Antiulcer Activity of a Traditional Pearl Preparation: Mukta Bhasma

Abstract

Mukta bhasma is unique herbo-mineral calcium containing preparation of Ayurvedic system of Indian traditional medicine. It is used traditionally as antipyretic, antiulcer and antacid. It is used in treatment of bone metabolic disorders associated with calcium deficiency. Mukta bhasma was evaluated for its antiulcer activity in experimental animals. The experimental paradigms used for antiulcer activity were cold restraint stress induced ulcer model and Diclofenac induced ulcer model in rats. It was observed that Mukta bhasma produced significant (P<0.001) 57.53% and 59.18% protection in cold restraint stress induced gastric ulcer and 54.19 % and 61.39% protection in Diclofenac induced ulcer at 60mg/Kg and 120 mg/Kg dose levels respectively, when compared with control. TBARS of stomach in ulcer induced rat was also reduced by Mukta bhasma. The results suggest that this ayurvedic preparation possess significant gastro protective and antiulcer activity in lower doses of therapeutic range and its effect is not dose dependent.

Full text

Research J. Pharm. and Tech.2 (2): April.-June. 2009
,
287
ISSN 0974-3618
www.rjptonline.org
RESEARCH ARTICLE
Antiulcer Activity of a Traditional Pearl Preparation: Mukta Bhasma.
Nitin Dubey
1
*, Nidhi Dubey
2
, RS Mehta
3
, AK Saluja
3
and DK Jain
1
1
College of Pharmacy, IPS Academy, Indore [M.P.]
2
School of Pharmacy, D.A.V.V., Indore [M.P.]
3
A. R .College of Pharmacy, Vallabh Vidyanagar [Gujarat], India.
*Corresponding Author E-mail:
nitindubeympharm@yahoo.com
ABSTRACT
Mukta bhasma is unique herbo-mineral calcium containing preparation of Ayurvedic system of Indian traditional
medicine. It is used traditionally as antipyretic, antiulcer and antacid. It is used in treatment of bone metabolic disorders
associated with calcium deficiency. Mukta bhasma was evaluated for its antiulcer activity in experimental animals. The
experimental paradigms used for antiulcer activity were cold restraint stress induced ulcer model and Diclofenac
induced ulcer model in rats. It was observed that Mukta bhasma produced significant (P<0.001) 57.53% and 59.18%
protection in cold restraint stress induced gastric ulcer and 54.19 % and 61.39% protection in Diclofenac induced ulcer
at 60mg/Kg and 120 mg/Kg dose levels respectively, when compared with control. TBARS of stomach in ulcer induced
rat was also reduced by Mukta bhasma. The results suggest that this ayurvedic preparation possess significant gastro
protective and antiulcer activity in lower doses of therapeutic range and its effect is not dose dependent.
KEYWORDS:
Ayurvedic formulation; calcination; Pearl, Biomineral, antiulcer.
INTRODUCTION:
Pearls are calcareous concretions formed as protection
against the foreign objects, either particles or minute
parasites, which penetrate inside between the mantle and
the shell of the mollusk. A fold of soft tissue envelops
the foreign particles and deposits layer after layer of
nacre on it to form a pearl. Wild pearl may occur in
spherical or irregular shapes in free state, or may be
found attached to the shell as ‘demi’ or ‘blister’pearls to
be cut out from the shells. They are obtained from pearl-
oysters which belongs to single genus, Pinctada Roding
(Pteriidae).
Many traditional ayurvedic formulations contains pearl
as an important active component .The pearl is used in
powdered or calcinated form. Mukta bhasma is an
unique oral ayurvedic preparation synthesized by
trituration and calcination of pearl according to method
described in classical texts. Mukta bhasma has been used
since long and is claimed to be very effective. Bhasma
are potent medicament given orally, in very small doses
(60-125 mg). However, the mechanism of action of
these unique preparations is not clearly understood and
very few scientific data on efficacy of the bhasma is
available
1
.
Received on 23.11.2008 Modified on 02.02.2009
Accepted on 12.03.2009 © RJPT All right reserved
Research J. Pharm. and Tech.2(2): April.-June.2009,;Page 287-290
A through investigation is needed for the bhasma with
regards to their elemental composition, organic content
and their therapeutic effects. Traditional preparation and
physicochemical characterization of Godanti bhasma
and kushta-e-sadaf preparation was reported in a study
by this lab
2,3
.
Anti ulcer effect of shanka bhasma was studied by
Pandit et al
4
. Synergistic effect of pearl preparation on
effectiveness of antibiotic was studied by Kulkarni et al
5
.Pharmacological evaluation of karpura shilajit bhasma
was also reported by Saleem et al
6
.
Mukta bhasma is not only a widely used marketed
ayurvedic preparation but it is also used as one of the
key ingredient of many popular ayurvedic preparations.
However, it appears that no scientific data on antiulcer
activity of mukta bhasma was reported despite its
traditional use; hence it was thought worth while to
investigate its antiulcer activity.
MATERIALS AND METHOD:
Raw Materials were procured by Bhardwaj
Pharmaceuticals Ltd. Indore, (M.P), India and
authenticated by experts of Govt. Astang Ayurvedic
medical college. Mukta perfect in shape, luster and
weight (known as Anie); mukta, not as perfect as Anie
(vadivu) and Mukta, failing in shape and luster
(Anatharie) were selected in equal proportions for
synthesis of bhasma. Jayanti patra consists of fresh and
dried leaf of Sesbania sesban (Linn.) Merr. ; Family

Research J. Pharm. and Tech.2 (2): April.-June. 2009
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Table 1: Antiulcer Effect of Mukta bhasma on cold stress induced ulcer tissue TBARS content in rats.
†Mean ± S.E.M.; number of animal used (n=8)
Fabaceae. The jayati patra swarus is juice obtained from
leaves of Sesbania sesban. Taruni parishruta jala (gulab
arka or rose water) is hydrosol obtained from steam
distillation of petals of Rosa Bucbi , family Rosaceae.
Drugs and chemicals: Injections of Ranitidine was
procured from local market. Other chemicals were of
analytical grade and procured commercially.
Animals: Wistar albino rats were housed in standard
cages at room temperature 22 ± 2 °C and 50±5% relative
humidity, under a light/dark cycle of 10/12 h, for 1 week
before the experiments. Animals were provided with
standard rodent pellet diet (Amrut, India), and water ad
libitum. The animals were deprived of food for 24 hrs
before experimentation, but had free access to drinking
water. All experiments were performed in the morning.
The study was approved by the institutional ethical
committee, (465/01/96/CPSCSEA) which follows the
guidelines of CPSCEA (Committee for the Purpose of
Control and Supervision of Experiments on Animals),
and complies with international norms of INSA.
Traditional Preparation of Mukta bhasma:
The Mukta bhasma was prepared under guidance of an
authentic traditional practitioner, in whose family these
bhasmas have been synthesized for a few generations.
Mukta bhasma was prepared by following the method
described in Ayurvedic texts
7
.
Mukta were first cleaned with hot water (Shodhana)
.Cleaned Mukta were immersed in juice of leaves of
Sesbania sesban (jayanti patra swarus) and boiled
(swedana)for three hours in specially prepared hanging
sealed earthen pot (dola yantra).. The shodhit mukta were
grinded to powder form in a mortar and pestle with rose
water (gulab ark) for eight hours (particle size 30 µm).The
mixture was pressed in form of cake and dried. Cakes
were placed in sealed earthen pot (sarava samputta) and
subjected to calcination (Marana) in a traditional furnace
(gajaputa), as described in ayurvedic literature at mean
temperature 650 ± 10.5°C, to obtain the intermediate
product The procedure was repeated two more times with
bhavana .Three batches of bhasma were prepared and
approved by two experts of Ayurveda unaware of the
procedure. Bhasma used for biological evaluation is a
mixture of all the three batches of bhasma prepared.
Biological evaluation:
Antiulcer studies:
Albino rats of Wistar strain of either sex weighing between
150-200g were used. Bhasma was prepared and
administered orally in 0.9%w/v sodium chloride solution
(saline) as vehicle at a dose of 60 and 125 mg/kg body
weight.
Cold restraint stress (CRS)-induced ulcers
8
:
Rats were divided in four groups consisting of eight
each. Group I and group II received bhasma prepared
using traditional method orally at a dose of 60 mg/kg
and 125 mg/kg body weight respectively. Group III
received Ranitidine orally at a dose of 20mg/kg body
weight. Control Group IV received vehicle only.
Animals were immobilized after one hour of dosing by
strapping the fore and hind limbs in restraint cage and
placed in a cold chamber (4±1°C). 3 hr later, the animals
were sacrificed under the influence of anesthetic ether;
the stomach was removed, washed carefully with 5.0 ml
of 0.9%w/v Sodium chloride and fixed on a cork plate.
Diclofenac induced ulcer
9
:
Rats were divided in four groups consisting of eight
each. All four groups received Diclofenac suspension
(80mg/kg body weight) in saline orally. After one hour
group I and group II received bhasma prepared using
traditional method orally at a dose of 60 mg/kg and 125
mg/kg body weight respectively. Group III received
Ranitidine orally at a dose of 20mg/kg body weight.
Control Group IV received saline only. 3 hr later, the
animals were sacrificed under the influence of anesthetic
ether; the stomach was removed, washed carefully with
5.0 ml of 0.9%w/v Sodium chloride and fixed on a cork
plate. The number and severity of ulcer per stomach, in
the glandular portion of the stomach was registered with
clinical microscope by a person unaware of the
experimental protocol.
The following arbitrary scoring system was used to
grade the incidence and severity of lesions:
0 = No ulcer, 1 = Superficial ulcer, 2 = Deep ulcers, 3 =
Perforations
The Mean Ulcer Index U
I
was calculated using following
formula: U
I
=U
N
+U
S
+U
P
X 10
-1
Where U
I
= Average of number of ulcer per animal.
U
N
= Average of severity score.
U
P
= Percentage of animals with ulcers.
Biochemical estimations: Gastric tissue lipid
peroxidation
10
were estimated in rats that developed
ulcers. The stomach homogenates were prepared in
chilled 0.15 M KCl and lipid per oxidation was
determined by estimating TBARS (thiobarbituric acid
reacting substances)
11, 12
.
Cold restraint stress induced Ulcer
Sr.
no.
Groups Dose
(mg/kg) Total no of Ulcers
†
Ulcer Index % protection TBARS
†
(nmol/mg protein)
1 Control (CRS) Vehicle 8.9±1.24 30.37 - 14.78 ± 0.39
2 Ranitidine 20 0.62±0.72 11.4 62.49 9.54 ± 0.48
3 MKT 60 1.12±0.25 12.4 59.18 12.71±0.27
4 MKT 120 1.25±0.12 12.9 57.53 10.65 ±0.63

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Table 2: Antiulcer Effect of Mukta bhasma on Diclofenac induced ulcer, tissue TBARS content in rats.
†Mean ± S.E.M.; number of animal used (n=8)
Figure 1: Antiulcer Effect of Mukta Bhasma .
Statistical analysis:
The statistical analysis was carried out using GraphPad
Prism, version 5.0. All in-vivo experimental results were
expressed as mean ± S.E.M. Data were analyzed using
two way analysis of variance (ANOVA) with Bonferroni
post test.
RESULTS:
The LD
50
of bhasma as per OECD guideline falls under
class IV with no signs of acute toxicity up to a maximum
dose of 2000mg/kg (data not shown). There were no
changes in normal behavioral pattern and no signs and
symptoms of toxicity and mortality were observed. So it
was thought worthwhile to proceed with classical dose
i.e. 60mg/kg and 125 mg/kg body weight.
Antiulcer studies: Mukta bhasma at the doses of 60
mg/kg and 125 mg/kg produced significant (P<0.001)
reduction in ulcer index as compared to control (Figure
1). It is interesting to note that Mukta bhasma at 125
mg/kg dose shows comparable activity to 60mg/kg dose
as difference was not significant at P>0.05 .The activity
was less than that of Ranitidine. The biochemical
estimation showed reduction in TBARS content of
stomach tissue in bhasma treated group (Table 1 and
Table 2).
DISCUSSION:
It is generally accepted that gastric ulcer results from an
imbalance between aggressive factors and the
maintenance of the mucosal integrity through the
endogenous defense mechanism
13
.Cold restraint stress
induced ulcer represents an unique ulcer model in
examining the cause, course, consequence and treatment
of peptic ulcer
8
. Pharmacological effects of Diclofenac
sodium are related to the inhibition of the conversion of
arachidonic acid to prostaglandins, which are the
mediators of the inflammatory processes. NSAID induced
ulceration causes accumulation of oxygen free radicals,
which play a crucial role in the pathophysiology of gastric
ulceration
14
. Oxygen derived free radicals cause lipid
peroxidation, which leads to membrane fluidity, resulting
in reduced membrane integrity of surface epithelial cells,
thereby causing gastric ulcers
15
. It has been found that
oxygen-derived free radicals are implicated in the
mechanism of acute and chronic ulceration and
scavenging these free radicals can play an appreciable role
in healing gastric ulcers
16
. The study revealed that the
ulcer severity and lipid peroxidation were aggravated
during cold resistance stress, which is also indicated by
TBRAS content under stress as compared to unstressed
rat, whereas inhibition of lipid peroxidation on Mukta
bhasma administration indicates the antilipid peroxidative
effect which could have prevented lipid peroxidation
mediated ulcerative damage to gastric mucosa. In this
study, we have observed protection offered by Mukta
bhasma in cold restraint stress induced gastric ulcers
(Fig.2) and Diclofenac induced ulcer in rat (Fig. 3).
Extensive experimental evidences indicate that certain
substances, through scavenging of free radicals, protect
the gastric mucosa
17
. The thiobarbituric acid reactive
substance (TBARS) was used as an indicator of lipid
peroxidation and free radical scavenging activity in
biological samples
11
.In the present study, Mukta bhasma
exhibits a potent anti-peroxidative effect. Hence, it can be
suggested from our study that Mukta bhasma provides
anti-ulcer activity in rats. It may act as gastric
cytoprotective agent by modulating scavenging of free
radicals. Further studies like, acids and
mucopolysaccharides estimations by pyloric ligated
models are required to establish the role of Mukta bhasma
in protection against gastroduodenal ulcer.
Diclofenac induced ulcer
Groups Dose (mg/kg)
Total no of
Ulcers†
Ulcer Index % protection TBARS† (nmol/mg
protein)
1 Control (DCL) 80 20.37±0.98 61.1 - 17.88 ± 0.46
2 Ranitidine 20 4±0.46 19.9 67.43 10.35 ± 0.33
3 MKT 60 7.87±0.81 28 54.19 11.97±0.84
4 MKT 120 5.87±0.53 23.6 61.39 9.87±0.74.

Research J. Pharm. and Tech.2 (2): April.-June. 2009
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Figure 2: Cold restraint induced Ulcer (CRS) in Rat (a)
Control (b) After Mukta Bhasma (120 mg/kg) treatment
CONCLUSION:
Mukta bhasma produces significant anti-ulcer activity in
rats. The results suggest that it may act as gastric
cytoprotective agent by modulating scavenging of free
radicals as Mukta bhasma exhibits a potent anti-
peroxidative effect. Its anti-ulcer activity was not dose-
dependent. The results suggest that Mukta bhasma
possess significant gastroprotective activity in lower
doses of therapeutic range.
Figure 3: Diclofenac induced Ulcer in Rat (a) Control (b)
After Mukta Bhasma (120 mg/kg) treatment
ACKNOWLEDGEMENT:
Authors are thankful to Bhardwaj pharmaceuticals,
Indore [M.P.] and Govt. Astang Ayurvedic medical
college, Indore for providing facilities and technical
inputs for synthesis of bhasma by traditional method
.Sincere thanks are due to Director, S.I.C.A.R.T., V.V.N.
(Gujrat ) for providing facilities for biological
evaluation.
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