Hepato-protective Effect of an Ayurvedic Formulation Prak-20 in CCl 4 Induced Toxicity in Rats: Results of Three Studies

Abstract

Prak-20 is a proprietary herbo-mineral ayurvedic formulation routinely used for years in the clinical practice of the first author in the treatment of liver ailments. Prak-20 is a judicious combination of nineteen herbs and Mandoor Bhasma. Three experimental studies were carried to evaluate its hepato-protective properties. In the first experiment, Prak-20 was compared vis-à-vis Liv52 in CCl 4 challenged Wister rats. The second study was a single blinded study of Prak-20 in CCl 4 challenged Wister rats. The third experiment was conducted to note the effect of different doses of Prak-20 in CCl 4 challenged albino rats. Serum alkaline phosphatase (ALP), serum alanine transaminase (ALT), serum aspartate transaminase (AST), along with liver histology was done in all animals at the completion of the first two studies. There was no significant elevation of ALT, AST and ALP levels despite CCl 4 exposure in Prak-20 treated animals. Similarly, the histology studies revealed that all animals of the treatment group had regenerating hepatocytes and no necrotic and degenerative changes were noticed in any animal. In the third experiment, ALT and AST levels were studied after 24 and 48 hours of experiment. There was 54 and 37 percent decrease in ALT, AST levels in CCl 4 plus Prak-20 (1.8 gm/d/rat) treated animals, respectively, as compared to CCl 4 treated animals. Prak-20 treated animals also had minimum necrotic changes after seven days. Based on the findings of aforesaid experiments which were carried at three centers, it may be concluded that Prak-20 is a potent hepato-protective herbo-mineral ayurvedic formulation. Further studies are required to understand its mechanism of action.

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International Journal of Pharmaceutical and Clinical Research 2010; 2(1): 23-27
23
Research Article
ISSN 0975 1556
Hepato-protective Effect of an Ayurvedic Formulation Prak-20 in CCl4
Induced Toxicity in Rats: Results of Three Studies
Vaidya Balendu Prakash1*, Arun Mukherjee2
1Ipca Traditional Remedies Pvt. Ltd., 142-AB, Kandivli Industrial Estate, Kandivli (West), Mumbai – 400067, India
2Uddan, A-59, Kailash Colony, New Delhi, India
ABSTRACT
Prak-20 is a proprietary herbo-mineral ayurvedic formulation routinely used for years in the clinical practice of the first
author in the treatment of liver ailments. Prak-20 is a judicious combination of nineteen herbs and Mandoor Bhasma. Three
experimental studies were carried to evaluate its hepato-protective properties. In the first experiment, Prak-20 was
compared vis-à-vis Liv52 in CCl4 challenged Wister rats. The second study was a single blinded study of Prak-20 in CCl4
challenged Wister rats. The third experiment was conducted to note the effect of different doses of Prak-20 in CCl4
challenged albino rats. Serum alkaline phosphatase (ALP), serum alanine transaminase (ALT), serum aspartate
transaminase (AST), along with liver histology was done in all animals at the completion of the first two studies. There
was no significant elevation of ALT, AST and ALP levels despite CCl4 exposure in Prak-20 treated animals. Similarly, the
histology studies revealed that all animals of the treatment group had regenerating hepatocytes and no necrotic and
degenerative changes were noticed in any animal. In the third experiment, ALT and AST levels were studied after 24 and
48 hours of experiment. There was 54 and 37 percent decrease in ALT, AST levels in CCl4 plus Prak-20 (1.8 gm/d/rat)
treated animals, respectively, as compared to CCl4 treated animals. Prak-20 treated animals also had minimum necrotic
changes after seven days. Based on the findings of aforesaid experiments which were carried at three centers, it may be
concluded that Prak-20 is a potent hepato-protective herbo-mineral ayurvedic formulation. Further studies are required to
understand its mechanism of action.
Keywords: Ayurveda, hepato-protective, herbo-mineral formulation.
INTRODUCTION
Liver play a major role in detoxification and excretion of
many endogenous and exogenous compounds. Any
impairment to its function may lead to serious implications
and even death. Management of liver diseases is still a
challenge to the modern scientific community. [1] There are
few conventional drugs that can stimulate liver function and
offer hepato protection or help in the regeneration of hepatic
cells. [2] Globally, many herbal preparations are used in the
treatment of liver disorders. [3] Phytoconstituents remain to be
a major contributor in the treatment of liver disorders.
Formulations available as remedy for hepatic disorders are
poly herbals without elucidating the efficacy of individual
herb of the formulation.
Prak-20 is a herbo-mineral ayurvedic formulation containing
nineteen herbs and Mandoor Bhasma. [4] The herbs used in
*Corresponding author: Vaidya Balendu Prakash,
Ipca Traditional Remedies Pvt. Ltd., 142-AB, Kandivli
Industrial Estate, Kandivli (West), Mumbai – 400067, India;
Tel: + 91 22 6647 4628 (off), + 91 9322492770 (M);
Fax: + 91 22 2868 6954
E-mail: balenduprakash@gmail.com
Prak-20 is well described in the ayurvedic treatises
mentioned under schedule Y of the Drug & Cosmetic Act. [5]
The hepato-protective effect of Prak-20 has been observed in
clinical practice of the first author for over two decades. This
formulation was also found to have anti-fibrotic properties [6]
and helped in fast recovery of acute viral hepatitis patients. [7]
The herbs used in Prak-20 have the following properties viz.
hepatoprotective, improves liver and kidney function, relives
hepatomegaly and splenomegaly, jaundice, cholagogue, anti-
nauseant, anti-spasmodic, dyspepsia, anti-pyretic, appetizer
and carminative. In this article, we report the observations of
three animal experiments which were carried at three centers
to study the hepato-protective properties of Prak-20 against
CCl4 induced liver injury.
MATERIALS AND METHODS
Composition of Prak-20
Prak-20 is a patent and proprietary ayurvedic medicine
prepared by Bharat Bhaishajya Shala Private Limited,
Dehradun, under Good Manufacturing Practices (GMP)
certificate issued by Department of AYUSH provincial
government of Uttarakhand. In house standard for raw
material and process quality control have been developed to

Prakash et al. / Hepatoprotective Effect of an Ayurvedic Formulation Prak-20........
IJPCR January-March, 2010, Vol 2, Issue 1 (23-27) 24
bring reproducibility of Prak-20. The composition of Prak-20
is given in the Table 1.
Experimental Animals: Adult rats (Wister / Swiss albino)
were kept in polypropylene cages with stainless lid with rice
husk bedding. Individual animal was identified by specific
marking and cages were identified with label pasted on cages
with relevant information. Animals were housed at a
temperature of 20 ± 2oC and relative humidity of 25 to 75 %.
A 12:12 light: dark cycle was followed. All animals had free
access to water and standard pelleted laboratory animal diet.
Ethical Clearance: Three experiments were done in rats to
evaluate the hepatoprotective potential in Carbon
tetrachloride (CCl4) induced hepato toxicity. The first
experiment was carried out at India Institute of Toxicology,
Mumbai. The second experiment was carried out at Fredrick
Institute of Plant Protection and Toxicology, Padappai and
the third study was carried out at G. B. Pant Hospital, Delhi.
Prior to the start of these experiments, clearance from the
Institutional ethical board was obtained from the respective
institutes.
Table 1: Composition of Prak-20
Common name Scientific name Proportion
Sunthi Zingiber officinale 13.88 mg
Maricha Piper nigrum 13.88 mg
Pippali Piper longum (fruit) 13.88 mg
Haritaki Terminalia chebulia 13.88 mg
Vibhitaki Termnallia bellirica 13.88 mg
Amalaki Emblica officinalis 13.88 mg
Chitraka Plumbago zeylanica 13.88 mg
Musta Cyperus rotundus 13.88 mg
Katuki Picrorrhiza kurroa 13.88 mg
Devadaru Cedrus deodara 13.88 mg
Vidanga Embellia ribes 13.88 mg
Kulu/Kushta Saussuria lappa 13.88 mg
Haridra Curcuma longa 13.88 mg
Daruharidra Berberis aristata 13.88 mg
Danti Baliospermum montanum 13.88 mg
Indrayav Holarrhena antidysentrica (seeds) 13.88 mg
Pippali mula Piper longum 13.88 mg
Trivrit Ipomoea turpethum 13.88 mg
Punarnava Boerhavia diffusa 27.77 mg
Mandoor Bhasma Ferric Oxide 250 mg
Experimental study I: The aim of this study was to evaluate
the hepatoprotective potential of Prak-20 vis-a-vis Liv52 in
CCl4 challenged Wister rats.
A total number of 32 animals were randomly divided into 4
groups. Except for the control group all the animals of other
groups received subcutaneous injection of 0.2 ml of CCl4
with 0.2 ml of liquid paraffin twice a week. The treatment
schedule is given below:
Group I: Normal control.
Group II: CCl4.
Group III: CCl4 plus oral administration of 30 mg Prak-20
per day for 13 weeks.
Group IV: CCl4 plus oral administration of 30 mg Liv52
per day for 13 weeks.
The animals were observed for 13 weeks. Their body weight
and food intake was taken weekly. All the animals were
sacrificed at the end of the experiment and a detailed
necropsy was performed with particular emphasis on Liver.
The livers were taken out and weighed and volume estimated
by the displacement method. Serum alkaline phosphatase
(ALP), serum alanine transaminase (ALT), Aspartate
transaminase (AST), was estimated by using Boehringer
Knoll Autoanalyser System 4010. Histopathology evaluation
of the liver of each animal was done after completion of the
experiment.
Experimental study II: This was a blinded study of Prak-20
coded as A (dark colour) and B (light colour) in CCl4
challenged Wister rats.
A total of 30 female Wister rats were used for this
experiment and were divided into 6 groups. The animals
received either liquid paraffin (10 ml/kg body weight) or 1
ml of 50 % (v/v) CCl
4 in 9 ml of liquid paraffin/kg body
weight i.p. every 72 hrs from day 0 to day 15. The treatment
schedule is given below:
Group I: Liquid paraffin from day 0 to day 15. From day
16 to day 30, the animals were given 10ml/kg
body weight of distilled water daily by oral
intubation.
Group II: CCl4 from day 0 to day 15.
Group III: CCl4 from day 0 to day 15. From day 16 to day
30, the animals were treated with Prak-20
(Code A) 30 mg/kg body weight by oral
intubation.
Group IV: Liquid paraffin from day 0 to day 15. From day
16 to day 30, the animals were given 30 mg/kg
body weight of Prak-20 (Code A) by oral
intubation.
Group V: CCl4 from day 0 to day 15. From day 16 to day
30, the animals were treated with Prak-20
(Code B) 30 mg/kg body weight by oral
intubation.
Group VI: Liquid paraffin from day 0 to day 15. From day
16 to day 30, the animals were given 30 mg/kg
body weight of Prak-20 (Code B) by oral
intubation.
Animal were sacrificed 24 hrs after the last treatment. Prior
to sacrifice, blood was collected and plasma was separated
for estimation of ALT, AST and ALP using a semi-auto
analyzer. The animals were then euthanized and the liver was
collected for histopathological examination.
Experimental study III: The aim of this study was to
evaluate the hepatoprotective effect of various doses of Prak-
20 in CCl4 challenged rats.
Seventy albino rats were used for this study. Twenty animals
were used as normal control. The rest 50 rats were randomly
divided into 5 groups. All these animals were challenged
with one subcutaneous injection of 0.5 ml of Carbon
tetrachloride (CCl4) per kg body weight. The details of
treatment schedule are mentioned below:
Group I: Normal control
Group II: CCl4
Group III: CCl4 + Prak-20 (40 mg/day/rat) for 7 days
Group IV: CCl4 + Prak-20 (600 mg/day/rat) for 7 days
Group V: CCl4 + Prak-20 (1.2 g/day/rat) for 7 days
Group VI: CCl4 + Prak-20 (1.8 g/day/rat) for 7 days
Blood samples were collected from all animals after 24 hrs
and 48 hrs after CCl4 administration and were analyzed for
ALT and AST. After 7 days the animals were sacrificed and
the liver was collected for histopathological examination.
Statistical analysis: The mean ± SEM were calculated for
each parameter. The statistical analysis of the results was
carried out with a SPSS 10.0 program based on an Analysis
of Variance (ANOVA) followed by the Dunnet’s test.
RESULTS
Experiment I:

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Table 2: Histology and Mean ± S.E. of various parameters studied in CCl4 challenged rats treated with Prak-20
Liver Weight Gr
oup
s N Body Wt.
(gm) Absolute
(gm) Relative
(%)
Liver
volume
(ml)
AKP
(IU/ml) ALT
(IU/ml) AST
(IU/ml) Histology
I 8 253.25 ± 9.4 8.27 ± 0.23 3.27 ± 0.09 8.5 ± 0.31 89.05 ± 6.46 10.93 ± 0.95 42.75 ± 1.12 Within normal limits
II 8 262.86 ± 12.14 12.96 ± 0.64* 4.83 ± 0.12* 13.0 ± 0.67* 151 ± 23.40 37.09 ± 5.13* 53.14 ± 5.51 Centro-lobular
necrosis with fatty
changes
III 8 234.86 ± 11.60 9.45 ± 0.65 4.0 ± 0.16* 9.29 ± 0.69 104.74 ± 20.0 14.42 ± 4.36 42.21 ± 2.28
Devoid of
degeneration of
hepatocytes in all
animals
IV 8 241.33 ± 11.46 10.15 ± 0.44 4.22 ± 0.11* 10.17 ± 0.37 92.73 ± 15.46 18.0 ± 4.71 39.75 ± 2.26
Moderate to severe
degeneration in 50%
and mild to moderate
regeneration in 50%
* Differs significantly [P<0.05] from the control group
Table 3: Effect of two coded Prak-20 formulation on liver enzymes (Mean ± S.E.) and liver histology in CCl4 challenged rats
Groups No. Treatment ALT (IU/L) AST (IU/L) ALP (IU/L) Histology
1 6 Control 75.9 ± 2.5 111.6 ± 3.77 60.1 ± 2.45 Within normal limits
2 6 CCl4130.5 ± 3.72* 150.1 ± 4.95* 86.0 ± 2.86* Diffused swelling of hepatocytes along with necrosis
3 6 CCl4 + Prak-20 (Code A) 84.0 ± 3.19 119.0 ± 3.31 65.2 ± 2.78 Significant improvement in necrotic condition
4 6 Prak-20 (Code A) 72.1 ± 2.37 120.1 ± 4.09 58.1 ± 1.77 Within normal limits
5 6 CCl4 + Prak-20 (Code B) 83.0 ± 3.19 118.0 ± 3.31 65.2 ± 2.74 Significant improvement in necrotic condition
6 6 Prak-20 (Code B) 73.2 ± 1.96 118.1 ± 2.58 57.1 ± 1.72 Within normal limits
* Differs significantly [P<0.05] from the control group
Table 4: Effect of different doses of Prak-20 on ALT / AST (Mean ± S.E.) and liver histology in CCl4 challenged rats
After 24 hrs of CCl4After 48 hrs of CCl4
Exp. Group
No. AST IU/L ALT IU/L AST IU/L ALT IU/L Histology done after 7 days of
CCl4 exposure
Control 20 29 ± 0.58#26 ± 0.93#38 ± 1.24#31 ± 0.71#Normal limits
CCl410 114 ± 2.76* 289 ± 6.49* 97 ± 1.38* 166 ± 1.71* Necrosis + 10
Inflammation +10
Fatty Liver + 10
CCl4+ Prak-20
(40mg/d/rat) 10 98 ± 1.79*#166 ± 1.73*#109 ± 1.63*#225 ± 1.57*#Necrosis + 9
Inflammation + 9
CCl4+ Prak-20
(600mg/d/rat) 10 108 ± 1.46* 152 ± 2.06*#114 ± 1.76* 148 ± 2.31*#Necrosis + 5
Inflammation + 4
Fatty Liver + 9
CCl4+ Prak-20
(1.2 g/d/rat) 10 99 ± 0.93*#158 ± 1.31*#58 ± 1.71*#72 ± 47*#Necrosis + 3
Inflammation + 2
Fatty Liver + 5
CCl4+ Prak-20
(1.8 g/d/rat) 10 53 ± 1.17*#50 ± 1.73*#62 ± 1.45*#70 ± 1.87*#Necrosis + 1
Fatty Liver + 2
* Differs significantly [P<0.05] from the control group
# Differs significantly [P<0.05] from the CCl4 treated group
All the animals were studied for the gain in body weight.
However, no significant variation in body weight was
observed in any group at the end of study. Two animals (25
%) in group IV and one animal each from group II and III
died during the experiment. All these animals died after 7
weeks of treatment.
Size, shape color and consistency of liver were almost
normal in the control group. The liver of animals in group II
showed slight to moderate fatty changes. In group III animals
no remarkable changes were noticed except the paleness of
liver was slightly less as compared to the CCl4 treated
animals of group II. In group IV there was no remarkable
changes and the livers were comparable to the livers of group
II animals. Significant increase in absolute liver weight and
liver volume was found in animals treated with only CCl4.
These changes though seen in animals treated with Prak-20
and Liv52, however, they differ significantly from that of the
control group (Table 2).
Significant increase in ALP levels and marked increase of
ALT and AST levels were found in animals treated with
CCl4. However, no significant changes in the level of these
enzymes were seen in animals treated with Prak-20 or Liv52.
Histopathological studies revealed that there was no
abnormality in animals of the control group. In group II:
three animals had typical centrolobular necrosis but not of
severe degree; centrolobular necrosis with fatty changes was
seen in three animals. In one animal necrosis of hepatocytes
was seen in group of cells scattered in the lobule. In group
III animals all the slide/section studied were devoid of
degenerative and necrotic changes. Regenerating hepatocytes
were seen in considerable number in almost all the areas of
section. However, degree of regeneration varied from slight
to moderate in different animals. Architectural pattern of the
liver appeared to return to normalcy in all the animals. In
group IV: two animals showed degenerative changes,
necrotic areas with slight to moderate regenerative
hepatocytes. Severe type of necrosis with degenerative
changes was noted in one animal. Least degenerative changes
and more active regenerative changes were observed in three
animals.
Experiment II
Identical results were obtained with both the coded A and B
Prak-20 formulation. The ALT, AST and ALP in plasma
were significantly elevated in animal treated with CCl4.
These enzymes reverted back to normal levels in animals
treated with Prak-20 (Table 3). The histopathological studies
indicated diffused swelling of the hepatocytes along with

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IJPCR January-March, 2010, Vol 2, Issue 1 (23-27) 26
necrosis in CCl4 treated animals. Significant improvement in
necrotic condition was noted in animals treated with Prak-20.
Experiment III
The ALT and AST levels were significantly elevated in
animals at 24 and 48 hrs after CCl4 exposure. However, these
enzymes were much lower in animals treated with Prak-20 at
a dose of 1.2 gm/day and higher (Table 4). The best effect
was seen in animals treated with 1.8 g of Prak-20 per day.
There were 54 and 83 percent reduction in ALT and AST
levels after 12 hrs and 37 and 58 percent reduction of ALT
and AST levels after 48 hrs, respectively, as compared to the
animals that received only CCl4. The histopathological study
indicated that in a 10 point scale the animals treated with
CCl4 had the maximum necrosis, inflammation and fatty
liver. The necrosis was much less in animals treated with
Prak-20 in dose of 600 mg/day and above (Table 4).
DISCUSSION
Prak-20 depicts strong hepatoprotection against CCl4 induced
liver injuries in three consequent experimental studies. It
brings significant reduction in liver enzymes towards
normalcy. Similarly, it not only arrests necrosis and
degeneration but also brings considerable regeneration of
hepatocytes.
Prak-20 is prepared using parts of 19 herbs and Mandoor
Bhasma (MB). Majority of the herbs have been studied
individually for their medicinal values such as, Picrorhiza
kurroa has shown choleretic hepatoprotective,
neuroprotective and immunostimulant properties. [8-9]
Curcuma longa is well known for its antioxidant,
hepatoprotective, anti-inflammatory, anticarcinogenic, and
antimicrobial properties, in addition it is also used in
cardiovascular disease and gastrointestinal disorders. [10]
Emblica officinalis is found effective against inflammation
[11], cancer, age-related renal disease [12], hepatitis [13] and
diabetes. [14] Fruits of Embelia ribes possess analgesic,
antipyretic, antibacterial, antifertility and anthelminthic
activities. It has also been reported to be useful in jaundice
and as a hepatoprotective agent. [15] Root of Piper nigrum is
used as cholagogue in obstruction of bile duct and gall
bladder.
Many studies so far have been conducted on establishing the
efficacy of individual medicinal plants and their properties.
In brief, 19 herbs used in Prak-20 have hepatoprotective,
immune-modulator, anti inflammatory, anti fibrotic,
improves liver and kidney functions, cholagogue, anti
nauseant, anti spasmodic, anti pyretic, appetizer and
carminative properties. Mandoor Bhasma is a major
constituent of Prak-20. It is a powerful haematenic and is
valuable in the treatment of hemolytic jaundice and
microlytic anemia. The main focus of action of MB in body
is liver. It is useful in the treatment of Pleeha vriddhi (Spleen
enlargement); Yakrit vriddhi (Liver enlargement); Kamala,
(Jaundice); Shotha (Oedema); Pandu, Raktaksaya (Anemia).
[4]
Ayurvedic Pharmacopeias is full of compound formulations
which are largely used by Ayurvedic physicians for centuries
in their respective clinical practice. Prak-20 is modified and
standardized form of a classical Ayurvedic formulation and
has been used by the first author for decades in his clinical
practice. This was used for treatment of patients suffering
from jaundice and hepatomegaly. However, the chemistry of
the finished product, maximum dose tolerance, safety and
mode of action were never known to the author. Therefore,
the first study was designed to revalidate the stated efficacy
of Prak-20 in liver disorders by studying its effect against
CCl4 induced liver damage in animal model. This study
showed cent percent regeneration of hepatocytes in Prak-20
treated animals after CCl4 challenge. The other two studies
were carried at different centers using the same toxin also
indicated a similar trend. Prak-20 has emerged as potent
hepatoprotective, anti-fibrotic, detoxifying and anti
inflammatory Ayurvedic formulation. [16] It has been licensed
for commercial manufacturing and is available in the market
as prescription.
A vast literature is available regarding the substances of
plant, mineral and animal origin used in the preparations of
Ayurvedic formulations. Ayurvedic pharmacopeia also
carries description of numerous Ayurvedic formulations and
its uses on human beings. And this practice started centuries
back and still continues.
In the aforesaid studies, authors tried to make an intervention
between the stated efficacies of a compound Ayurvedic
formulation within the ambit of modern research
methodology of evaluation of a formulation. The findings
credibly establish that the structured Ayurvedic formulations
used in the treatment of liver disorders have strong hepato-
protection activities. This opens a platform to design and
carry more interventional, experimental and clinical studies
to understand and development of intrigue composite
Ayurvedic formulations.
CONCLUSION
Prak-20 has shown potential hepatoprotective activity against
the CCl4 induced liver damage in animal model. The
observed effect of Prak-20 could be because of the
synergistic effect of the various herbs used along with MB.
The clinical use of Prak-20 as a hepatoprotector is
substantiated in these experimental studies. However, more
experimental studies and clinical trials are required to know
the exact potential of this formulation.
ACKNOWLEDGEMENT
We like to thank Mr. Rajiv Gulati, the then Product Manager
of Ranbaxy India Limited for initiating and sponsoring the
first experimental study on Prak-20 at the Indian Institute of
Toxicology, Mumbai. Authors also acknowledge Fredrick
Institute of Plant Protection and Toxicology, Padappai for
carrying out the second experimental study on their own
behest and utilizing their own resources. A special thanks to
Prof. S K Sarin for conducting the third experimental study at
G B Pant Hospital, Delhi. Lastly, we acknowledge the
contribution of Dr. Sanjoy K Pal and Ms. Megha Prakash for
drafting and editing of the manuscript.
REFERENCES
1. Sunilson JAJ, Muthappan M, Das A, Suraj R. Hepatoprotective
activity of Coccinia grandis leaves against carbon tetrachloride
induced hepatic injury in rats. Int J Pharmacol 2009; 5: 227 - 227.
2. Guntupalli M, Chandana V, Pushpangadan P, Shirwaikar IA.
Hepatoprotective effect of rubiadin, a major constituent of Rubia
cordifolia Linn. J Ethnopharmacol 2006; 103: 484 – 490.
3. Samudram P, Rajeshwari H, Vasuki R, Geetha A, Moorthi PS.
Hepatoprotective activity of bi-herbal ethanolic extract on CCl4
induced hepatic damage in rats. Asian J Biochem 2008; 3: 308-314.
4. Mandura Bhasma. In The Ayurvedic Formulary of India - Part I.
Government of India, Ministry of Health and Family Welfare,

Prakash et al. / Hepatoprotective Effect of an Ayurvedic Formulation Prak-20........
IJPCR January-March, 2010, Vol 2, Issue 1 (23-27) 27
Department of Indian Systems of Medicine & Homeopathy, New
Delhi. pp 237 - 238.
5. Schedule Y of the Drug and Cosmetic Act, India. Available from:
http://www.docstoc.com/docs/928490/Schedule-Y-of-the-drugs-
and-cosmetics-act-India. [Assessed on 5th October, 2009]
6. Gupta RC, Prakash B, Mukherjee A, Sarin SK. Double blind
randomized trial of Prak-20, an ayurvedic drug in acute viral
hepatitis. In the proceedings of the IX Biennial Scientific Meeting
of The Asian Pacific Association for the Study of the Liver, held at
Kuala Lumpur, Malaysia from 26 – 29 January 1994. FP 12/1.
7. Sharma G, Ramanamurthy B, Abdullah M, Malhotra V, Mukherjee
A, Prakash B, Sarin SK. Efficacy of Prak-20 an India ayurvedic
medicine, as an antifibrotic drug in a murine model of chronic liver
disease. Gastroenterology 1992; 102 (Suppl - 2): A886.
8. Karmuse P, Prakash VB, Indu G. Benchmarking of mandoor
Bhasma for industrial production. In abstract book of Symposium
on Safety Profile of Rasaushadhies, held at Banaras Hindu
University, Varanasi from 06 – 07 February, 2009. p 72.
9. Feroz Khan Z, Asdaq SMB, Prasanna Kuamar SR. Effect of few
Indian medicinal herbs on carbon tetrachloride induced hepatic
injury in animals. Int J PharmTech Res 2009; 1 (3): 579 – 587.
10. Anonymous. Curcuma longa (turmeric), Monograph. Altern Med
Rev 2001; 6 (Suppl): S62 – S66.
11. Ganju L, Karan D, Chanda S, Srivastava KK, Sawhney RC,
Selvamurthy W. Immunomodulatory effects of agents of plant
origin. Biomed Pharmacother 2003; 57 (7): 296–300.
12. Yokozawa T, Kim HY, Kim HJ, et al. Amla (Emblica officinalis
Gaertn.) attenuates age-related renal dysfunction by oxidative
stress. J Agric Food Chem 2007; 55 (19): 7744–7752.
13. Tasduq SA, Kaisar P, Gupta DK, Kapahi BK, Maheshwari HS
Jyotsna S, Johri RK. Protective effect of a 50% hydroalcoholic fruit
extract of Emblical officinalis against anti-tuberculosis drug
induced liver toxicity. Phytother Res 2005; 19(3): 193-197.
14. Rao TP, Sakaguchi N, Juneja LR, Wada E, Yokozawa T. Amla
(Emblica officinalis Gaertn.) extracts reduce oxidative stress in
streptozotocin-induced diabetic rats. J Med Food 2005; 8 (3): 362–
368.
15. Tabassum N, Agarwal SS. Hepatoprotective activity of Embelia
ribes against Paracetamol induced acute hepatocellular damage in
mice. JK-Practitioner 2003; 10 (1): 43-44.
16. Prak-20: A herbo-mineral compound. Vaidya Chandra Prakash
Cancer Research Foundation, Dehradun, 2009.