active hypotensive agents among methanolic
and 30 mg/kg showed 17—20% fall in MABP, comparatively extracts of different parts of plant. They showed comparable
low intensity of action for 1.7ꢃ1.2 min and 2.2ꢃ1.2 min re- 50% reduction in blood pressure with maximum duration of
spectively. BCBMM—the other fraction of BCBM, showed effect at 30 mg/kg. Methanolic extract of stem bark (BCB) is
hypotensive effect of 31% and 65% at the dose of 3 mg/kg found more active than stem (BCS) (Table 2). In each of
and 15 mg/kg. Hypotensive action of BCBMM lasted for these extracts activity lasted for 1—3 min (Table 2).
1
.6ꢃ0.2 min and 3.7ꢃ1.1 min respectively at corresponded
Acetylcholine used as positive control caused 58.3ꢃ3.1%
doses. VLC of BCBMM resulted in highly potent fractions (meanꢃS.E.M., nꢄ4) fall in mean arterial blood pressure at
BA, BC and BD, moderately active BE and inactive fraction the dose of 1 mg/kg. Solvents (saline and 10% tween-80)
BB (Table 1). BC and BD showed comparable potency of hy- used as negative control did not cause any significant change
Table 1. Effect of Extracts, Fractions and Pure Compounds from Bombax ceiba Stem Bark on Mean Arterial Blood Pressure (MABP) of Rat
MABP
(mmHg)
before
MABP
(mmHg)
after
%
MABP
Fall in
S.
No.
Dose
(mg/kg, i.v)
Duration
(min)
Name
(mmHg)
treatment
treatment
1
.
.
BCBP
Petroleum ether extract of B. ceiba stem bark)
Lupeol
10
5
117.3
49.5
57.8ꢃ18.7
0.7ꢃ0.3
(
2
130.3
127.1
115.0
128.8
121.9
111.3
124.2
128.3
118.3
127.7
138.6
127.8
124.1
134.2
113.3
117.4
143.3
128.3
128.5
73.2
61.6
65.7
88.4
59.3
88.7
102.3
88.3
40.8
91.4
59.2
104.3
70.4
133.5
86.7
42.0
96.1
34.2
91.2
43.8ꢃ10.9
51.5ꢃ4.3
42.9ꢃ6.2
31.1ꢃ12.1
51.4ꢃ10.5
20.3ꢃ4.2
17.6ꢃ3.1
31.2ꢃ5.3
65.5ꢃ9.7
28.4ꢃ2.0
57.3ꢃ15.0
18.4ꢃ6.7
43.3ꢃ16.1
0.5ꢃ0.02
23.5ꢃ1.7
64.2ꢃ7.7
32.9ꢃ5.1
73.3ꢃ10.5
29.0ꢃ11.7
1.7ꢃ0.6
2.8ꢃ1.8
1.8ꢃ1.1
0.8ꢃ0.3
1.4ꢃ0.1
1.7ꢃ1.2
2.2ꢃ1.2
1.6ꢃ0.2
3.7ꢃ1.1
0.9ꢃ0.3
0.4ꢃ0.2
0.2ꢃ0.01
1.4ꢃ0.1
—
1
5
3
4
.
.
Lupeol acetate
BCBM
3
10
30
10
30
3
(Methanolic extract of defatted stem bark)
5.
6.
7.
8.
9.
BCBM-50
(Extraction of marc with 50% MeOH)
BCBMM (Filtrate from BCBM)
1
5
BCBMI (Residue from BCBM)
BA
10
3
0
10
3
0
BB
BC
30
10
1
1
1
0.
1.
2.
0.5ꢃ0.3
2.1ꢃ0.2
0.2ꢃ0.3
0.8ꢃ0.1
0.3ꢃ0.02
3
0
BD
BE
10
3
0
30
Values shown represent meanꢃS.E.M. of four determinations. Number of animals used for each dose is two.

44
Vol. 26, No. 1
Table 2. Effect of Methanolic Extract of Different Parts of Bombax ceiba on Mean Arterial Blood Pressure (MABP) of Rat
MABP
(mmHg)
before
MABP
(mmHg)
after
%
MABP
Fall in
S.
No.
Dose
(mg/kg, i.v)
Duration
(min)
Name
(mmHg)
treatment
treatment
1.
2.
3.
4.
BCS (Extract of stem)
BCB (Extract of stem bark)
BCF (Extract of flowers)
BCP (Extract of pulp)
10
138.1
110.9
117.6
116.7
136.1
127.2
119.1
112.7
117.8
89.2
79.1
69.8
120.7
58.9
14.7ꢃ4.3
19.5ꢃ6.4
32.7ꢃ1.6
40.2ꢃ6.2
11.3ꢃ2.4
53.7ꢃ25.5
15.7ꢃ1.9
50.4ꢃ18.2
1.8ꢃ0.6
1.1ꢃ0.6
1.0ꢃ0.3
2.4ꢃ0.8
1.2ꢃ0.2
2.7ꢃ0.6
1.5ꢃ0.2
2.5ꢃ0.8
3
0
10
3
0
10
3
0
10
100.4
55.9
3
0
Values shown represent meanꢃS.E.M. of four determinations. Number of animals used for each dose is two.
Table 3. Effect of BCBMM at 1000 mg/kg/d on Weights (g) of Vital Or-
gans in Mice
Weight of
Weight of
Weight of
heartꢃS.E.M.
liverꢃS.E.M. kidneysꢃS.E.M.
Control
Treated
0.14ꢃ0.2
0.15ꢃ0.001
ꢅ7.14*
1.23ꢃ0.5
2.05ꢃ0.12
ꢅ66.67**
0.31ꢃ0.04
0.37ꢃ0.04
ꢅ19.35**
%
Change
Values represent meanꢃS.E.M. of three determinations. ∗ pꢂ0.01, ∗∗ pꢂ0.001.
in blood pressure of rat.
Hypotensive Activity of BCBMM through Oral Route
Animals treated orally with BCBMM at the dose of 200
mg/kg/d showed MABP at 105.3ꢃ2.8 mmHg while that of
control was found at 121.3ꢃ1.2 mmHg. Hence drug caused
Fig. 1. Heart from Control Mice (ꢆ40)
13.2% reduction of MABP by oral route which is significant
as pꢂ0.01.
Toxicology BCBMM caused expiry of two male and one
female mice at the dose of 100 mg/kg/d. Male mice showed
17% loss in body weight while female mice showed compar-
atively less reduction of only 7%. None of these mice showed
any change in their physical activities. At 1000 mg/kg/d, all
five male mice were expired just after two doses. Females
were more resistant and last female mice died after six doses.
All these animals become inactive after first dose and some
adopted statue like position before death. Control group did
not show any mortality or significant change in their general
behavior or physical activities.
Tissue Analysis Three of mice died as a result of oral
administration of BCBMM at 1000 mg/kg/d, showed 67% in-
crease in weight as compared to control liver. Colour of liver
got fade and light yellow patches were clearly seen. Heart
and kidneys did not show any morphological change, but
their weights were significantly increased by 7% and by 19%
respectively as compared to control values (Table 3).
Histopathology
Fig. 2. Heart from Mice after Treatment (ꢆ40)
Heart Nuclei seems to be disintegrated or disappeared.
Muscle fiber architecture was disturbed and areas of cellular
infiltration were also present (Figs. 1, 2).
Liver Most of the hepatocytes were swollen, hydropic
(ballooning degeneration) and often binucleated (Figs. 3, 4).
The condition indicates that BCBMM might have caused
liver cell necrosis resulting in liver cell derangement.
Kidneys Cortical region was full of fibrous and granular
tissues. Most of the proximal and distal convulated tubules
were destructed. The adjacent tubules were separated from Fig. 3. Liver from Control Mice (ꢆ40)

January 2003
45
Fig. 4. Liver from Mice after Treatment (ꢆ40)
Fig. 5. Cortical Region of Kidney from Control Mice (ꢆ10)
Fig. 6. Cortical Region of Kidney after Treatment (ꢆ10)
Fig. 7. Glomeruli after Treatment (ꢆ20)
Fig. 8. Medullary Region of Kidney from Control Mice (ꢆ10)
Fig. 9. Medullary Region after Treatment (ꢆ10)
one another by edematous interstitial tissue (Figs. 5, 6).
Glomerular architecture appeared disturbed. There is patchy
diffuse infiltration of glomerular tuft with eosinophilic mater-
ial and is fibrinoid in appearance (Fig. 7). In medullary re-
gion of kidney the architecture of papillary ducts and loop of
Henle (thick and thin segment) appeared disturbed while lin-
ing epithelium of papillary ducts showed necrosis (Figs. 8,
9).
CONCLUSION
Lupeol and fraction BCBMM have emerged as potent hy-
potensive constituents of Bombax ceiba stem bark while new
dimeric glycoside shamimicin was found devoid of any activ-
ity at the dose of 15 mg/kg. Intravenous administration of
15 mg/kg of BCBMM caused 65% fall in blood pressure of
rats for 4 min. While oral administration of 200 mg/kg/d for
five days showed significant hypotension of 13.18%. Low
percentage of hypotension (though significant) at higher dose
of 200 mg/kg/d through oral route as compared to intra-
venous administration indicates that active ingredient of
BCBMM has low absorbance in blood either due to its very
hydrophilic nature or biotransformation. Chemical and phar-
macological evaluation of BCBMM (which is underprogress)
may lead to active constituents which might be able to show
significant bioavailability. Toxicology of BCBMM deter-
mined 1000 mg/kg/d as lethal dose (LD100) killing all mice
by affecting their vital organs including heart, kidneys and
liver. Dose at 100 mg/kg/d is also not safe in mice as it

46
Vol. 26, No. 1
caused expiry of three animals. It is interesting to note that
00 mg/kg/d of BCBMM did not show any mortality in rats,
although sample size was quite small (only three). Hence it
may be assumed that low doses of BCBMM (smaller than
331—334 (1999).
5
6
)
)
Mukherjee J., Roy B., J. Ind. Chem. Soc., 48, 769—770 (1971).
Buckingham J., “Dictionary of Natural Products,” Champan and Hall
Scientific data division, London, 1992, 3, 3698.
Ahmad V. Q., Bano S., Mohammad F. V., Planta Med., 521—523
(1985).
2
7)
8)
9)
100 mg/kg) can be useful in reducing blood pressure safely.
Sholichin M., Yamasaki K., Kasai R., Tanaka O., Chem. Pharm. Bull.,
However, in order to determine exact effective and safe dose
of BCBMM, some more experiments in different species of
animals at different doses are required.
28, 1006—1008 (1980).
Nikiema J. B., Vanhaelen-Fastre R., Vanhaelen M., Fontaine J., Graef
C. D., Heenen M., Phytother. Res., 15, 131—134 (2001).
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