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“PREPARATION, PHYSICO- CHEMICAL ANALYSIS OF
SHANKHA NABHI BHASMA AND EVALUATION OF ITS HEPATO
PROTECTIVE ACTIVITY, -AN EXPERIMENTAL STUDY”.
BY
DR. JAYASHREE S. Dissertation Submitted to the Rajiv Gandhi University Of Health Sciences,
Karnataka, Bangalore.
In partial fulfillment of the requirements for the degree of
AAYYUURRVVEEDDAA VVAACCHHAASSPPAATTII ((DDOOCCTTOORR OOFF MMEEDDIICCIINNEE))
IN RASASHASTRA
Under the guidance of
Dr. M.C. Patil M.D. (Ayu)
Professor & H.O.D. Dept. of Rasashastra
and
Co-guidance of
Dr. G.N. DANAPPAGOUDAR M.D.(Ayu) Asst. Professor Dept. of Rasashastra
POST GRADUATE DEPARTMENT OF RASASHASTRA D.G M. AYURVEDIC MEDICAL COLLEGE AND RESEARCH CENTER,
GADAG – 582103 2005 - 2008
Rajiv Gandhi University Of Health Sciences, Karnataka, Bangalore.
DECLARATION BY THE CANDIDATE
I here by declare that this dissertation / thesis entitled “Preparation, Physico-
Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato
protective Activity - An Experimental Study” is a bonafide and genuine research
work carried out by me under the guidance Dr. M.C. Patil M.D.(Ayu), Professor &
H.O.D. Post graduate department of Rasashastra. and under the Co-guidance of Dr.
G.N. Danappagoudar M.D.(Ayu), Asst. Professor Post graduate department of
Rasashastra, D.G.M.A.M.C, PGS & RC, Gadag.
Date: Place: Gadag. Dr. Jayashree S.
SHRI D. G. MELMALAGI AYURVEDIC MEDICAL COLLEGE, POST GRADUATE DEPARTMENT OF RASASHASTRA.
CERTIFICATE BY THE GUIDE
This is to certify that the dissertation entitled “Preparation, Physico-
Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato
protective Activity - An Experimental Study” is a bonafide research work done by
Dr. Jayashree S. in partial fulfillment of the requirement for the degree of “Ayurveda
Vachaspathi M.D (Rasashastra)”, Under Rajiv Gandhi University of Health
Sciences, Bangalore, Karnataka.
Date: Guide Place: Gadag.
Dr. M.C. Patil M.D.(Ayu) Professor & H.O.D. Dept. of Rasashastra, Post Graduate Research Center D.G.A.M.C. Gadag
SHRI D. G. MELMALAGI AYURVEDIC MEDICAL COLLEGE,
POST GRADUATE DEPARTMENT OF RASASHASTRA.
CERTIFICATE BY THE Co - GUIDE
This is to certify that the dissertation entitled “Preparation, Physico-
Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato
protective Activity - An Experimental Study” is a bonafide research work done by
Dr. Jayashree S. in partial fulfillment of the requirement for the degree of “Ayurveda
Vachaspathi M.D (Rasashastra)”, Under Rajiv Gandhi University of Health
Sciences, Bangalore, Karnataka.
Date: Co Guide
Place: Gadag. Dr. G.N. Danappagoudar, M.D. (Ayu) Asst. Professor,
Postgraduate department of Rasashastra. D.G.M.A.M.C. Gadag.
J.S.V.V. SAMSTHE’S
ENDORSEMENT BY THE H.O.D AND PRINCIPAL OF
THE INSTITUTION
This is to certify that the dissertation entitled “Preparation, Physico-
Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato
protective Activity - An Experimental Study” is a bonafide research work done by
Dr. Jayashree S. under the guidance of DR. M.C. Patil M.D. (Ayu), Professor & H.O.D,
Postgraduate department of Rasashastra and co-guidance of DR. G.N.
Danappagoudar M.D. (Ayu), Asst. Professor, Postgraduate department of Rasashastra,
in partial fulfillment of the requirement for the post graduation degree of “Ayurveda
Vachaspati M.D. (Rasashastra)” Under Rajiv Gandhi University of Health
Sciences, Bangalore, Karnataka.
DR. M.C. Patil M.D. (Ayu) Dr. G. B. Patil. Professor & H.O.D. Principal. Post graduate department of Rasashastra. D.G.M.A.M.C, GADAG. D.G.M.A.M.C, GADAG. Date: Date: Place: Gadag Place: Gadag
COPYRIGHT
Declaration by the candidate
I hereby declare that the Rajiv Gandhi University of Health Sciences,
Karnataka shall have the rights to preserve, use and disseminate this dissertation /
thesis in print or electronic format for academic / research purpose.
Date: Signature of Scholar
Place: Gadag
Dr. Jayashree S.
© Rajiv Gandhi University of Health Sciences, Karnataka.
ACKNOWLEDGEMENT
I salute to Lord Vighneshwara and His Holiness Shri Abhinava Shivanand
swamiji to have bestowed their blessings throughout my carrier.
I acknowledge my parents Shri Shantaveerappa Pattanshetti and Smt.
Annapurana S. Pattanshetti being my inner strength to achieve every milestone in my
life.
With pleasure I express my full respect and regards to my Grand Mother Smt.
Gangamma Mahajanshetter and Doddamma Smt. Gouramma Yadavannavar who
made me proficient and gave constant support and encouragement.
I am always at remembrance of my life partner Dr. Anilkumar V and his
family members who are spirit behind my enthusiasm and for their needful support.
With deep sense of pleasure, I acknowledge my gratitude to my beloved guide
Dr. M. C. Patil MD (Ayu) Professor and HOD, PG Dept of Rasashastra, DGMAMC,
Gadag, for his scholarly guidance, supervision, creative criticism, constant
encouragement and high inspiration at every stage of this work.
My gratitude is greatest towards my co-guide Dr. G. N. Danappagoudar MD
(Ayu) Lecturer, PG Dept. of Rasashastra, DGMAMC, Gadag, who gave me timely
advises and suggestions during the entire period of this effort.
I express my sincere obligations to our beloved Principal Dr. G. B. Patil, PGS
& RC, DGMAMC, Gadag, for his encouragement and provision of amenities required
during this toil.
I offer my sincere thanks to Dr. R.K. Gacchinmath, Professor and H.O.D, UG
Dept of Rasashastra, DGMAMC, Gadag, for his constant support and valuable
directions.
I wish to convey my regards to Dr. Jagadeesh G. Mitti, Lecturer, PG Dept of
Rasashastra, DGMAMC, Gadag, for his enormous co-operation.
I owe my heartfelt thanks to Dr. Dilipkumar B, Asst. Professor, PG Dept of
Rasashastra, DGMAMC, Gadag, for his critical views and precious suggestions.
I express my earnest gratitude to Dr. G.S. Hiremath, Dr. Varadacharyulu, Dr.
Avvanni, Dr. Prushottamacharyulu, Dr. K.S.R. Prasad, Dr. R.V. Shettar, Dr. S.N.
Belawadi, Dr. Rajshekar, Dr.Kuber Sankh, Dr Shashikant B. Nidagundi & Dr M.D.
Samudri for their great co-operation.
I
I am greatful to Dr. Shreevatsa MD (Ayu), Lecturer, TGMAMC, Bellary, Dr.
Anjaneya murthy, Dr. Gopi Krishna for their inspiration and constant support
through out my carrier.
With pleasure I extend my sincere gratitude to Dr S.D. Yarageri RMO, Dr. B.G
Swamy, Smt. P.K. Belwadi, Smt. Sarangmath, Tippanagoudar, Kallangoudar,
Biradar, Smt. Ekbote, Smt. A.C. Patil, Shri Shankar Belwadi, Shamshad, Mangala &
Manju for their co-operation and help during the study.
I extend my gratitude to Shri V.M. Mundinmani, Shyavi and Karur for
providing the required books during the study.
I am deeply indebt to Dr. Sharnu Angadi, Dr. Pattanshetti, Dr. B.Y. Ganti, Dr.
Pradeep, Dr. Sobagin, Dr. Shakuntala, Dr. Anita, Dr. Suvarna, Dr. Anand, Dr. Teggi
for their friendly affection.
I ackwoledge my sincere thanks to Shri Shivakumar Inamdar for his statistical
work, Dr. Revati, Shri Sarfaraz, Shri Angadaki, Subhash for their kind co-operation
and help in analytical study.
I am blessed to have precious supervision, pinpoint suggestion and constant
help of Shri. Shivakumar Inamdar Lecturer, Dept. of Pharmacology, J.T. College of
Pharmacy, Gadag & Dr. Shashikant B. Nidagundi M.D. (Ayu), Lecturer, Dept. of
Dravyaguna, DGMAMC Gadag who guided and accompanied me during
experimental study.
I am grateful to Shri Chaitrakumar (Sadguru Computers) for his kind co-
operation and immense help to complete the dissertation work.
With great pleasure I offer my recognition to my friends Dr. Ashwini V, Dr.
Rudrakshi P.D, Dr. Suma J, Dr. M. Kattimani, Dr. Prasanna Mathad, Dr. Sunita
Sundaran, Dr. Shri Mukund for their friendly affection and amiable attitude during
my study period without which it would never be complete.
I offer my sincere thanks to my friends and colleagues Dr. Madhushri, Dr.
Ashok, Dr. Siba Prasad, Dr. Shivaleela, Dr. Kamalakshi, Dr. Sulochana, Dr.
Payappagoudar, Dr. Budi, Dr. Prasanna, Dr. Amnish for their immense help and
affection.
I am also thankful to my junior friends Dr. Shivakumar, Dr. Ravindra, Dr.
Kavita, Dr. Anupama, Dr. Sarvamangala, Dr. M. Hiremath, Dr. Deepa, Dr. Praveen,
Dr. Jadhav, Dr. Ghorpade, Dr. C.C. Hiremath, Dr. Jaya, Dr. Kala, Dr. Savita, Dr.
Mukta Y, Dr. Mukta H, Dr. Shailej, Dr. Natraj, Dr. Uday Ganesh, Dr. Adarsh, Dr.
II
Joshi, Dr. Veena, Dr. Vijayalakshmi, Dr. Sanjay, Dr. Trupti for their support and
affection.
I am happy to express my regards to my in-laws, Shri Veerappa. M & Smt.
Chitravati. C for their moral and cordial support during the study.
It would be my privilege to convey my regards to families of Mr. Mallikarjun
and Mr. Timmaraj for their affection and care during my study.
I express my affectionate love to my Sister Rajeshwari S. Martur & Brother
Basavaraj Pattenshetti for their moral and cordial support during my study.
I am ever thankful to my family members Mr. Ashok Yadavannavar and Smt.
Anita Yadavannavar, Mr. Ulavappa Kondikoppa & Smt. Drakshayani Kondikoppa
who stood with me all the way at my turmoil.
I thank my kiths and kins Rashmi, Smita, Prajval, Sangamesh for their
affection.
I am very much greatful to all the lecturers, house surgeons, Hospital staff and
non teaching staff for their timely assistance in completion of this work.
I express my thanks to all those who have helped me directly and indirectly
with apologies for my inability to identify them individually.
I dedicate this work done as partial fulfillment of postgraduate degree to my
ever remembering respectful parents and my husband Dr. Anilkumar V.
Dr. Jayashree S.
Date:
Place:
III
LIST OF ABBREVIATIONS USED
R T Rasatarangini
A P Ayurveda Prakash
R R S Rasa Ratna Samucchaya
R S S Rasendra Sara Sangraha
BRRS Brihat Rasaraja Sundara
B P Bhava Prakasha
KN Kaideva Nighantu
SN Shaligram Nighantu
R N Raja Nighantu
Sh.N Shodala Nighantu.
M N Madanapala Nighantu
B R Bhaishajya Ratnavali
Y R Yoga Ratnakara
C S Charaka Samhita
S S Sushruta samhita
Sha. S Sharangdhara Samhita
AH Ashtanga Hridaya
M N Madhava Nidana
CCl4 Carbon tetrachloride
SGOT Serum Glutamic – Oxaloacetic Transaminase
SGPT Serum Glutamic – Pyruvic Transminase
ALP Alkaline Phosphatase
T-Bil Total Bilirubin
S.Alb Serum Albumin
SNB Shankhanabhi Bhasma
SNB – I Shankhanabhi Bhasma in 1 Karsha Matra
SNB – II Shankhanabhi Bhasma in 2 Ratti Matra
‘+’ Mentioned
‘- ’ Not Mentioned
IV
ABSTRACT
BACKGROUND
In the present Scenario change in life style of human beings like excessive
alcohol consumption, smoking, stress, irregular food habits have increased the
prevalence of Yakrit vikara.
Liver plays a mojor role in detoxification and excretion of many endogenous
and exogenous compounds, any injury (due to systemic drugs, food preservatives
agrochemicals and addiction to alcohol) to it or impairment of its function may lead to
many complications on one’s health.
There is no rational therapy available for treating liver disorder and is still a
challenge to the modern medicine. Modern medicine have little to offer for alleviation
of hepatic ailments where as most important representatives are of natural origin.
Before evaluating efficacy of any drug, it is essential to carry out an
experimental study and to find out potent therapeutic form.
Objectives:
1. Preparation of Shankhanabhi Bhasma.
2. Physico-chemical analysis of Shankhanabhi Bhasma.
3. Evaluation of Hepato-protective activity of Shankhanabhi Bhasma.
Methods:
Pharmaceutical study:
1) Shankhanabhi shodhana according to Rasatrangini 12th chapter sholka no.10.
2) Shankhanabhi marana according to Rasatarangini 12th chapter
sholka no.17-19.
V
Analytical study
Shankhanabhi bhasma is subjected to physico-chemical analysis like
organoleptic characters, determination of pH values, loss on drying at 1100C, loss on
ignition, determination of total Ash, Acid insoluble ash, water soluble extractive,
alcohol soluble extractive, finess of particles, particle size, flow property, flow rate,
Solubility text, Estimation of Ca, CaCO3, Fe, Mg, S and NPSTest.
Experimental study:
Shankhanabhi bhasma was evaluated for Hepato-protective activity against
carbon tetrachloride (CCl4) induced Liver toxicity in albino rats. The trial drug was
administered in two different doses and biochemical parameters and histopathological
reports was observed, recorded and statistically analysed.
Results:
The Shankhanabhi bhasma has shown significant hepato protective effet (with
“p” value <0.001) by reduction in elevated serum enzyme levels such as serum
glutamic oxaloacetic transminase (SGOT), serum glutamic pyruvic transaminase
(SGPT), alkaline phosphatase (ALP), Total bilirubin and Serum Albumin. These
biochemical observations were supplemented by histopathological examination of
Liver sections.
Interpretation and Conclusion:
1. The dravyas which are mentioned in the classical procedure of Shankha
shodhana and marana convert the Shankhanabhi into quick absorbable form
and induces the disease curing property.
2. Ayurvedic bhasma pareeksha and modern physico-chemical analysis are
confirmation tests for the complete formation of bhasma and its genuinitity.
VI
3. Shankhanabhi Bhasma is one of the ideal Hepato protective drug which has
been proved experimentally by reducing the LFT values and histopathological
report.
Keywords:
Shankhanabhi Bhasma, Analytical study, Yakritodara, Hepatoprotective
activity, Carbon tetrachloride, Significant results.
VII
TABLE OF CONTENTS
Sl. No.
Name of Topic & Sub Topics Page No.
1 INTRODUCTION 1-5
2. OBJECTIVES 6
3. REVIEW OF LITERATURE
Drug Review
Disease Review
7-32
33-57
4. MATERIALS AND METHODS
Pharmaceutical Study
Analytical Study
Experimental Study
58-68
69-84
85-90
5. RESULTS 91-101
6. DISCUSSION 102-115
7. CONCLUSION 116
8. SUMMARY 117-118
9. BIBLIOGRAPHY 120-128
10. ANNEXURE – SHLOKAS
VIII
LIST OF TABLES
Sl No
Tables Page N0
01 Showing full name of texts mentioned about Shankha 9 02 Showing full name of Nighantu’s mentioned about Shankha 9 03 Showing Synonyms of Shankha according to various texts 11-12 04 Showing Shodhana procedure of Shankha according to
various Rasa classics 16
05 Showing Rogaghnata of Shankha according to various texts 17-18 06 Showing classical yogas of Shankhanabhi and Shankha 18-19 07 Showing brief introduction of animal of Shankha belonging
to Phylum mollusca 20
08 Showing correlation between Ranjakapitta and Bile 37 09 Showing Nidana of Pleehodara and Yakritodara according to
various texts 38
10 Showing Roopa of Pleehodara and Yakritodara according to various texts
38-39
11 Showing principle alterations of hepatic morphology produced by some commonly used drugs and chemicals.
51-52
12 Showing Laboratory Evaluation of Liver disease 57 13 Showing quantity of liquid during preparation of kanji. 60 14 Showing the changes in wt. of shankhnabhi before shodhana
and after shodhana. 62
15 Shwoing physical features of raw and shodhita shankhanabhi 63 16 Showing drugs used for shankhanabhi marana. 64 17 Showing Time and Temperature of gajaputa 1 66 18 Showing Time and Temperature of gajaputa 2 67 19 Showing Physical features of Shankhanabhi bhasma after
each puta. 68
20 Showing the loss of Shankhanabhi during its marana. 68 21 Showing Analysis of Shankhanabhi bhasma by ancient
method. 70
22 Showing Experimental Protocol. 87 23 Showing Summary of Biochemical values of all groups 91 24 Showing Intermediate calculation, ANOVA table SGOT 94 25 Showing one way analysis of variation (ANOVA) 94 26 Showing Intermediate calculation, ANOVA table SGPT 95 27 Showing one way analysis of variation (ANOVA) 95 28 Showing Intermediate calculation, ANOVA table ALP 95 29 Showing one way analysis of variation (ANOVA) 96 30 Showing Intermediate calculation, ANOVA table T- bil 96 31 Showing one way analysis of variation (ANOVA) 96 32 Showing Intermediate calculation, ANOVA table Albumin 97 33 Showing one way analysis of variation (ANOVA) 97 34 Showing the comparison of effect of toxic and natural group
with treated groups (By means of t values). 100
IX
LIST OF GRAPHS:
Sl. No Graphs Page No 1 Mean SGOT of all the groups 92 2 Mean SGPT of all the groups 92 3 Mean ALP of all the groups 93 4 Mean T-Bil of all the groups 93 5 Mean Serum Albumin of all the groups 94 6 Comparison between Biochemical parameters of
G2 and G3 97
7 Comparison between Biochemical parameters of G2 and G4
98
8 Comparison between Biochemical parameters of G2 and G5
98
9 Comparison between Biochemical parameters of G3 and G4
99
10 Comparison between Biochemical parameters of G3 and G5
99
11 Comparison between Biochemical parameters of G4 and G5
100
LIST OF PHOTOGRAPHS:
Fig.No Photographs 1 Shankha 2 Shankhanabhi 3 Pachana in Kanji 4 Pieces of Shankhanabhi after shodhana 5 Marana of Shankhanabhi 6 Shankhanabhi pieces after I Gajaputa 7 Mardana of Shankhanabhi bhasma with Kumari swarasa 8 Chakrikas of Shankhanabhi 9 Shankhanabhi bhasma after II Gajaputa 10 Suspensions of Shankhanabhi bhasma 11 Grouping of animals 12 Weighing of Rat for selection 13 Administration of CCl4 0.7 ml /kg i.p 14 Administration of trial drug orally. 15 Opening of abdomen for liver isolation. 16 Phatomicrograph of Normal Healthy liver (G1) 17 Phatomicrograph of Damaged liver with CCl4 (G2) 18 Phatomicrograph of Toxic control/ Natural recovery liver (G3) 19 Phatomicrograph of liver (G4) treated with SNB – I 20 Phatomicrograph of liver (G4) treated with SNB – II 21 Alcoholic extraction of Haridra 22 Haridra paper prepared 23 Haridra paper with standard spot of Shankhanabhi bhasma
X
XI
Introduction
1
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
INTRODUCTION
Ayurveda is science of life and its aim is for prevention and cure of diseases
and also attainment of chaturvidha Purushartha i.e Dharma, Artha, Kama, Moksha.
Though we can prevent diseases just by following sadvritta and regimen of
Dinacharya, Ritucharya etc. yet for the cure of diseases, drugs are essential.
In Ayurveda, acharyas have explained different charyas to maintain the health.
The diseases are manifested mainly because of neglecting these routines.Change in
life style of human beings like excessive alcohol consumption, smoking, stress,
irregular food habits have increased the prevalence of Yakrid vikara.
From the time immemorial mankind has been in search for plant, animal and
other materials that could be used to take care of the pains, deformities and diseases
that affect some of the unfortunate members of the society.Among them one of the
major problem is the hepatic disorders.
Liver is the largest organ in the body, whatever taken orally or parenterally,
eventually it has to pass through the liver, either by portal or systemic circulation and
gets altered, destroyed or detoxified by it. In this process of protecting the body from
harmful substances, liver cells themselves get destroyed and are usually regenerated
with the same speed provided, the toxins are not in abundant quantities and
continuously pass through it.
Liver plays a pivotal role in regulation of physiological process like
metabolism and excretion. It is constantly endowed with the task of detoxification of
xenobiotics, environmental pollutants and chemotherapeutic agents. Liver diseases are
mainly caused due to an exposure to toxic chemical substances like antibiotics,
analgesics, antipyretics, chemotherapeutics, peroxidised oil, carbon tetrachloride,
chlorinated hydrocarbons and also due to chronic alcoholism, viral infections and auto
Introduction
2
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
immunity disorders. Most of the chemotoxic chemical effect on liver cells mainly by
inducing lipid peroxidation and other oxidative damages takes place in liver.
It has been estimated that about 90% of the acute hepatitis are resulting due to
viruses. The major viral agents involved are hepatitis B,C,D,E & G of these, hepatitis
B infection often results in chronic liver diseases and cirrhosis of liver. It has been
estimated that approximately 14-16 million people are infected with this kind of virus.
In India alone over 2,00,000 people die due to liver disorders in every year.
Consumption of spirits/ liquors is the prime cause for liver damage, drug toxicity is
another cause for liver disorders. On survey studies around 2-3% of Indian population
are carrying hepatitis B & C type of viruses, worldwide these figures are increasing
with an alarm.
In Ayurveda also Acharyas have given utmost importance to the yakrit. It is
the seat of Ranjaka pitta, Bhutagni, Rakadhara kala and mula of Raktavaha srotas.
Any damage to liver ultimately disturbs the digestive system, which is the main
causative factors of all the diseases according to Ayurveda.Among the
Ashtamahagadas the udara roga is one. Among the eight udara rogas pleehodara and
yakritodara are described. In the derangement of yakrit and pleeha, raktakshaya and
other symptoms are manifested.Thus liver and spleen plays vital role in udara and
raktavikaras.
In spite of the tremendous advances made in modern medicine, no effective
and safe hepato-protective medicines are available. Hence an effective and safe
hepato guard formulations are need of the hour. About 80% of the world population
relay on the use of Ayurvedic medicaments which has the treasure of unique
Rasoushadhis.
Introduction
3
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
In present scenario several hepato-protective drugs as well as compound
formulations explained in classics should be pharmacologically evaluated for their
efficacy, safety profile & world wide acceptance.
Drugs used in Ayurveda can be broadly classified into three groups as herbal,
animal and minerals.
1) Herbal Drugs:
In vedic literature and in Ayurvedic classics mostly vegetable drugs were
prescribed for the treatment of different categories of ailments. Very few animal
products and still fewer metals and minerals are described in those texts.
During the fifth century B.C and thereafter the important branch of Ayurveda
namely shalyatantra was viewed as a form of hinsa or violence.
Ahinsa or non-violence was the cardinal rule of the religion prevalent in those
days. The religion which was adopted by the rules and subjects alike, discouraged the
practice of surgery and it was almost legally banned. Therefore the physicians in that
had to find alternative for curing their obstinate surgical conditions. This provided
development of Rasashastra. Metals and Minerals were processed and extensively
used therapeutically in Bhasma form in that time.
According to classics therapies are of three categories viz;
a) Asuri - which includes surgical therapies.
b) Manushi – which is performed by the use of decoctions etc of vegetable drugs.
c) Daivi - which is performed by the administration of metallic and mineral
preparations. The succeeding one are superior to the preceding categories of
therapies.
2) Mineral Drugs:
a) These are usefull in obstinate and incurable conditions.
Introduction
4
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
b) Mineral remedies are therapeutically effective even when administered only in
a small dose (unlike vegetable products which are generally required to be
administered in a much larger dose).
c) These mineral products are not unpalatable (unlike some of the vegetable
remedies which are sometime very unpalatable because of bitter, astringent
and pungent taste)
d) Mineral products produce their therapeutic effects instantaneously (unlike
vegetable products which take larger time because they have to pan through
the process of digestion and metabolism) before they become therapeutically
active.
3) Drugs of Animal origin-
The drugs of animal origin like Sudha varga dravyas do posess actions
like minerals. Many drugs have been mentioned in sudha varga, among them
Shankha is one.
Shankha is a shell of an aquatic insect called Turbinella pyrum. These
shells are available in various sizes and colours, probably according to the
habitat. In India these shankha are available almost throughout its vast sea
shore, but in different sizes and in various shades.
Shankha is the base for therapeutically active drug called shankha bhasma
which is effective medicine in various diseases such as Amlapitta, Shula, Grahani,
Atisara, Yakrit vruddi, Pleeha vruddi etc. Calcium, apart from forming an essential
constituent of bones, plays an important role in body homeostasis and disturbances in
calcium metabolism are associated with derangement of various cellular functions.
Shankhanabhi bhasma is one of the cost effective Rasoushadhi commonly
mentioned both for pleeha and yakrit vikaras in Ayurvedic classics. With this
Introduction
5
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
perspective the study is taken to evaluate the Hepatoprotective activity of
Shankhanabhi bhasma.
Plan of suudy:
The entire study was done in following chapters:
1. Introduction : Involves need of the present study, emphasis on its importance
and plan of the whole study.
2. Review of Literature: The detailed classical Ayurvedic literature and relevant
modern literature about the drugs Shankha, Nimbuka, Kumari, Kanji and
about the disease Yakrit vikara were reviewed.
3. Methodology:
a. Pharmaceutical study: In this, preparation of Kanji, method of
shodhana of shankhanabhi, marana of shankhanabhi is incorporated.
b. Analytical study: This includes physico-chemical analysis of
shankhanabhi bhasma.
c. Experimental study: This includes evaluation of hepato protective
activity of Shankhanabhi bhasma with nimbu swarasa in albino rats.
4. Results: In this part the obtained results are systematically presented, which
include data related to biochemical parameters, data related to
histopathological examination and data related to response to treatment.
5. Discussion: In this chapter observation, findings and results of various studies
have been found out with possible explanation for its effects.
6. Conclusion: The essence of the whole study is mentioned in this chapter.
7. Summary: It contains the information of the overall work in a nut shell.
Objectives
6
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
AIMS AND OBJECTIVES
1.Preparation of Shankhanabhi Bhasma.
2.Physico-Chemical analysis of Shankhanabhi Bhasma.
3.Evaluation of Hepato protective activity of Shankhanabhi Bhasma.
Drug review
7
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
DRUG REVIEW
Rasaoushadhis by virtue of their unmatched properties like Alpamatra,Shigra
phaladayi,Rasayanas, Bahu upayoga etc,have been leading the field of chikitsa since
time immemorial. Shankhanabhi Bhasma along with Jambeera swarasa is a unique
combination to treat Yakrit vruddi and Pleeha vruddi.
In Yogaratnakara under Udara roga chikitsa, Shankhanabhi Bhasma with
Jambeera swarasa is mentioned for Yakrit vruddi1.
The author of Bhaishajya ratnavali also explained the combination of .
Shankhanabhi Bhasma with Jambeera swarasa under Pleehayakrit roga chikitsa
prakarana to treat Yakrit vruddi2.
This combination is also mentioned in texts like Vangasena in
Udararogadhikara3, Rasa Chikitsa4 and Bharata Bhaishajyaratnakara5.
Drug : Shankhanabhi Bhasma.
Anupana : Jambeera swarasa.
SHANKHA :
As Shankhanabhi is the part of Shankha itself, review of the same is done.
Aurvedic literature of Shankha :
1) Utpatti : Relation with God
Shankha is related with Lord Dhanwantari as it has been held by one of the
hands of God.
2)Historical aspect :
a) Vedic kala- In Atharva veda we get references of Shankha many times.According
to it Shankha is mangalakaraka on dharana6 and krumighna on sunada7.
b) Mahabharata : In Mahabharata and Bhagwata granthas also we get references of
Shankha.
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c) Samhita kala : Brihatrayee’s
1. Charaka samhita-
Charaka explained shankha in 27th chapter of sutrasthana under Varishaya varga8.
Also mentioned it in various places as a medicinal use.
2. Shushruta samhita-
Shushruta explained Shankhanabhi in 11th chapter Ksharavidhi adyaya in preparation
of madyama kshara9. Further he has used Shankha churna for lomashatana10.
Shankhanabhi Bhasma is extensively used in many yogas in uttaratantra to cure
various eye diseases in the from of Netra varti’s11.
3.Ashtanga sangraha-
In Ashtanga sangraha shankha is used as anjana paatra12 and in treatment raktapitta
and various netra rogas.
4.Ashtanga hridaya-
In Ashtanga hridaya shankha is used in the treatment of shvitra13 and various netra
rogas.
Laghutrayee’s :
1.Sharangdhara samhita-
Sharangdhara explined the shankha shodhana by giving bhavana with jambeera
swarasa and drying in sunlight14. Shankha bhasma has been used in many yogas and
lepas.Putapak yoga has also been explained15. Shankhanabhi Bhasma is used in
preparation of various netra varti’s.
2.Bhavaprakasha samhita-
The synonyms,guna and doshaghnata of shankha has been explained under dhatu-
upadhatu varga16.
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Table-1 Showing Full name of texts mentioned about shankha
Sl.No Full name of texts mentioned about shankha
1. Ayurveda prakasha.
2. Brihat rasaraja sundara.
3. Bhava prakasha.
4. Rasarnava tantra.
5. Rasamrutam.
6. Rasendra chintamani.
7. Rasendra chudamani.
8. Rasa hrudaya tantra.
9. Rasa jalanidhi.
10. Rasa kaumudi.
11. Rasa kamadhenu.
12. Rasa tarangini.
13. Rasa paddati.
14. Rasa ratna samucchaya.
15. Rasa prakasha sudhakara.
16. Rasendra sara sangrha.
17. Yogaratnakara.
Table-2 Showing Full name of Nighantu’s mentioned about shankha
Sl.No Full name of Nighantu’s mentioned about shankha
1. Bhava prakasha Nighantu.
2. Dhanwantari Nighantu
3. Kaideva Nighantu.
4. Madanapala Nighantu.
5. Raja Nighantu.
6. Shaligrama Nighantu.
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Bhaishajya Ratanavali:
Acharya Govinda Das Sen has explained the shodhana and marana of
Shankha. Shaodhana by swedana in Kanji for one hour and by swedana in Jambeera
swarasa.
He explained common process of Jambeera swarasa swedana for both shankha
and shankhanabhi (Khulwaka) in a single sutra17. He has mentioned the shankha
gunas and indicated for udara roga18. Author has explained many yogas containing
shankha and shankhanabhi.
Yoga Ratanakara:
In yoga ratnakara shankha has been mentioned under Koshasta varga of
Anupa mamsa19. Many yogas of shankha are explained. Shankhanabhi bhasma is a
content for many Netra varti’s.
Rasa Grantha:
In Rasa Shastra Shankha is included for the first time in Rasarnava in “Shukla-
Varga”20 & then in all granthas under “Sudha-Varga”.
Shukla Varga:
vÉÑYsÉ uÉaÉïÈ xÉÑkÉÉMÔüqÉï vÉÇZÉ zÉÑÌ£ü uÉUÉÌOûMüÈ| (UxÉhÉïuÉ – 5/40)
Based on Shukla varnata these drugs are classified in one group.
Sudha Varga:
As they contains lime (Sudha), they are included in sudha varga. Almost all
drugs in shukla varga contains lime and majority of them are of aquatic orgin &
shankha is one of them.
Properties:
1. These drugs contains high percentage of calcium.Many of them are formed
from Kshariya jala of samudra & they are alkaline in nature.
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2. Almost all drugs are hard in nature. Usually they are formed by the
skeleton of marine animals.
Other drugs in sudha varga:
1. Shukti
2. Kapardika
3. Pravala
4. Kukkutanda twak
5. Mruga Shrunga
6. Godanti
7. Badari Pashana
8. Khatika
9. Shambuka
10. Kurma Prushta
11. Ajasthi
12. Mukta
Table No. 3 Showing Synonyms of Shankha21,22,23,24,25,26,27,28,29,30
Synonyms R.T A.P RRS RSS BRRS BP KN SN RN M.N
Arnobhava - - - - - - - - + -
Bahunada - - - - - - - - + -
Deerghanada + - - - - - + - + -
Dhaval - - - - - - + - + -
Dirghanisvana - - - - - - + - - +
Drushtidravi - - - - - - - - - -
Haripriya - - - + - - - - + -
Jaladhara - - - - - - - - - +
Jalaja - - - - - - + - + -
Kamboja + - - - - - - - - -
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Kambu + + + + + + + + + +
Kshudra - + + - + - - - -- -
Kutilanta - - - - - - - - + -
Mahanada - - - + - - - - + -
Mangalaprada - - - - - - - - + -
Pavana dhwani - - - + - + - + + -
Putha - - - - - - - - + -
Samudraja + + + - + + - + - -
Shankha + + + - - + + + + +
Shankhaka + - - - - - - - - -
Shankhanaka + + - + - - - - - -
Shrivibhushana - - - - - - - - - -
Strivibhushana - - - - - - + - - -
Sunada + - - - - + - + + -
Suswara - - - - - - + - + -
Trirekha + - - - - - - - -- -
Varibhava - - - - - - - - - -
Varichara - - - - - - + - - -
Varija - - - - - - - - - +
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NIRUKTI31
Based on Nada / Sound
Shanka - zÉÉqrÉÌiÉ AzÉÑpÉqÉxqÉÉÌSÌiÉ |
Pecifies ashubhata
xÉqÉÑSìÉå°uÉeÉliÉÑÑÌuÉzÉåwÉÈ |
Special kind of Jantu found in samudra.
Deerghanada - SÏbÉÉåï SÕUaÉÉå lÉÉSÉåSxrÉ | ( UÉ. ÌlÉ)
Produces deep sound.
Pawana dhwani - mÉÉuÉlÉÈ mÉÉÌuɧÉeÉlÉMüÉå kuÉÌlÉrÉïxmÉ | (UÉ. ÌlÉ)
Produces devine sound.
Mahanada - qÉWûÉgcÉxÉÉæ lÉÉSgcÉåÌiÉ | qÉWûÉzÉoSÈ
CSqÉÎ°È xÉqÉÇ mÉëÉmiÉÉ rÉå MåüÍcÉSè kÉëÑuÉeÉ…ûqÉÉÈ |
mÉëÌuÉzÉliÉÉå qÉWûÉlÉÉSÇ lÉSÎliÉ pÉrÉmÉÏÌQûiÉÉ || (UÉ. ÌlÉ)
Produces strong sound.
Bahunada - oÉWÒûqÉïWûÉlÉç lÉÉSÈ zÉoSÉå rÉxrÉ | (UÉ. ÌlÉ)
Produces many sound.
Based on Swaroopa:
Dhavala - μÉåiÉaÉÑhÉrÉÑYiÉåå | (AqÉU)
White in colour
Kutilanta - MÑüOûÌiÉ uÉ¢Çü uÉëeÉiÉÏÌiÉ |
Curved inside
Based on Origin:
Kambu - MüqooÉgcÉ¢üzÉUcÉÉrÉaÉSÉÍxÉcÉqqÉï
urÉaÉëæÌWïûUhrÉqÉmÉpÉÑeÉæËUuÉ MüÍhÉïMüÉUÈ ||
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A shell, one marked with three lines.
Kamboja - mÉgcÉlÉSÇ xÉqÉÉUprÉ qsÉåcNûɬͤÉhÉmÉÔuÉïiÉÈ |
MüqoÉÉåeÉSåzÉÉå SåuÉåÍzÉ |
Collected from the kamboja desha.
Varibhava - uÉÉËUhÉå lÉå§ÉeÉsÉÉmÉ pÉuÉÌiÉ mÉëpÉuÉiÉÏÌiÉ |
Found in water.
Jaladhara - kÉUiÉÏÌiÉ kÉUÈ | eÉsÉxmÉ kÉUÈ |
Found in water.
Varija - uÉÉËUÍhÉ eÉÉrÉiÉå CÌiÉ | (CÌiÉ WåûqÉcÉlSìÈ)
Originated from water
Varichara - uÉÉËUrÉÑ cÉUiÉÏÌiÉ |
Moves in water.
Vernacular Names32:
Latin Name : Xanchus pyrum or turbinella Rapa
Sanskrit : Shankha
English : Conch shell
Hindi : Shankh
Kannada : Shankha
Gujrati : Shankha
Marathi : Shankha
Telugu : Sankham
Bengali : Sankh
Tamil : Sankha
Chemical Composition : Calcium Carbonate (CaCo3)
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Source33
Shankhas are of different varities according to its animal. These are widely
dispersed in marine water. It can occur in all the sea shores.
It is the outer covering of “Mollusca group” of aquatic animal which are seen
in sea. It is collected from the sea and put in boiling water. The animal which is
present inside dies and the outer portion, shankha is obtained. It is sold in market.
Chanks are gregarious and exclusively marine animals occurring in large
numbers on muddy sand bottom thirteen metres in depth in Tamilnadu shores and
Andaman waters.
Grahya Lakshana34:
In Rasatarangini, grahya lakshanas of shankha have been described as
Vrutta : It should have round shape.
Snigdha : It should have snigdha sparsha
Sukshma mukha : Having small mouth (opening at apex)
Nirmala : Having clean surface.
Chandusundara : Being like moon looks beautiful.
Dirghakaya : It must be large
Guru : It must be heavy
Sunada : It must produce good sound.
It is of two types according to classics35
1) Dakshinavarta : It is rare and not preferred for preparation of Bhasma.
It is mangalyakaraka & krimighna
2) Vamavarta : It is found abundantly in all sea shores. It is preferred
for preparation of Bhasma.
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Shodhana of shankha:
Shodhana of shankha is done by doing pachana in Dola yantra containing
below mentioned liquids for particular time. Shankha pieces are first kept in pottali.
Which is suspended in Dola yantra for pachana. After complete cooling shankha
pieces are removed from pottali and washed with warm water.
Table No. 4 Showing Shodhana of Shankha36,37,38-42,43,44,45-46
S.L.
no
Procedure & Liquids used RRS AP RT BRRS Sh.S BR
1. Swedana in Kanji + + + +
2. Pachana in Kanji for 1 Yama +
3. Swedana in Jambeera swarasa +
4. Pachana in Jambeera swarasa
for 4 Yama
+
5. Made fine powder, Bhavana is
done with Jambeera swarasa
and kept one day in sunlight for
drying.
+
6 Pachana in Jayanti patra
swarasa for 1 Yama
+
7 Pachana in Tanduliya patra
swarasa for 1 Yama
+
8 Pachana in Nimbuka swarasa
for ½ Yama
+
Almost all Granthas have mentioned shodhana of Shankha in Amala Dravyas.
In the present study procedure of shodhana in kanji is selected.
Shankha Marana47:
Pieces of shodhita shankha are dried well, then they are placed in sharava,
closed with other sharava & sandhibhandhana is made. After complete drying it is
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subjected to Gajaputa. After swangasheeta the sharava is removed and the pieces are
collected, powdered and given bhavana with kumari swarasa & chakrikas are made
and they are once again subjected to Gajaputa such 2-3 putas will yield good white
bhasma of shankha.
The pieces of shodhita shankha are put into the fire and subjected for samyag
laghu puta till they becomes bloomed.48,49
One pala of shodhita shankha along with half masha of tankana is heated in
blind crucible to get shankha bhasma.50
Pharmacological Properties51:
Rasa : Katu Rasa (Kshara)
Guna : Laghu, Sheeta
Veerya : Sheeta
Karma : Grahi, Balya, Vilekhana, Agni deepana, Vishagnhna, Varnya,
Hridya.
Dosha karma : Tridosha shamaka
Rogaghnata :
Table No. 5 Showing Rogaghnata of Shankha bhasma52,53,54,55,56,57,58
Rogaghnata R.T A.P R.S.S B.R.R.S R.N Y.R B.R
Amlapitta + - + - + - -
Grahani + + - + + + -
Parinamashoola + - - - - - -
Tarunyapidika + + - + - + -
Netra pushaphara - + - - + + -
Gulma - - + - - - -
Swasa - - - - - - -
Meha - - + - - - -
Udara shoola - - + - - - -
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Sarva ruja - - - - - - +
Udaramaya - - - - - - +
Matra59 : 2 Gunja (250 mg).
Anupana60 : Nimbu – Swarasa,
Trikatu churna or according to disease.
Table No. 6 Showing Classical Yogas of Shankhanabhi and Shankha
S.L.
No
Vishishta Yoga Indications Reference
1 Muktadi choorna Hikka, Swasa, Kasa, Netra
rogas
Cha.Chi. 17/125
2 Usheeradi choorna Raktapitta, Tamaka swasa,
Trushna etc
Cha. Chi 4/73
Y.R. Raktapitta Chi / 45-46
3 Shankhapishti Prelepa Visarpa Cha. Chi 21/82
4 Shankha Pravaladi Varti Sarva Akshi Roga Cha. Chi. 26/246
5 Lomashatana yoga Loma shatana Su. Chi. 1/105
6 Anjana varti Kaphaja Netra Roga Su. Chi. 12/8
7 Ksharajana Balasagrathita Su. Chi. 12/33
8 Shankadyanjana Arma pidika Su. U. 15/25
9 Rasakriya Kaphaja Timira Su. U. 17/43
10 Lekhana putapaka Kaphaja netraroga Su. U. 18/24-25
11 Revati Pratisheda pradeha Revatighaha badha Su. U. 31/6
Y.R Baharoga Chi./4
12 Shankha dravaka (1) Pleehodara etc R.T. 12/35-39
13 Shankhodara Rasa Grahani etc R.P.S. 8/26-28
14 Hemagarbha Pottali Rasa Mandagni shwasa etc R.P.S. /80-83
15 Lokanatha Rasa Cures 29 diseases with
different Anupana.
R.P.S. /89-93
16 Krutrima pravala nirmana R.P.S. 11/133
17 Kaphaketu Rasa Peenasa, swasa, shiroroga B.R.5 /743-745
18 Vishamajwarataka loha Jwara, Pleeha, Yakrit B.R.5. /1162
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19 Agnisuno Rasa Udara B.R.5. 8/519
20 Shankha vati (1,2,3) Agnimandya B.R.5. /187
21 Mahashankha vati (4) Pleeha, Udara B.R.5. /202,204
22 Ratnagarbha pottali Rasa Rajayakshma, Udara B.R. 14/182
Y.R. Rajayakshma /1-6
23 Sarvangasundara Rasa Rajayakshma B.R . 14/195
24 Shankha choorna Yakrit shula B.R. 30/77
25 Vaishwanara louha Shula B.R. 30/137
26 Shankhanabhi bhasma Pleeha, Yakrit B.R. 30 /112-116
27 Chandrodaya varti (1&2) Netraroga B.R. 64/203
Y.R. Netraroga Chi,
Sha. U. 13/75-77
28 Chandraprabha varti Netraroga B.R.64/203
29 Pradarantaka louha Netraroga B.R 64/72 (R.S.S)
30 Sutikaharo Rasa Sutika roga B.R. 69/99
31 Dantodbhedagadantaka
Rasa
Dantaroga B.R 71/123
32 Trilokyachintamani Pleeha, Jalodara, Rasayana B.R 73/136
Y.R Rajayakshma / 1-12
33 Sutashekhara Rasa Amlapitta, Ajeerna B.R Anubhuta/223
Y.R Amlapitta Chi/1-5
34 Pratapalankeshwara Rasa Sutika roga B.R.Anubhuta /266
35 Hemagarbha pottali Kasa Y.R. Kasa Chi /1-7
Sha. M. 12/107
36 Muktadi mahanjana Netraroga Y.R. Netraroga chi/1-3,
(B.P)
37 Panchabana Rasa Pleehodara, Jalodara
Vajeekarana
Sha.U. 13. Vajeekarana/1-4
38 Tuttadi Rasakriya Netraroga Sha.U. 13/87-88
39 Krishna sarpa vasanjana Netraroga Sha.U. 13/105
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MODERN DESCRIPTION
Origin of Shankha (Shell) 61
According to Zoology, Shankha is exo-skeleton formed by secretion of
Gastropoda class snails which belongs to phylum mollusca.
Table No. 7 Showing Brief introduction of their animals
Phyllum Mollusca
Class Gastropoda
Subclass Prosobranchiata (Strepto neura)
Order Pectini branchiata (Mono tocardia)
Sub order Tenio glossa
Tribe Platypoda
Family Fila
Species Globosca or Xancus Pyrum or
Turbinella pyrum.
There are thousands of species of Gastropoda class.
Structure of shell:
The typical Gastropoda shell is a conical spire composed of tubular whorls &
containing visceral mass of the animal.
Vertical section of shankha showing its part:
1) Whorls : The term whorl is complete revolution of the “Spiral cone”
2) Apex : The Apex is topmost or initial whorl which is the first to be
formed.
3) Aperture : It is the opening of the lowermost end of spire.
4) Collumela : It is the central axis to which whorls of spiral closing applied
together coiling round it.
5) Sutures : It is the line that separates several whorls.
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Starting at apex which contains smallest & oldest successive larger whorls are
called about central axis called collumella. The larger whorl eventually terminates at
opening or Aperture from which head and foot of living animal protrude.
The collumella itself is called Shankhanabhi.
Identification of Right & left handed shells:
A shell possesses a Right handed spiral when the aperture open to the right of
columella if shell is held with spire up and aperture facing observer and left handed
opens to left.
Most of Gastropodas are right handed. Few are left handed and some species
have both Right handed and left handed individuals.
Varieties of shells:
Gastropoda shells display an infinite variety of colours, patterns, shapes. In
considerable number of gastropodas, the shell is conspisously spiralled only in
juvenile stage. The coil nature disappears with growth and adults shell represents a
single large expanded whorl.
Size of shells:
The height of shell is the distance between the apex and the lower margin of
aperture. In some species of these little snails it does not exceed 1.5 mm. The largest
shell which may reach a height of 2 ft. are found.
Chemical composition of shell:
According to modern science a typical Gastropoda shell is composed of three
layers, an outer peri-ostracum, a middle pre-mastic layer, a inner nacreous layer.
Periosteoum is thin & composed of horney organic material called concolin.
The two layers consists of calcium carbonate.
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Calcium Carbonate62:
Calcium carbonate occurs in large quantities in nature as chalk, marble and
lime stone. However enough CaCO3 is observed to cause systemic and renal effects
but it has mainly considered to be the non-systemic antacid.
Ref: B.S. Bahl & G.D. Sharma – Inorganic chemistry uses of CaCo3, P-470.
Absorption And Excretion63:
CaCO3 Ca+2 + CO3-2
H2O+ H2CO3 H2O + CO2
The Calcium cations formed in reaction and present as the water soluble
calcium chloride salt can be either absorbed or precipitated as the insoluble calcium
phosphate salt in the intestine or as insoluble calcium soaps from the hydrolyzed
glycerides resulting from digested food. Calcium excretion varies directly with the
creatinine clearance.
Preparation64
It is obtained in the laboratory by the action of soluble carbonate on a calcium
salt or by passing CO2 through lime water.
CaCl2 + Na2CO3 CaCO3 + 2NaCl
Ca (OH)2 + CO2 Ca CO3 + H2O
Untowards Effects65
Constipation and chalky taste of calcium carbonate are clinical disadvantages.
Nausea is an occasional complaint, mere seriously infrequent instances of
hypercalcimia with alkalosis, caleinosis and azotemia occur during chronic calcium
carbonate usage.
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Contraindications
Patients with renal disease, history of calculi, gastro intestinal haermorrhage,
hypertension or dehydration and electrolyte imbalance due to excessive vomiting.
Properties66
1) It is soluble in water containing carbon dioxide forming calcium bicarbonate.
CaCO3 + H2O + CO2 Ca (HCO3)2
2) It is fine white, odorless, tasteless, microcrystalline powder which is stable in
air.
3) It is insoluble in alcohol, water and dissolves with effervescence in diluted
hydrochloric & diluted Nitric acids.
Uses:
1) It is used as Diuretic, Emmenogogue, Astringent, Antacid, Local sedative and
Antiseptic.
2) For the manufacture of lime.
3) As a flux in the smelling of ores.
4) In the preparation of tooth pastes and face powders.
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Review of Researches carried out on Shankha Bhashma67:
1) PL-31 Adarsh kumar /1984
Shankha Bhasma Nirman Evam Amlapitta par Ahdyayan.
2) Pune – Tilak – 901 Kulkarni (Ms) Manisha M /1995
A comparative study of Shankhanabhi Bhasma and Shanka Bhasma (Avarta)
from the point of view of their chemical composition.
3) JM- 554 Tank ZG /2000.
A Pharmaco – clinical study of Shankha Bhashma alone and Shankha
Bhashma along with Amalaki churna in the management of Amlapitta.
4) JM – 563 Rameshchandra A/ 2002
Shankha Bhashma evan Yuvana – Pidika lepa ka Nirmanatamaka evam
Yuvana – pidaka vyadhi par prabhavatmaka Adyayan.
5) Belgaum – 109 Benade shakhar V / 2003
Comparative analytical study and standardization of different samples of
Shankha and shankha Bhashma.
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REVIEW OF LITERATURE OF KUMARI
Classification
Kula : Rasona Kula
Family : Liliaceae
Latin name : Aloe vera
English name : Indian aloe
Synonyms68:
1) Kumari
2) Grita kumari
3) Gruhakanya
4) Deerghapatrika
5) Ajara
6) Veera
7) Taruni
8) Rama
9) Kapila
10) Sthaladala
11) Mata
12) Mandala
13) Akshayakaraka
14) Atipittala
Botanical Description69:
Shrubs : 30-60 cms high
Leaves : 1-2 ft in length
2-4 fingers in circumference.
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The margins of the leaves have spikes. Leaves contains a thick and
mucilaginous juice.
Once the shrub becomes old a straight stalk emerges from its centre which
bears red flowers in cluster. The plant flowers and fruits at the end of winter.
Kala bol:
The solid gum obtained after boiling the juice of kumari is called as aliabol,
kala bol, kumarisara, musabbar etc. It is called aloes in English.
Chemical composition70
1) Aloin ie anthraquinone glycoside i.e barboloin as crystalline glucosidee.
2) Aloe amodine
3) Resins : 16 to 63%
4) Water soluble substances 50%
Properties71:
Guna : Snigdha, picchila
Rasa : Tikta, madhura
Veerya : Sheeta
Vipaka : Katu
Prabhava : Bhedan
Kala bol : Laghu, Ruksha, Teekshna, Ushna
Dosha : Tridosha shamaka
Dhatu : Rasayani, Rasa, Rakta, Meda, Shukra vardhaka
Mala : Purisha bhedaka, Arthavajanaka.
Uses: Gulma, Pleeha Yakrit vruddi, Kaphajwara, Visphota, Vishahara, Kushtagna, Swasa, Pittaja kasa. Aloe is used as a purgative, effect is mainly on colon. Upayukta Anga: Juice from leaves, Kala bol.
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REVIEW OF LITERATUER OF JAMBEERA
Classification
Kula : Jambeera kula
Family : Rutaceae
Latin Name : Citrus limon
Synonyms72
1) Jambeera
2) Dantashata
3) Jamba
4) Jambeera
5) Jambala
Botanical Description73
Small tree : Strangling, bushy, 3-4m high with thorny branches.
Leaves : Ovate, Petiole margined or winged
Flowers : Small, white or pinkish, sweet-scented.
Fruit : Oblong or ovoid, bright yellow, rind thick, pulp acid, pale
yellow.
Macroscopic character74:
Colour : Fresh outer surface, bright green yellow, internally white.
Dried : Outer surface is yellow and inner surface pithy white
Odour : Strong, fragnant, aromatic & characteristic
Taste : Bitter
Distribution
Cultivated /grown in U.P, Maharashtra, Tamil Nadu, and Karnataka. Found
wild in the north-west regions of India upto 1300m.
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Major Chemical constituents75
The Volatile oil of the drug contains mainly limonene (about 90%), citral (about 4%)
and other aromatic compounds like geranyl acetate and terpineol.
Pharmacological properties76
Rasa : Amla
Guna : Guru, Teekshna
Veerya : Ushna
Vipaka : Amla
Doshaghnata : Vatashleshmahara
Uses/ Rogaghnata: Shoola, Kasa, Chardi, Trushna, Ama, Asyauairasya, hritpeeda,
Vanhimandya, Krumiroga.
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REVIEW OF LITERATURE OF KANJI
Names in different languages:
Sanskrit : Kanji
English : Sour gruel
Method of preparation77:
The liquor prepared by the fermentation of manda obtained by boiling
kulmasha i.e broken masha or dhanya i.e raktashali or yavachurna etc is called kanji.
Pharmacological properties78:
Guna : Laghu, Teekshna, Ushna, Sparshat sheeta
Veerya : Ushna
Karma : Dahanashaka, Klamahara, Deepana, Pachana, Bhedi, Rochana,
Koshtashuddikara, Bastishodhaka, Rechaka, Amahara
Doshaghnata : Vatakaphahara
Rogaghnata : Trushna, Shula, Ajeerna, Jwara.
Use : For swedana of parada
For shodhana of Rasa dravya’s.
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PHARMACEUTICAL PROCEDURES AND YANTRAS USED IN PRESENT
STUDY
Shodhana79:
The process by which the malas or doshas of rasadravyas are eliminated is
called as shodhana. Shodhana is accomplished by applying various methods like
mardana, swedana etc.
By shodhana rasadravyas becomes soft and brittle for further steps.Shodhana does not
mean the mere elimination of impurities but also it is potentiation making the drugs
biologically acceptable.
Main objectives of shodhana are :
■ Detoxification
■ Therapeutic potentiation
■ Making rasadravya suitable for next process.
In present study shankhanabhi shodhana is done by pachana in dolayantra and
marana by subjecting to gajaputa.
Pachana :
The process of pachana (boiling) of any rasadravya along with kshara or any
dravya swarasa is called as pachana.By doing pachana the mala or impurities loose
their adhesiveness.
Dolayantra80:
In present study dolayantra is used for pachana of shankhanabhi.
At the neck of earthen pot two holes are made and a iron stick is inserted.The
pot is half filled with required liquid and pottali made of three fold cloth containing
rasadravya is suspended such that it is four angula above from the bottom of pot and
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immersed in the liquid.Then the pot is subjected to mrudvagni.This apparatus is
known as dolayantra.
Marana :
qÉÉUrÉiÉå lÉzrÉiÉå pÉxqÉÏ¢üÏrÉiÉå CÌiÉ |
Marana means “killing” and converting a metal into irreversible and
final form i.e bhasma.
Definition:
The processes by which a metals, minerals or any hard substance is
subjected to soaking, drying and ignition to convert into bhasma is known as
Marana.This marana process converts metals into fine state of smaller molecules
and makes them so light as to be highly absorbable and assimilable after oral
administration.
1) Marana is process by which metal looses its original state (metallic) & still
retains its originality (medicinal value)
2) By marana process drug is converted into a biologically acceptable form.
Stages of marana:
1) Mardana: The shodhita dravya is put into khalva and mardana is done with
swarasa of specified plants or kashayas of drugs mentioned for a particular mineral or
metal.Mardana is done for a specific period.
2)Chakrika: The small cakes are made which are called as chakrika.The size
and thickness of cakes depends on the heaviness of the drugs.The heavier the drug the
thinner are the cakes.These cakes are dried well under sunlight.
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3)Sharava and Sandhibandhana : These chakrikas are placed in one single
layer in shallow earthen plate called as sharava.It is closed with same another
plate.The edge is sealed with clay smeared cloth in seven consecutive layers and
dried.This process is called sandhibandhana.
4)Puta : The puta is the measure of heating arrangement meant for preparing
various kinds of bhasmas of maharasa,uparasa,loha,etc.We should not give more or
less quantity of heat.
Gajaputa: Its an arrangement of heating in a pit 90cms (one Rajahasta) in
length,breadth and depth.
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DISEASE REVIEW
Review of yakrit in Ayurveda
In Ayurveda yakrit is considered as one of the Koshtanga and it is a matruja
avayava formed from samana vata, dehoshma and rakta. (Arunadatta)81
The origin and development of the word Yakrit82
rÉ qÉç A M×ü : rÉ xÉÇrÉqÉ MüUÉåÌiÉ
The meaning is that the liver controls various physiological events.
(Ayurved & Hepatic disorders)
In veda yakrit is called as Takima or yakna.
(Shabda stoma mahanidhi).
The other synonyms are 83:
Kalakhanda – Dalhana on Su.Sa.Sha.4/25
Jyotisthana / Agnisthana – Su.Sa.Sha.4/57-58
Raktadhara – Dalhana on Su.Sa.Sha.4/17
Raktashaya – Su.Sa.Sha.5/9,
Yakritpinda - Parishadya shabdartha shareera.
Origin of Yakrit:
In garbhavastha yakrit & pleeha are formed from shonita. (Su.Sha.4/25)
Position of Yakrit84:
The anatomical site of the yakrit is mentioned as below and right to the
hridaya,which is hard in texure.
Site of Blood85:
Sthana of Shonita is Yakrit, Pleeha.
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Raktavaha Srotas86:
Raktavaha Srotas are two in number & their mula is yakrit,pleeha &
raktavahini dhamanis.
Yakrit87: An Abdominal organ
Yakrit is one among 15 koshtanga’s.
Site of Ranjaka pitta88:
The pitta present in yakrit & pleeha gives ranjana.
Raktadhara kala89:
It is the second kala present inside the muscles,within which shonita is present,
especially in siras localized in yakrit and pleeha.
Just as milky sap flows out when trees with milky sap are cut ( or either bark
is bruised), similarly when muscles are cut , blood flows out quickly in great quantity.
Purishadhara kala90:
This kala extends throughout the koshta and yakrit.Its function is to separate
the faeces from food at the level of unduka (the caecum).
The functions of yakrit are mentioned as follows91
It is the moola of the rakta vaha srotas
It is the seat for the ranjaka pitta
Primary seat for the formation of the rakta
Gives gati to the rakta
Serves as a seat of raktadhara kala.
The mula of raktavah srotas is considered as yakrit and pleeha. Also it is
mentioned that when rasa reaches yakrit and pleeha it gets coloured and then it is
called as rakta. From above description it follows that yakrti, rakta have an Samavaya
relation. Therefore for every vitiation of Rakta there will also be derangement in
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functions of yakrit and vice versa92. The involvement of pitta in this pathology should
also be considered as rakta and pitta bear Asraya and Ashrayi bhava and also acha
pitta is derived from yakrit. In many disease conditions, where Rakta involvement or
yakrit involvement is explained in ayurveda. From the description of the digestion
process mentioned in ayurveda it is evident that yakrit also takes an important role in
digestive process. Like this in conditions such as agnimandya, aruchi, hrallasa,
uttklesha, ama etc, the involvement of yakrit should be considered.
Agni vyapara and Yakrit:
13 types of Agni carry out the process of digestion according to Ayurveda.
Jatharagni paka leads to the breakdown of different proximate components of the food
and renders them fit for absorption. Bhutagni paka processes and converts the
nutrients absorbed from adhoamashaya as pre- homologous of substances, which are
meant finally to be utilised for the upachaya, or building up of the sthayidhatus.
Shri Dwarakanathji (Digestion and metabolism in Ayurveda. Edition-First,
1971) has concluded that the bhutagni paka takes place in the Liver from its
anatomical and physiological relationship to the koshta. This clearly indicates the role
of Yakrit in production of “Ama” in its impaired condition and hence produces aruchi,
agnimandya, and ajeerna etc conditions.
Further according to Dwaraknathji, eventhough the metabolism of asthayi
dhatu into sthayi dhatus takes places in dhatus itself, but yakrit has an important role
in action on dhatwagnis also.93
Yakrit in Raktapitta:
Both rakta and pitta are mutually vitiated in this disease. Dravatwa, ushnatwa
in rakta and pitta will be increased, and it is clearly mentioned by Charaka, that yakrit
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pleeha and raktavahi siras are affected in this disease94 (adhistana). While explaining
raktapitta chikitsa, Sushruta advised consume Aja yakrit (raw) along with pitta.95
Yakrit in Pandu roga:
Pandu roga has been counted both in pittaja and raktaja diseases. Pandu
manifests because of raktakshaya. There may be sanga in rasavaha, which inhibits the
nourishment to rakta etc dhatus96. For the formation of rakta, proper functioning of
rakta dhatwagni and ranjaka pitta is necessary and it takes place at yakrit and pleeha.
If ranjaka pitta and rakta dhatwagni are deficit there will be alparakta formation
leading to raktalpata. Functional deficiency/vitiation of Yakrit is mentioned in this
disease indirectly97.
Yakrit in Kamala:
When the person suffering from pandu, intakes more pittaja substances, the
excessively increased pitta burns rakta and mamsa leading to kamala98. Yakrit is root
of rakta vaha srotoses. In this disease the increased mala pitta vitiates rakta and hence
produces the kamala. It is also evident from investigations that there will be functional
and structural derangement of yakrit in this disease. The symptoms like haridranetra,
twak,nakha, anana, raktapeetashakranmutra, bekhavarna, daha, avipaka, aruchi etc
mentioned in koshta shakhashraya kamala are similar to the disease jaundice99.
In second variety of this disease shakhashrita kamala, there will be sanga of
pitta by kapha, the symptom mentioned such as tilapista nibha varchas etc, are similar
to obstructive jaundice. The investigations suggest that there will be sanga of pitta in
yakrit, which moves to shakhas to produce the disease condition100.
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Toxins and Yakrit:
Garavisha, a special type of toxin described in Ayurveda affects liver &
hepatomegaly may take place.
The reason for hepatomegaly is that the blood has special affinity towards
toxins.The toxins immediately get spread in the body through blood.101
Table No. 8 Showing Correlation Between Ranjakapitta And Bile.
Ranjaka pitta Bile
Site Yakrit/Liver Liver
Derived from Pitta Breakdown products of
haemoglobin
Function Imparts colour to purisha, rasa Imparts colour to stool,
helps in emulsification of
fats
Obstruction results in Tila pishta nibha varchas Clay coloured stool
Yakrit in Udara roga: -
In this condition the direct involvement of Yakrit is mentioned. There will be
enlargement of Yakrit in this disease (sparsha gamya –kathinavastha). As the
Agnimandya is the root cause of all the Udaras, the functional derangement of Yakrit
can be inferred, because it takes part in digestion process102. All the nidana, linga and
chikitsa etc. mentioned for the disease Plehoodara should be taken similar for
Yakritoodara. All the types of Udara rogas mentioned if neglected would turn to
Jalodara. Yakritodara can be taken, as one of the ajatodakavastha of Jalodara, which if
neglected becomes Jalodara along with morphological and functional changes in
Yakrit, and this condition resembles ‘Ascites’.
All the measures described in Pleehodara are to be adopted in
Yakritodara103,104,105,106. The causative factors of Pleehamaya are just applicable to
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Yakritamaya also. The description of Yakrudulyudara in all the text of Ayurveda is
mixed up with that of pleehodara.
Dalhana while commenting on Sushruta Nidana Sthana 7/16 categorically
stated that Pleehodara itself as Yakrudulyudara. But Bhavamishra describes
Yakrudulyudara as a variety of Pleehodara.
Table No. 9 Showing Nidana of Pleehodara and Yakritodara:107,108,109,110,111
Nidana C.S S.S A.H M.N Y.R
Ashitasya atiyana sevana + +
Atichesta + +
Ativyavaya + +
Bharavahana + +
Adhwa + +
Vamanavyadhi karshana + +
Vidahi bhojana + +
Abhishyandi bhojana + +
Ajeerna + + +
Malina anna sevana + + +
Mala sanchaya + + +
The etiological factors in modern science also given more importance to the
diet such as exposure to toxic chemical substances, contaminated food, consumption
of spirit/liquors and through drug injury, is at large producing liver disorders.
Table No. 10 Showing Roopa of Pleehodara and Yakritodara:112,113,114,115,116
Roopa C.S S.S A.H M.N Y.R
Dourbalya + +
Aruchi +
Avipaka +
Varchograha +
Mutragraha +
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Tama pravesha + +
Angamarda +
Chardi + +
Murcha + +
Angasada +
Kasa + +
Gourava +
Admana +
Daha +
Shwasa + + + + +
Mrudujwara + +
Anaha +
Agninasha +
Karshya +
Asyavairasya + +
Parvabheda +
Koshta vata shula + +
Aruna varna udara + +
Vivarna udara + +
Neelarajimudara +
Haritarajimudara +
Haridrarajimudara + +
Katinya +
Dakshinaparshwa parivruddi + + +
Seedati + + +
Mandagni + + +
Kaphapitta lingopadruta + + +
Atipandu + +
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Doshasambanda Vivechana in Pleehodara and Yakritodara117:
If the Yakritodara is associated with -
Udavarta, Ruja, Anaha it is Vataja
Moha, Trushna, Daha, Jwara it is Pittaja
Gourava, Aruchi, Katinya it is Kaphaja
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LIVER DISEASES - A MODERN APPROACH118
Liver plays a pivotal role in regulation of physiological processes. It is
involved in several vital functions such as metabolism, secretion and storage. Further
more detoxification of a variety of drugs and xenobiotics occur in liver. Liver diseases
are mainly caused due to an exposure to toxic chemical substances like antibiotics,
chemotherapeutics, peroxidised oils aflatoxin, carbon tetra chloride, chlorinated hydro
carbons, varied infections, auto immuno disorders and also due to chronic alcoholism.
Most of the hepato toxic chemicals damage liver cells mainly by inducing lipid
peroxidation and other oxidative damages takes place in liver.
It has been estimated that about 90% of the acute hepatitis is resulting due to
viruses. The major viral agents involved are Hepatitis A, B, C, D, E & G. Of these
Hepatitis B infection often results in chronic liver diseases and cirrhosis of liver.
Consumption of spirits/liquors is the prime cause for liver damage in India, so
also been through drug injury is at large producing liver disorders. On survey, studies
around 2-3 % of Indian population are carrying Hepatitis B or C type of viruses.
World wide these figures are increasing with an alarm. (Ref: Indian Journal of
Pharmacology 1999; 31:166-175)
Physiology and its affections119
It is the largest organ in the body weighing from 1200 gm- 1500 gm. It
occupies the whole of the right hypochondriac region and greater part of the epigastric
region; some times it may even extend to the left hypochondriac region. It is roughly
pyramidal or wedge shaped with the base towards the right and apex towards the left.
The liver is both exocrine and endocrine in nature. The exocrine part secretes
bile, which is carried by bile duct and endocrine part secretes plasma proteins, glucose
and heparin.
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Its various function interrelates to each other. This becomes particularly
evident in clinical abnormality of the liver because many of its function are
distributed simultaneously but in different combinations, depending upon the nature
of the disorder.
The basic function of the liver can be divided into:
1. Its vascular functions for storage and filtration of blood.
2. Its metabolic functions concerned with the majority of the metabolic system of the
body.
3. Its secretary and excretory functions that are responsible for forming the bile that
flows through bile ducts into the gastrointestinal tract.
Physiologic Anatomy of the Liver120
The basic functional unit of the liver is the liver lobule, which is a cylindrical
structure several mm in length and 0.8-2mm in diameter. The human liver contains
50,000-1,00,000 individual lobules.
The liver lobule is constructed around a central vein that empties into the
hepatic veins and hence into the inferior vena cava. The lobule itself is composed
principally of many hepatic cellular plates that radiate centrifugally from the central
vein like spokes in a wheel. Each hepatic plate is 1-2 cells thick and between the
adjacent cells lie small bile canaliculi that empty into bile ducts in the fibrous septa
separating the adjacent liver lobules.
Also in the septa are small portal venules that receive their blood from the
portal veins. From these venules blood flows into flat, branching hepatic sinusoids
that lie between the hepatic plates, and then into the central vein. Thus the hepatic
cells are exposed continuously to portal venous blood.
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In addition to the portal venules, hepatic arterioles are also present in the
interlobular septa.
Functions of the Liver121
Synthetic: The liver synthesizes
1. The plasma proteins (except the immunoglobulins). The plasma albumin is
especially important in this respect. Hepatic failure, after a couple of days,
produces hypoalbuminemia and edema. γ- globulins (immunoglobulins) are
however, not synthesized by the liver (they are synthesized by the B-
lymphocytes).
2. Some blood clotting factors, like fibrinogen, prothrombin, factors ‘V’, the liver
synthesize ‘VIII’ and ‘X’. Hepatic failure thus leads to haemorrhagic disorders.
The liver also manufactures some anti-coagulants. Hepatic failure can thus lead
to intravascular clotting too.
3. The liver also synthesizes many of the enzymes, the alkaline phosphatase and
transaminases are notable examples. After damage of the liver cells, the
concentration of the glutamic oxalocetic transaminase and glutamate pyruvate
transaminase of serum (SGOT/AST and SGPT/ALT) rise considerably, as these
enzymes are released from the damaged hepatic cells. Estimation of the SGOT
and SGPT, thus are often done in cases of hepatic damages, for diagnostic and
prognostic purposes.
4. Urea synthesis also occurs in the liver. Blood urea level therefore may fall in
liver failure (provided kidneys are working satisfactorily).
5. Liver synthesizes cholesterol. Serum cholesterol values may fall in liver failure
(whereas in obstructive jaundice, serum cholesterol concentration, recall, rises).
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Detoxification
Liver detoxifies many drugs by one of the two mechanisms.
1. The drug is made more soluble, so that this more soluble derivative can be
eliminated via bile or urine, after conjugation with glucuronic acid.
2. Inactivation (by oxidation or reduction) of the drug by the liver.
In a liver failure, even a standard dose of morphine or barbiturate thus, can
produce signs of overdose and indeed these two drugs are contraindicated in liver
failure. Liver also metabolizes alcohol (CH3CH2OH), first by oxidizing it into
CH3CHO, which is then further metabolized in the mitochondria of the liver cell.
Hormone Inactivation
Liver inactivates many important hormones like cortisol, aldosterone, insulin,
glucagon, testosterone and thyroxin.
Storage
Liver stores some glycogens and some vitamins (notably A and B12).
Bile Secretion
Bile acids (colic acid and related products) are synthesized in the liver and
released into the biliary channel as their salts. Conjugation of bilirubin, the bile
pigment, with glucoronic acid also occurs in the liver.
Metabolic Functions
Liver participates in all the three metabolisms namely-
a) Carbohydrates – Glycogenesis, glycogenolysis and glycogenogenesis all occur in
the liver
b) Protein – In connection with protein metabolism, the liver performs the following
functions:
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It synthesizes the plasma proteins, blood clotting factors, enzymes, lipo
proteins, urea etc. As a result when the liver fails, the amino acids remain unutilized.
In massive hepatic damage, rising amino acid concentration of the blood and ‘amino
aciduria’ result. Further, because of the fact that urea synthesis occurs in the liver,
liver is the organ, which removes ammonia from the body. So, in liver failure blood
NH3 rises and this probably constitutes the most dangerous development.
c) Fat metabolism- β oxidation occurs in the liver, the FFA, which reaches the liver
from adipose tissue via blood, is also esterified to form triglycerides in the liver. The
synthesis of saturated fatty acids from the ‘active acetates’ occurs in the liver. In liver
failure, the fat may not be sufficiently removed for lack of adequate quantities of
lipoproteins or, lack of β oxidation may cause accumulation of fatty acids in the liver.
Therefore excess accumulation of the fat in the liver (fatty liver) can occur in hepatic
insufficiency. Some drugs (e.g. Carbon tetrachloride) may cause fatty liver by this
way.
Most of the body tissues (liver, testes, brain, aorta, intestine) can synthesis
cholesterol from active acetate, but for practical purposes the liver (and to some extent
the intestine) are the most important organs for cholesterol synthesis. Acute hepatic
failure is thus, usually associated with fall of plasma cholesterol values.
Antibacterial Action
The intestine harbors many bacteria and the venous blood from the intestine
(portal vein) contains bacteria, but these bacteria are removed while the blood passes
through the sinusoids of the liver. The sinusoids are lined by Kupffer’s cells, which
destroy these bacteria. In severe liver damage, bacterial invasion of the body from the
intestinal bacteria can occur, as the Kupffer’s cells no longer trap them.
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Classification of Liver Diseases122
Majority of liver disorders cannot be classified accurately into a disease
pattern, because in many instances the etiology and pathogenetic mechanism are
obscure. As a consequence, one finds an abundance of labels and names applied to
hepatic disorders. Some individuals use the term hepatitis to imply viral infection,
others simply to note evidence of hepatic inflammation. One finds ambiguity in the
use of the words acute, sub acute and chronic. Chronicity should refer to continuing or
recurrent disease (i.e. duration). Activity should refer to evidence of the presence of
perpetuation of liver cell injury; this is most readily identified on biopsy by the degree
of hepatocellular necrosis and by serum transaminase elevations.
Because of the difficulties involved in defining the perfect etiology of many
types of liver disorders, in most instances the process is best defined and described by
an examination of the morphology characters of the lesion. Therefore a morphologic
classification of liver diseases as out lined. Classification present more practical than
one based on etiology.
I. Parenchymal
a) Hepatitis (viral, drug-induced, toxic)
i) Acute
ii) Chronic [persistent or ‘active’ (aggressive)]
b) Cirrhosis
i) Laennec’s (portal, nutritional, and ‘alcoholic’)
ii) Post necrotic
iii) Biliary
iv) Hemochromatosis
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v) Rare types (e.g. Wilson’s disease, Galactosemia, cystic fibrosis of pancreas,
α1- antitrypsin deficiency)
c) Infiltrations:
i) Glycogen
ii) Fat (neutral fat, cholesterol, gangliosides, cerebrosides)
iii) Amyloid
iv) Lymphoma, leukaemia
v) Granuloma (e.g. sarcoidosis, tuberculosis)
d) Space-occupying lesions:
i) Hepatoma, metastatic tumour
ii) Abscess (pyogenic, amoebic)
iii) Cysts (polycystic disease, Echinococcus)
iv) Gummas
e) Functional disorders associated with jaundice
i) Gilbert’s syndrome
ii) Crigler-Najjar syndrome
iii) Dubin-Johnson and Rotor syndromes
iv) Cholestasis of pregnancy and benign recurrent cholestasis
II. Hepatobiliary
a) Extra hepatic biliary obstruction (by stone, stricture, or tumour)
b) Cholangitis
III. Vascular
a) Chronic passive congestion and cardiac cirrhosis
b) Hepatic vein thrombosis (Budd-Chiari syndrome)
c) Portal vein thrombosis
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d) Pylephlebitis
e) Arteriovenous malformations.
Toxic And Drug Induced Hepatitis123
The liver plays a central role in the metabolism of a large number of organic
and inorganic chemicals and drugs. Drug induced hepatitis is an inflammation of the
liver with symptoms similar to viral hepatitis, but one difference it is caused by
medication not a virus.
Toxic liver injury produced by drugs and chemicals may virtually mimic any
form of naturally occurring liver diseases. Liver injury may follow the inhalation,
ingestion, or parenteral administration of a number of pharmacologic and chemical
agents. These include industrial toxins (eg. Carbon tetrachloride, Trichloro ethylene,
and yellow phosphorous), the heat-stable toxic bicyclic octapeptides of certain species
of Amanita and Galerina heptotoxic mushroom poisoning and more commonly,
pharmacologic agents used in medical therapy.
In fact any patient presenting with liver diseases or unexplained jaundice is
thoroughly questioned carefully about exposure to chemicals used in work or at home
and drugs taken by prescription or brought “over the counter”.
As the major drug metabolising and detoxifying organ in the body, the liver is
subject to potential damage from an enormous array of therapeutic and environmental
chemicals. Injury may result (1) from direct toxicity (2) via hepatic conversion of
xenobiotic to an active toxin. (3) through immune mechanism, usually by the drug or
a cellular protein in to immunogen.
It has been estimated that about 90% of acute hepatitis is resulting due to
viruses. Consumption of liquors and also through drug injury is the major cause of
liver diseases.
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In general, two major types of chemical hepatotoxicity have been recognised;
(1) Direct toxic type and (2) Idiosyncratic type.
Direct toxic hepatitis occurs with predictable regularity in individuals exposed
to the offending agent and is dose dependent. The latent period between exposure and
liver injury is usually short (often several hours), although clinical manifestations may
be delayed for 24 –48 hours. Agents producing toxic hepatitis are generally systemic
poison are converted in the liver to the toxic metabolites. The direct hepatotoxins
result in morphologic abnormalities that are reasonably characteristic and
reproducible for each toxin. For eg. Carbon tetrachloride and trichloro ethylene
characteristically produced a centri lobular zonal necrosis. Yellow phosphorous
poisoning typically results in periportal injury. Amanita phalloids usually produced
massive hepatic necrosis.
The toxicity produces by the direct hepatotoxins may go unrecognized until
jaundice appears.
In Idiosyncratic drug reactions the occurrence of hepatitis is usually infrequent
and unpredictable, the response is not dose dependent, and it may occur at any time
during or shortly after exposure to the drug. Extra hepatic manifestations of
hypersensitivity such as rash, arthralgias, fever etc. – with Idiosyncratic hepatotoxic
drug reactions is immunologically mediated.
In most cases, even idiosyncratic reactions represent direct hepatotoxicity but
are caused by drug metabolites rather than by the intact compound. Even the
prototype of idiosyncratic hepatotoxicity reactions, halothane hepatitis and isoniazid
hepatotoxicity, associated frequently with hyper sensitivity manifestations are now
recognized to be mediated by toxic metabolites that damage liver cells directly. In
selected cases, the drug or its metabolite has been shown to bind to a host cellular
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component forming a hapten; the immune response to this “neoantigen” is postulated
to play a role in the pathogenesis of liver injury. Therefore, some authorities
subdivide idiosyncratic drug hepatotoxicity into hypersensitivity (allergic) and
“metabolic” categories.
Idiosyncratic reactions lead to a morphologic pattern that is more variable than
those produced by direct toxins; a single agent is often capable of causing a variety of
lesions, although certain patterns tend to predominate. Depending on the agent
involved, idiosyncratic hepatitis may result in a clinical and morphologic picture
indistinguishable from that of viral hepatitis (eg.halothane) or may simulate
extrahepatic bile duct obstruction clinically with morphologic evidence of cholestasis
and minimal hepatocellular damage (eg. Chlorpromazine). Morphologic alterations
also may include bridging hepatic necrosis (eg. Methyldopa) or infrequently hepatic
granulomas (eg. Sulfonamides).
Not all adverse hepatic drug reactions can be classified as either toxic or
idiosyncratic in type. For eg. Oral contraceptives, which combine estrogenic and
progestational compounds, may result in impairment of hepatic tests and occasionally
in jaundice. They do not produce necrosis or fatty change, manifestations of
hypersensitivity are generally absent and susceptibility to the development of oral
contraceptive induced cholestasis appears to be genetically determined.
It should be noted that (1) the injury may be immediate or take weeks to
months to develop. (2) It may take the form of overt hepatocyte necrosis cholestasis
or insidious onset of liver dysfunction. Most important, drug induced chronic hepatitis
is clinically and histologically indistinguishable from chronic viral hepatitis, and
hence serologic markers of viral infection are critical for making the distinction.
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Because drug-induced hepatitis is often a presumptive diagnosis and many
other disorders produce a similar clinicopathologic picture, evidence of a causal
relationship between the use of a drug and subsequent liver injury may be difficult to
establish, which lead to a high frequency of hepatic impairment after a short latent
period. Idiosyncratic reactions may be reproduce, in some instances, when rechalenge,
after an asymptamatic period, results in a recurrence of signs, symptoms and
morphologic and biochemical abnormalities. Rechalenge however, is often ethically
unfeasible, because severe reactions may occur.
Table No. 11. Showing Principle Alterations Of Hepatic Morphology Produced
By Some Commonly Used Drugs And Chemicals.124
Principal
Morphologic Change
Class of Agent Example
Cholestasis Anabolic steroid,
Anti-inflammatory
Antibiotic
Oncotherapeutic
Oral contraceptive
Methyl testosterone
Sulindac
Erythromycin estolate,
rifampcin
Busulfan, tamoxifen
Norethynodrel with
mestranol.
Fatty liver Antibiotic
Antiviral
Oncotherapeutic
Tetracycline
Dideoxynucleosides
Asparaginase,
methotrexate
Hepatitis Anticonvulsant
Antihypertensive
Anti-inflammatory
Anti fungal
Phenytoin, carbamazine
Methyldopa, captopril
Indomethacin, Ibuprofen
Fluconazole, Ketocanazole
Toxic (necrosis) Metal
Hydrocarbon
Yellow phosphorous
Carbon tetrachloride
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Analgesic Acetaminophen
Granulomas Anti arrhythmic
Anti biotic
Quinidine
Sulfanomides
Symptoms
There are similar symptoms in drug induced hepatitis and in viral hepatitis
include jaundice, fatigue, loss of appetite, abdominal pain, dark urine, and elevated
enzymes. In case of allergic drug reactions, generalized fever, rash and elevated white
blood cell count may occur.
Classification of Heptatoxic Agents
Hepatotoxins can be classified into the following classes depending on the
source of the toxin. They are,
1. Natural Origin – eg. Tannic acid, Aflatoxins, Pyrrelidizone alkaloids, Gyrometrin,
Amatoxins, Microcystin LR.
2. Synthetic Origin – (a) Toxins of clinical significance eg. Paracetomol,
Sulfonamides, PAS, Rifampicin, Iproniazid, Isinazid, Ethanol, Anabolic and
Controceptive steroids etc.
(b) Toxins having pathologic singificance eg. Chloroform, Phosphorous,
Tetrachlorethane, Ethionine etc.
(c) Toxins used as a common lab models eg. Carbon tetrachloride, Paracetamol,
Galactosamine. (Ref: Zimmaman H.J.et.al.,1998).
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Hepatomegaly125
Causes
1 Infections
Viral: viral hepatitis,Yellow fever,Infectious mononucleosis,lassa fever
Spirochetal : weils diseases,Syphilis,relapsing fever.
Bacterial : Typhoid,pneumonia, brucelosis,tuberculosis.
Protozoal : Amoebiasis, malaria, kala-azar.
Parasitic : Schistosomasis, echinococcus,clonorchiasis.
Fungul : Actinomycosis, Histoplasmosis.
2. Toxic hepatitis
Carbon tetrachloride, arsenic, phosphorus, cincophen, sulphonamides,
chloropromazine, methylteststerone, halothane.
3. Degenerative - fatty infiltration and early cirrhosis.
4. Congestive hepatomegaly
a. General-Cogestive cardiac failure, tricuspid regurgitation, constrictive
pericarditis
b. Local- Portal hypertention(cirrhosis), hepatic vein thrombosis.
5. Tumors and cysts
a. Primary: benign and malignant hepatoma, : benign and malignant cholangioma,
fibroma, sarcoma, hemangioma.
b. Secondary: Direct due to spread by contiguity or embolic metastatic.
Cysts- Polycystic disease, solitary cyst (parasitic or non parasitic), malignant
pseudocysts.
6. Biliary obstruction.
Gall stones, strictures of bile ducts.
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7. Storage disorders:
Amyloidosis, glycogen storage disase, haemochromatosis.
8. Myeloid metaplasia:
Secondary carcinoma of bone, multiple myeloma.
9. Genetic abnormalities:
Sickle cell disease.
10.Reticulosis:
Hodgkins disease,leukemia.
Carbon Tetrachloride126
During the first quarter of this century, CCl4 was found to produce hepatic injury
in man and experimental animals. The intervening years have seen thousands of
reports devoted to this agent. Indeed, it is the most extensively studied of the
hepatotoxins. Poisoning which CCl4 has been a widely used model to study the
pathogenesis and character of hepatic necrosis and the effect of induced hepatic injury
or hepatic function. In the course of unraveling the mechanism by which it produces
fatty liver. CCl4 has served to clucidate the pathogenesis of fatty metamorphosis
induced by other etiologic factors. While it can lead to damage to a number of tissues,
it is particularly damaging to the liver and kidneys of many species.
Single dose of CCl4 in mammals results in acute centrilobular necrosis and
steatosis in the liver. With in a few minutes there is injury to the endoplasmic
reticulum, which leads to functional defects of the hepotocyte and multiple
biochemical manifestation of hepatic injury. Prolonged administration of CCl4 can
lead to cirrhosis and hepatic carcinoma. Most of the acute and chronic hepatic injury
appears to be due to the metabolites.
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CCl4
PLASMA MEMBRANE
DISTORTED INTRACELLULAR ENVIRONMENT
ENDOPLASMIC RETICULUM
METABOLITE CCl3
MITOCHONDRIA LYSOSOMES INJURY
PROTEIN SYNTHESIS
LIPOPROTEIN
PERIPHERAL FAT DEPOTS
STEATOSIS
NECROSIS DISRUPTION LIPID-PROTEIN LINKAGE
LIPID EXIT
In This Experimental Study Carbon Tetrachloride – Induced As Hepatotoxic
Agent.
Chart Showing A suggested mechanism of production of the spectrum of liver
manifestation
+20 Free Radicals
(Ref: Zimmaman H. J. et.al, 1998).
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INVESTIGATIONS IN LIVER DISEASES127
Liver function tests can be classified as :
a. Tests of the excretion by the liver
b. Evaluation of synthesis in liver
c. Evaluation of enzyme activity.
Liver function tests are most often employed to determine
a. The presence of liver disease
b. The type of liver disease
c. The extent and progression of liver disease.
Many liver function tests are based on a wide variety of biochemical reactions,
such that the clinician can select combination of tests that often measure different
aspects of hepatic function. Many tests however are still empiric and semi
quantitative and no single test is universally helpful in diagnosis.
Serological lab investigations like -
Serum Bilirubin, Serum Albumin and globulins, Serum Enzyme Assays, Serum
Alkaline phophatase, Transaminasis [Amino transferases]
Others tests like; urine bilurubin, urine urobilinogen, Lactic dehydogenase (LDH),
other enzymes GGT, OLT, Serum proteins, immunoglobulins, clotting factors,
Serum ammonia, Blood lipids.
Radiologic procedures – Cholecystography, and Cholangiography, Angiography,
Radioisotope liver scans, Portal and Hepatic vein manometry, Percutaneous
needle biopsy of the liver, Peritoneoscopy, Leparotomy.
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Table.No. 12 Showing Laboratory Evaluation of Liver disease128
Test Category Serum Measurement
Hepatocyte integrity Cystolic hepatocellular enzymes
Serum aspartate aminotransferase (AST)
Serum alanine aminotransferase (ALT)
Serum lactate dehydrogenase (LDH)
Biliary excretory function Substances secreted in bile
Serum bilirubin
Total: Unconjugated plus conjugated
Direct: Conjugated only
Delta: Convalently linked to albumin
urine bilubin
Serum bile acids
Plasma membrane anzymes.
(from damage to bile canaliculus)
Serum alkaline phosphates
Serum r – glutamyl transpeptidase
Serum s – nucleotidase
Hepatocyte function Proteins secreted into the blood
Serum albumin
Prothrombin time
(factors V, VII, X, Prothrombin, fibrinogen)
Hepatocyte metabolism
Serum ammonia
Aminopyrine breath test
(hepatic demethylation)
Galactose elimination
(Intravenous injection)
Methodology
METHODOLOGY
Methodology can be studied under three headings,
1) Pharmaceutical study.
2) Analytical study.
3) Experimental study.
Pharmaceutical Study
The study involves proper identification, collection, processing of raw
drugs and preparation of Shankhanabhi Bhasma.
The rationale of this branch is to make available the effective, safe, and
suitable medicine. It is evident that samskara given to the drug will change the
quality and also acts in a different manner when mixed with other drugs. Timing
of medication and anupana also direct the medicine to act in a different ways.
Study design
This section includes major steps,
Step 01: Identification and Collection of Shankha.
Step 02: Shodhana of Shankhanabhi.
Step 03: Marana of Shankhanabhi.
Date of Commencement:
Date of Completion:
Ref: Rasatarangini- Dwadasha Taranga.
Method:
Step 01: Identification and Collection of raw drugs.
Date of Commencement: 25.12.06
Date of Completion: 25.12.06
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Methodology
An important part in the preparation of medicament lies in the proper
identification and procurement of the raw materials. This determines the quality of
drugs. So this section of the study deals with the same.
Preparation of Shankhanabhi Bhashma involves
Key ingredient: Shankhanabhi
Samskarartha Prayojya Ghatakas: Kanji, Kumari
Special request was made to the herbo mineral drug shop dealer to get the
particular quality drugs and those were screened for classical grahya lakshanas
and selected.
Pratical No. 1
Name of the practical : Preparation of Kanji
Reference : Bhavaprakasha. Sandhana varga /1
Date of commencement : 07-11-2006
Date of completion : 20-11-2006
Materials required : Gas stove, steel vessel with lid, sieve, cloths, dry husk.
Drugs used : a) Shali (Wheat) – 1 kg
b) Water - 16 liters
Dhoopanartha Prayojya Ghatakas – Guggulu, Jatamamsi, Karpura, Musta each5gms
Procedure:
• Shali is cleaned and washed twice with water.
• Then to it 16 parts of water is added and kept on agni at at 11 AM. Mandagni
is maintained throughout the procedure.
• One part is evaporated and three parts is retained, so when 12 liters remained
the gas is turned off at 1.30 PM.
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protective Activity - An Experimental Study”
Methodology
• Left their itself for cooling upto 4.00 PM.
• Then its covered with the lid and tied with the cloths and kept in dry husk for
fermentation.
• On the 3rd day vessel is opened and filtered through mesh. The thicker portion
is discarded.
• The manda portion is kept in the plastic jar of capacity 10 litres which is
fumigated before and closed with lids. Once again jar is kept in husk.
• From 10th day of filtration the Kanji was suitable for the purpose of shodhana.
Table No. 13 Showing quantity of liquid during preparation of Kanji
Quantity of Liquid Loss
Initally 16 ltrs -
After boiling for 2 ½ hrs 12 ltrs 4 ltrs
On filtration on 3rd day
obtained
3 ltrs 9 ltrs
Kanji obtained 3 ltrs -
Observation
1) After boiling for 2 ½ hr the rice particles were found broken.
2) On 3rd the hissing sound was absent, on filtration the manda obtained is thick
& white in colour.
3) On 14th day it has became thin.
End Product
Quantity obtained - 3 ltrs
Colour - White
Odour - Amla gandha
Taste - Amla
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protective Activity - An Experimental Study”
Methodology
Consistency - Thinner.
Precaution
1) Strictly sterilization should be maintained.
2) Plastic jar used for storage should be fumigated well before with Guggulu,
Jatamamsi, Karpura, Musta.
Pratical No. 2
Name of the Practical : Shankhanabhi shodhana
Reference : Rasatarangini 12/10
Date of Commencement : 10-01-2007
Date of completion : 10-01-2007
Materials required :
1) Dolayantra – For Dolayantra we had taken
a) Earthen pot of Capacity – 4 litres
Shape – Round
b) Iron rod
c) Cotton thread
d) Drava – Kanji
e) Cotton cloth
2) Khalwa yantra
Drugs used:
1) Shankhanabhi – 250gms
2) Kanji – 3250ml
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Methodology
Procedure:
• As Shankha is very hard, the tiles cutting machine is used to remove its
avartha and Sharkhanabhi’s are obtained.
• These Shankhanabhi’s are made into small pieces of size 0.5 – 1cm by
pounding in Khalva yantra.
• Earthern pot was taken as mentioned above, Kanji is added upto 2/3rd of the
pot.
• Shankhanabhi pieces were put in cotton cloth and tied with thread to make
pottali.
• This pottali was suspended in Kanji by tying it to the iron rod.
• The Dolayantra was subjected for heating on mandagni which was maintained
throughout.
• Required amount of kanji was added frequently in Dolayantra to maintain
liquid level.
• The procedure was continued for three hours.
• After three hours, the flame was turned off and liquid was allowed to cool.
• After Swangasheeta the pieces of Shankhanabhi were removed from the
pottali and washed with hot water by rubbing with brush.
• These pieces were then kept for drying.
Table No. 14 showing the change in wt of Shankhanabhi before shodhana and
after shodhana.
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Dravya Quantity of Shankhanabhi
before shodhana (in gms)
Quantity of
Shankhanabhi after
shodha (in gms)
Wt loss
(in gms)
Shankha nabhi 250 244 6
protective Activity - An Experimental Study”
Methodology
Table No. 15 Showing Physical Features of Raw & Shodhita shankha nabhi
Properties Raw Shankhanabhi Shodhita Shankhanabhi
Colour Off white Grownish white
Form Carbonate Carbonate
Lustre Dull Bright
Odour Odourless Amla
Structure Massive Massive
Taste Kshariya Kshariya
Transparency Opaque Opaque
Touch Hard, Sharp Hard, Sharp
Observations:
1) As Shankha is very hard, its not possible to get Shankhanabhi by pounding.
Hence tiles cutting machine was used to remove its avartha.
2) Procedure followed is pachana of shankhanabhi in kanji Therefore pottali is
immersed fully in kanji throughout.
Precautions:
1) The pottali was hanged such that it moved freely in the pot. Distance of 4
angula from the bottom of the pot to the pottali was maintained.
2) Mandagni was maintained throughout.
3) To avoid overflow of Kanji, twice the foam was removed.
Practical No. 3
Name of the Practical : Shankhanabhi marana
Reference : Rasatarangini 12/17-19
Date of Commencement : 12-01-2007
Date of Completion : 09-02-2007
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protective Activity - An Experimental Study”
Methodology
Materials Required:
• Sharava : Two of equal size
• Seven white cotton cloths : Length – 50 cms
Breadth – 6 cms
• Gopichandan Mruttika
• Khalwayantra, Tulayantra, Pyrometer
• Cowdung cakes.
Table No. 16 Showing Drugs Used
Dravya Mana
Shodhita Shankhanabhi 244 gms
Kumari Swarasa 60 ml
Procedure:
• Shodhita Shankhanabhi pieces were kept in sharava and was closed by another
Sharava of same diameter.
• The white cloth of 6cm in breadth & 50cm in length was taken and wet
gopichandan mruttika was applied on it.
• Sandhibandhana was made by applying this cloth to sharava without leaving
any gap & kept for drying for 12 hrs.
• Next day another layer of cloth was smeared & dried, similarly it took 7 days
for 7 layers.
• The pit was digged measuring 30 angulas in length, breadth and height.
• 700 vanopalas were kept in the pit and sharava samputa was placed over it.
Again 300 vanopalas were kept over the Sharava samputa.
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protective Activity - An Experimental Study”
Methodology
• Then subjected to Gaja puta by igniting the fire.
• After Swangasheeta the Sharava was taken outside and smeared mud was
removed carefully.
• Shankhanabhi pieces are collected & powdered well and bhasma pareeksha
was done.
• As it did not passed bhasma pareeksha, it was once again subjected to gajaputa
for the second time.
• Kumari swarasa bhavana was given, for 244 gms of Shankhanabhi powder 60
ml of kumari swarasa was added & mardana was done for 8 hrs.
• When it became paste like, chakrikas were prepared and kept for drying.
• After drying chakrikas are kept in sharava in one layer and sharava samputa
was done and second gajaputa was given same as that of first puta. After
swangasheeta bhasma is collected from sharava & powdered.
• Now the bhasma passed the bhasma siddi lakshanas.
• Shankhanabhi bhasma is preserved in air tight container.
Observations
1) After first gajaputa, shankhanabhi colour changed to grayish white which on
doing mardana turned to white colour.
2) After second gajaputa, shankhanabhi bhasma is obtained in white colour.
3) Essential bhasma pareekshas were observed like rekhapurnata, varitara etc
after second gajaputa.
4) After each gajaputa loss in quantity was found.
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Methodology
Table No 17 showing Time and Temperature of Gajaputa 1 in 0C.
Temperature was measured by using Pyrometer which was placed in the middle of
pit near sharava.
Date: 22/01/07
Time Temp. Time Temp.
9.00 Am. 38°C 1.45 Pm. 393° C
9.15 Am. 42° C 2.00 Pm. 358° C
9.30 Am. 50° C 2.15 Pm. 326° C
9.45 Am. 592° C 2.30 Pm. 298° C
10.00 Am. 638° C 2.45 Pm. 270° C
10.15 Am. 808° C 3.00 Pm. 245° C
10.30 Am. 895° C 3.15 Pm. 230° C
10.45 Pm. 960° C 3.30 Pm. 210° C
11.00 Pm. 995° C 3.45 Pm. 190° C
11.15 Pm. 912° C 4.00 Pm. 172° C
11.30 Pm. 882° C 4.15 Pm. 154° C
11.45 Pm. 850° C 4.30 Pm. 134° C
12.00 Noon. 780° C 4.45 Pm. 118° C
12.15 Pm. 685° C 5.00 Pm. 98° C
12.30 Pm. 615° C 5.15 Pm. 77° C
12.45 Pm. 555 °C 5.30 Pm. 62° C
1.00 Pm. 515° C 5.45 Pm. 52° C
1.15 Pm. 470° C 6.00 Pm. 37° C
1.30 Pm. 435° C
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Methodology
Table No. 18 Showing Time and Temperature of Gajaputa 2
Date : 06/02/07
Time Temp. Time Temp.
9.45 Am. 33°C 1.45 Pm. 521° C
10.00 Am. 34° C 2.00 Pm. 469° C
10.15 Am. 621° C 2.15 Pm. 400° C
10.30 Am. 584° C 2.30 Pm. 325° C
10.45 Am. 806° C 2.45 Pm. 264° C
11.00 Am. 896° C 3.00 Pm. 214° C
11.15 Am. 950° C 3.15 Pm. 191° C
11.30 Am. 962° C 3.30 Pm. 154° C
11.45 Am. 938° C 3.45 Pm. 135° C
12.00 Noon 912° C 4.00 Pm. 121° C
12.15 Pm. 827° C 4.15 Pm. 109° C
12.30 Pm. 767° C 4.30 Pm. 99° C
12.45 Pm. 697° C 4.45 Pm. 92° C
1.00 Pm. 630° C 5.00 Pm. 85° C
1.15 Pm. 592° C 5.15 Pm. 60° C
1.30 Pm. 549° C 5.30 Pm. 40° C
Precautions
1) One layer of shankhanabhi pieces was made in sharava so that they would get
sufficient uniform heat. Similarly chakrikas are placed in one layer for second
gajaputa.
2) While giving Bhavana, Kumari Swarasa was added till the Shankhanabhi
powder became semi-solid and mardana was done.
3) While doing Sandhibandhana, no gap was left. Mruttika (Gopichandan)
selected for sandhibandhan was sticky. After every layer of mud smear,
sharava samputa was kept for drying for atleast 12 hrs.
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Methodology
4) Agni was created by putting flame at four corners.
5) Sharavasamputa should be removed only after complete cooling, so its left in
the pit for 48 hrs and collected.
Table No. 19 Showing Physical features of Shankhanabhi Bhasma after each
puta:
Properties After 1st puta After 2nd puta
Colour Grayish white White
Form Carbonate Carbonate
Luster Dull Bright
Odour Odourless Odourless
Structure Brittle pieces Powder
Taste Kshariyata+++ Kshariyata+
Transparency Opaque Opaque
Touch Smooth Smooth
Table No. 20 Showing Quantity of Shankhanabhi before & after Marana
Before Marana Gaja putas After Marana Loss
244gms 1st Gajaputa 232gms 12gms
2nd Gajaputa 221gms 11gms
Total weight loss after marana 23gms
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Methodology
Analytical Study
The Rasoushadhies mentioned in Ayurvedic Pharmacopoeia should be
analysed for physical & chemical properties to confirm the genuinity & safety before
administration in human beings. Hence it becomes obligatory to adopt modern
analytical methodology for better understanding and interpretation of physico-
chemical changes occurred during the process.
In the present study the sample is collected at the completion of the
preparation of sankhanabhi bhasma & subjected to analysis.
Aims & Objectives
• To analyze the physico-chemical properties of shankhanabhi bhasma.
• To carry out quantitative estimation of Ca, CaCO3, Fe, Mg, S in shankhanabhi
bhasma.
Materials
• Ancient parameters were conducted at PG Dept., of Rasashastra,
D.G.M.A.M.C, Gadag.
• Modern physical & chemical tests were conducted at Bangalore Test House –
Bangalore, K.L.E. Society’s college of Pharmacy Gadag.
• N.P.S. Test was done at P.G. Dept. of Rasashastra, D.G.M.A.M.C, Gadag.
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Methodology
1) Ancient Parameters
Table No. 21 Showing Analysis of Shankhanabhi Bhasma by Ancient method
OBSERVATION AND RESULT Sl.
No.
TEST
Shankhanabhi Bhasma
1 Varna Shweta
2 Gatarasatvam Nirasa (slightly alkaline)
3 Sparsha
(Slakshnatvam and
Mrudutvam)
Mrudutva and Slakshnatva was felt by simple
touch with finger tips
4 Gandha Nirgandha
5 Rekhapurnatva The Bhasma was rubbed in between first finger
and thumb. It penetrates into the furrows of the
fingers - Positive
6 Varitaratva A small amount of Bhasma was carefully
sprinkled in beaker full of water. It was found
that total portion of Bhasma was floating on the
water surface - Positive
7 Nischandratvam The Bhasma observed in bright sunlight. It
was not having any lusture – Positive
8 Kshariyata Bhasma when placed on tongue Kshariyata of
the bhasma was not found – Negative (slightly
alkaline)
2) Modern Parameters
The study has been divided into two parts
1) Physical analysis
2) Chemical analysis
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Methodology
1) Physical analysis
a) Organoleptic characters
Colour : White
Odour : Odourless
Touch : Fine
b) Analysis
Determination of pH Value:
Procedure: The pH value of the sample was determined by a digital pH meter.
One gram of Shankhanabhi bhasma was weighed accurately and dissolved in 100ml
of water and pH was noted in the digital pH meter.
Result : pH = 9.8
Loss on drying at 1100C
Procedure: Two grams of Shankhanabhi bhasma was weighed in a silica crucible and
dry in a hot air oven at 1100C till a constant weight is obtained. The difference in the
two weighings gives the loss on drying & then the percentage of loss on drying was
calculated.
Result : 0.24%
Loss on Ignition:
Procedure: Weigh a silica curucible previously ignited for one hour at a temperature
not exceeding 5000C and cooled in a desicator. Transfer to the crucible accurately
weighed sample. Weigh the crucible accurately. Place the loaded crucible in the
muffle furnance and ignite the crucible to 5000C, until constant weight is indicated.
Calculate loss on ignition with reference to the air dried drug.
Result: 32.26%
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Methodology
Determination of Total Ash:
Procedure : Take about 2 gms accurately weighed, ground drug in a previously
traced silica dish, previously ignited and weighed. Scatter the ground dry in a fine
even layer on the bottom of the dish. Incinerate by gradually increasing the heat not
exceeding dull red heat (4500C) until free from carbon. Cool and weighed. Then the
percentage of ash with reference to the air dried drug was calculated.
Result: Total ash: 61.3%
Determination of Acid Insoluble Ash:
Procedure : Boil the ash obtained in the process described under determination of
total ash for 5 minutes with 25ml of dilute hydrochloric acid. Collect the insoluble
matter on an ashless filter paper. Wash with hot water and ignite. Weigh it and
calculate the percentage of acid insoluble ash with reference to the air dried drug.
Result: 0.53%
Determination of water soluble extractive :
Procedure: Macerate about 5 grams of air dried drug with 100ml of chloroform water
in a closed flask for twenty four hours, shaking frequently during six hours and
allowing to stand for eighteen hours. Filter this and pipette 25ml of this liquid and
evaporate to dryness in a tared flat bottomed dish and dry at 1050C, to constant
weight. Calculate the percentage of water soluble extractive with reference to air dried
drug.
Result: water soluble extractive : 9.62%.
Determination of Alcohol soluble extractive:
Procedure : Macerate about 5 grams of the air dried sample with 100ml of ethanol in
a closed flask for twenty four hours, shaking frequently during six hours and allowing
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Methodology
to stand for eighteen hours. Filter rapidly taking precautions against loss of solvent
evaporate 25ml of the filterate to dryness in a tared flat bottomed dish and dry at
1050C, to constant weight and weigh. Calculate the percentage of alcohol soluble
extractive with reference to the air dried drug.
Result: 1.85%
Determination of Fineness of particles:
Procedure: The degree of coarseness or fineness of a powder is differentiated and
expressed by the size of the mesh of the sieve through which the particle is able to
pass.
A suitable quantity of the sample is weighed and transferred to the set of
sieves shaken in a sieve shaken for about 30minutes and the residue on each sieve is
weighed separately.
Result: 125 micron
Passes through sieve No. 120
Determination of size of particle:
Procedure : It can be possible to use the ordinary microscope for particle size
measuring in the range of 0.2 micrometer to about 100micrometer. According to
microscope method the fine powder was sprinkled on the slide covered with covering
slip and placed on a mechanical stage. In initial standardization of micrometer was
carried out by coinciding the lines of both oculo micrometer, stage micrometer and
standardized by using the formula.
SM / OM x 10 = m
In the next step, the stage micrometer was removed and the mounted slide was
placed on a mechanical stage and focused.
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Methodology
The particles are measured along an orbitarily chosen fixed lines covered by
the particles using the oculo micrometers. The size of the partical was calculated
using the standard value.
Result : Size of particle : 51.20 Micrometer.
Determination of Flow property:
Procedure : Angle of repose : It is the maximum angle that can be obtained between
the free standing surface of a powder heap and the horizontal plane i.e tan θ = 2h/D
Where D is the diameter of the circle & ‘h’ is the height of the powder heap.
Angle of repose by which we can analyse either the powder having very good flow
property, good property or a bad flow property. This test involve the hollow cylinder
half is filled with the sample with one end sealed by transparent plate. The cylinder is
rotated about its horizontal axis until the powder surface cascades. The curved wall is
lined with sand paper to prevent preferential slip at this surface. If the value comes
between 200-400 indicates reasonable flow potential.
Result : Flow property :
Angle of repose = 27.40
Determination of Flow Rate:
Procedure : A simple indication of the ease with which a material can be induced to
flow is given by application of a compressibility index “I”
I = [1-V/V0] x 100
Where ‘V’ is the volume occupied by sample of the powder after being subjected to a
standardized tapping procedure.
V0 = Volume before tapping procedure.
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Methodology
In this procedure one measuring cylinder is taken and is filled with sample. The level
of the sample should be noted. Then at a height of 2 cm continuous 10 tapping should
be done, after that the level of the sample in the cylinder is once again noted and the
value “I” is calculated with respect to the Vo and V value. If the “I” is below 15%
usually having good flow rates.
Results : Flow rate: 15 %
Solubility:
Procedure: About one gram of the sample was weighed and dissolved in 10 ml of the
solvents. When the sample did not dissolve, an excess of solvent by 10 ml quantity up
to 100 ml was added and noted that the sample was sparingly soluble in water and 1
M Hydrochloric acid (1 gm of sample in 100 ml of water) and slightly soluble in
chloroform (1 gram of sample in 600 ml to 1000 ml of chloroform) and soluble in
alcohol (1 gm of sample in 600ml to 1000ml of alcohol).
Shankhanabhi bhasma is soluble with effervescence in dil Hydrochloric acid.
Results:
Alcohol – Sparingly soluble.
Water - Sparingly soluble.
Chloroform - Insoluble.
Dilute HCl – Soluble with effervescence
2. Chemical analysis
Determination of Calcium:
Regents:
Ammonium Oxalate – Saturated solution
Methyl Red indicator – Dissolve 0.5 g of Methyl Red in 100 ml of 95% Alcohol.
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Methodology
Dilute Acetic acid
Dilute Ammonium Hydroxide
Dilute Sulphuric acid: Add acid to water slowly and with constant stirring. Cool and
make up to volume.
0.1N Potassium Permanganate (KMnO4)
0.01 N Potassium Permanganate – Working standard:
Dilute 10 ml of 0.1 N KmnO4 solution to 100 ml with water (1 ml = 0.2 mg of
Calcium).
Prepare fresh solution before using.
Procedure
Pipette an aliquot (20 to 100 ml) of the ash solution obtained by dry ashing to
a 250 ml beaker. Add 25 to 50 ml of water if necessary. Add 10 ml of saturated
Ammonium Oxalate solution and 2 drops of Methyl Red Indicator. Make the solution
slightly alkaline by the addition of dilute ammonia and then slightly acidic with a few
drops of acetic acid until the colour is faint pink (pH 5.0). Heat the solution to the
boiling point. Allow to stand at room temperature for atleast 4 hours or preferably
overnight. Filter through whatman No. 42 paper wash with water till the filtrate is
oxalate free (since HCl has been used for preparing the original ash solution, it is
convenient to test for the absence of chloride using AgNO3). Break the point of the
filter paper with platinum wire or pointed glass rod. Wash the precipitate first using
hot dilute H2SO4 from wash bottle into the beaker in which the calcium was
precipitated.
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Methodology
Then wash with hot water and titrate while still hot (temperature 70 to 800C)
with 0.01 N KmnO4 to the first permanent pink colour. Finally, add filter paper to
solution and complete the titration. Calculated as
Calcium (mg/100g) = Titre X 0.2 × Total volume of Ash solution × 100
Volume taken for × weight of sample
Estimation taken
If the KmnO4 standard solution is not exactly 0.01 N, use the following expression.
Calcium (mg/100g) = Titre × Normality of KmnO4 ×20 ×Total Volume of ash soln×100
ML of Ash solution × Weight of the sample
Taken for estimation taken for ashing
Result: 38.2 %
Determination of calcium carbonate
Procedure: Weigh accurately appropriate quantity of the sample and dissolve in 3ml
of dilute hydrochloric acid and 10 ml of water. Boil for 10 minutes. Cool, dilute to
50ml with water. Titrate width 0.05M Disodium edetate to within a few ml of the
expected end point, add 8ml of Sodium Hydroxide solution (saturated solution) and
0.1g of the calcon mixture and continue the titration until the colour of the solution
changes from pink to full blue colour. Each ml of 0.05 M Disodium edetate is
equivalent to 0.0020 g of calcium.
Result : 95.3 %
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Methodology
Determination of Iron:
Reagents:
1. Nitric acid
2. Hydrochloric acid
3. Dilute Sulphuric acid
4. 10% w/v solution of Ammonium Thiocyanate in water
Standard Solution:
1000 ppm Iron Stock Solution
Sample Solution:
To accurately weighed sample, add 1 ml of Nitric acid and 3 ml of Hydrochloric acid
and heat on a low heat until the sample is dissolved. Dilute to 100 ml and filter.
Procedure:
To 2 ml each of sample and standard solutions, add 5 ml of Ammonium Thiocyanate
Solution, dilute to 25 ml dilute sulphuric acid. Measure the absorption of both the
solutions at 520 nm against reagent blank and result is calculated by comparison.
Result: 0.06 %
Determination of Magnesium
Reagents:
1. Ammonia-Ammonium Chloride solution
2. 0.05 Disodium Edetate (EDTA)
3. Solochrome Black T Indicator
Procedure:
Weigh accurately about 0.3 g, dissolve in 50 ml of water, add 10 ml of strong
Ammonia Ammonium Chloride solution and titrate with 0.05 M Disodium Edetate
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protective Activity - An Experimental Study”
Methodology
(EDTA) using 0.1 g of Solochrome Black – T indicator, until a blue colour is
obtained.
Each ml of 0.05 M Disodium Edetate is equivalent to 0.002916 g of Mg
Calculation:
% of Magnesium
= Volume of EDTA x Actual M of EDTA x 0.002916 x 100 Molecular weight of magnesium
Weight of Molecular weight of magnesium hydroxide Sample taken x 0.05 M
Result = 0.14 %
Determination of Sulphur:
Eschka Mixture – Mix two parts by weight calcined magnesia with one part of
anhydrous sodium carbonate.
Procedure
Cover the bottom of a 50 ml crucible with 0.5 gm of Eschka’s mixture. Weigh
accurately the appropriate quantity of the sample material and mix it immediately
with 2gms of Eschka’s mixture and put evenly on the previously weighed Eschka’s
mixture. Level the contents by tapping gently on a bench. Cover this uniformly with
0.5gm of Eschka mixture. Place crucible in the muffle furnace. Raise the temperature
from room temperature to 8000C +250C in about one hour and then heat for further 90
minutes. Transfer the ignited mixture as completely as possible from the crucible to a
beaker containing 25 to 30 ml of water. Wash out the crucible thoroughly with about
50 ml of hot distilled water and add the washings to the contents of the beaker. Add
carefully sufficient quantity of concentrated hydrochloric acid to dissolve the solid
matter, warming the content of the beaker to effect solution. Boil for 5 minutes to
expel carbon dioxide. Add drop wise from a pipette; warm 5% Barium chlorine
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Methodology
solution. Stir the solution constantly during the addition. Allow the precipitate to
settle for a minute or two.
Then test the supernatant liquid for complete precipitation by adding a few
drops of Barium chloride solution. If a precipitate is formed, add slowly a further 3 ml
of the
reagent allow the precipitate to settle as before and test again, repeat this operation
until an excess of Barium Chloride is present. When an excess of the precipitating
agent has been added, keep the covered solution hot, but not boiling for an hour
(steam bath) in order to allow time for complete precipitation. The precipitation
should settle and a clear
supernatant liquid should be obtained. Test the latter with a few drops of Barium
chloride solution for complete precipitation. If no precipitate obtained, the Barium
sulphate is ready for filtration.
Filter the solution through an ash less filter paper (Whatmann No. 42). Wash the
precipitate with small portion of hot water. Dry the paper and place it in a silica or
porcelain crucible, previously ignited to redness and cooled in desiccators and
weighed. Gradually increase the heat until the paper chars and volatile matter is
expelled. Do not allow the paper to burst into flame as mechanical loss may thus
ensue. When charring is complete, raise the temperature of the crucible to dull redness
and burn off carbon with free excess of air. When the precipitate is white ignite the
crucible at red heat for 10-15 minutes. Allow the crucible to cool in air, transfer it to
desiccators and when cold, weigh the crucible and contents. Repeat until constant
weight is attained.
A blank is necessary. Calculate the percentage of sulphur converting Barium
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protective Activity - An Experimental Study”
Methodology
sulphate X 0.1374.
Result: 1.05 %
Numburi Phased Spot Test: for identification of Bhasma
Based on Namboori Phased Spot Test, one can confirm the sample. This test
is based on the nature of solution it gives when dissolved in water is acidic, basic and
neutral.
The following observations are to be made and noted in the following order.
1. Name of the Bhasma.
2. On heating (Heat Treatment): Observe for the following changes.
i) Liberation of any fumes: nature of the fumes i.e gasses, smoke or
moisture.
ii) Odour: Pleasant/unpleasant smell.
iii) Change of colour: Charring or any change in colour.
3. On Wetting: (Wet Treatment): Observe for the following changes.
i) Endothermic changes: The lower part of the test tube becomes cool
instantaneously after adding distilled water (absorption of heat).
ii) Exothermic changes: The lower part of the test tube becomes
instantaneously hot after adding distilled water (liberation of heat)
4. Absorption of distilled water: When equal quantities of samples are treated
with equal volume of distilled water some samples are treated with equal
volume of distilled water some samples exhibit more absorbing nature, For
example, when 0.25 gm of a sample is taken and 4 ml of distilled water is
added to prepare a solution, in the case of some samples all the distilled water
is absorbed. But in others some distilled water (in solution form) is seen as a
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protective Activity - An Experimental Study”
Methodology
supernatant layer. Settling time: This is another observation to be made when
distilled water is added to the fine powder of the samples and shaken. In the
case of some samples the contents settle down very slowly and in some very
rapidly.
Hence micro test tubes of equal size and caliber are to be taken for
preparing solutions.
In the context of this new technique (N.P.S.T) and methodology evolved to
identify bhasmas of Sudhavarg, with minute differences in overall chemical
reactions, it is necessary to study the organoleptic properties of standard
pravala Bh (Shakha). Therefore some relative standards of these bhasmaas are
to be laid down particularly with reference to their solubility time and time
taken for fading away of the spots.
Relative Standards:
Solubility: The amount of distilled water absorbed by the known quantity of
pravala bhasma (Shakha) is termed as ‘NORMAL QUANTITY’. So also when
any bhasma of Sudhavarga possessing same weight as that of pravala bhasma
(Shakha) absorbs same amount of distilled water as above is termed as
‘NORMAL’.
Settling Time: The time taken by the contents Pravala bhasma (Shakha) of the
test tube to settle down after shaking with distilled water is termed as ‘NORMAL
SETTLING TIME’. Similarly when any bhasma of Sudha Varga possessing same
weight as that of pravala bhasma (Shakha) takes the same time to settle down is
termed as ‘NORMAL SETTLING TIME’.
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Methodology
Fading away Time: The time taken by a spot of standard pravala bhasma
(Shakha) to fade away on Haridra paper is termed as ‘RAPID’. Thus the fading
away of spots of various bhasmas of Sudhavarga are described as ‘SLOW’ and
‘VERY SLOW’ as the case may be when compared with the fading away time for
standard pravala bhasma (Shakha).
Procedure:
The procedure can be divided into
1. Preparation of Haridra paper
2. Preparation of supernatant fluids of samples
1. Preparation of Haridra paper
Air dried rhizomes of Curcuma longa (Haridra) were reduced to coarse
powder and 50g of powder was mixed thoroughly with 100ml of rectified spirit and
this was kept in a airtight bottle at a cool place. To prevent the vaporization of spirit,
airtight bottle should be used. After 5 days decant the spirit extract of haridra into
another air tight bottle.
Sheets of Whatmann No. 1 filter paper were cut into pieces of convenient size. These
papers were uniformly dipped in 50% Haridra color solution about one minute for
each side. Try to avoid excess stain on the paper.
Glass plates of convenient size were cleaned and impregnated papers were kept on the
glass plates from one end to another and in such a way that there should be no air
bubbles between glass sheets and impregnated papers. Once the front side of the
papers was dry, the rear side of the papers was brought upside and is again dried on
glass plates. After drying, the papers were kept in between two glass sheets, which
prevent the deposition of foreign dust particles on the paper.
2. Preparation of supernatant fluid of the samples
0.5 gm of Shankhanabhi bhasma was taken into a semi micro test tube and
heated it on a spirit lamp till the tip of the lower end of the test tube becomes red hot.
Then the test tube was allowed to cool.
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Methodology
After cooling the test tube 1 ml of distilled water is added to it, shaked well
and allowed to settle. The sample became clear within five minutes. After settling, the
supernatant fluid was taken in a dropper and put 2-3 drops on the Haridra paper.
Immediately pink coloured spot will appear.
Results: Organoleptic Properties
On Heating
Liberation of Fumes : Nil
Charring : Nil
Odour : Nil
On Wetting
Solvent : Distilled water
Exothermic : Present
Endothermic : Nil
Colour of the solution : Colourless
Absorption : Normal
Settling Time : Normal
Namburi phased spot test observations
1st Phase:
A deep pink solid spot appears with immediate formation of more deep pink
margin and wet periphery. By the end of 1st phase the central space become light pink.
2nd Phase:
By the end of 2nd phase the margin merges with the central space.
3rd Phase
By the beginning of 3rd phase the wet periphery fades away and spot starts
fading away after 24 hours.
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Methodology
Experimental Study
Aims and objectives
To evaluate the hepatoprotective activity of Shankhanabhi bhasma on albino
rats.
Materials and methods:
Animals : Albino rats (150-200gms)
Drug and chemicals : Gum acacia
: Jambeera swarasa
: Carbon tetrachloride
: Shankhanabhi bhasma
Equipments : Tuberculin syringe (1ml)
: I.V. Canula tube
: Weighing balance.
Selection of Animals:
Colony bred albino rats of either sex were utilized for present study. The
reason for selecting albino rats is that the regeneration of liver after hepatic
damage/partial hepatectomy is almost complete within a week.
The animals were obtained from the animal house, J.T. College of Pharmacy,
Gadag. Albino rats of either sex weighing between 150-200 gm breeds in animal
house were selected for the study. They were housed individually in polypropylene
cages with paddy husk bedding in well-ventilated rooms. The rats were kept under
observation for seven days with standard laboratory diet. After which they were
examined for their normal health and then subjected to experimental study.
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Methodology
30 animals were selected, which have been separated into 5 groups. Each
group with six animals were kept in separate cages after proper labeling for identity.
Drugs and chemicals used in our study :
1. Shankhanabhi bhasma : Is prepared in P.G.department of Rasashastra,
DGMAMC,Gadag.
2. Carbon tetrachloride : Is used as hepatotoxic agent. Carbon tetrachloride has
been supplied by S.d. fine – chem limited, Mumbai.
3. Gum acacia : This is the dried gummy exudates of stems and barks
of Acacia or other African species of Acacia. It has no
pharmacological action129 and is used as a suspending
agent for the oral administration of the trial drug in 1%
strength. Gum acacia has been supplied by liberty
pharmaceuticals, Mumbai.
4. Distilled water : Used for the preparation of suspension of trial drug.
And it was supplied by Dept. of Pharmacology, J.T.
College of Pharmacy, Gadag.
5. Jambeera swarasa : Is taken as anupana for trial drug.
Equipments
1. Tuberculin syringe : (1ml capacity) is used as a support to needle for oral
administration according to dose calculated with
respect to weight of animals.
2. I.V. Canula tube : Used for safe and convenient oral or intragastric
administration of test drug.
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Methodology
Method
The experimental model suggested by Watanabe and Takita (1973) was
adopted.
Albino rats of either sex weighing between 150-200 gms were kept in separate cages
and labeled as
Group 1 – Control
Group 2 – Intoxicated control – Liver damage
Group 3 – Intoxicated control – Natural recovery
Group 4 – Test drug (1)
Group 5 – Test drug (2)
Table No. 22 Showing Experimental Protocol:
Group Pre-treatment
dose/route
Duration in
days
Days of
withdrawal
of blood
Purpose
G 1 Vehicle 1-5 6th Control
G 2 CCl4 0.5ml/kg i.p 1-5 6th Liver damage
G 3 CCl4 0.5ml/kg i.p
[no drug]
1-5 11th Natural recovery
CCl4 0.5ml/kg i.p 1-5
G 4
G 5
Drug Shankhanabhi
bhasma (1 karsha)
Drug Shankhanabhi
bhasma (2 ratti)
Were given orally
6-10
11th Curative
For identification, rats were marked with different colours as head,
body, tail, head body, head tail, body tail in each group.
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Methodology
Dose Determination130
Carbon Tetrachloride
Carbon Tetrachloride (CCl4) was given at the dose of 0.7ml/kg, intra
peritoneal (i.p) for first five days to induce liver damage.
Shankhanabhi bhasma
By the given human dose, rat dose is calculated based on the conversion formula
Rat dose per 200gms body weight = 0.018 × human dose.
Human dose of Shankhanabhi bhasma is studied in two doses as 1 Karsha
(11.5 gms) and 2 Ratti (250 mg) as per different references in classics.
If the human dose of SNB is taken as 1Karsha = 11.5 gm
The rat dose per 200 gms body weight = 0.018 x 11.5 gms
= 0.207 gms = 207 mg
If the human dose of SNB is taken as 2 Ratti = 250 mg
The rat dose per 200 gms body weight = 0.018 x 250 mg
= 4.5 mg
Experimental Procedure
Animals were divided into five groups. Each group consist of six animals.
Group 1 (Control/Normal):
To this group Jambeera swarasa with distilled water was given orally from 1st
day to 5th day. Blood samples were drawn on the sixth day to estimate the
biochemical analysis. The animals were sacrificed on the same day for the
histopathological observations of the liver.
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Methodology
Group 2 (Intoxicated Control – Liver Damage) Toxicated Group:
Carbon tetrachloride (CCl4) 0.7ml/kg i.p administered for 5 days. Blood
samples were withdrawn on the sixth day and Biochemical analysis were carried out.
Animals were sacrificed for histopathological studies to assess the extent of the liver
damage.
Group 3 (Natural Recovery) Intoxicated Control Group:
Animals were administered with CCl4 0.7ml/kg i.p for 5 days. No drugs were
administered for next 5 days and kept as intoxicated control group, from 6th to 11th
day Jambeer swarasa with distilled water is given orally. Blood samples were drawn
only on 11th day for Biochemical Analysis and the animals were sacrificed for
histopathological studies (to assess the natural recovery).
Group 4 (Curative group) Treated with SNB -I:
Animals administered with CCl4 0.7ml/kg i.p for 5 days were followed by
Shankhanabhi bhasma (207mg) with Jambeera swarasa orally for 5 days that is from
6th to 10th day. Blood samples were drawn on the 11th day and the biochemical
analysis were done. On the same day, these animals were sacrificed for
histopathological study (to assess the curative effect of the drug).
Group 5 (Curative group) Treated with SNB-II :
Animals administered with CCl4 0.5ml/kg i.p for 5 days were followed by
Shankhanabhi bhasma (4.5 mg) with Jambeera swrasa orally for 5 days that is from
6th to 10th day. Blood samples were drawn on the 11th day and the biochemical
analysis were done. On the same day, these animals were sacrificed for
histopathological study (to assess the curative effect of the drug).
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato
89
protective Activity - An Experimental Study”
Methodology
Experimental Parameters:
For the present study following investigations are taken as parameters:
1. Biochemical changes in blood.
2. Histopathological studies.
Biochemical Parameters:
Blood samples were withdrawn from albino rats at different intervals that are
on 6th day for 1st and 2nd group while on 11th day for the remaining three groups (3rd,
4th, and 5th). The serum enzyme activity was estimated by standard bio-chemical
procedure using an auto-analyzer for all the groups.
Following enzyme levels were estimated for the study.
1. Alkaline phosphatase.
2. SGOT (Serum glutamate oxalacetate transaminase)/AST
3. SGPT (Serum glutamate pyruvate transaminase)/ALT
4. Total serum bilirubin.
5. Serum albumin.
Histo-pathological Studies:
Animals were sacrificed on the day of withdrawal of blood from all the five
groups and liver was isolated, sliced and washed with saline. Then it was preserved in
10% of formalin, for histopathological studies.
Routine staining procedures using haematoxylin and eosin dye were done in
the histopathological studies.
Then the sections were observed under microscope for histopathological
changes in liver architecture and their photomicrographs were taken.
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato
90
protective Activity - An Experimental Study”
Results
91
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Results of Experimental Study
The results of the present study are based on the bio-chemical values like
Alkaline Phosphatase, Serum Glutamic Oxalacetate Transminase (SGOT), Serum
Glutamic, Pyruvate Transaminase (SGPT), Serum Total Bilirubin, Serum albumin and
also Histopathological changes (microscopic) present in the section of the liver
sample of all animals.
Table No. 23 Showing summary of Biochemical values of all groups
Bio-chemical Parameters (mean & ± SEM) Group
No
of
Ani
mal
s
Drug and
Dose
Duration
of
Treatment
in days
SGOT SGPT ALP T-Bil Albumin
G1
Control
6 1-5 214.0
±2.45
76.40
±3.67
250.0
±0.450
0.733
±0.049
4.90
±0.310
G II
CCl4
6 CCl4
0.7 ml/kg
1-5 548.0
±23.50
139.7
±3.71
426.0
±2.41
1.93
±0.098
2.50
±0.046
G III
Natural
recovery
6 CCl4
0.7 ml/kg
1-5
6-10
No drug
543.0
±56.80
137.5
±5.61
417.6
±1.70
1.85
±0.042
2.80
±0.240
CCl4
0.7 ml/kg
1-5 G IV
Treated
with
SNB
6
SNB-I
207mg/
200gms
6-10
257.0
±8.67
84.60
±4.72
314.7
±3.80
0.96
±0.033
4.40
±0.190
CCl4
0.7 ml/kg
1-5
G V
Treated
with
SNB
6
SNB-II
4.5mg/
200gms
6-10
501.0
±78.45
98.30
±2.71
410.2
±2.71
1.55
±0.042
3.00
±0.420
Results
92
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Graph No. 1
214
548 543
257
501
0100200300400500600
IU/L
G1 G 2 G 3 G 4 G5
Groups
Mean SGOT of all the groups
SGOT
Graph -2
76.4
139.7 137.5
84.698.3
020406080
100120140
IU/L
G1 G 2 G 3 G 4 G5Groups
Mean SGPT of all the groups
SGPT
Results
93
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Graph -3
250
426 417.6314.7
410.2
0100200300400500
IU/L
G1 G 2 G 3 G 4 G5
Groups
Mean ALP of all the groups
ALP
Graph -4
0.733
1.93 1.85
0.96
1.55
0
0.5
1
1.5
2
mg/dl
G1 G 2 G 3 G 4 G5
Groups
Mean T. Bil of all the groups
T. Bil
Results
94
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Graph - 5
4.9
2.5 2.8
4.4
3
012345
gm%
G1 G 2 G 3 G 4 G5
Groups
Mean Serum Albumin of all the groups
Albumin
Table No. 24 Intermediate calculation, ANOVA table SGOT
Source of
variation
Degrees of
freedom
Sum of squares Mean square
Treatments 4 6408 16028
Residuals 25 3004 1201
Total 29 9412
F = 13.332
Table No. 25 One way analysis of variation (ANOVA)
Comparison Mean difference t value P value
G2 vs G3 5.00 0.111 P > 0.05
G2 vs G4 291.00 6.502 *** P < 0.001
G2 vs G5 47.00 1.05 P > 0.05
G3 vs G4 286.00 6.39 ** P < 0.01
G3 vs G5 42.00 0.938 P > 0.05
G4 vs G5 -244.00 5.45 ** P < 0.01
Non significant (P > 0.05)
** Medium significant (P < 0.01)
*** Highly significant (P < 0.001)
Results
95
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Table No. 26 Intermediate calculation, ANOVA table SGPT
Source of
variation
Degrees of
freedom
Sum of squares Mean square
Treatments 4 21077 5269.3
Residuals 25 2649.8 105.99
Total 29 23727
F = 49.714
Table No. 27 One way analysis of variation (ANOVA)
Comparison Mean difference t value P value
G2 vs G3 2.200 0.523 P > 0.05
G2 vs G4 55.10 13.11 *** P < 0.001
G2 vs G5 41.40 9.85 *** P < 0.001
G3 vs G4 52.90 12.58 *** P < 0.001
G3 vs G5 39.20 9.32 *** P < 0.001
G4 vs G5 - 13.70 3.26 P > 0.05
Non significant (P > 0.05)
*** Highly significant (P < 0.001)
Table No. 28 Intermediate calculation, ANOVA table ALP
Source of
variation
Degrees of
freedom
Sum of squares Mean square
Treatments 4 14566 36416
Residuals 25 920.54 36.822
Total 29 146585
F = 988.89
Results
96
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Table No. 29 One way analysis of variation (ANOVA)
Comparison Mean difference t value P value
G2 vs G3 8.40 3.39 P > 0.05
G2 vs G4 111.30 44.32 *** P < 0.001
G2 vs G5 15.80 6.37 ** P < 0.01
G3 vs G4 102.90 41.53 *** P < 0.001
G3 vs G5 7.40 2.98 P > 0.05
G4 vs G5 - 95.50 38.55 *** P < 0.001
Non significant (P > 0.05)
** Medium significant (P < 0.01)
*** Highly significant (P < 0.001)
Table No. 30 Intermediate calculation, ANOVA table T-bil
Source of
variation
Degrees of
freedom
Sum of squares Mean square
Treatments 4 6.849 1.712
Residuals 25 0.510 0.020
Total 29 7.359
F = 83.930
Table No. 31 One way analysis of variation (ANOVA)
Comparison Mean difference t value P value
G2 vs G3 0.08 1.42 P > 0.05
G2 vs G4 0.966 16.57 *** P < 0.001
G2 vs G5 0.383 6.57 *** P < 0.001
G3 vs G4 0.883 15.14 *** P < 0.001
G3 vs G5 0.300 5.14 * P < 0.05
G4 vs G5 - 0.583 10.00 *** P < 0.001
Non significant (P > 0.05)
* Medium significant (P < 0.05)
*** Highly significant (P < 0.001)
Results
97
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Table No. 32 Intermediate calculation, ANOVA table Albumin
Source of
variation
Degrees of
freedom
Sum of squares Mean square
Treatments 4 27.048 6.762
Residuals 25 11.049 0.442
Total 29 38.097
F = 15.299
Table No. 33 One way analysis of variation (ANOVA)
Comparison Mean difference t value P value
G2 vs G3 - 0.30 1.105 P > 0.05
G2 vs G4 - 1.90 7.00 *** P < 0.001
G2 vs G5 - 0.50 1.842 P > 0.05
G3 vs G4 - 1.60 5.895 P > 0.05
G3 vs G5 - 0.20 0.736 * P < 0.05
G4 vs G5 1.40 5.158 ** P < 0.01
Non significant (P > 0.05)
* Medium significant (P < 0.05)
** Medium significant (P < 0.01)
*** Highly significant (P < 0.001)
Graph No- 6
548543
139.7137.5
426417.6
0
100
200
300
400
500
600
Mea
n va
lue
SGOT SGPT ALP
1.931.85
2.52.8
0
0.5
1
1.5
2
2.5
3
T-Bil Alb
G1G2
Comparison between Biochemical parameters of G2 and G3
Biochemical Parameters
Results
98
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Graph No- 7
548
257
139.784.6
426
314.7
0
100
200
300
400
500
600
Mea
n va
lue
SGOT SGPT ALP
1.93
0.96
2.5
4.4
0
1
2
3
4
5
T-Bil Alb
G2G4
Graph No- 8
548501
139.798.3
426410.2
0
100
200
300
400
500
600
Mea
n va
lue
SGOT SGPT ALP
1.931.55
2.5
3
0
0.5
1
1.5
2
2.5
3
T-Bil Alb
G2G5
Comparison between Biochemical Parameters of G2 & G4
Biochemical Parameters
Comparison between Biochemical Parameters of G2 & G5
Biochemical Parameters
Results
99
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Graph No- 9
543
257
137.584.6
417.6
314.7
0
100
200
300
400
500
600
Mea
n va
lue
SGOT SGPT ALP
1.85
0.96
2.8
4.4
0
1
2
3
4
5
T-Bil Alb
G3G4
Graph No- 10
543501
137.598.3
417.6410.2
0
100
200
300
400
500
600
Mea
n va
lue
SGOT SGPT ALP
1.851.55
2.83
0
0.5
1
1.5
2
2.5
3
T-Bil Alb
G3G5
Comparison between Biochemical Parameters of G3 & G4
Biochemical Parameters
Comparison between Biochemical Parameters of G3 & G5
Biochemical Parameters
Results
100
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Graph No- 11
257
501
84.698.3
314.7
410.2
0
100
200
300
400
500
600
Mea
n va
lue
SGOT SGPT ALP
0.961.55
4.4
3
0
1
2
3
4
5
T-Bil Alb
G4G5
Table No. 34 Showing the comparison of effect of toxic & natural group with
treated groups
(By means of t values)
Parameters G2 vs
G3
G2 vs G4 G2 vs
G5
G3 vs G4 G3 vs
G5
G4 vs G5
SGOT 0.111 6.502 *** 1.05 6.39 ** 0.938 5.45 **
SGPT 0.523 13.11 *** 9.85 *** 12.58*** 9.32*** 3.26
ALP 3.39 44.92 *** 6.37 ** 41.53*** 2.98 38.55***
T- Bil 1.42 16.57 *** 6.57 *** 15.14*** 5.14 * 10.00***
Alb 1.105 7.00 *** 1.842 5.895 0.736 * 5.158**
- Non significant table value (P > 0.05)
* - Medium significant table value (P< 0.05)
** - Medium significant table value (P< 0.01)
*** - Highly significant table value (P<0.001)
Comparison between Biochemical Parameters of G4 & G5
Biochemical Parameters
Results
101
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Statistical Analysis
The analysis of experimental data,using Anova for significance of the
difference between averages of the different groups reveals
1.Comparing G2(Liver damage) and G3(Natural recovery) there is no significant
difference in their effect at levels indicated in the table but a little reduction in the
values of G3 is observed except for Alb.
2.Comparing G2(Liver damage) and G4(Treated with SNB-I) there is a significant
difference in their effect at levels indicated in the table for all tests.
3.Comparing G2(Liver damage) and G5(Treated with SNB-II) there is a significant
difference in their effect at levels indicated in the table except for SGOT and Alb.
4.Comparing G3(Natural recovery) and G4(Treated with SNB-I) there is a significant
difference in their effect at levels indicated in the table except for Alb.
5.Comparing G3(Natural recovery) and G5(Treated with SNB-II) there is a significant
difference in their effect only for SGPT, medium significant difference in their effect
for T.bil and Alb, where as no significant difference in their effect for SGPT and ALP
as indicated in the table.
6.Comparing G4(Treated with SNB-I) and G5(Treated with SNB-II) there is a
significant difference in their effect at levels indicated in the table for all values
except for SGPT.
The above analysis shows that both SNB-I and SNB-II are effective in
treatment. Among the two SNB-I is more effective in treatment.
Discussion
102
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
DISCUSSION
The present topic deals with the interpretation of the materials, which are
explained in previous chapters.
Shankhanabhi
Rasarnava of 10th century, which is considered as one of the oldest text, is the
first to include the Shankha in ‘Shukla varga’ may be because of its white colour.
It is the central axis of the shankha. Shankha is the exoskeleton of the
“mollusca” group of aquatic animals. It is collected from the sea and put in the boiling
water, the animal present inside will die and the outer portion, shankha is obtained. It
has an external lustrous yellowish brown horny layer beneath it has a thick layer
chiefly formed of calcium carbonate.
As shankha produces devine deep sound it may be called by synonyms like
Sunada, Deerghanda, Mahanada, Pawana dhwani. By the place of its origin it may be
explained as Kambu, Kamboja. To indicate its aquatic origin it may be explained as
Jalaja, Varija, Varichara, Varibhava, Jaladhara. To denote its morphology it may be
said as Trirekha, Kulilanta and to stress on its mythological relation it may be said as
Haripriya, Mangalaprada. Kambu is the only common synonym explained by all the
authors.
Bhaishajya Ratnavali is the only text, which has mentioned the synonym of
Shankhanabhi as “Khulvaka”.
Two varieties of shankha viz Dakshinavarta and Vamavarta have been
explained by the texts Rasaratna samucchaya, Ayurveda Prakasha, Brihat Rasaraja
Sundara, Rasatarangini and Shaligrama nighantu.
Their classification is based on the direction of the attachment of the whorls
around the axis.
Discussion
103
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Authors after explaining the varieties of shankha, have given importance to
Dakshinavarta shankha and have narrated its properties. Only Rasataranginakara has
clearly mentioned that the vamavarta shankha should be opted for the preparation of
Bhasma.
As the Dakshinavarta shankha is rarely available & sacred, probably to
preserve that variety it is secured from Bhasmeekarana. On the other hand as the
vamavarta shankha is easily available and has the same medicinal potency, it is
selected for the medicinal purpose.
Shankha with the grahya lakshnas has been accepted for this study because of
its less impurities and is ideal one for therapeutics, according to the texts. Then by
removing the avarthas, shankhanabhi are obtained.
Shankhanabhi is then made into small pieces by pounding and subjected to
shodhana. Shodhana is adopted to reduce its toxicity / kshareeyata and also to deal for
therapeutic purpose.
Shodhita shankhanabhi is subjected to marana process by giving gajaputa.
Shankhabhasma is sheetala, kshreeya, amalapittanashaka, agnimandyahara,
balya, grahi, grahanikahara, parinamothashulaghna, tarunyapidikahara,
vishadoshahara, varnya as explained by Rasataranginikara.
Kumari
Bhavaprakasha has explained its properties as bhedani, netrya, rasayani,
brumhana, balya, vrushya, vatahara and vishahara. Further he has indicated it in
gulma, pleehayakrit vruddi, kapha jwara, granthi, agnimandya, visphota, pithaja roga,
raktaja roga & twak roga.
Discussion
104
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Jambeera
Bhavaprakasha has explained its properties as vatashleshma hara and indicated
in shula, kasa, kaphotklesha, chardi, trishna, amadosha, asyavairasya, hriutpeeda,
vanhimandya, krimiroga.
Shankhanabhi Bhasma
Shankhanabhi bhasma along with Jambeera swarasa is a unique drug indicated
for yakritodara in Yogaratnakara, Bhaishajyaratnavali and other Rasa classics. The
dose of the drug is 1 karsha pramana.
Discussion on Pharmaceutical study
As a first phase of Pharmaceutical processing, the raw drugs selected were
subjected for quality assessment to assess the grahya lakshanas, as only the best raw
drug selected can give the best expected result and the efficacy of the medicine is
dependent on the genuinity of raw drugs.
The Rasa dravyas being vijateeya they are subjected to shodhana samskara
which make them sajateeya, that is to say that they are converted into homogenous to
the body system. The shodhana samskara has diversified meanings which not only
aim at making dravya sajateeya but also act in many ways like increasing the potency
of drug, reducing toxicity & enhancing the medicinal property of drug.
Shankhanabhi shodhana
Before going for shodhana shankhanabhi are made into small pieces of 0.5 – 1
cm by pounding so that all the surface of the drug comes in contact with the
liquid used. These pieces are tied in the four fold cotton cloth and pottali is
made.
Discussion
105
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Procedure of dolayantra pachana in kanji for 3 hrs is selected for shodhana.
Only Rasataranginikara has explained to do pachana.
Reasons may be
1) As swedana is time consuming
2) Shankha is basically hard drug so pachana is preferred as it makes soft &
brittle.
3) Here the drug comes in contact with the liquid media used which may
enhance the therapeutical properties.
4) May help in quicker absorption.
Dolayantra is the ideal yantra used for doing pachana. The pottali was
immersed in dolayantra in such a manner that it will not touch the bottom of
the vessel, otherwise their may be chance of burning of cloth and
shankhanabhi pieces come out.
During the process of shankanabhi shodhana in kanji by pachana in
dolayantra, many physico-chemical changes can be infered. Physical
impurities will be removed, making the dravya more bright, clean and clear.
This process makes the dravya more fragile with a view to reduce it to a fine
powder form by the process of marana. Dravya releases certain undesired
chemical constituents in the liquid, taking certain required chemical
constituents of the liquid and because of which certain chemical changes take
place. Pachana is done with acids. As it is chemically active and potent, the
chemical reaction taking place will be more significant, so due to pachana in
kanji probably chemical changes would be oxidation, reduction,
neutralization, salt formation, sublimation and formation of complex
conjugates etc., might have happened upon Shankhanabhi.
Discussion
106
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Effects of Amla Rasa
While describing the properties of amla rasa, Acharya Charaka has given
(Ch.Su.26)
1) Bhuktam Apakarshayati
2) Kledayati
3) Jarayati
This means it softens the substance. Therefore kanji helps in reduction of
hardness & particle size.
Then while describing atiyoga of rasa, he has mentioned that atiyoga of
Amlarasa produces laxity, this clearly indicates Amlarasa is having
dissociative property by which it softens the drug under purification.
As shankha is of marine origin, basically it is kshareeya dravya. So the use of
Kanji as shodhana dravya helps in neutralization.
After shodhana, changes were observed in shankhanabhi, dull white
shankhanabhi changed to bright white.
Shankhanabhi Marana
Marana is a procedure adopted to convert the heterogeneous material in to
homogenous substance and converting it in to nano particles. The puta adopted in
the present study was gajaputa, which exerts up to 10000C.
Shodhita shankhanabhi is then subjected to marana process. For shankha
marana both open and closed puta are explained by Acharyas. Open puta
method is explained in Rasaratna samucchaya and closed puta in
Rasatarangini. Here closed puta is selected to check the contamination of drug.
Discussion
107
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244 gms of Shodhita shankhanabhi pieces are placed in sharava in one layer &
sharava samputa is made and subjected to Gajaputa with average temperature
of 7810C
As the nature itself Agni moves in upward direction, hence 750
vanopalas are placed below the sharava and 250 above.
After swangasheeta, sharava is collected as the swangasheeta period
will allow the drug for complete paka.
After the first Gajaputa, the colour of shankhanabhi pieces changed from
white to grey and weight loss of 12 gms was observed and they become so
brittle that they can be broken with hands and can be easily powdered in
khalwayantra.
Shankhanabhi pieces were powdered, mardana was done with kumari swarasa
and chakrikas were prepared.
Trituration is a process, which allows effective combination of
different constituents of a particular preparation and divides it into finest
particles, thus increasing its assimilative power and therapeutic effect.
Bhavana makes the particles finer by ‘Sanghatha bhedana’ effect. It potentiate
the Dravya and in augmentation of different types of therapeutic values.
Kumari swarasa having sheeta veerya, is able to reduce the teekshnata of the
drug.
By doing mardana with kumari swarasa particle size will be reduced
considerably.
As its picchila, serves as a binding agent and helps in formation of
chakrikas.
Discussion
108
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Kumari has vatakaphahara, shwasahara property and due to the good
purgative effect, it also counteracts the constipating effect of calcium
carbonate.
Chakrikas are kept in sharava in one layer, sharavasamputa is made and
subjected to second Gajaputa.
During Shankhanabhi marana, chakrikas were found to be more
advantageous due to the better agni paka, availability of more surface area and
hence maximum amount of dissociation of particles took place.
Chemically, Shankhanabhi is Calcium carbonate, and it undergoes
thermal decomposition at 500-6000c or 9320F and the chemical reaction
occurring during Shankhanabhi marana can be explained as below-
On heating, CaCO3 dissociates into calcium oxide with liberation of CO2
CaCO3 CaO + CO2↑.
The reaction is reversible and to avoid it, CO2 must be swept off. In
some procedures CO2 escapes leaving the CaO alone and CaO on exposure to
atmosphere catches the water molecule and forms the Calcium oxide hydrate,
CaO.H2O.
Marana done in closed condition have little chance of escaping of CO2
through the minute pores of the Sharava. So, in the present study dissociated
CaCO3 might have combined with CO2 to reform CaCO3 and also CaO when
exposes to atmosphere, it readily absorbs moisture and CO2 to form calcium
carbonate
CaO + H2O →Ca(OH)2
Ca(OH)2 + CO2→CaCO3 + H2↑
Discussion
109
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So, the left out CaO might have react with atmospheric moisture and
CO2 to form calcium carbonate, hence major composition of Shankhanabhi
bhasma will be CaCO3 and very less concentration of calcium oxide may be
present.
Marana is an endothermic reaction in energy supplied in the form of
heat.
The temperature recording during Shankhanabhi marana in
Gajaputa was done with an intention of giving pyrometric objectivity to the
pharmaceutical process. Temperature was recorded by placing the pyrometer
vertically at the junction of upper 1/3rd and thermocouple placed near the
Sharava Samputa. No much difference were observed in average temperature
of two Gajaputas.
The obtained bhasma is analysed.
Shankhanabhi bhasma passed the bhasma pareeksha like rekhapurnata,
shlakshnata, varitara etc tests and there was weight loss of 15 gms.
There was weight loss after every Gajaputa, might be due to reduction
in the particle size and also due to procedure like mardana.
Discussion
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Discussion On Analytical Study
This part exposes the hidden facts about the final product when it was
critically
analysed with the help of physical and chemical parameters.
Discussion on Organoleptic characters
The change in organoleptic characters after shodhana is because of removal of
external impurities of Shankhanabhi by acidic media of kanji, which causes increasing
in brightness and decreasing in weight.
Discussion on ancient parameters: The colour of Shankhanabhi bhasma is white in
colour which is similar as explained in classics i.e. ‘Kundavajjayate bhasma”. Sparsha
is smooth and soft, and odourless. Sample fulfilled rekhapurnatwa and varitaratwa
which are bhasma siddi Lakshanas.
Discussion on modern parameters:
pH: report showed that pH was 9.8, recommends that the final product is slightly
alkaline. Possibly, this property may not irritate the mucous membrane of the GIT
during
its absorption. It may be due to the alkaline nature of the Shankha.
Loss on drying (at 1100C)
0.24 % reveals the presence of negligible amount of moisture in Shankhanabhi
bhasma. Reduction in moisture content reduced the chance of microbial
contamination, decomposition due the undesired chemical changes. Moisture content
of Shankhanabi bhasma shows the rare chance of bacterial and fungal growth, less
hygroscopic, least drug deterioration and contamination. Hence, the shelf life of
prepared Shankhanabhi bhasma in the present study is more.
Discussion
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“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Total ash: In Pharmaceutical preparation the amount of Ash which is left after
processings represents the inorganic residue. Shankhanabhi bhasma was evaluated for
total ash value and it was found to be 61.3 % which implies the inorganic constituents
add the left 38.7% in Shankhanabhi bhasma was in organic bioacceptable form.
Acid insoluble ash: is 0.53% suggests that the quantity is less than total ash. The low
acid insoluble ash values facilitates the easy absorption of drug.
The water soluble extractive : is 9.62%
Alcohol soluble extractive : is 1.85% this shows the absorption of Shankhanabhi
bhasma in the gut.
Particle size: is 51.20 micrometer this shows the particle size are fine in nature,
which is able to enter into the small capillaries, the particle size is proportional to
optimal biological activity.
Marana reduces the particle size of particular drugs. Particle size of the
Shankhanabhi bhasma is reduced in successive putas supporting to their
Rekhapoornatva and Varitaratva lakshanas.
Reduction in the particle size during marana may be due to the following
reason: Solid crystal at a rest has packed particles which are closed together in a
lattice force and vibrate in their fixed portion. But when temperature increases, the
particle grain (crystalline area) increase and vibrate more strongly, occupies more
spaces this causes solid to expand. Due to increase in intra-atomic distance,
electrostatic forces get weakened. Due to continuous heating, particles get enough
energy to break forces holding them together and to get reduce in their sizes.
Flow property:
As the drug is in powder form it is tested for its flow property. This analysis
makes us to know weather any adjuncts are essential for proper flow of drug during
Discussion
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Capsule or Tablet preparation. Flow property was identified by “Angle of Repose
(Tanθ)” and Flow rate by “Compressibility index (I)”. Shankhanabhi bhasma has a
good Flow property with Tanθ =27.40 and Flow rate I=15%. Hence it can be said that
it does not need any adjuncts in capsule preparation.
Solubility:
It is sparingly soluble in alcohol and water means that 1 gm of Shankhanabhi
bhasma is soluble in 100-1000 parts of the solvents. It is found to be insoluble in
chloroform. Shankhanabhi bhasma is soluble with effervescence in dil Hcl.
Discussion on Quantitative tests:
The quantitative estimation of Ca, CaCO3, Fe, Mg & S in Shankhanabhi
bhasma were 38.2%, 95.3%, 0.06%, 0.14% and 1.05% respectively, which were
within the standard limits given for Shankha bhasma by Pharmacopeal standards for
Ayurvedic medicines. This infers that there is no much difference between
Shankhanabhi bhasma and Shankha bhasma in their chemical composition.
NPST
Namboori’s phased spot test showed no marked difference of Shankhanabhi
bhasma when compared with the Pravala bhasma (Shakha) standards.
The sample showed the exothermic reaction and the absorption and settling
time was also normal.
Shankhanabhi bhasma shows a characteristic formation of solid Pink spot with
wet periphery in N.P.S. Test on Haridra paper.
Only way to differentiate them was by “fading away time”. Shankhanabhi
bhasma spot starts fading away slowely than that of pravala bhasma shakha.
The 3rd phase spot is taken as standard spot.
Discussion
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Discussion on Experimental Study
By seeing all the guna karma of trial drug one can assess that it may help in
condition where the total digestion and metabolism is hampered.
Shankhanabhi bhasma is having specific action like deepana, lekhana,
vishaghna, balya etc. Certainly this may help for removal of toxic substances and has
stimulating action on the liver (Siddaprayoga sangraha)
Shankhanabhi bhasma by the virtue of its kshara guna is beneficiated in Yakrit
and Pleeha vruddi (SPS and OGDS)
As basically yakritodara is agnimandyajanya vikara, which is tridoshaja and
kledayukta, Shankhanabhi bhasma kshara by its Tridosha shamaka, agnivardhaka
gunas overcomes the samprapti.
The protection of liver can be attributed by the role of major application of the
drug on pitta dosha (O.G.D.S).
Shankhanabhi bhasma is having vishanashakha property. Therefore the
toxicity of the liver due to any reasons shall be corrected by Shankhanabhi bhasma.
As the present drug shankhanabhi bhasma is mentioned under
pleehayakridroga chikitsa adhikara, its hepatoprotective activity was screened
carefully using the experimental model suggested by Watanabe and Takita.
CCl4 is used as hepato toxic agent. Proper selection of animals, groupings and
experimental protocol are explained in the methodology (page - 87) The dose
determination, experimental procedure and experimental parameters are given in
(Page – 88 -90) methodology in detail. Shankhanabhi bhasma in two different doses
(SNB-I ancient- 1 karsha, SNB-II recent- 2 ratti) with Jambeera swarasa are given in
the form of suspension, by converting it to the animal dose with the help of standard
converting formula.
Discussion
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“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Discussion on experiment
Biochemical and histopathological observations shown that these two groups
treated with SNB- I and SNB- II are significantly efficient in protecting the liver.
To reveal the synthesis efficacy of the liver the Alkaline phosphate, SGOT,
SGPT, Total bilirubin, Serum albumin and histopathological study of liver were
analyzed by comparing between the groups like Group 2 (Toxic) with Group 3
(Natural Recovery) Table no – 34.
The Group-2 and Group-3 compared with the treated groups, i.e in treated
groups Group 4 (SNB- I) Group 5 (SNB –II).
In Group I (Control) the Alkaline phosphatase, SGOT, SGPT, Total bilirubin,
Serum albumin and the normal histopathology of liver was observed. In Group 2
(Intoxicated/ liver damage) all the biochemical values Alkaline phosphatase, SGOT,
SGPT, Total bilirubin were highly increased except the Serum albumin which is
decreased. [The t-values corresponds to control group and G2 at 5% level (p<0.001)
CCl4 is effective in producing liver damage] (Table No. 91). The histopathology study
of liver of this group showed massive fatty changes, necrosis, ballooning
degeneration, and broad infiltration of the lymphocytes and kupffer cells around the
central vein and the loss of cellular boundaries. It indicates that in course of CCl4
administration leads to functional defects of the hepatocytes and multiple biochemical
variation took place.
In comparing between Group 2 (Intoxicated/ liver damage) and in Group 3
(Intoxicated control / Natural Recovery) to assess the natural recovery, the LFT
values slightly came down except Serum albumin which is slightly increased. [Group
3 is not significant at P>0.05] (Table no. 91). In histopathological report also there is
loss of architecture and cell necrosis (perivenular) extending to the central zone. The
Discussion
115
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
cell necrosis with inflammatory collections is more prominent in the central zone than
around central vein.
For assessing the curative effect of two trial drugs, the two separate groups of
rats were taken, CCl4 was administered to the rats for 5 days. Then for the next 5
days trial drug were given to assess the curative effect when compared with Group 2
and Group 3 the LFT values were increased in intoxicated groups except Serum
albumin which decreased. In the treated groups (G4 & G5) the LFT values decreased
considerably. The LFT values of Group 4 came very nearly to normal values. [highly
significant with P<0.001 (Table no. 91). The LFT values of Group 5 also decreased
considerably but are not near to the normal values. [medium significnacy with P<0.01
(Table no. 91).
The Histopathology study of both Group 4 and Group 5 showed well brought
out central vein, hepatic cell with well preserved cytoplasm, prominent nucleus and
nucleolus in the liver section.
All the biochemical values of all groups were showed in tabular form and also
the comparison between the groups by doing the statistical analysis and using the bar
diagram is shown in chapter of result.
Conclusion
116
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
CONCLUSION
1) Shankhanabhi is an aquamarine Calcium Carbonate compound categorized
under sudha varga used in medical practice since ancient times.
2) Dolayantra Kanji Pachana method is good operative procedure for Shodhana
of Shankhanabhi.
3) Shankhanabhi marana by Kumari swarasa bhavana requires 2 Gajaputas.
4) The quantitative analysis has shown normal limits of CaCO3 , Mg, Fe and S,
when compared with the normal limits of Shankha bhasma.
5) Shankhanabhi bhasma has shown good hepatoprotective activity on Albino
rats which is statistically significant (P<0.001).
By comparing biochemical, histological and statistical analysis both SNB-I
and SNB-II have significant therapeutic effect on hepatoprotective activity.
Among the two drugs when it is analysed statistically, the hypothesis of equal
effective is rejected only for alkaline phosphate at 5% and accepted for the all
other parameters at any level. But the observations reveal that the values are
closer to the normal values for the SNB-I group than the SNB-II.
Therefore on the basis of analysis it is concluded that SNB-I is more effective
than SNB-II.
6) By this it can be speculated that any drug which reverses the hepatic damage
induced by CCl4 can certainly protect liver parenchyma against other
hepatotoxins.
7) In the classics Jambeera swarasa is mentioned as Anupana of Shankhanabhi
bhasma, which implies that ancient Vaidyas were also aware of the fact that
Jambeera swarasa will neutralize the remaining alkaline property present in
bhasma and will reduce its irritant effect.
Summary
117
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
SUMMARY
The present dissertation work entitled “Preparation, physico-chemical analysis
of Shankhanabhi bhasma and evaluation of its Hepato-protective activity –
Experimental study” contains topics introduction, review of literature, methodology
which embraces pharmaceutics, analytical study, experimental study, observations,
results, discussion and conclusion.
Importance of Liver, Need for the study and plan of the study is discussed in
introduction.
Aims and objectives:
Preparation of Shankhanabhi Bhasma.
Physico-chemical analysis of Shankhanabhi Bhasma.
Evaluation of Hepato protective activity of Shankhanabhi Bhasma.
The review of literature relating to the research topic was done from both
Ayurvedic and modern texts. The literature pertaining to the trial drug Shankhanabhi
is described. The Hepatoprotective activity of the trial drug was particularly referred
according to Yakriduttejaka, Deepana, Lekhana, Vishaghna, Tridoshashamaka,
Kshara properties of the drug on different references available in the texts.
Hepatoprotective activity being the object of the experimental study, the
importance of Liver, its functions and involvement in various diseases from both the
Ayurvedic and Modern literatures are elaborately explained under this chapter.
Information found sporadically in these literatures were compiled and presented under
appropriate heading.
In Methodology, Pharmaceutical study includes identification and collection
of raw drug, preparation of kanji, Shankhanabhi shodhana and Shankhanabhi Marana.
Summary
118
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
The analytical study was done for Shankhanabhi bhasma. In that the physico-
chemical analysis like organoleptic characters, determination of pH values, loss on
drying at 1100C, loss on ignition, determination of total Ash, Acid insoluble ash,
water soluble extractive, alcohol soluble extractive, finess of particles, particle size,
flow property, flow rate, Solubility text, Estimation of Ca, CaCO3, Fe, Mg, S and
NPST were done.
In experimental study, selection of animals, mode of administration of drug,
experimental protocol and experimental parameters were mentioned.
Under the heading of results the data obtained from the study conducted is
presented with the help of graphs and statistical analysis is done. The readings and the
statistical analysis of Alkaline phosphatase, SGOT, SGPT, Total bilirubin and Serum
albumin were dealt. Histopathological findings of all groups in comparison to the
biochemical reports were discussed.
Discussion was done on Pharmaceutical study, Analytical study and
Experimental study. The rationality behind the Shodhana, Marana and Probable mode
of action of Shankhanabhi bhasma were also discussed.
On the basis of results finally this study concluded that Shankhanabhi bhasma
by virtue of its properties does the samprapti vighatana and subsidies the Yakritodara
and is a highly significant Hepatoprotective drug.
Summary
119
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
Scope of the study
Analytical estimation can be done at various stages of the preparation of
Shankhanabhi bhasma.
Shankhanabhi bhasma prepared by different shodhana procedures and their
efficacy in yakritodara can be studied.
Shankhanabhi bhasma can be compared with other bhasmas mentioned for
yakrit pleeha roga experimentally.
Further the present study can be extended for clinical evaluation.
Bibiliography
120
“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
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68) Bhavamishra, Bhavaprakasha Nighantu, Guduchyadi varga, sloka 230, edited by Bramashankara mishra and Rupalayi vaisya: 5th ed, Varanasi: Choukamba Sanskrit series office: 1969, P.419.
69) Ibid chapter Guduchyadi varga, P 419.
70) Ibid chapter Guduchyadi varga, P 419.
71) Ibid chapter Guduchyadi varga, shloka 230, P 419.
72) Ibid chapter Amradi phala varga, Shloka 135, P 594.
73) Ibid chapter Amradi phala varga, Shloka 135, P 595.
74) C.K. Kokate, Pharmacognosy, 12th ed. Pune: Nirali prakashana; 1999, Drugs
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75) Pharmacognosy, C.K. Kokate et al, chapter 11, 12th ed,; Nirali Prakashan; Pune; 1999: P. 286
76) Bhavamishra, Bhavaprakasha Nighantu, Amaradi phala varga, sloka 135,
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83) Sushruta, Sushruta samhita sutrasthana chapter 4, sloka 57-58, Ambikadatta
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84) Sushruta, Sushruta samhita shareerasthana chapter 4, sloka 30, Ambikadatta shastri: 14th ed, Varanasi: Chawkhambha publication: 2003, P.32.
85) Sushruta, Sushruta samhita shareerasthana chapter 4, sloka 10, Ambikadatta
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86) Sushruta, Sushruta samhita shareerasthana chapter 9, sloka 11, Ambikadatta shastri: 14th ed, Varanasi: Chawkhambha publication: 2003, P.71.
87) Sushruta, Sushruta samhita chikitsasthana chapter 2, sloka 12, Ambikadatta
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89) Sushruta, Sushruta samhita shareerasthana chapter 4, sloka 10, Ambikadatta
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90) Sushruta, Sushruta samhita shareerasthana chapter 4, sloka 15, Ambikadatta shastri: 14th ed, Varanasi: Chawkhambha publication: 2003, P.30.
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94) Agnivesha, Charaka samhita, Nidana sthana Chapter 2, Sloka 4, edited by Dr. Brahmananda Tripathi, reprint edition 2003, Choukhamba surabharati Prakashana Varanasi. P 497.
95) Sushruta Acharya Sushruta samhita, Uttarardha, sutrasthana Chapter 45
Shloka 30, edited by Ambikadatta shastri Reprint edition 2005, Chawkahmbha samskrita samsthana, P 307.
96) Agnivesha, Charaka samhita, chikitsastana Chap 16, Sloka 4, Dr.
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97) Vagabhata, Astanga hrdaya, Vol-II chikitsasthana, chapter 13, sloka 1, Proff.
K.R. Shrikantha Murthy editor: 5th ed, Varanasi: Choukhambha krishnadas Academy 2003, P.517.
98) Agnivesha, Charaka samhita, chikitsasthana Chapter 16, Sloka 34, edited by
Dr. Brahmananda Tripathi, reprint edition 2003, Choukhamba surabharati Prakashana Varanasi. P 419.
99) Ibid shloka 35-36, P 419.
100) Ibid shloka 124, P 430. 101) Vagabhata, Astanga hridaya, Vol-II chikitsasthana, chapter 35, sloka 50-52, Proff. K.R. Shrikantha Murthy editor: 5th ed, Varanasi: Choukhambha krishnadas Academy 2003, P.336. 102) Sushruta Acharya Sushruta samhita, Nidanasthana Chapter 7, Shloka 16, edited by Ambikadatta shastri Reprint edition 2005, Chawkahmbha samskrita samsthana, P 258. 103) Agnivesha, Charaka samhita, chikitsasthana Chapter 13, Sloka 88, edited by Dr. Brahmananda Tripathi, reprint edition 2003, Choukhamba surabharati Prakashana Varanasi. P 397. 104) Vangasena, Vangasena samhita, Chapter 42, Shloka-221, 1st edition, Varanasi: Chowkhambha Sanskrit series office: 2004, P-616. 105) Shri Govindadas, Bhaishajya Ratnavali Chapter 41st, shloka 51, Kaviraj Shri Ambikadatta shastri editor, 18th ed, Varanasi: Chaukhambha Sanskrit sansthana: 2005, P.775. 106) Yogaratnakara, Uttarardha, chapter udaroroga chikitsa, Shloka 1, editor Laxmipati shastry, 6th ed, Varanasi: Choukamba Sanskrit sansthana: 1997, P.114.
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107) Agnivesha, Charaka samhita, chikitsasthana Chapter 13, Sloka 12-15, edited by Dr. Brahmananda Tripathi, reprint edition 2003, Choukhamba surabharati Prakashana Varanasi. P 382. 108) Sushruta Acharya, Sushruta samhita, Nidanasthana Chapter 7, Shloka 14, edited by Ambikadatta shastri Reprint edition 2005, Chawkahmbha samskrita samsthana, P 257. 109) Vagabhata, Astanga hridaya, Nidanasthana, chapter 12, sloka 22, Proff. K.R. Shrikantha Murthy editor: 5th ed, Varanasi: Choukhambha krishnadas Academy 2003, P.117. 110) Sri Madavakara. Madhava nidanam with madhukosha Sanskrit commentary by sri vijayarakshita and srikantha data, vidyotini hindi commentary by sri sudarshana sastri, Purvardha, chapter 35, edited by yadunandana upadhyaya; 26th ed, Varanasi, Chaukhambha Sanskrit sansthana: 1996.Shloka 16 P. 38. 111) Yogaratnakara, Uttarardha, chapter udaroroga nidina, Shloka 17, editor Laxmipati shastry, 6th ed, Varanasi: Choukamba Sanskrit sansthana: 1997, P.104. 112) Agnivesha, Charaka samhita, chikitsasthana Chapter 13, Sloka 37, edited by Dr. Brahmananda Tripathi, reprint edition 2003, Choukhamba surabharati Prakashana Varanasi. P 387. 113) Sushruta Acharya, Sushruta samhita, Nidanasthana Chapter 7, Shloka 15, edited by Ambikadatta shastri Reprint edition 2005, Chawkahmbha samskrita samsthana, P 257. 114) Vagabhata, Astanga hridaya, Nidanasthana, chapter 12, sloka 23-27, Proff. K.R. Shrikantha Murthy editor: 5th ed, Varanasi: Choukhambha krishnadas Academy 2003, P.117. 115) Sri Madavakara. Madhava nidanam with madhukosha Sanskrit commentary by sri vijayarakshita and srikantha data, vidyotini hindi commentary by sri sudarshana sastri, Purvardha, chapter 35, Yadunandana upadhyaya editor; 26th ed, Varanasi, Chaukhambha Sanskrit sansthana: 1996.Shloka 17 P. 38. 116) Yogaratnakara, Uttarardha, chapter udaroroga nidina, Shloka 18-19, editor Laxmipati shastry, 6th ed, Varanasi: Choukamba Sanskrit sansthana: 1997, P.104. 117) Sri Madavakara. Madhava nidanam with madhukosha Sanskrit commentary by sri vijayarakshita and srikantha data, vidyotini hindi commentary by sri sudarshana sastri, Purvardha, chapter 35, Yadunandana upadhyaya editor; 26th ed, Varanasi, Chaukhambha Sanskrit sansthana: 1996.Shloka 18 P. 39. 118) Human Physiology Vol I, Chapter 10 metabolism, Dr. C.C. Chatterjee editor 11th edition, reprint 2000, Calcutta : Medical Allied Agency, P 652-653. 119) Ibid
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120) Ibid P 655 – 663
121) Harrison, Principles of Internal medicine, Vol -2, Part 11, Section 2, 14th edition, Anthony S. Fauci et al editors, McGraw- Hill Book Co-Singapore: 1998: P 1663. 122) Ibid P 1692. 123) Aspi F. Golwalla & Sharukh. A. Golwalla, Medicine for Students, Chap 1, 21st edition, Dr. A.F. Golwalla Express court, Church gate, Mumbai :2005: P 72-73. 124) Harrison, Principles of Internal medicine, Vol -2, Part 11, Section 2, 14th edition, Anthony S. Fauci et al editors, McGraw- Hill Book Co-Singapore: 1998: P 1664. 125) Ibid P 1663. 126) Dr. Vinayak Bhat, Anti Hepatato toxicity activity of Vasa moola 2002 March, A.L.N. Rao Ayurvedic medical college Koppa, RGUHS. P 48-49. 127) Harrisons principles of Internal medicine part 11, 14th edition Harisons M.C Graw- Hill, Book Co-singapore, Part 11 Section 2, P 1663. 128) Vinay Kumar et al, Robbins Basic Pathology, Chap 16, 7th edition, Harcourt (India) Pvt. Ltd: New-Delhi: 2003, P 518. 129) R.S. Satoskar et al, Pharmacology and Pharmaco Therapeutics, section 1, chap 1, 16th edition, R.R. Satoskar editor, Mumbai Popular Prakashan: 1999, P 4. 130) Fundamentals of experimental pharmacology, M.N. Gosh, 3rd ed, Calcutta : Hilton & Company : 2005, P 192.
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ANNEXURE
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