Hepatoprotective rs018 gdg

“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

Ayurmitra

TAyComprehended

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-


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




Dr. Jayashree S. in partial fulfillment of the requirement for the degree of “Ayurveda

Vachaspathi M.D (Rasashastra)”, Under Rajiv Gandhi University of Health


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




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


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


98


99


99


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


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


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


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


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.

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Objectives

6


AIMS AND OBJECTIVES

1.Preparation of Shankhanabhi Bhasma.

2.Physico-Chemical analysis of Shankhanabhi Bhasma.

3.Evaluation of Hepato protective activity of Shankhanabhi Bhasma.

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Drug review

7


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 mÉÉÌuÉ§ÉeÉlÉMüÉå 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 qsÉåcNûÉ¬Í¤ÉhÉmÉÔuÉïiÉÈ |

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.

Disease review

51


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

Disease review

52


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).

Disease review

53


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.

Disease review

54


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.

Disease review

55


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).

Disease review

56


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.

Disease review

57


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

“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato

58

protective Activity - An Experimental Study”

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.


59


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


60


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


61


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.


62

Dravya Quantity of Shankhanabhi

before shodhana (in gms)

Quantity of

Shankhanabhi after

shodha (in gms)

Wt loss

(in gms)

Shankha nabhi 250 244 6


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


63


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.


64


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.


65


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


66


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.


67


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


68


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.


69


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


70


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%


71


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


72


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.


73


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.


74


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.


75


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.


76


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 %


77


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


78


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


79


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


80


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


81


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’.


82


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.


83


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.


84


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.


85


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.


86


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.


87


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.


88


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).


89


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.


90


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Results

91


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


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


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


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


Table No. 26 Intermediate calculation, ANOVA table SGPT

Source of

variation

Degrees of

freedom


Treatments 4 21077 5269.3

Residuals 25 2649.8 105.99

Total 29 23727

F = 49.714



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



Table No. 28 Intermediate calculation, ANOVA table ALP

Source of

variation

Degrees of

freedom


Treatments 4 14566 36416

Residuals 25 920.54 36.822

Total 29 146585

F = 988.89

Results

96




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




Table No. 30 Intermediate calculation, ANOVA table T-bil

Source of

variation

Degrees of

freedom


Treatments 4 6.849 1.712

Residuals 25 0.510 0.020

Total 29 7.359

F = 83.930



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


* Medium significant (P < 0.05)


Results

97


Table No. 32 Intermediate calculation, ANOVA table Albumin

Source of

variation

Degrees of

freedom


Treatments 4 27.048 6.762

Residuals 25 11.049 0.442

Total 29 38.097

F = 15.299



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


* Medium significant (P < 0.05)



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


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




Results

99


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





Results

100


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)



Results

101


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.

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Discussion

102


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


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


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


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


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


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


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


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

110


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

111


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

112


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

113


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

114


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


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.

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Conclusion

116


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


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


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


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

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ANNEXURE

Shlokas of zÉ„¡ûlÉÉpÉÏ pÉxqÉ

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