Rasapushpa antimicrobial rs014-gdg

By

DR. ANITHA. H.

Dissertation Submitted to theRajiv Gandhi University Of Health Sciences,

Karnataka, Bangalore.

In partial fulfillment of the requirements for the degree of

AYURVEDA VACHASPATHI

In

RASASHASTRA

Under the guidance of

DR. M.C. PATIL M.D. (Ayu)

Under the co-guidance of

DR. JAGADEESH MITTI. M.D. (Ayu)

DEPARTMENT OF RASASHASTRA,POST GRADUATE STUDIES AND RESEARCH CENTER,

SHRI D. G. MELMALAGI AYURVEDIC MEDICAL COLLEGE,GADAG – 582103.

2004-2007

PREPARATION, PHYSICO-CHEMICAL ANALYSIS OFPREPARATION, PHYSICO-CHEMICAL ANALYSIS OFPREPARATION, PHYSICO-CHEMICAL ANALYSIS OFPREPARATION, PHYSICO-CHEMICAL ANALYSIS OFPREPARATION, PHYSICO-CHEMICAL ANALYSIS OF

RASAPUSHPA AND ITS ANTIMICROBIAL ACTIVITYRASAPUSHPA AND ITS ANTIMICROBIAL ACTIVITYRASAPUSHPA AND ITS ANTIMICROBIAL ACTIVITYRASAPUSHPA AND ITS ANTIMICROBIAL ACTIVITYRASAPUSHPA AND ITS ANTIMICROBIAL ACTIVITY

Ayurmitra

TAyComprehended

Rajiv Gandhi University Of Health Sciences, Karnataka,Bangalore.

DECLARATION BY THE CANDIDATE

I here by declare that this dissertation entitled

“Preparation, Physico-Chemical Analysis of Rasapushpa and its

Antimicrobial Activity.” is a bonafide and genuine research work car-

ried out by me under the guidance of Dr. M.C. Patil, M.D.(Ayu),

(Rasashastra), Professor and H.O.D, Post graduate department of

Rasashastra and under the co-guidance of Dr. Jagadish. Mitti. M.D.(Ayu),

Lecturer, Post graduate department of Rasashastra.

Date:Place: Gadag.

Dr. Anitha. H

CERTIFICATE BY THE GUIDE

This is to certify that the dissertation entitled “Preparation,

Physico-Chemical Analysis of Rasapushpa and its Antimicrobial Activity.”

is a bonafide research work done by Dr. Anitha. H in partial fulfillment of

the requirement for the degree of Ayurveda Vachaspathi. M.D (Rasashastra).

Date:

Place:Gadag.

Dr. M.C. Patil, M.D. (Ayu)

Professor & H.O.D. Department of Rasashastra,

D.G.M.A.M.C, Gadag.

SHRI D. G. MELMALAGI AYURVEDIC MEDICAL COLLEGE,POST GRADUATE DEPARTMENT OF RASASHASTRA.

CERTIFICATE BY THE CO- GUIDE


Physico-Chemical Analysis of Rasapushpa and its Antimicrobial Activity.”

is a bonafide research work done by Dr. Anitha. H in partial fulfillment of

the requirement for the degree of Ayurveda Vachaspathi. M.D (Rasashastra).

Date:

Place:Gadag.

SHRI D. G. MELMALAGI AYURVEDIC MEDICAL COLLEGE,POST GRADUATE DEPARTMENT OF RASASHASTRA.

Dr. Jagadeesh Mitti. M.D. (Ayu)

Lecturer Department of Rasashastra,

D.G.M.A.M.C, Gadag.

J.S.V.V. SAMSTHE

D.G.M. AYURVEDIC MEDICAL COLLEGEPOST GRADUATE STUDIES AND RESEARCH CENTRE

DEPARTMENT OF RASASHASTRAGADAG –582103

CertificateI here by certify that this dissertation entitled “ PREPARATION, PHYSICO-

CHEMICAL ANALYSIS OF RASAPUSHPA AND ITS ANTIMICROBIAL ACTIVITY” is

bonafide and genuine research work done by Dr. Anitha. H in partial fulfillment of the requirement for

the degree of Ayurveda Vachaspati (M.D. in Ayurveda) in Rasashastra of Rajiv Gandhi University of

Health sciences, Bangalore, Karnataka under my Guidence.

Associate-GuideDate: Dr. K.Krupanidhi. M.Sc, M.Phil, Phd

Lecturer,Place: Gadag DEPT OF MICROBIOLOGY & BIOTECHNOLOGY,

S.C.S. COLLEGE OF PHARMACY,

ENDORSEMENT BY THE H.O.D AND PRINCIPAL OF

THE INSTITUTION


Physico-Chemical Analysis of Rasapushpa and its Antimicrobial

Activity.” is a bonafide research work done by Dr. Anitha. H under the

guidance of Dr.M.C.Patil, M.D.(Ayu), Professor & H.O.D, Postgraduate

department of Rasashatra and Under the co-guidance of Dr. Jagadish

Mitti M.D.(Ayu), Lecturer, Post graduate department of Rasashastra.

Dr. G. B. Patil, Principal.

D.G.M.A.M.C, Gadag.

Dr. M.C. Patil, M.D. (Ayu)

Professor & H.O.D. Department of Rasashastra,

D.G.M.A.M.C, Gadag.

Date:

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 in print or electronic format for academic /

research purpose.

Date:

Place:Gadag.

© Rajiv Gandhi University of Health Sciences, Karnataka.

Dr. Anitha. H

ACKNOWLEDGMENT

By the grace of Lord Dhanvantari and blessings of Shri Raghavendra Swamiji,

it is pleasure to express my full respect and regards to my parents

Shri Raghavendrachar and Smt. Pushpa, and my husband Shri Gururaj Joshi, who

made me to proficient and gave constant support and encouragement.

With deep sense of Pleasure, I express my obligation to my honourable

guide Dr. M.C. Patil MD (Ayu) HOD, PG Dept. of Rasashastra, DGMAMC, Gadag, for

his scholarly guidance, supervision, creative criticism, and high inspiration at every

stage of this work.

I express my deep gratitude to beloved Principal Dr. G.B. Patil, Principal

DGMAMC, Gadag, for his encouragement and providing all necessary facilities for

this research work.

My gratitude is greatest towards my co-guide Dr. Jagadeesh. Mitti, 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 thanks to Dr. K.Krupanidhi. M.Sc, M.Phil, Phd Lecturer, Dept. of

Microbiology & Biotechonology, S.C.S. College of Pharmacy, Harpanahalli, for his

valuable directions.

I owe my heartfelt credit to Dr. Dilipkumar B, Asst Professor, PG Dept of

Rasashastra DGMAMC, Gadag, for his critical views and precious suggestions.

I ofer my sincere thanks to Dr. R.K. Gaccchinmath, professor and HOD, UG

Dept of Rasashastra, DGMAMC, Gadag, for his constant support.

I wish to convey my regards to Dr. G.N. Danappagoudar MD (Ayu) Lecturer PG

Dept of Rasashastra, DGMAMC, Gadag, for his creative criticism and

encouragement.

I express my sincere thanks to Dr. Varadacharylu M.D. (Ayu), Dr. Mulagund M.D.

(Ayu), Dr. G. Purushothamacharyalu M.D. (Ayu) for their constant support.

I express my gratitude to Dr. R.V. Shettar M.D. (Ayu), Dr. Samudri

M.D.(Ayu) for their encouragement as well as suggestions for this research work.

I am grateful to Dr. K.S.R. Prasad M.D. (Ayu), Dr. Shivramudu M.D. (Ayu), Dr. S.S.

Doddamani M.D. (Ayu), Dr. R.V. Shettar M.D. (Ayu), Dr. Kuber sankh M.D. (Ayu), Dr. Santosh

I

Belvadi M.D. (Ayu), Dr. Sashikanth Nidagundi M.D. (Ayu), and other P.G. Staff for their

constant encouragement.

I extend my gratitude to Shri V.M. Mundinmani and Surebanu for providing

the required books during the study.

With great pleasure I offer my recognition to my friends Dr. Suvarna P.

Nidagundi, Dr. Sharanabasappa S. Angadi, Dr. Anand H and Dr. Shambulinga Teggi

for their friendly affection and amiable attitude during my study period without which

I would never be complete.

I offer my sincere thanks to my friends Dr. B.Y. Ghanti, Dr. Pradeep, Dr.

Sobagin, Dr. Shankuntala and Dr. M.S. Hiremath for their kind co-operation and help.

I offer my sincere thanks to my friends Dr. Santoji, Dr. Koteshwara, Dr, V.S.

Hiremath, Dr. Jaggal, Dr. R.B. Pattanshetty, Dr. for their immense help and affection.

I am also thankful to my junior friends Dr. Rudrakshi, Dr. Suma Jamakhandi,

Dr. Jaya, Dr. Kattimani, Dr. Shivakumar, Dr. Ravindra, Dr. Anupama Bijjal, Dr.

Sarvamangala, Dr. Kavitha for their support and affection.

I am grateful to Shri Chaitrakumar (Sadguru computers) for his kind co-

operation and immense help to complete the dissertation work,.

I express my regards to my in-laws, Shri C.L. Joshi, Smt. Shanta, for their

moral and cordial support during the study.

It would be my privilege to convey my love to my brother Krishnamurthy, and

Co-brothers Ramesh Deshpande, Koteshwara, and my sisters Sunita, Poornima,

Bhavani for their moral support during this study.

I express my thanks to all those who had 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

remembering respectful parents and my husband Shri Gururaj Joshi.

Dr. Anitha H

II

TABLE OF CONTENTS

S.L.NO. INDEX PAGE. NO

1. Introduction 1-3

2. Objectives 4

3. Review of Literature 5-102

4. Methodology 103-138

5. Results 139-140

6. Discussion 141-155

7. Conclusion 156-157

8. Summary 158-160

9. Bibliography 161-174

10. Annexure

I. Slokas of Rasapushpa II. Slokas of Hingula

VI

ABSTRACT

Krimis are responsible for manifestation of oupasaigika rogas. Oupasaigika

rogas can be considered as infectious diseases and krimis as microorganisms.

Infectious diseases continue to take very high rank as cause of death, which accounts

for ten million persons each year. Hence it is necessary to control these

microorganisms.

Our treatises have described variety of treatment options for management of

krimis. Rasapushpa is one such preparation, which has got properties such as twak

doshahara, bhutavishapaha, krimighna etc. Considering these indications, in a present

study, antimicrobial activity of Rasapushpa was carried out.

Objectives

The present study was planned with the following objectives:

1. Preparation of Rasapushpa.

2. Physico- chemical analysis of Rasapushpa.

3. To evaluate antimicrobial activity of Rasapushpa.

Methods

Pharmaceutical study

1. Hingula shodhana according Rasatarangini (9/16,17)

2. Hingulotha parada according to Rasatarangini (5/39)

3. Preparation of Rasapushpa according to Rasatarangini (6/29,30,31)

Analytical study

Rasapushpa was subjected to physico-chemical analysis i.e. organoleptic

characters, loss on drying, solubility, fineness of particles etc. proportion of Hg and Cl

in Rasapushpa was estimated.

IV

Experimental study

• Disc diffusion method was selected to evaluate the antimicrobial activity of

Rasapushpa.

• Organisms selected for the study include 2 G(+)ve and 2 G(-)ve bacterias and

2 fungi.

• Activity was conducted in the microbiology department pharmacy college,

Harapanahalli.

Results

Results were expressed in terms of zone of inhibition. Vernier Calipers was

used to calculate zone of inhibition.

Rasapushpa has shown less activity for all bacterias compared to standard drug

Cefataxime in both concentrations (5o mg/ml, 100mg/ml).

Rasapushpa has shown significant zone of inhibition for fungi in both concentration

compared to standard “Fluconazole”

Interpretations and conclusion

• Asthasamskaras is a difficult and time taking procedure. Hence Hingulotha

Parada is taken for the preparation of Rasapushpa. It is devoid of naga, vanga

and saptakanchukadi doshas.

• Urdwapatana method used for Satwapatana of Hingula is a easy method to

obtain Parada.

• Because of specific pharmaceutical procedure and minimal dose efficiency of

Kupipakwa method, it was selected for Rasapushpa nirmana.

• Rasapushpa is chemically identified as mercurous chloride.

• Rasapushpa has got significant antimicrobial activity.

Key words

Kupipakwa Rasayana, Rasapushpa, Kramagni, Analytical study, Anti

microbial activity, Disc diffusion method and Significant results.

V

LIST OF TABLES Table No Tables Page No.

1 Showing the synonyms of Hingula according to different authors.

5

2 Showing Hingula included under following category by different authors:

9

3 Showing the bheda of Hingula 10 4 Showing the rasa of Hingula according to different

texts. 12

5 Showing doshagnata of Hingula according to different authors.

12

6 Shows the properities of Hingula according to different granthas.

13

7 Showing the synonyms of parada: 19 8 Showing the ores of parada along with their chemical

composition. 21

9 Showing the varities of Parada depending on the colour and their karma also specified.

22

10 Showing the varieties of Parada depending upon the place of origin.

22

11 Naisaragika Dosha and its effect on sharira. 23 12 Yougika Dosha and its effect on sharira 23 13 Oupadhika (Saptakanchuka) Dosha and its effect 23 14 Showing samanya shodhana of parada according to

different authors: 25

15 Showing vishesha shodhana of parada according to different authors:

26

16 Showing synonyms of kasisa according to different Acharyas.

36

17 Showing classification according to different granthas. 37 18 Showing varities of kasisa according to orgin. 38 19 Showing varities of kasisa according to Colour. 38 20 Showing various procedures for purification of Kasisa. 39 21 Showing Rasa of kasisa according to different

acharyas. 39

22 Showing synonyms of saindhava lavana 44 23 Showing the karma of saindhava lavana 46 24 Showing the Rogaghnata of saindhava 46 25 Showing the Matra of Rasapushpa in different

diseases. 69

26 Showing the distinguishing features of Rasapushpa and Rasakarpoora.

70

27 Showing laxanas sthana and prabhava of different krimis.

76

28 Showing the difference between Gram positive and Gram negative bacteria.

80

VII

29 Showing properties of Fluconazole 102 30 Observations made during Hingula shodhana. 106 31 Showing the results of Hingula shodhana 107 32 Temperature recorded during the procedure. 110 33 Physical examination of Ashodhita and

Shodhita Kasisa. 114

34 Showing the observations made during the procedure 120 35 Showing the yield of Rasapushpa in different

practicals. 124

36 Showing the composition of nutrient broth. 134 37 Showing the composition of Potato dextrose agar. 135 38 Showing the composition of nutrient agar. 136 39 Showing the efficacy of standard and trial drugs

against Gram positive and Gram negative organisms 139

40 Showing efficacy of standard and treated drug against fungus.

140

41 Showing the yield of Rasapushpa. 150

LIST OF PHOTOGRAPHS

S.L. No. Photographs 1 Ingredients and Preparation of Rasapushpa 2 Antimicrobial Activity

FLOW CHART AND GRAPH

S.L. No. Flow chart and Graph 1 Graph showing the Temperature and Hour

readings of Rasapushpa. 2 Flow chart of Classification of Kupipakwa

Rasayana.

VIII

Introduction

INTRODUCTION

Ayurveda is an age-old science of Indian system that is based on its own

fundamentals. It represents totality of life and provides complete knowledge to

maintain holistic balance of body and mind. Ayurveda and its medicines are serving

the needs of ailing humanity since many centuries.

Rasashastra is an important branch of Ayurveda, which is pioneered by

Nagarjuna. This shastra is related to metals and minerals. It is believed that

Rasashastra is an expansion of Rasayana therapy of Ayurveda. Rasaoushadhis have

good preventive, curative and rejuvenating potential.

Rasayogas are classified on the basis of samskara given to them as Kharaliya

Parpati, Kupipakwa and Pottali Rasayana. Among these pharmaceutical preparations

Kupipakwa Rasayana are important as they exhibit fast action in small dose.

Historically their manufacturing and therapeutic uses have started since 12th

century AD onwards. Sri Dundukanth the author of Rasandra chintamani was the first

intellectual to introduce this type of preparation in Ayurvedic therapeutics in view of

their high potency, and least toxicity in treating almost all types of ailments.

In Kupipakwa Rasayana agni plays an important responsibility in changing

Physico- chemical properties of drug by duration and pattern of heating.

Based on use of Gandhaka in the preparation of Kupipakwa Rasayana, they

are called as sagandha and nirgandha kalpanas. The two important nirgandha

preparations mentioned in Rasa literature are Rasapushpa and Rasakarpoora.

Nirgandha preparations are most neglected part of Rasashastra. No much

research work was done in this direction. Hence for present research work,

Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity

1

Introduction

Rasapushpa was selected as trial drug on virtue of its importance in therapeutics as

well as skill involved in its pharmaceutical preparations.

Rasapushpa is a sagni, nirgandha moorchana of Parada prepared by adopting

bahirdhuma procedure. For the preparation of Rasapushpa, Hingulotha Parada along

with equal quantities of Kasisa and Saindhava lavana are mentioned.

Rasapushpa is being indicated in Vrana dosha, Bhootavisha, Hikka, Phiranga,

and Visuchika and is told as Parama Virechanakara. Most of the indications specified

are caused by microorganisms.

Microorganisms such as bacteria, viruses, fungi and parasites are present

everywhere in atmosphere and on body surfaces. They are responsible for large

number of infectious diseases in human beings. Infectious diseases continues to take

very high rank as cause of death which accounts for ten million persons each year. If

the infections becomes chronic, it can do serious damage to vital organs such as liver,

lungs, kidney intestines etc. especially in immouno- compromised hosts. The worst

part of infection is its transmission to other people directly or indirectly which leads to

an epidemic.

We get references about microorganisms since vedic period. In vedic period

many terms used for it such as Krimi, Bhuta, Rakshasa, Pisacha, Yatudhana etc.

In Ayurvedic literature also many references regarding krimis are found. They

have recognized a number of diseases as sankramaka or oupasaigika. The terms

bhutopasaraga indicates invasion of organisms. The terms prasangat,

gatrasamsparashtwa etc denotes the mode of spread of infective organism.

To avoid the ill effects of organisms, appropriate treatment is necessary. Many

antibiotics such as pencillins, cephalosporins, etc are heavily prescribed by

contemporary medical science. But these drugs are having some adverse effects on


2

Introduction

the body such as gastric irritation, destruction of gastric flora, nausea, vomiting,

anaphylactic reaction causing even death.

Apart from these, development of newer antibiotic resistant strains of

microorganisms is a major existing problem. Hence there is a need for establishment

of new antimicrobial compound.

Looking at the therapeutic indications of Rasapushpa mentioned in our

classics, in a present study a modest attempt was made to study antimicrobial activity

of Rasapushpa.

Previous work Published in relation to the subject

Standardization of Rasapushpa and its pharmaco-chemical study w.r.s.t. its

antimicrobial activity by Dr. Shailesh Sahadeo Nawkar, Gujarat Ayurvedic

University, Jamnagar.


3

Objectives

The Present study was planned with the following aims and objectives.

Preparation of Rasapushpa.

Physico-Chemical analysis of Rasapushpa.

Evaluation of antimicrobial activity of Rasapushpa.


4

LIST OF ABBREVIATIONS

⇒ A. H. – Ashtanga Hridaya.

⇒ A. P. – Ayurveda Prakasha.

⇒ A. S. – Ashtanga sangraha.

⇒ B.R.R.S – Brihat Rasa Raja Sundara.

⇒ C. S. – Charaka Samhita.

⇒ D. N. – Dhanvantari Nighantu.

⇒ K. N. – Kaideva Nighantu.

⇒ M.P.N. – Madana Pala Nighantu.

⇒ R.A. - Rasamritam.

⇒ R.A.N. - Rasarnava.

⇒ R.C. - Rasendra chudamani.

⇒ R.J.N. - Rasa Jala Nidhi.

⇒ R.N. - Raja Nighantu.

⇒ R.K. - Rasa Kamadhenu.

⇒ R.P.S. - Rasaprakasha Sudhakara.

⇒ R.R.S. - Rasa Ratna Samucchaya.

⇒ R.S.S. - Rasendra Sara Sangraha.

⇒ R.T. - Rasa Tarangini.

⇒ S.S. - Sushruta Samhita.

⇒ R.P. - Rasapushpa.

III

Review of Literature

Hingula

Introduction

Hingula is a compound of Parada and Gandhaka. It is obtained from mines as

a natural mineral and prepared artificially also1. This is the chief source of mercury

since ancient times to till date. Anandakanda named Hingula as Rasagarbha and it is

Rasagandha sambhava according to Rasarnava. Parada extracted from Hingula is said

to be equivalent to ashtasamskarita Parada.

Synonyms2, 3,4

Various paryayas are described by different acharyas.

Table No.1 showing the synonyms of Hingula according to different authors.

Synonyms RAN RJN RSS AP RA RT DN KN

Hingulam + - - + - - - -

Hingul - + - - - + - -

Hingula - - + + + + - -

Ingula - - - - - + - -

Hingulaka - - - - - - - +

Mleccha + + - + + + - +

Rakta - - - + + + - +

Gandhika + - - - - - - +

Suranga - + - + - + - +

Chitranga - + - - - + - +

Churnaparada + + - - - + + -

Rasodbhava - - - - - + + -

Rasasthana - - - - - + + -

Ranjana - - - - - + - -

Kupishirshaka - - - - - + - -

Raktakaya - - - - - + - -

Hamsapada - - - - + + - +


5


Darada + + + + + + - -

Shukatunda - - + - - - - -

Jati - - - - - - - +

Rasagandha

samboota

+ - + - - - - -

Daitya Raktaka - - + - - - - -

Mani Ragaja + - - - - - - +

Rasagarbha - - + + - - + -

Parvata - - - - - - - +

Divya - - - - - - - +

Vernacular Names 5

Scientific Name - Red Sulphide of Mercury

English - Cinnabar

Sanskrit - Hingula, Darada

Hindi - Singraph

Bengali - Hingula

Marathi - Hingula

Gujarathi - Hinguato

Assami - Janjapher

Pharsi - Singraph

Kannada - Ingulika

Telegu - Inguikamu

History

The Possible references regarding Hingula, since ancient period to present day

are as follows:


6


Vedic Period

There are no references about Hingula in Vedic, Prevedic, and Upanishad

period.

Samhita Kala

No references of Hingula found in Charaka Samhita, Sushruta Samhita,

Ashtanga sangraha and in Hridaya,

Kautilya Arthashastra 6

The author of Kautilya Arthashastra, “Chanakya” has mentioned Hingula in

his text for the first time. He mentioned it for testing of swarnadi dhatus. Use of

Hingula as a medicine was not described by him.

Nighantu Period

In Dhanvantari, Raja and Kaideva Nighantu, references of Hingula is found in

bhoumadhatu varga.

Rasa Kala

Rasendra mangala

The oldest text of Rasashastra mentioned first time about shodhana and

therapeutic usage of Hingula. Author has used Hingula for the preparation of loha

bhasma. He described Parada as a satwa of Hingula and has used the word Darada for

Hingula.

Rasahridaya tantra

Acharya Bhagavata Govindapada has mentioned about Hingula.

Rasarnava

Acharya has included Hingula in maharasa varga. In his text, he discussed

about synonyms, varities and used the term “ Rasagandha sambhootam” for


7


Hingula.Use of this term suggests that they were aware of chemical composition of

Hingula.

Rasaratnakara

Author discussed almost all aspects of Hingula and also mentioned artifical

preparation of Hingula.

In other Rasagranthas such as RRS, RPS, RSS, R.Chu etc., and in naveena

rasa granthas like RT, RA, Siddabhaishajaya manimala etc, we find detail description

about Hingula.

Origin

Mythological story described that Hingula is a virya of lord Shiva which was

received by god Agni but due to its high intensity he vomited it, this vomited material

fell in darada desha and become mixed with earthy materials, and called by name

Hingula.

Occurance7, 8

It is obtained from mines as a natural mineral and also prepared artificially

It can be found at most many places all over the world i.e. Spain Italy, Russia,

Yogoslavia, Jechoslovia, Germany, Japan, China, USA, Austria, Nepal etc.

But no deposit of cinnabar can be detected in India. Hence artificial Hingula is

prepared in Surat and Calcutta.

The Hingula what we get from market is artificially prepared.

Preparation of Artificial Hingula

Reference of artificial Hingula preparation is found since Rasaratnakara

period. Thereafter many Rasa texts mentioned artificial preparation of Hingula.

Following are the ratio of Gandhaka and Parada according to different

acharyas.


8


Ratarangini 9

42 parts of Parada and 8 parts of Gandhaka subjected to paka in mrudanga

yantra.

Ayurveda Prakash10

1 part of Parada and 4 parts of Gandhaka, subjected to pachana in loha patra,

after paka, 1/10th part of Manashila added and triturated well. Then it is filled in

kachakupi and subjected to pakakarma (mrudu, madyama, and teevragni) in valuka

yantra.

Inclusion of Hingula

Different authors of various Rasagranthas have included Hingula

under various titles.

Table No.2 showing Hingula included under following category by different authors:

Sl.No. Category Rasagranthas

1 Rasa Rashridaya Tantra 11

2 Maharasa Rasarnava12, Rasakamadhenu

3 Uparasa Anandakanda,

Rasendra Sara Sangraha13,

Brihat Rasa Rajasundara14,

Ayurveda Prakasha15

4 Sadharana rasa Rasajalanidhi16

Resendrachintamani17,

Rasa Ratnasamuchaya18,

Rasaprakash sudhakara 19,

5 Rasadhatu Rasamruta20 Yogartnakara

Hingula Bheda

No description about varities of Hingula is available in Rasahridaya tantra but

we get reference of Hingula bheda in other texts.


9


Table No. 3 showing the bheda of Hingula

Name of Grantha Charmara Shukatunda Hamsapada Anya

Anandakanda + + + -

Rasendra Chudamani 21 - + + -

Ayurveda Prakash 22 + + + -

Rasa Ratna Samuchaya 23 - + + -

Rasaprakash Sudhakara 24

- + + -

Rasatarangini 25 - - - Kritrima

khanija

Rasamrita 26 - - +

Grahya Hingula Laxanas27

The laxanas of ideal varity of Hingula is as follows:

• Japakusuma Varnabha : Resembles the color of hibiscus flower

• Peshane sumanoharaha : When triturated its colour becomes

beautiful

• Mahojwala : Reflects specially hibiscus when exposed

to sunlight.

• Bharapurna : Heavy in weight

• Sheweta rekha : Having silvery streaks

• Pravalabha : Resembles like that of pravala.

According to Rasendra Sara Sangraha Hingula possess Bimbiphala samana rakta

varna.


10


Ashuddha Hingula sevana dosha, Tasya chikitsa28

Ashuddha Hingula sevana may produce many grave symptoms such as

1. Andhata

2. Kshinata

3. Klama

4. Bhrama

5. Moha

6. Prameha

It is treated similar to the ashuddha Parada bhakshanajanya dosha, shuddha

Gandhaka should be administered for 2 months29.

Shodhana

Various shodhana methods are explained in different classics as follows:

• Hingula should be kept in kushmanda khanda, pottali prepared and swedana

done in Lakucha swarasa Portia dola yantra. 30

• Subject Hingula to 7 bhavanas of Ardraka swarasa and lakucha swarasa 31

• Subject Hingula to 7 bhavanas Ardraka swarasa. 32

• Hingula should be subjected to bhavana with mahisha dugda and any amla

rasa dravya for 7 times. 33

• Hingula subjected for 7 bhavanas of Nimbuswarasa 34

Hingula Properties

Rasa

Various opinions are available regarding the rasa of Hingula.


11


Table No.4 showing the Rasa of Hingula according to different texts.

Granthas Madhura Tikta Kashaya Katu

Rasarnava + + - -

Paradasamhita - + + +

Ayurveda Prakash - + + +

Rasendra Purana - + + +

Dhanvantari Nighantu + + - -

Raja Nighantu + + - -

Guna35

Most of the Rasa Acharyas have considered Hingula as ushna gunayaukta

dravya.

Virya and Vipaka

No Rasa text has mentioned about virya and Vipaka of Hingula. Though

Dhanvantari nighantu is the text of dravyaguna vijnana, has mentioned that Hingula is

having ushna virya and katu vipaka.

Doshakarma36, 37,38,39,40,41

Even though almost all the authors agree with the tridoshaghna karma of the

Hingula, some authors have mentioned kaphaghna or kaphapittagna action of

Hingula as well.

Table No. 5. Showing doshagnata of Hingula according to different authors.

Dosha RPS RRS R.Chu B.R.R.S AP RT RA RJN

Tridoshaghna + + + - - - + +

Kaphapittagna - - - + + - - -

Kaphaghna - - - - - + - -


12


Karma42

Sarva doshaghna, Agnivardaka, Rasayana, Balya, Medya, Garavishanashaka,

Netrarogaghna, Ruchikaraka, Loha Ranjana.

Vyadhi Prabhava 43

Pramehaghna, Jwaragna, Hridrogahara, Kushtaghna, Amlapittahara,

Kamalahara, and is also useful in Aruchi, Amavata, Pleehavruddi, Mandagni,

Sandivata, Hrillasa.

Table No.06 Shows the prosperities of Hingula according to different granthas.

Properties RPS RRS RC AP BRRS RT RJV RS RRSarvadoshaghnata + + + KP KP KV + - + Deepana + + + - + - + - + Atirasayana + + + - + - + - + Sarvarogahara + + + - - - + - + Vrishya - + + - - - + - + Jaranartha - + - - + - + - - Mehahara - - - + + + + + - Kushtagna - - - + + + + + - Aruchihara - - - + + - + + - Medya - - - + + + + + - Balya - - - + + + + + - Agnivardhaka - - - + - + + + - Netrarogahara - - - + + + - + - Hrillasa - - - + + - - - - Jwaragna - - - + + - - - - Kamalahara - - - + + + - - - Pleehari - - - + + + - - - Amavatari - - - + + + - - - Garavisha nashaka - - - + - + - - - Dehakantikara - - - - - + - - - Lohamaranartha - - - - - - - - - Dravanartha + - - - - - - - -

Matra

½ to 1 ratti [ 62.5mg to 250mg ]


13


Anupana

Maricha, Guda, Pippali, Guduchi swarasa, Madhu, Ardraka swarasa, Tambula

swarasa.

Satwapatana 44,45,46

Hingula satwa is Parada; hence Hingula is considered as chief source of

parada. Rasa can be extracted through various procedures viz patana, nadayantra etc.

most popular and common method of extraction of Parada is through urdwapatana. It

is described in most of the granthas viz Ayurveda Prakash, Rasatarangini, Rasendra

Sara Sangraha etc.

Marana47, 48

Generally marana is not advised for Hingula. Shodita Hingula can be used for

the preparation of yogas, however elaborate procedure for marana has been described

in Ayurveda Prakash, Brihat RasaRajaSundara etc.

Vishishta Yoga

1) Ananda Bhairava Rasa 49

2) Hinguleshwara Rasa 50

3) Kaphaketu Rasa 51

4) Hingulad Rasasindoora 52


14


Cinnabar53, 54

Cinnabar is important ore of mercury, when used as a pigment it is called

Vermillion. Nearly all mercury in the world is obtained from cinnabar. It occurs both

in crystalline and massive forms. The ore is a red crystalline mass that is easily

distinguished from all the red minerals by its peculiar shade of colour and its great

weight.

General Information

Chemical formula - Hgs

Molecular Weight - 232.66 gm

Composition - Mercury (Hg)-86.221 and sulphur (S) –13.781

Locality - Alma den, Spain

Synonyms - Cinnabre, zinnober

Verities

Verities are made according to colour and percentage of Hgs present in it.

1) Cinnabar Native

This is one of the most important ore of mercury. It contains 95% mercury

Sulphide and other impurities like carbon, silica, etc. It is bright and dark red in

colour.

2) Hepatic Cinnabar

When percentage of Carbon impurities is higher in cinnabar, its colour

becomes darker like liver colour; such ore is called hepatic cinnabar.

3) Meta Cinnabar

In this type muddy dust is present in more percent which makes its colour still

darker almost to black shade.


15


4) Coral ore

This ore specially occurring in Germany and Italy. When mercury sulphide in

coral ore its separated, it is rosy in colour. It contains about 5% mercury.

Physico Chemical Property

Cinnabar is granular, massive or earthy, it some times occurs beautifully

crystallized in small complex and highly modified hexagonal Crystals. Usually the

crystals are rhombohedral or prismatic. It is cohineal red in colour often inclining

brown. Its streak is bright red. Adamantine to dull luster and perfect prismatic

clevage.

Transparency - Opaque or translucent

Hardness - 2 – 2.5

Specific gravity - 8.09

Lustre - Adamantine

Occurance - Generally it occurs due to the volcanic activity, also

available near hot springs. Important places of occurrence

are Spain, Western States of USA, Mexico.

Solubility:

It is insoluble in water and acids but dissolves in aquaragia and forms mercuric

chloride.

In the presence of a strong oxidizing agents like potassium chloride, it forms mercuric

chloride

Roasting

When Unconcentrated ore is roasted in air, Cinnabar is oxidized to mercuric oxide


16


and sulphur dioxide is released and mercuric oxide so forms decomposes to give

mercury and oxygen.

2Hgs+302 2So2+2Hgo

2Hgo 2Hg+O2

The mercury thus obtained will be pure in nature.

Mercury sulphide reacts with concentrated potassium Sulphide solution to give a

complex thiosalt.

Hgs + K2S K2Hgs2

On sublimation mercuric sulphide becomes red.

Extraction of Parada from Hingula

In ancient days the only source of mercury was Hingula. Since olden days it is

accepted that Hingulotha Parada is pure and devoid of sapta kanchuka doshas and

believed to posses the property similar to Gandhajeerna Suta55. In Rasa Ratanakara

also it is advised to use Hingulotha Parada for all purposes.

History

Its references are not found in Prevedic, Vedic and Samhita period.

Its references are found since Rasarnava period. After, it has been explained in all

Rasa granthas viz Rasaprakash Sudhakara, Rasendra Sara Sangraha, Anandakanda,

Ayurveda Prakash, Rasatarangini etc. Vidyadhara yantra, Urdwapatana yantra etc. are

used for the extraction of Parada.

Hingulad Rasa karshana Vidhi

Prior to extraction of parada, Hingula should be subjected to shodhana.

Many procedures are described in Rasa texts for extraction of Parada from Hingula.


17


They are following:

1) Urdwapatana yantra process 56

2) Adhapatana Yantra process 57

3) Tiryakapatana yantra process 58

4) Nada yantra process 59

Urdwapatana Method

This method is more commonly followed for extraction of mercury. In this

process Hingula is triturated with Nimbu swarasa and made into chakrikas & kept in

sunlight for drying. They after being dried are kept in earthen pot covering with

another pot of same size and sealed at the joint with cloth and mrittika in such a way

so as to make it air tight. Then heat the lower pot and cool the upper pot with wet

cloth so that mercury fumes can condense in the upper pot. Then allow the

urdwapatana Yantra (Damaruyantra) to cool by itself and open the pot and collect the

mercury. If some amount of unburnt Hingula is remaining repeat the process and

collect the mercury.

Superiority of Hingulotha parada60

Parada extracted from Hingula is considered to be best because it is free from

various types of doshas. Hence, the same does not need any further samskar and could

be used even without subjecting to asthasamskaras. It is claimed that, Parada so

extracted is capable of performing all the actions attributed to it. According to

Rasaprakash, Sudhakara” Hingulakrshta Parada possesses all the properties, which are

seen in gandha jarita parada. Thus it is considered superior.


18


Parada

In Rasashastra, Parada has been given an elevated place. It is claimed that, it

not only cures the diseases and senility but also responsible for moksha. Its mere

touch, vision of Parada can make the man free from all sins and bless him with

punya61. Mythological it is believed the Parada is originated by Lord Shiva. Parada

has got amalgamating quality because of which it carries the properties of

amalgamated materials. Hence it is successfully used as potent medicine in Ayurvedic

therapeutics. It is a shining, silvery white metal, liquid at ordinary temperature. It is

three times heavier than water. Pure mercury does not possess taste and smell62.

Synonyms of parada63, 64

Many synonyms of Parada are explained by different Rasacharayas depending

on factors such as its swaroopa, gati, dehavada, dhatuvada etc.

Table No.7. Showing the synonyms of parada

Sl No.

Property Synonyms

1 Swaroopatmaka Galadroupyanibham, Mahavahni, Mahateja, Suvarna.

2 Dharmika / Devatamaka Trinetra, Trilochana, Deva, Dehaja, Prabhu Rudraja, Lokesh, Vijendraja, Budha, Rajaswala, Shiva, Shivaveerya, Skanda, Harateja, Harabija, Shivabija, Pavana, Lokanath.

3 Gatyatmaka Khechara, Chapala, Chala, Dhurtaka. 4 Dehavadatmaka Amrita, Jaiva, Dehada, Paramamrita Parata, Parada,

Mrityu nashana, Rasayana, Rasayana shreshta, Jaitra.

5 Dhatuvadatmaka Divyarasa, Maharasa, Rasa, Rasendra, Rasesh, Rasottama, Rasadhatu, Rasaraja, Rasanath, Siddadhatu, Soota, Sootaka, Sootaratha, Mishraka.

6 Vishishta guna Ananta, Amara, Yashade, Soubhagya, Sukshma, Kalikantaka.

7 Darshanika Adyatmika Jeeva, Jaiva, Divya, Achintya.


19


Vernacular Name 65,66

English - Mercury

Latin Name - Hydrargyrum

Sanskrit - Parada

Hindi - Para

Bengali - Para

Gujarathi - Para

Assami Jivaka, inul Hayata

Parsi - Simaba, Jivah

Telegu - Padarasamu

Kannada - Padarasa

Historical Review67,68,69,70

In Indian Alchemy Parada is considered as one of the important drug. It has

got ability to amalgamate with most of the metals. Indian history says that Parada is

being used as a medicine since 6000 years.

Vedic period

The references of Parada are not available in prevedic, vedic and Upanishad period.

Samhita Kala

References of Parada are found in Charaka Samhita. Sushruta Samhita and

Ashtanga Hridaya. In Samhita Parada is used externally for lepanartha and its internal

administration is not found.

Koutilya Arthashastra

We find reference of Parada in Koutilya Arthashastra. In Swarna bheda

prakarana author has explained Rasavidda as one of the types of Swarna.


20


RasaKala

In all Rasagranthas we find the references of Parada. Even the old granthas

such as Rasendra Mangala, Rasarnava etc. include Parada as their major content.

Most of Rasa granthas have described every aspect of Parada i.e., Parada parayaya,

Parada utpatti, Parada dosha, Shodhana, different samskaras, marana etc.

Occurance71

In Rasaratna Samucchaya, it is mentioned that in ancient times mercury was

found mainly in Darada desha and also in Himalayas in small amounts. But now a

days it is obtained mainly from the mines of Spain, America, Italy, Australia, British,

China, Russia, Japan and Africa as Cinnabar or Metacinnabar.

It occurs in 2 forms.

1. Native 2. Ore form

Ores of Mercury72

Generally mercury is found in the form of ores, the most important are

cinnabar and Metacinnabar, which are in sulphide forms.

Table No. 8 Showing the ores of parada along with their chemical composition.

Sl No. Ores Chemical Composition

1 Cinnabar Hgs 2 Metacinnabar Hgs 3 Calomel Hg2Cl2 4 Living stonite 2sb2C3Hgs 5 Montrodyte Hgo 6 Falh ore 7 Bassenite 8 Gwadal kajrite 9 Living ore of mercury 10 Caronine ore of mercury 11 Brick ore of mercury

Bhedas

Bhedas of Parada described in various texts are based on following factors:


21


1. Depending on the color.

2. Depending on the place of origin.

01. Depending on the Color 73

As soon as Parada is taken out of kupa, it has been classified into 4 varities on

the basis of its color and its luster.

Table No.9 Showing the varities of Parada depending on the color and their

karma also specified.

Sl.No. Types Color Varna Karma 1 Shweta White Brahmana Shwetakarma 2 Rakta Red Kshatriya Therapeutics 3 Peeta Yellow Vaishya Used in alchemy or to prepare gold. 4 Krishna Black Shudra Used in Maintaining health

Depending on the Place of origin 74,75,76,77

Table No. 10 showing the varieties of Parada depending upon the place of origin.

Sl.No. Varieties Color Impurities Uses 1 Rasa Rakta Which is free from all

types of impurities Rasayana

2 Rasendra Peeta Free from impurities Rasahara 3 Suta Ishatpeeta Impurities present Rogahara 4 Mishraka Mayura

Chandrika Varna

Impurities present Saravasiddi dayaka

Parada dosha 78,79

Parada is highly reactive and it readily mixes with other dhatus. Hence

probability of impurities present in it is more. These are explained under the heading

of Parada doshas in our classics. If are not eradicated by shodhana process can lead to

many diseases and even death. Some authors described the doshas or blemishes of

Parada collectively. In other granthas viz Rasa Ratna Samuchaya, doshas have been

considered in 3 groups:


22


1. Naisargika Dosha

2. Yougika Dosha

3. Oupadhika Dosha (Sapta Kanchuka Dosha)

Table No.11 Naisargika Dosha and its effect on sharira.

NAISARGIKA DOSHA Sl.No. Dosha Prabhava

1 Visha Mrityu 2 Agni Santapa 3 Mala Moorcha

Table No. 12 Yougika Dosha and its effect on sharira

YOUGIKA DOSHA Sl.No. Dosha Prabhava 1 Naga Jadya, Adhmana,

Kushta 2 Vanga Jadya, Adhmana,

Kushta

Table No. 13 Oupadhika (Sapta kanchuka) Dosha and its effect

Sl.No. Dosha Prabhava 1 Bhumija Kushta 2 Girija Jadya 3 Varija Vata sanghata 4 Two Naga Dosha Vividha dosha

vikara 5 Two Naga Dosha Vividha dosha

vikara

Though Oupadhika doshas like Bhumija etc are also called sapta kanchuka, it

is unclear why again different sapta kanchuka doshas have been described. This may

lead to assumption that they are two different sets of doshas.

The names of sapta kanchuka doshas are as follows

Bhumija : Parpati.

Girija : Patini.


23


Varija : Bhedi.

Two Naga dosha : Dravi, Malakari.

Two Vanga dosha : Andhakari, Dhwankshi.

Sapta kanchuka doshas are claimed to be responsible for various disorders,

according to the Subhodini tika on Rasaratna Samucchaya.

Parpati : Dries up the skin making it scaly.

Patini : Cracking of the skin.

Bhedi : Loose stools.

Dravi : Liquefying the dhatus.

Malakari : Aggravation of malas.

Andhakari : Blindness.

Dhwankshi : Hoarseness in the voice.

Parada Grahya Swaroopa

Bahirujwala : Externally Parada is shining

Madhyahna surya pratimam : It looks like mid-day sun.

Antah neelavarna : Internally it has bluish tinge.

Effect of Ashuddha parada sevana 80

It is said in the texts that impure mercury if used internally may produce

various diseases in the body viz Vidaha, Krimi, Kushta, Agnimandya, Aruchi, Chardi,

Jadya and even Mrityu.

Thus it may cause serious ill effects and hence purified Parada should be used

for medication. It is also said that shuddha Parada is like an amrita and ashuddha is

like poison.


24


Shodhana81

For elimination of Naisargika, Yougika and Oupadhika doshas, Parada shodhana is

very important. Shodhana of parada has been broadly divided into 2 divisions.

1. Samanya and Vishesha Shodhana.

2. Group of Eighteen special procedures i.e. Ashtadasha Samskar.

When Parada is to be used to prepare medicine to combat diseases, the first set of

shodhana is useful. If it is to be used for Rasayana karma then the Ashtadasha

samskar has to be followed.

Any Rasa karma has to be conducted in shubha nakshatra and shubha dina,

worshiping Lord Shankara and Bhairava, because the studies have shown that those

particular days, time and worshipping will impart the efficacy in medicine. Hence it

should be necessarily followed.

Samanya Shodhana82, 83

Different procedures are explained by various granthas for Samanya shodhana

of Parada.

TableNo.14 Showing Samanya shodhana of Parada according to different

authors:

S.L.No. Shodhana dravyas used Procedure used

1 Sudharaja, Mardana done for 3 days,

Vastraghalana

2 Lashuna, Saindhava Mardana for 7 days in taptakhalva,

Prakshalana.

3 Tambula swarasa, Ardraka

swarasa, Ksharatraya

Mardana for 3 days, Prakshalana.

With amla dravya.

4 Kumari swarasa,

Chitraka, Raktasarshapa,

Brihati, Triphala kwatha.

Mardana for 3 days prakshalana

with kanji.


25


Vishesha Shodhana84

Different acharyas have explained specific measures to eradicate the specific

doshas present in the Parada.

Table No.15 Showing Vishesha shodhana of Parada according to different

authors

Sl.No. Dosha Shodhana dravyas

used

Procedure used

1 Naga dosha Gruhadhuma, Ishtika

choorna, Haridra

choorna, Urna bhasma

Mardana for 1 day,

prakshalana with

kanji

2 Vanga dosha Indrayava Ankola

churna, Haridra

churna,

Mardana,

prakshalana with

kanji.

3 Agni dosha Chitrakamula churna

or Triphala churna,

parada

Mardana

4 Mala dosha Aragwada churna or

Kumari swarasa,

Mardana,

prakshalana

5 Chapala dosha Krishnaduttura

panchanga, parada

Mardana,

prakshalana

6 Visha dosha Triphala churna or

Chitrakamula churna

Mardana,

prakshalana

7 Giri dosha Trikatu churna Mardana

vastra ghalana

8 Asahya agni

dosha

Gokshura churna Mardana

Ashtadasha samskara85

The eighteen special procedure of shodhana of Parada can further be classified

into a sub group called as Astha samskara. It will have both properties i.e. Vyadhi


26


nashana and rasayana. Remaining 10 procedures are specially utilized in dhatuvada.

They are as mentioned below.

Rasayanartha Dhatuvadartha

1) Swedana 5) Patana 9) Gaganabhakshana 14) Sarana

2) Mardana 6) Rodhana 10) Charana 15) Ranjana

3) Moorchana 7) Niyamana 11) Garbhadruti 16) Kramana

4) Uttapana 8) Deepana 12) Bahyadruti 17) Vyadha

13) Jarana 18) Bhakshana

Pharmacological and Therapeutic properties of Shodita Parada86

Rasa : Shadrasa

Guna : Snigdha, sara

Virya : Ushna

Vipaka : Madhura

Karma : Yogavahi, Rasayana, Ativrishya, Balya, Vajikara,

Drushitibala prada, Vayasthapaka, Bhukti, Muktiprada,

Pushtikara, Deepana, Ayushkara, Agni Sandhukshana,

Dehasiddikara, Lohasiddikara, Ropana, Krimighna.

Dosha Prabhava : Tridoshaghna

Vyadhi Prabhava : Krimi, Kushta, Akshiroga, Kshaya, Tridosha roga, Papaja

roga Sarvarogahara.

Parada Marana87

Different acharyas have described several procedures of Parada marana.

Method: Kajjali is prepared with 2 palas of Parada and 1 pala of Gandhaka, then

bhavana given with swarasa or kashaya of Parada maraka gana oushadhies and dried


27


later it is kept in musha and sandhi bandhana made. Parada bhasma prepared by

bhoodara yantra method.

Matra88,89

Parada can be administered for Vyadhi nashanartha as well as for Rasayanartha.

Mruta parada : 2 ratti

Swarna jarita parada : ½ ratti

Vaikranta jarita parada : ½ ratti

Vajra jarita parada : 1/4 ratti

Parada yoga : 1 ratti

Anupana

It has to be suggested according to Vyadhi.

Patyapatya

Purana godhuma, Shalianna, Godugda, Gritha, Dadhi, Hamsodaka, Mudga, Yusha are

included under patyavarga

Kakarshtaka gana, Masha, Kulutha, Anupa mamsa, Guru, Vishtambhi, Amla ahara

sevana, etc are included under apatya varga.

Parada Yogas

Kajjali90

Makaradhwaja91

Rasaparpati92

Hemagarbha Pottali Rasa93


28


Mercury94, 95,96,

1. English Name :Quick silver

2. Latin Name : Hydrargyrum

3. Chemical Formula : Hg.

2) Mercury is the only metal which is liquid at room temperature. It is shining, silvery

white heavy liquid easily divisible into globules. It is extremely mobile, readily

volatises on heating. It is 13.5 times more dense than water and 1.2 times heavier than

lead.

It exists in three forms metallic, mercurous and mercuric. Metallic mercury is

(Hg2+) also known by the name quicksilver, i.e. a liquid metal having a bright silver

luster. It exists in nature as metal itself and also found in sulphide form (Cinnabar).

Metallic mercury is not poisonous if taken orally because it is not absorbed. It

vaporizes even at room temperature to an extent sufficient to permit inhalation to

toxic amounts. Mercury depresses cellular enzymatic mechanisms by combating with

sulphahydral groups.

Physical Properties

Colour : Silvery white.

Atomic No : 80

Atomic Weight : 200.61

Specific Gravity : 13.59

Freezing Point : -390C

Boiling point : 357.250C


29


Simple tests of pure mercury

1. Boiling point of mercury is 357.250C. When metallic impurities are present, its

boiling point changes to lower temperature.

2. Pure mercury does not stick to a clean glass, on the contrary impure mercury

leaves behind its track on the clean glass.

3. Impure mercury when shaken for some time in an open air, it forms a thin film

of blackish powder over its surface. This is due to oxidation of the metallic

impurities. If mercury is pure, this does not occur.

Chemical Properties

Effect of Air:

At ordinary room temperature, with low or high humidity, mercury is not at all

affected chemically. If it is heated in an open air gradually up to its boiling point, it

reacts with oxygen present in the atmosphere to form oxide of mercury.

Effect of water on mercury

Water at any temperature has not any chemical effect on mercury.

Effect of Acids:

Hydrochloric acid, dilute or concentrated does not cause any change in

mercury chemically. Concentrated sulphuric acid also does not bring about any

change in mercury, however it produces sulphur dioxide gas when used in hot and

concentrated form.

Hg+2H2SO4 Hgso4+2H2o+SO2

Concentrated nitric acid reacts with mercury, to produce mercury nitrate and nitrogen

oxide

Hg+6HNo3 Hg (NO3)2 + No + 3H2o + No2

Effects of Alkalies:


30


Alkalies may it be concentrated or dilute, hot or cold do not have any effect on

mercury

5) Halogen compounds:

Halogen and halogen compounds i.e Iodine, Bromide, Flourine, Chlorine and

their compounds do have effects on mercury to from Iodides, bromides fluorides and

chlorides.

Hg + Cl2 HgCl2

Amalgam Formation

Mercury forms alloys with many metals and those are called amalgams.

Metallic properties of such amalgams are very useful in different industries and

medicines.

Absorption, Distribution and Excretion:

The absorption, distribution and excretion of mercury and its compounds vary

considerably with the chemical form of the metal.

The soluble inorganic mercurial (Hg2+) readily gain access to the circulation

when taken by mouth, although a considerable portion of ingested Hg2+ may remain

fixed to the alimentary mucosa and the intestinal contents. Insoluble inorganic

mercurous compounds, such as calomel (Hg2Cl2), may undergo some oxidation to

soluble, absorbable compounds. In suitable vehicles, inorganic mercurials may be

reduced to elemental mercury and deposited as a grey to bulish pigment. In the blood,

Hg+2 is first fixed to α globulins and erythrocytes but later shifts to albumin, from

which it is redistributed tissues with a half-time of about 15 days. Within few hours

the mercury is found in tissues in the following approximate order of decreasing

concentration. Pancreas, kidney, liver, spleen, blood, bone marrow, upper respiratory

and buccal mucosa, intestinal wall (especially colon), skin, salivary glands, heart,


31


skeletal muscle, brain and lung. There is some evidence that mercuric salts can be

stored in bones. Some tissues have a lower capacity bind to mercury than to others so

that distribution may be dose dependant. Also the distribution changes over the course

of time. In the kidney, mercury is found primarily in the proximal tubules.

Excretion of mercury starts immediately after absorption, mainly by way of

kidney and colon, and to a lesser extent via the bile and saliva. Small amounts are also

excreted in volatile elemental form, through both lungs and the skin. Most of the

mercury is excreted within 6 days after administration but traces may be detected

from months to even years.

The absorption, distribution and excretion of the mercury of organic

mercurials is determined by Physico-chemical factors and the extent of in-vivo

conversion to inorganic mercury. Methyl mercury compounds, the most important of

the environmental mercury contaminants, are lipid soluble and are rapidly and almost

completely absorbed from the gastrointestinal tract.

Therapeutic uses

The inorganic mercuric salts, as well as certain organic compounds of

mercury, are employed chiefly as antiseptics and preservatives. Also certain mercury

compounds are effective parasiticides and fungicides when locally applied. Certain

complex organic mercurial compounds are employed as diuretic. Mercurous chloride

is an absolute cathartic. The metal is of historical interest in syphilo therapy. The

specific actions of mercury are:

1. Antiseptic

2. Anti syphilitic

3. Diuretic

4. Cathartic


32


Mercury exhibits its therapeutic manifestation in a following manner:

• Mercury ions are strong protein precipitants and act as antiseptic.

• They stimulate secretory activities of many glands such as salivary, intestinal

etc.

• They interfere with reabsorption of fluid by the intestine (cathartic action)

• The mercurial diuretics act primarily to inhibit water reabsorption by the

tubules of the kidney and therefore interfering with reabsorbing function of the

tubules.

Other uses of mercury

• It finds use in thermometers, barometers, manometers, and high vaccume air

pumps.

• It is used in extraction of gold and silver by amalgamation process.

• Mercury is used in preparation of its alloys with other metals called amalgams.

• Amalgams of tin, silver and gold are used in the dentistry.

Mercury poisoning

Mercury is highly toxic, its ingestion may lead to a manifestation of many symptoms

and even death.

Acute poisoning

Acute poisoning usually results from the oral ingestion of highly dissociated

inorganic preparations, although it may also result from inhalation of vapours of

elemental mercurial ointments applied tropically. Symptoms produced may vary from

one individual to other.

Metallic taste, swollen and grayish appearance of the mouth, pharynx, and

gastric mucosa, intense pain in the affected tissues, vomiting, nausea, severe, profuse,


33


bloody diarrhea, oliguria, circulatory collapse, uremia, gangrenous colitis may be

observed.

Chronic poisoning

Chronic mercury intoxication can result from wide variety of industrial,

agricultural, and domestic exposures. It also occurs among those who have taken

internally for a prolonged period of excessive doses of mercury compounds.

The symptoms produced are gingivitis, stomatitis, loosening of the teeth,

salivation, metallic taste, colitis, retrogressive renal damage, loss of appetite,

nutritional disturbances, anemia, hypertension and peripheral neuritis. The CNS is

especially involved, as evidenced by behavioral changes, mental depression,

irritability, blushing, insomnia, intention tremors, occasionally hallucinations.

Fatal dose : 1 – 4 gms.

Fatal period : 3 – 5 days.

Treatment

• A source of sulphahydral-rich protein such as milk or raw eggs, is introduced

into the stomach.

• Copious lavage is performed with 5% solution of sodium formaldehyde

sulphoxylate.

• This provides an excellent local antidote. It reduces bivalent mercuric ion to

the much less soluble mercurous form.

• Intramuscular dimercaprol or a penicillamine is given to chelate the mercury

and accelerate its excretion. Prompt therapy with metal-chelating agents

affords the kidney almost complete protection from the toxic effects of

mercury.


34


• Fluid, electrolyte and cardiac abnormalities and shock must be corrected

• Hemodialysis may be required to relieve uremia.

Line of treatment remains same for chronic poisoning also. But response is

slow to therapy and patient may remain ill health for years.


35


Kasisa

Kasisa is considered under Uparasa varga. It is a compound of iron and

sulphur, it being a sulphate of Iron. It is made artificially and found in native form

also97. Artificially it is produced by the reaction of sulphuric acid on iron98. Now days

it is mostly made artificially.

Synonyms99, 100,101,102

Different paryayas of Kasisa are listed by various acharyas depending on its

origin action, properties and specific characteristics.

Table No.16 Showing synonyms of Kasisa according to different Acharyas.

Paryayas R.S.S3 A.P4 R.A5 R.T6

Kasisa + + + +

Kashisha - + - +

Dhatukasisa + + - -

Pushpa kasisa - - - +

Panshukasisa - - - +

Pamshukam - + - +

Dhatu Ranjaka + - - -

Khechara + - + -

Khaya + - + +

Vernaculars Names103

English : Green vitriol

Hindi : Kasis

Marathi : Hira kas

Bengali : Hira kas


36


Gujarati : Hiro kas

Arabic : Jajarujhar

Parsi : Jajs obja

Telegu : Annabedhi

Classification of Kasisa104, 105,106

Kasisa is explained under different titles by our acharyas.

Table No. 17 Showing classification according to different granthas.

Sl.No. Class Granthas

1 Uparasa Rasachintamani, Rasarnava, Rasendra Purana,

Rasahridaya Tantra, Rasaratna Samucchaya,

Brihat RasaRajaSundara,

RasendraSaraSangraha, Rasaprakash sudhakara.

2 Upadhatu Sharangadhara Samhita, Rasatarangini

3 Dhatu Varga Rasamrita

In Charaka Samhita, Kasisa is described under Bhouma gana, and in ushakadi

gana in Sushruta Samhita, while in Ashtanga Hridaya it is described as Parthiva

dravya.

Occurance107

It is found in native form as well as made artificially.

It is available naturally resulting from decomposition of Iron pyrites by the

action of atmospheric moisture and found in small quantities where iron pyrite occurs.

Malenterite, which is available naturally (not abundantly) found in USA, Bavaria,

Sweden, Spain and Bihar, Punjab in India.

Artificial preparation

Kasisa is prepared in laboratories by adding sulphuric acid to iron fillings.


37


Varities of Kasisa

Different varieties are explained in different granthas based on its origin and

color.

Table No. 18 Showing varities of Kasisa according to origin.

Reference Churna

kasisa

Pushpakasisa Pamshu

kasisa

Valu kasisa

R.T 108 + + -

A.P 109 - + + -

RRS 110 - + - +

R.Ch 111 - + - +

R.PS 112 - + - +

Table No. 19 Showing varities of Kasisa according to Color.

Reference Shweta

kasisa

Peeta

Kasisa

Krishna

kasisa

Harita

kasisa

Rakta

kasisa

RAN 113 + + + -

AK + + + - +

RP 114 + + + - -

BRRS115 + + + - -

RJN116 + + + + -

Grahya Kasisa117

Of all different verities of Pushpakasisa, which is told as peeta varnaja, is

grahya and it is used for different therapeutic purpose.

In Rasamrita, Yadavaji has mentioned that Pushpakasisa is one which is

artificially prepared and is the ideal for medicinal purpose.

Shodhana of Kasisa118, 119,120,121

Different shodhana methods are explained in various texts.


38


Table No.20 Showing various procedures for purification of Kasisa.

Sl.No Dravya used in

Shodhana

Procedure used Yantra

used

Reference

1 Nimbu swarasa Bhavana for a

day

Khalva BRRS, RJ.N,

RP

2 Jambiri swarasa Bhavana for a

day

Khalva RSS

BRRS, RP

3 Bhringaraj swarasa Bhavana for 3

times

Khalva RP, RJN, RA

4 Bhringaraj swarasa Swedana for 3

hrs

Dolayantra R.T, RPS,

BRRS, R.P, A.P

5 Bhringaraj swarasa Klinna RRS, R.C, RJN

6 Stri shonita Bhavana Khalva BRRS, R.P,

RRS, RJN,

7 Panchapitta Bhavana Khalva RP, BRRS,

RRS

8 Kasamarda rasa Bhavana Khalva RAN

Pharmacological and Therapeutic properties of Kasisa

Slight difference of opinion exists among different acharyas regarding Rasa of

Kasisa.

Table No.21 Showing Rasa of Kasisa according to different Acharyas.

Rasa RPS RRS RKD BRRS AP RA RJN RT

Amla + + + + + - + -

Kashaya + + + + + + + +

Tikta - - - - + - - -

Guna122 : All Rasa granthas mentioned it as Ushnagunayukta.

Rasendra Sara Sangraha has mentioned Snigdha guna

also


39


Virya123 : Ushna.

Vipaka : Katu.

Dosha prabhava124 : Vatahara, kaphahara.

Karma125 : Netrya, Keshya, Kesharanjaka, Pataranjaka, Balya,

Kandughna, Vishahara, Rajapravartaka, Jwaragna.

Vyadhi Prabhabva126 : Rasavad gunakaraka, Mootra kriccha, Ashmari,

Shwitra, Pandu, Kamala, Kshaya, Vrana , Pleehari

Pittaja vikara, Apasmara and Netra rogahara.

Kasisa Marana127, 128

Many methods of Kasisa marana are explained in different Rasa texts.

I Method

Kasisa Sukshma churna is prepared, 7 bhavanas with Kanji is given and

subjected to laghuputa. Rakta varna bhasma is obtained, which is again triturated with

Nimbu swarasa and laghuputa given. This process to be repeated till Kasisa loses its

amlata. By this sarvadosha rahita, vimala bhasma of Kasisa is obtained.

II Method

Kasisa should be given bhavana with Nimbu swarasa. Chakrika prepared and

dried in sunlight, they are sealed inside sharava samputa and heat is applied using

10kg of vanopalas. The procedure is repeated till Kasisa bhasma becomes free from

sour taste and acquires red color like Gairika.

Kasisa is a ferrous sulphate, which can be used after its shodhana only. It does

not need marana process. However some texts have recommended its marana. Its red

colored bhasma can be prepared within three to four putas. By subjecting it to

bhasmikarana process the ferrous sulphate changes to ferric or ferrous oxide, which


40


imparts red colour to it. From the therapeutic point of view there is not much

difference between shodhita and marita Kasisa.

Kasisa Bhasma Guna129

Kasisa bhasma has properties similar to Loha bhasma but it is specially

indicated for Kushta and Shwitra.

Matra of Kasisa130

Shodhita Kasisa 23 : ½ - 2 ratti

Kasisa Bhasma 24 : Loha bhasma samana matra is indicated.

Anupana

Triphala churna

Madhu

Satwapatana131

The satva of Kasisa can be obtained by adopting the similar process of Tuvari

Satwapatana, i.e. it should be triturated with Kshara and Amla dravyas and heated

intensively to obtain satwa.


41


Ferrous Sulphate132, 133

English Name : Green vitriol

Chemical Name : Ferrous Sulphate

Chemical Formula : Feso4 7H20

Atomic Weight : 278.0

It is the hydrated salt, which contains 20% iron. It consists of pale, bluish-greenish

crystals or granules. In moist air crystals of ferrous sulphate rapidly oxidize and

become coated with a brownish yellow basic ferric sulphate and must then not be used

for medicinal purposes. The drug is odorless and has a saline, astringent taste.

Ferrous sulphate may be obtained by dissolving scrape iron in dilute sulphuric

acid.

Solubility: It is completely soluble in 1 in 1:5 of water 1 in 0.5 of boiling water

and practically insoluble in alcohol. The British pharmacopoeia specifies that a 5%

solution in water has a PH of 3.0 to 4.0.

It is usually dispensed as pills or tablets coated to protect them from moisture. The

salt is mixed with glucose or lactose to protect it against oxidation. Ferrous sulphate

contains about 60mg of Iron in 300mg. Ferrous sulphate syrup contains 40mg of salt

(8mg of Iron) in each millimeter. The average adult dose is 2 tsp thrice daily for

children who weigh from 15-35 kg, 1tsp twice daily for children who weigh from 15-

35 kg, 1tsp twice or thrice daily is advocated.

Properties and uses: Light green crystals of ferrous sulphate lose water and turn

brown on exposure to air due to oxidation.

4Feso4 + 2H2o + o2 4Fe (OH) SO4 Basic Ferric sulphate.

On heating it decomposes as follows


42


2FeSO4 Fe2 O3 + SO2 + SO3

With Nitric oxide, ferrous sulphate turns black due to the formation of nitro

ferrous sulphate, FeSO4 NO

Reducing property: It is a good reducing agent, for e.g. it declorises acidified

potassium permanganate and turns into acidified potassium dichromate green.

With ammonium sulphate, it gives ferrous ammonium sulphate (Mohar’s salt), Fe

(NH4)2 SO4 6 H2O. This is not oxidized so readily as ferrous sulphate and is therefore

used in volumetric analysis in preference to ferrous sulphate.

Uses: It is extensively used in Iron deficiency anaemia. Ferrous sulphate is largely

used in manufacture of blue-black ink and as mordant in dyeing and tanning

industries.

Toxicity: Ferrous sulphate rarely proved fatal to adults but it has produced fatal

poisoning in children under the age of 4 years who took proprietary sugar coated

ferrous tablets mistaking them for sweets. Each tablet contains 180mg of FeSO4 2mg

of CuSo4 and 2mg of manganese sulphate. It is believed that iron released from tablets

has a necrotising effect on stomach and intestine and this disturbs the normal mucosal

block and releases ferritin into the stream and allows toxic quantity of iron to be

absorbed.


43


Saindhava Lavana

Saindhava lavana is mineral, which is obtained from Punjab mines134. It is

literally means produced in sindh or the country along the Indus. The term is applied

to rock salt, which is regarded as best of salts135.

Synonyms, 138,139,140,141.

Table No. 22 Showing synonyms of Saindhava lavana

Synonyms RT RM RN KN DN MPN

Saindhava + + + + + +

Sindhu lavana + - - + + -

Sindhootha + - - + + -

Sindhudeshaja + - - - - -

Sindhu Bhava + - - - - -

Sindhumanjhaja + - - - - -

Sheeta shiva + + + + - -

Nadeya + + - + + -

Shilatmaka + - - + - -

Shiva + - + + + -

Vashira + - - - - -

Manimantha - + + + - +

Sindhuja - + + - + +

Shivatmaja - - + - - -

Pathya - - + - - -

Vimalavara - - - + - -

Lavanavara - - - + - -

Doutheya - - - + - -

Pututhama - - - + - +


44


Vernacular Names142

Sanskrit : Saindhava

English : Rock salt

Chemical Name : Sodium chloride

Hindi : Sendha namak

Bengali : Saindhavanun

Marathi : Sende luna

Gujarathi : Sindha luna

Kannada : Uppu

Classification: Lavana skandha

Varities 143

Acharya Yadavaji Trikramaji has mentioned two varities of Saindhava lavana

1. White

2. Red

Source144

Found in nature in extensive beds mostly associated with clay and calcium

sulphate. To obtain it, holes are dug into these rocks which soon become filled up

with salt water; the water is then evaporated and the salt is left ready for use.

Characters145

It is found in small white crystalline grains or transparent cubes. It is brownish

white externally and white internally. It has a pure saline taste and burns with a

yellow flame.

Pharmacological Properties146, 147,148,149,150

Rasa : Lavana, Madhura.

Guna : Snigdha, Laghu.


45


Virya : Sheeta.

Dosha Prabhava : Tridoshaghna.

Karma

Table No. 23 Showing the karma of Saindhava lavana

Karma RT RM MPN DN KN RN

Hridya + + + + - -

Vrishya + + + + + +

Netrya + + + + + +

Pachaka + - + - + -

Deepaka + + + + + +

Avidahi - + - + - -

Sukhada - + - - - -

Rogaghnata151, 152,153,154

Table No. 24 Showing the Rogaghnata of Saindhava

Rogaghnata RT RM RN DN

Netra roga + + - +

Vrana + + + +

Vibhanda + + + +

Aruchi - + - +

Importance of Saindhava Lavana

1) Used in diet as well as in medicine.

2) Saindhava lavana is best among all the lavanas for internal use.

3) In any preparation when any particular type of lavana is not mentioned in

literature, then Saindhava lavana has to be considered.

4) Used in treatment of Aruchi, Netra roga, Hridroga, Vrana and Vibhanda.


46


Vishishta yogas

1) Nareekelalavana

2) Rasakarpoora

3) Rasapushpa

MODERN VIEW155

The body contains 250gm of Sodium Chloride, which is an essential

constituent of the body as well as the chief mineral constituent of the blood serum. It

helps to maintain the water and salt balance of the tissues, which is regulated by the

posterior pituitary antidiuretic and adrenocortical hormones. Any change in the

osmotic tension causes the movement of fluids and diffusion of salts in the cellular

tissue. Sodium metabolism is intimately related to the concentration of sodium,

potassium and chloride in the blood. Its deficiency causes retention of potassium and

diminution of sodium. The balance in the blood is kept uniform, and some stored in

the tissue as reserves, but the surplus water and salt passes out through kidneys

producing diuresis. It is therefore essential that the necessary supply of this should be

introduced either with the food itself, or as an addition to the food.


47


Haridra156,157

In vedic literature Haridra is extensively described. Haridra is one of the most

commonly used herb both externally and internally. Sushruta and vagbhatacharya

quoted it as best Pramehaghna dravya.

Gana

Charaka : Tikta Skanda, Kandughna, Vishaghna, Shirovirechana.

Sushruta : Haridradi, Mustadi, Shleshma samshamana.

Kula : Haridra kula

Family : Zingiberaceae

Vernacular Names

Sanskrit : Haridra

English : Turmeric

Latin : Curcuma Longa

Hindi : Haldi

Punjabi : Haldi

Bengali : Halud

Gujarati : Halder

Konkani : Halad

Tamil : Manjal

Telegu : Pasapu

Kannada : Arishina

Synonyms

Haridra, Harita, Jayanti, Kanchana, Nisha, Krimighna, Yoshit priya, Rajani


48


Botanical Description

Annual shrub, rhizome grows under ground, leaves are 30-40 cms in length

and they smell like ginger. The petiole is long and broad like leaves and smells like

mango. Flowers are yellow in colour and grows at the tip. The stalk of flower is 12-16

cm long. Fruits are oval with deep yellow pulp. The flowering of plant occurs in the

beginning of rainy season.

Microscopic Structure of Haridra

Externally drug is yellowish brown in colour with characteristic odour and

slightly bitter. White long variety is the cylindrical and short branched. The fracture is

horny and internal surface is orange.

Microscopic Characters

The transverse section of turmeric rhizomes shows the outer most 4 to 6 layers

of brick shaped parenchymatous cork. The cortex consists of thin walled rounded

parenchymatous cells containing scattered vascular bundles.

Oleoresin cells with brownish contents are also observed throughout the

ground tissue. Endodermis is well marked and starch grains (5 to 15 inches in

diameter) are abundant.

It contains volatile oil, starch, resin and yellow colouring substance known as

curcuminodis. The chief component of curcuminodis is known as curcumin. Chemical

constituent are known to vary as per geographical locations and curcumin content is

reported to vary from 1 to 10%.

Geographical Source

India accounts for as much as 90% of total out put of the world. Tamilnadu

and Andhra pradesh together contribute about 70% of Indian production. Kerala also


49


produces large quantity of turmeric. It is very superior in quality and is exported on

large scale.

Properties

Rasa : Tikta, Katu

Guna : Ruksha, Laghu

Virya : Ushna

Vi Paka : Katu

Doshaghna : Tridoshaghna

Karma : Varnya, Kushtaghna, Vrana shodhana, Vrana Ropana,

Rakta prasadana, Raktasthambana, Krimighna, Vishaghna,

Rechaka, Anulomaka, Lekhana, Kandughna.

Vyadhi Prabhava : Aruchi, Vibhanda, Kamala, Jalodara, Krimi, Sheetapitta,

Raktasrava, Kushtaghna, Shwasahara, Pramaehaghna.

Prayojya Anga : Kanda

Matra : Swarasa : 10-20 ml

Churna : 1-3 gm

Vishishta yoga : Haridra Khanda.


50


Nimbuka 158,159

For present study Nimbu swarasa is used for shodhana of Hingula and Kasisa.

It is an important dravya of Amla varga. In Rasa classics, it is utilized for shodhana

and marana of various metals and minerals.

Latin Name : Citrus acid.

Family : Rutaceae.

Kula : Jambeer.

Gana : Charaka : Phala varga, Amla Varga

Sushtuta : Phala Varga

Vagbhata : Phala Varga

Synonyms

Amla Jambeer, Dantaghata, Nimbuka, Rochana, Vahni, Jantumari, Deepya,

Raj nimbooka, Shodhana, Dantasatha, Vahnibeeja, Amlasara

Vernacular Names

English : Lemon

Hindi : Nimbu

Kannada : Nimbehannu

Telegu : Nimma chettu

Description

A straggling, bushy, small tree 3-4 m high with thorny branches, leaves –

ovate, petiole- margined or winged, flowers – small, white or pinkish, sweet scented,

Fruit- oblong or ovoid usually with a nipply shaped extremely bright yellow, rind-

thick, pulp – pale yellow and acidic.


51


Chemical Composition

Fruit juice contains 7 to 10% of Citric acid, 4% Phosphoric acid, 10.9% of Sugar,

Cellulose, Vitamin A and Vitamin C, 76% citrin, 7.8% Citrol, and Sulphuric acid.

Distribution: It is available throughout India.

Pharmacological and Therapeutic Properties

Rasa : Amla, Katu.

Guna : Laghu, Tikshana.

Virya : Ushna.

Vipaka : Madhura.

Dosha Prabhava : Vata, Kapha shamaka.

Karma : Deepana, Pachana, Rochaka, Anulomaka, Krimighna,

Chakshushya.

Vyadhi Prabhava : Agnimandya, Trishna, Udarashoola, Chardi, Aruchi,

Vibhanda, Visuchika, Amlapitta, Gulma, Vataroga.

Part Used: Fruit

Dosage: Fresh Juice, 10-20ml


52


Kupipakwa Vidhi

Kupipakwa is a unique pharmaceutical procedure mentioned in Rasashastra.

Rasaoushadhis prepared by this method or considered wide range of therapeutic

activity.

Etymology of Kupipakwa Rasayanas160

“Kupi Iti Kacha Kupi, Pakwam Iti Angina Pakwam, Rasayana Paradasya,

Ayanam Sthanam, Arthat Kupayam Agninam Pakwam Yadrasayanam Tat

Kupipakwa Rasayanam”

Kupipakwa Rasayana is also known as sindura kalpana. Kupipakwa rasayana

is composed of 4 words, Kupi, Pakwa, Rasa, and Ayana. Thus it can be defined as a

Rasayana product prepared from parada in a glass bottle by applying heat in a valuka

yantra.

Historical Background

There is no reference of Kupipakwa Rasayana found during Prevedic Vedic

and Samhita period.

Rasakala

Rasarnava

In this text different types of Gandhaka Jarana and Parada marana procedures

have been mentioned. But no reference of Kupipakwa Rasayana is found.

Rasa Hridaya Tantra

The References of valuka yantra found RHT.The non-availability kachakupi,

lead them to use sharava or mushas for Gandhaka Jarana process.


53


Rasendra Chudamani

The author has mentioned about valuka yantra, but not about Kupi

preparations

Rasaprakash sudhakara

Acharya Yashodhar (13th Century AD) for the first time quoted Rasasindura

kalpana by the name of Udayabhaskara rasa. At the same time he mentioned the

method of Rasakarpoora preparation by the name Ghanasara rasa. He mentioned

sikatayantra for the preparation of Udayabhaskara rasa.

Rasa Ratna Samuchaya

The author Rasa Vagbhata has mentioned very clearly about valuka yantra and

kachakupi.

Later on Rasachintamani, Rasakamadhenu, RasendraSaraSangraha, Ayurveda

Prakash, Rasatarangini have described different Kupipakwa rasayana.

Overall after critical study one comes to the conclusion that, the process of

Gandhaka Jarana mentioned in RHT developed and came in light as Kupipakwa

Rasayana.

Importance

The doctrines of Rasashastra have given more importance to Kupipakwa

Rasayana than other kalpana. Because of its below mentioned properties.

Potency of these drugs can be retained for longer period.

It can be administered in minimal dose.

More potent as compared to other herbal preparations.

Easy for administration.

When mixed with other drugs, it reduces their dosage.

It exhibits quicker action.


54


Chemical bonding present in Kajjali, Parpati, Pottali and Kupipakwa Rasayana

is stronger and stronger respectively.

Classification

Kupipakwa Rasayanas are classified based on use of Gandhaka in the

preparation, mode of preparation and by the site of collection of finished product.

Types according to

Ingredients Method of preparation Site of finished

Product Sagandha Antardhuma Kanthastha (Prepared with the use (Corck is applied in (The finished product of Gandhaka) the beginning) deposited at the neck) Makardhwaja Rasasindura Rasapushpa, Sameerapannaga Rasa Hinguliya Manikya Rasa Nirgandha Bahirdhuma Talastha (Prepared without the (Corck is applied after (The product is use of Gandhaka) evaporation of fumes) collected at the Rasapushpa, Shila Sindura bottom of kupi) Rasakarpoora Malla Sindura Swarnavanga Samirapannaga Ubhayastha (Finished product

obtained from both sites) Makaradhwaja

Procedure of Kupipakwa Rasayana161

Whole procedure can be studied under three headings

a) Purva karma

b) Pradhana karma

c) Paschat karma


55


a) Purvakarma

During Purvakarma following points are to be considered.

i) Collection of Appropriate Instruments

ii) Purification of Ingredients

iii) Preparation of Prematerial

iv) Preparation of Kupi

v) Filling of kupi with Prematerial

Bhatti (Bhrashtri) 162

The height and width of the Bhatti should be 18 angulas shaped like an Ant

hill with a hollow space of 5 Gulpha (20”) inside and should have many holes in its

lower portion. There should be an opening for introducing fuel, of about 12 angulas.

In the Bhatti heat of the burning fuel should properly reach the center as well as

surrounding the valuka yantra. There should be sufficient ventilation inside the

furnace. An outlet for the fumes should be there from inside the flame should go up

rather coming down. Bhatti can be made with the fireproof bricks, which minimizes

the loss of heat and fuel consumption.

Fuel used: wood, coal, electricity etc.

Valuka yantra163

It should have narrow base and wide mouth depending on the size of kupi (i.e.,

1” taller than kupi) should be prepared with the 2 handles the circumferences of

valuka yantra should fit exactly over the hole of Bhrashtri. According to Rasendra

Chudamani, it should fill Adhaka sand and have a central hole of 2 to 2.5 cm at the

bottom, which should be closed with Abhraka patra before keeping the kupi during

heating. According to Yadavaji Trikamji Acharya the depth of the vessel should be 1

vitasti pramana.


56


Valuka

Natisthoola and clean valuka should be filled into the valuka yantra. At first 2-

3 cm of sand is spread over Abhraka patra which is kept covered over the central hole,

over which Kajjali filled kupi is kept and remaining part of Valuka yantra should be

filled with Valuka till the neck of kupi.

Kupi 164

Synonyms of Kupi are Kupika, Siddha etc.

During ancient period they used to prepare sinduras in Andha mushas or Kupi made

with Hema, Tara, Ayas, Mrittika. Any material can be used but they should sustain

intense heat. After 10th century when glass bottles were invented, it was utilized for

medicine preparations. Now days beer bottles of 650ml capacity with the neck 1-1.5

inches in length and moderate thickness with variable varna are used.

Kupadamitti

Aim: To enhance the heat susceptibility of Kupi, also to strengthen the glass bottle.

So that it cannot break even when inside vapour pressure increases.

Because of above cited reasons, Kupi, which is used for oushadhi nirmana,

should be covered with the cloth smeared with mud.

Mrittika

Valmeeka mrittika or potters mud can also be used. It is advised to prepare

kapadamitti from husk- 2 parts + Cotton – 1Part + Mud 3 Parts and fibers grinded and

kept soaked in water for 7 days and then used to cover the Kupi. Now-a-days

Gopichandana or clay is used for this purpose. The mud smeared cloth applied to the

Kupi from bottom to mouth and should be well dried, whole length of the Kupi can be

applied with kapadamitti as it prevents breakage of Kupi during heating.


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Cloth: Usually cotton cloth is used.

For the base of Kupi – Circular piece

For the surrounding – Rectangular shape

From neck to mouth – A piece having broad base and a narrow top.

Different Shalaka

Iron rods can be used, thin rod (about 0.5cm diameter) for cold Shalaka test,

thick rods (about 1-1.5cm) for hot Shalaka test.

Cork

Corking material is called Mudra. In Kupipakwa Rasayana procedure, after

complete evaporation of fumes and cessation of flame, kupi’s mouth is closed with

cork and is called mudrana or corking. For this purpose any sticky substance which

gets hardened with further heating and which can properly fit the mouth of kupi is

used.

Madana Mudra

Made with latex of Udumbara and Vata, Lac, Kantaloha, Gorochana and Atasi

taila. It is sticky and slightly hard.

Hata Mudra

Made with glass powder, Brick powder, Tankana, Mandura rubbed with latex

of Vata, Udumbara and Arka for one day.

Now-a-days cork made of Gairika or Ishtika is plugged into mouth of the

bottle and wrapped with the cloth smeared with Gopichandana.

Pyrometer

In Kupipakwa Rasayana, heating schedule is the most important factor.

Through pyrometer, one can regulate the heat supplement for the preparation. The


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purpose of using pyrometer is to record the temperature of sand by keeping the

sensors in valuka yantra.

Purification of Ingredients

All the raw materials should be identified first for its genuinity. Every raw

material should be purified according to the method prescribed in classics. Purified

ingredients must be tested according to samyak shuddha laxanas described in texts.

Preparation of Kajjali 165

Here the term Kajjali can be used for pre-material for making Kupipakwa

rasayana. Equal quantity of Hingulotha Parada, Gandhaka taken and mardana should

be done without using any liquid till the mixture becomes shlakshana choorna.

Prematerial of Rasapushpa is ash in colour. The colour of Prematerial used to

prepare Kupipakwa rasayana depends on the ingredients used for e.g.:- Red color in

Hingulad Rasasindura, white in Rasakarpoora etc.

Filling of Prematerial into Kupi

The Kupi should be filled up to 1/3rd part by Kajjali, so that there would be

enough space inside the kupi for melting and boiling of Kajjali and also for the

sublimation of compound, which is going to be condensed and deposited at the neck

of the kupi. Such kupi should be kept exactly at the center of valuka yantra.

Pradhana Karma

• Temperature monitoring

• Heating pattern/schedule

• Shalaka sanchalana

• Observation of fumes and flames

• Corking of mouth of kupi (mukha mudrana)

• Self cooling (Swanga Shitalikarana)


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Temperature Monitoring

Ancient parameter: Traditionally following tests were in practice-

Cotton, dry grass test: Cotton or dry grass kept on the valuka catches fire and burns,

then it is considered to beTivragni.

Rice test: when a paddy or maize put on valuka, it puffs up – Teevragni.

Modern parameter: Now-a-days pyrometer, thermocouples, thermometers

are used for measuring the temperature.

Heating pattern 166

Few signs and standards of different heating stages of Kupipakwa Rasayanas

are mentioned by ancient scholars for deciding proper pachana of the ingredients

through kramagni paka.

It has to be considered in the following way:

1) In terms of duration of heating

2) In terms of temperature.

The term duration indicates the time for maintenance of kramagni and the term

temp indicates the temperature limit in each stage of kramagni.

Kramagni pattern of Rasapushpa

Mrudu Agni : 100-1500 C (Initial stage)

Madhyamagni : 150 - 2500 C (Middle stage)

Teevragni : 250 - 3500 C (End stage)

Kramagni pattern for sagandha Kalpana (Rasasindura)

Mrudu Agni : 150-2000 C (Intial stage)

Madhyamagni : 350 - 4000 C (Middle stage)

Teevragni : 550 - 6000 C (End stage)


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I stage mrudu Agni (150-2000 C) (Stage of Liquefaction of kajjali)

During this stage, material in the kupi completely melts which may be

ascertained by inserting cold Shalaka into the kupi.

This heat should be maintained for prescribed time to allow chemical reactions

to start with.

II Stage Madhyamagni (350 - 4000 C) (Stage of profuse fumes & boiling of

Kajjali)

♦ This stage commences from complete melting of Kajjali and lasts till initiation

of sindura compound formation.

♦ Liquefied Kajjali starts boiling

♦ Boiling of Kajjali can be ascertained by inserting cold iron rod in the kupi or

by visualizing through light.

♦ The care should be taken that heating should not be strong otherwise boiling

Kajjali may come out of kupi and may catch fire which may leads to breaking

of kupi.

♦ Profuse fumes of sulphur from the kupi are present.

♦ Deposition of fumes at the neck of kupi may block it, hence it ought to be

frequently removed by inserting tapa shalaka into the kupi mouth.

♦ Moderate heat for prescribed period has to be maintained to ensure burning of

extra sulphur in the product.

III Stage–Teevragni (550-6000 C) (Stage of appearance of flame and corking of

kupi mouth)

• This stage commences from formation of sindura compound and lasts up to

the completion of Jarana of Gandhaka. The process of formation of sindura

occurs in the middle stage, it means when Kajjali is in boiling stage (Honey


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comb like appearance), chemical changes occurs and as a result formation of

new compound takes place which is called as sindura kalpa. Afterwards as

heating persists, this newly formed compound sublimates and gets condensed

at the neck and mouth of the Kupi.

• At the end of middle stage sulfur fumes catches fire and it takes form of flame.

At this stage, flame slowly grows up and diminishes when process of

Gandhaka Jarana completely finishes.

• When extra sulfur burns out completely, flame disappears and this indicates

the completion of Gandhaka Jarana.

• Redness starts appearing at the bottom of the kupi when seen through torch

light which gets more brightened (Suryodayavat), this assures formation of

sindura.

• When copper coin is placed on the kupi mouth, due to mercury fumes, which

is getting evaporated, it becomes white.

Shalaka Sanchalana167

During this procedure cold and hot shalakas are being used. Ayurveda

prakashakar mentioned the use of tapa shalaka for clearing the mouth of Kupi from

the deposition of sulfur fumes. Iron rods of different size and shape were used. Tapta

shalaka is used for clearing the sublimated sulfur deposited at the neck region of kupi,

otherwise sulfur may block the mouth of kupi and fumes may increase, the inside

pressure and there may be chances of bursting of kupi.

Sheeta shalaka is used especially for noting the state of Kajjali, whether it is in

powder form or in boiling state or in sublimating compound state.


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Observation of Fumes and Flames 168

All the characteristics of fumes like color, smell etc must be observed. It

differs according to the ingredients.

• Colour : yellowish, orange, blush or white etc.

• Odour : Sulphurous, Arsenical etc.

• Quantity : Mild, Moderate profuse/dense etc.

Flame

This is also an important factor while preparing Kupipakwa Rasayana. Time

of starting of flame, its height, colour and its duration are the important features.

These important features depend on ingredients used.

Corcking of Kupi Mouth 169

It is important to decide the proper time of corcking, which is a difficult task.

For proper assessment few tests are to be conducted such as

absence of flame

Absence of fumes

Copper coin test should be positive

Appearance of Redness at the bottom of Kupi.

Dry grass test should be positive.

If a Sheeta Shalaka is introduced into the Kupi a white dense fume appears

suggests completion of Gandhaka Jarana. Sheeta shalaka gets coating of

different coloured powder according to different compounds. e.g., blackish in

Rasasindura, white coating in Rasapushpa etc. This is called positive sheeta

shalaka test and is confirmatory before corcking.


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Before corking, 2-3 inches of sandy layer should be moved aside from the

neck region to make it cool as the sublimating compound can get well

condensed at the neck portion of the kupi.

Method of Corcking 170

Cork made up of ishtika, wood or mud is kept over the mouth of the Kupi and

then it is covered with the cloth smeared with clay/ multani mitti. For sealing purpose

Chalk powder (Khatika) Guda (Jaggery), Madhu (Honey), etc can be used.

There is controversy regarding heating after corking, some opine to continue

heating as per textual reference for complete period and then wait for self cooling and

while some opine that heating should be discontinued after corking and allow yantra

to get self cooled.

Swanga Shitalikarana

After heating for a prescribed period, Bhatti is left for self cooling in order to

avoid breaking of Kupi by sudden exposure to cold air.

Paschat Karma

Removal of kupi from valuka Yantra

Breaking of kupi

Collection of final product

Examination of final product

Removal of Kupi from Valuka yantra

Sand should be removed from valuka yantra and then kupi is taken out

carefully. Then kapadamitti layers are carefully scraped out, Kupi is cleaned with wet

cloth. Level of product inside the kupi is observed and marked.


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Breaking of Kupi 171

A thread soaked in Kerosene or spirit is tied just below (2-3cm) the level of

the product and fire is set, Kupi is kept horizontal and rotated so that whole thread

gets completely ignited. Little cold water is sprinkled over it or kupi can be wrapped

with wet cloth, then kupi breaks into two halves at desired level. The product, which

is either talastha or kantastha, is collected, powdered and stored well in airtight

container.

Examination of Final Product

The judgment about the colour and shape of the crystals of sindura can be

made by ingredients of Kajjali. Similarly smell and color of flame are the basis for

determination of sindura compound going to be formed. Chemical analysis,

crystallographic study and clinical study are the confirmatory evidences of sindura.

Thus in nutshell, during Kupipakwa Rasayana kalpana cardinal features has to be kept

in mind are

○ Preparation of kupi

○ Preparation of kajjali

○ Filling of kajjali in kupi

○ Maintenance of kramagni type of heating pattern

○ Clearance of mouth of kupi with tapta shalaka

○ Examination of completion test of sindura kalpana

○ Corking of mouth of kupi

○ Breaking of Kupi and collection of final product

In short this description is all about Sagandha, Bahirdhuma Kupipakwa

Rasayana. But in the preparation of Antardhuma Kupipakwa, kramagni should be


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given up to mentioned duration. Corking of kupi mouth should be done initially

before subjecting it to agni.

In case of preparation of Nirgandha Kupipakwa Rasayana, heating pattern

should be kramagni as described before and corking should be done after complete

expulsion of water vapours from kupi and positive sheeta shalaka test. Heat required

for the nirgandha preparation is comparatively less.

Matra 172

Due to variation in individuality, on the basis of treatment, compatibility,

acuteness of disease age, climate etc. The dose will differ from one individual to

another. Generally dose should be decided according to particular yoga and rogi

avastha.

Rasapushpa173,174

Rasapushpa is nirgandha type of Kupipakwa Rasayana. It is first described by

Acharya Sadananda Sharma in his text Rasatarangini. Rasendra Sara Sangraha is the

first text in which nirgandha type of Kupipakwa kalpana is described. Overall it can

be said that origin of nirgandha yogas are found in Rasa classics from 16th century AD

onwards.. Till date three most common nirgandha yogas found in Rasa classics viz

Mugdha rasa, Rasakarpoora and Rasapushpa.

Acharya Sadananda Sharma was first person who clearly differentiated

Rasapushpa from Rasakarpoora. The author has described three different methods of

preparation of Rasapushpa, but on careful assessment they can be classified into two

types viz Antardhuma and Bahirdhuma.

The Paryayas of Rasapushpa are:

Rasakusuma, Rasasuma and Sudhanidhi Rasa.


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Ingredients

Hingulotha Parada

Shodhita Kasisa Each 5 Tolas

Saindhava lavana

These ingredients are mentioned for Bahirdhuma Kupipakwa method of

preparation, while for Antardhuma (Damaruyantra) method an additional fourth

ingredient viz Shuddha Sphatika (5tolas) is mentioned.

I Procedure

Hingulotha Parada, Shodhita Kasisa and Saindhava lavana 5 tolas each should

be properly weighed. These ingredients are to be triturated well in Khalva yantra till it

becomes shlakshana choorna. Then keep this nischandra churna into lower pot of the

Damaruyantra and on the upper earthen pot a hole should be made at the center for

evacuation of water vapours. Then heat it on a very mild fire for two yamas. When all

water vapours are expelled out, hole has to be sealed with cork. When it becomes

swangasheeta, sandibandhana is removed and chandrama sadrusha (white coloured)

Rasapushpa deposited in the upper pot is collected.

II Procedure (Kupipakwa method)

Hingulotha Parada, Shuddha Kasisa and Saindhava lavana in equal quantities

taken, these are triturated well with the help of Khalva yantra till it becomes

shlakshana choorna. This Prematerial is filled in the kupi (having applied 7 layers clay

smeared cloth) and Placed in the middle of sikatayantra. Kramagni is given for two

yama. After swangasheeta, Rasapushpa sublimated at the upper part of the Kupi is

collected. While giving heat when all water vapors are expelled out, corking should

be done and sand at the neck portion of kupi is removed, so that Rasapushpa can


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deposit there. As this compound is obtained at the neck of Kupi like a flower, it is

called by the term “Rasapushpa”

III Procedure (Damaruyantra)

Here, the Pushpakasisa is heated on mandagni till the evaporation of watery

portion. Similarly purification of Sphatika is done, then Kasisa, vahnibharjita

Sphatika, Hingulotha Parada, and Saindhava lavana in quantity of 5 tolas each is

mixed in a mortar and triturated effectively to make it shlakshana churna. Obtained

nischandra churna is kept in Damaruyantra and is heated on very mild fire for 2

yamas (i.e., 6 hours). When it becomes swangasheeta, white, shiny, sharadindu

samana Rasapushpa is collected from the upper pot.

Properties of Rasapushpa

It has got following properties

Pittahara

Mootrala

Vranadoshahara

Parama virechanakara

Bhootavishapaha

Swasthikaranakara

Jaleeyamsha Vishoshanam

Malapitta sarakam

Visuchika krimihara

Hikka nashaka

Phirangahara

Cures jalodara very quickly


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Rasapushpa Matra

The matra of Rasapushpa in different diseases in different age group are

represented in below mentioned table.

Table No.25 Showing the Matra of Rasapushpa in different diseases.

Sl. No.

Purpose Matra

1 Virechanartha 21/2 Ratti 2 Virechana for children ¼ Ratti 3 Hikka ½ Ratti 4 Phiranga ¼ Ratti 5 Visuchika ¼ Ratti

Therapeutic uses of Rasapushpa

Rasapushpa in the dose of 21/2 ratti when given with svarjika kshara starts purgation

after two yamas (6hours)

In the initial stage of Visuchika, if Rasapushpa is given with water in the dose of 1/4th

ratti kills Pathogenic bacteria of Visuchika

Acharya adds here precautionary sentence that the bheshaja who does not have

thorough knowledge of Rasashastra should never prescribe Rasapushpa especially for

longer duration.

Test for genuinity of Rasapushpa

To examine the purity of Rasapushpa, take a clean iron pan, put a drop of

water on it then add a pinch of Rasapushpa on that drop, after a moment remove the

drop and if there is no blackening at drop site, then Rasapushpa sample should be

understood as genuine.

Formulations of Rasapushpa

• Chandanadivatika

• Rasapushpa Malahara

• Rasapushpadya Malahara


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Other nirgandha Kupipakwa kalpana described in Rasa classics is

Rasakarpoora. It is one of the mercurial preparations prepared with different

ingredients and Pharmaceutical procedures. These formulae reveal that it may be

prepared from Parada with Sulphate compounds like Gandhakamla, Kasisa, Tutta,

Sphatika etc. on this basis according to Shri P.C Ray, Rasapushpa and Rasakarpoora

Preparation methods are of mix compound (History of Hindu chemistry)

There exists close resemblance between the two compounds. In Parada Vijnana, Dr.

Dwivedi has also mentioned few tests for differential identification of Rasapushpa

and Rasakarpoora. Out of these color test could not be said as significant in the

differential identification point of view as we look at the words mentioned to express

the color of Rasapushpa and Rasakarpoora.

Rasakarpoora – Karpura Sannibham, Kundendu Sannibham, Galadroupya nibham and

Hirakavat etc.,

Rasapushpa – Nihar prabham, Kundavat, sharadindu Sannibham, Pushapavat.

These words are not very much distinguishing.

Other important tests are as follows according to V.M Dwivedi

Table No. 26 Showing the distinguishing features of Rasapushpa and

Rasakarpoor.

Sl.No Test Rasapushpa Rasakarpoora

1. Solubility Con.Nitricacid 1/16th part in cold water,

more than this in hot water

Alcohol

Ether

2 Precipitation in liquid

Ammonia

Black precipitation White precipitation

3 Chemical composition Hg2Cl2 HgCl2


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Mercurous Chloride175

English Name : Calomel

Chemical Name : Hydragyri subchloride or mercurous chloride.

Chemical Formula : Hg2Cl2

It is known by the name subchloride of mercury. It is insoluble in water and its

insolubility is the greatest bar to its toxicity. It is used as purgative, as it is non-toxic

for human consumption in therapeutic dosage. It has been known since the early

middle ages. It is prepared by the process of sublimation.

Preparation

• It is prepared by subliming a mixture of mercuric chloride and metallic

mercury in appropriate proportions.

HgCl2 + Hg Hg2Cl2

The sublimate is treated with cold and dilute nitric acid to remove any metallic

mercury and then washed thoroughly to remove any mercuric chloride.

• By adding an excess of hydrochloric acid or any other chloride to any soluble

mercurous salt, e.g. mercurous nitrate.

Hg2 (No3)2 + 2HCl Hg2Cl2 + 2HNo3

A salt can be obtained as a sublimate when mixture of mercurous sulphate and sodium

chloride is heated.

Hg2SO4 + 2Nacl Na2SO4 + Hg2Cl2

Properties

Appearance : Amorphous heavy powder

Taste : Tasteless

Colour : White


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Molecular weight : 236.1

Sublimating point : 4000C

Impurities : Mercuric chloride

Action : Purgative, diuretic.

Fatal dose : 1.5 to 2gm / 70kg

Solubility : Practically insoluble in water, alcohol, and cold dilute

acids.

Identification

o Blackens by contact with dilute ammonia solution or with solutions of alkali

hydroxides.

o It becomes yellow when triturated or compressed.

o Volatises when strongly heated.

o Stable in air but gradually darkness when exposed to light.


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Krimi

The word Krimi has very much potential in Ayurvedic literature. In the

etiology of many diseases microbial relation played very important role. In Ayurveda

visible (drushya) and invisible (Adrushya) organisms that affect living and non-living

things are described. The term krimi has been used in broader sense i.e. it includes all

pathogenic and non-pathogenic organisms covering wide range of infection and

infestation. These infectious diseases caused by krimi are explained under the title of

Oupasaigika rogas, which spread through contact with patients and through other

routes.

Those that spread in the form of epidemics are called janapadowansa rogas.

This theory dealt within Charaka Vimanasthana, this results due to contamination of

air, water, place and soil. We can prevent ourselves from clutches of disease by

following the measures put in Charaka Vimanasthana. During this epidemic the

normal characteristics of air, water, and soil changes and can be perceived if observed

keenly.

Acharya Sushruta has asked to perform puja, homa, dana, meditation, tapa,

faith in god and vacate the place where epidemics is present.

Etimology176

The term is derived from the root ‘kram’. It denotes the meaning “ Brahame

samprasarane” i.e. which moves about.

Historical Background of Krimi

We get abundant material about krimi since Veda Kala, especially in Atharva

Veda.


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Atharvaveda177

Author has described both drushya (visible) and adrushya (invisible) krimis.

They are present everywhere and can flourish on mountains forests, plants, animals,

water and other liquid things. They make their way inside our body by means of food,

water, air and injury.

Microbes may invade the uncooked, semicooked, and cooked foods and

contaminate them. If an individual consumes a food material, it has potential to harm

the body.

Krimis capable of producing sutikaroga and balaroga also mentioned.

Atharveda has given solution to combat the krimis; they can be destroyed by

administering different oushadhies, and by chanting mantras and by performing homa.

Samhita Kala

Charaka Samhita178

As per Samhita literature, it can be assessed that Acharyas has definite

knowledge about micro organisms and their pathogenecity For this one has only refer

to krimis of blood (Raktaja krimi).

According to Charaka krimi are found in blood vessels (vahinyo dhamanyah).

They are of different shape and size i.e. of microscopic size (anavaha), circular, or

disc like (vitta) etc.

Sushruta Samhita179

Acharya Sushruta has enumerated the reasons for transfer of infection such as

prasangat, gatrasamsparshana, sahabhojana, mala, lepa etc. Many diseases spread by

above said routes, The Acharyas like Dalhana and some other later on endorsed this

view of Sushruta. There is also a clean indication that some organisms gain entry

through injury.


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Acharya has mentioned Rakshoghna gana, which includes group of plants

possessing Krimighna properties. These dravyas keep the environment free from

krimi. At the time Rakshoghna dhoopana have been used for sterilization of rooms,

kitchens, rasayanashala, shastragara.

Ashtanga Hridaya180

Acharya Vagbhata has asked to keep the bed, pillows, clothes, etc of the

patients in direct sunrays to disinfect them.

From this survey one may confidently summarize that the knowledge of

microbes, infectious diseases, and epidemics was nothing alient to ancient India.

Microbial pathology was precisely codified in many Ayurvedic classics precisely.

Krimi Bheda181

20 types of krimis are explained under the heading of Malaja, Purishaja,

Kaphaja and Raktaja krimis.

Krimi

Malaja Purishaja Kaphaja Raktaja

Bahya Abhyantra Kakeruka Antarada Keshada

Makeruka Udarada Lomada

Yooka Leliha Hridayachara Lomadweepa

Pipilika Sashoolaka Churava Sourasa

Sousarada Darbapushpa Jantumatara

Sougandhika Oudumbara

Mahaguda


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Krimi Laxanas and Their Prabhava182

Table No. 27 Showing laxanas sthana and Prabhava of different krimis.

Bheda Laxanas Sthana Prabhava

Mala Anu, Bahupadayukta,

Krishna or Shukla varna.

Kesha, Shmashru,

Loma

Kandu, Kota,

Pidaka.

Purishaja Sukshama, Vritta,

Deergha akara Shyava,

Neela, Harita and Peeta

varna.

Pakwashaya Atisara, Karshya,

Parushya,Kandu

Lomaharsha,

Pandu,

Adhogudamukha,

Agnimandya,

Kaphaja Some are Sthoola,

Sukshma, Vritta, Pruthu

in akara, they have got

hairs on their body, white

in color

Amashaya Hrillasa, Jwara

Pandu, Moorcha,

Chardi, Angamarda,

Anaha Shirshoola,

Hridroga.

Raktaja Some are Anu, Vrittakar,

Apada (no leg), Sukshma

(invisible), Rakta or

Krishna varna

Dhamani Kandu, Kushta

Visarpa, Toda

Pidaka, Harsha,

Jwara.

Among these krimis, Raktaja krimis are told as very minute and invisible to naked

eye, which is expressed by the term “Kechid darshana”. The opinions of all Acharyas

are same regarding this issue.


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As for the treatment of this disease is considered, it consists of the removal of

worms, eradication of the source of origin/breeding ground and prophylaxis against

causative source. The removal may be manual as for organism like lice. In some

locations it may be done by evacuative measures including Nasya, Vamana,

Virechana and Niruhabasti. The source is eradicated by the administration of kashaya,

katu and tikta medications and other agents opposite to kapha. Prophylaxis is

accomplished by personal conduct, which scrupulously avoids the contact with

nidana.

Apart from these, for infected wound treatment, Ayurveda advocates

fumigation with Krimi hara dravya in order to kill disease-causing microorganisms

locally. Also the fumigation was a routine process for cleansing shalyagara, sutikagar,

and oushadhagar with krimihara dravya.

These substantiating evidence are enough to say that disease-causing

microorganisms of present era can be equated with krimi of old Indian system of

medicine.

For the convenience of present study, we have selected most prevalent disease

causing organisms in the clinical practice.


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Microorganism

Microorganisms occur in large numbers in most natural environments and

bring about many changes. Some desirable and others undesirable. The diversity of

their activities ranges from causing diseases in humans. Microorganisms are the major

causative factors for many infectious diseases.

Historical Background183

History is the story of achievements of men. Many important contributions

were made by people whose names have been forgotten and whose accomplishments

have been lost in the longer and deeper shadows. Long before the discovery of

microorganisms certain processes happened by their life activities such as

fermentation of wine juice, milk, yeast etc. were known to mankind.

Varo and Columella (first century BC) suggested that invisible organisms inhaled or

ingested caused diseases.

Kircher (1659) could find minute worms in the blood of plague patient.

• Antonivan Leeuwen hoek (1683) could give description of various types of

bacteria. He also invented simple microscope.

• Robert hook, a contemporary of Leeuwen hook developed compound

microscope in 1678, as microbes are not visible to unaided.

• Von plenciz (1762) proposed that each disease was caused by a separate agent

Augstino Bassi (1835) proposed that, some diseases are caused by fungus.

• Pasteur (1857) proposed that different types of fermentation was associated

with different kind of microorganisms, He introduced techniques of

sterilization and developed steam sterilizer, hot air oven, and autoclave.


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• Robert Koch perfected bacteriological technique He introduced staining

techniques and also methods of obtaining bacteria in pure culture using solid

media.

• Koch is remembered for his isolation of the bacteria that cause anthrax and

tuberculosis. For this important contribution to the creation of science of

microbiology won him the 1905 Nobel Prize. New agent of infectious diseases

continue to emerge, e.g.: HIV (identified in 1980) the outbreaks of plague in

1994, cholera in 1995, and dengue hemorrhagic fever in 1996.

Though of relative short duration, the history of microbiology is filled

with thrilling achievements. This science has won many battles with microorganisms

and learned to control them.

These agents of human infectious diseases belong to 5 major group of

organisms viz bacteria, fungi, protozoa, helminthes and viruses.

Bacteria184

Bacteria’s share a unique place in the world of living organisms. Bacteria’s are

considered as prokaryotes, which means primitive nucleus.

Size and Shape and Arrangement

Bacteria’s are very small, most being approximately 0.5 to 1.0μm in diameter. The

shape of bacterium is governed by its rigid cell wall. Typical bacterial cells are

spherical (cocci), straight rods (bacilli) or rods that are helically curved (spirilla).

Although most bacterial species have cells that are of a fairly constant and

characteristic shape, some have cells that are pleomorphic i.e. that can exhibit a

variety of shapes.

Bacterial cells are usually arranged in a manner characteristic of their

particular species. Hence the arrangement of bacteria is important for example, certain


79


cocci occurs in pairs (diplococcic), some in chains (streptococci) and others in grape

like clusters (staphylococci). These arrangements are determined by the orientation

and degree of attachment of the bacteria at the time of cell division.

Gram positive and Gram negative Bacteria185

G (+) ve and G (-) ve bacteria can be identified by their cell wall structure.

Cell wall is the outer most component common to all bacteria.

It is elastic and porous and is freely permeable to solute molecules of less than 10,000

molecular weight. It is about 10-25 nm in thickness and shares 20 to 30 percent of dry

weight of cells.

The structure, chemical composition and thickness of the cell wall in gram

positive and gram-negative bacteria differ as follows:

The peptidoglycan layer is much thicker in G (+) ve bacteria than in G (-) ve bacteria.

Some G (+) ve bacterias also have layer of teichoic acid out side the peptidoglycan

whereas G (-) ve bacterias do not.

In contrast, the G (-) ve organisms have a complex outer layer consisting of lipo

polysaccharide, lipoprotein, and phospholipid lying between the outer membrane

layer and the cytoplasm membrane in G (-) ve bacteria is the periplasmic space, which

is the site of enzymes in some species, (e.g.: β−lactamases) that degrade penicillin and

other β−lactam drugs.

Table No.28 Showing the difference between Gram positive and Gram negative

bacteria.

Sl.No. Features G (+) ve G (–) ve

1 Thickness 15 to 23mm 10 to 15mm

2 Variety of amino acid Few Several

3 Aromatic and sulfur Absent Present


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containing amino acids

4 Lipids Low 2 to 4% High 15 to 20%

5 Teichoic acids Present Absent

Staphylococcus Aureus186, 187,188

Staphylococci are present everywhere and are spherical G (+) ve organism.

They are non-motile and non-sporing and can grow aerobically or anaerobically. They

are 0.5 to 1.5μm. They grow characteristically in aggregates like a bunch of grapes.

Most staphylococci are non pathogenic and are called Staph albus, because

they usually produce white colonies on culture. Staph albus cause infection only if the

resistance of the patient is lowered and even then, their virulence is low.

Most Staphylococci have harmless as commensals. The pathogenic

Staphylococci are called Staph aureus. They produce golden colonies on culture.

They are parasites occurring on the skin and mucus membrane of humans.

Three species of Staphylococci are human pathogen viz-Staph aureus, Staph

epidermis, and Staph saprophyticus, among these Staph aureus is by far the most

important. It has several important cell wall components and antigens, which include

protein A, teichoic acids and surface receptors. Lesions produced by Staphylococcus

may be external or internal. A common manifestation of its infection is production of

pus i.e organism is pyogenic.

Transmission

These are ubiquitous in the human environment and in the normal human

flora. Staph aureus is often found in the nose and some times on the skin especially in

hospital staff and patients. Additional sources of Staphylococcal infection are

shedding from human lesions and fomites contaminated by these lesions. Disease


81


production is favored by a heavily contaminated environment (Family with boils) and

compromised immune system.

Pathogenesis

Staphylococci cause disease both by producing toxins and by multiplying in

tissue and causing inflammation. The typical lesion of Staph aureus infection is an

abscess.

Several important toxins and enzymes are produced by Staph aureus

(enterotoxin Toxic shock syndrome toxin).

Clinical findings

The important clinical manifestation caused by Staph aureus can be divided

into 2 groups. Inflammatory and toxin mediated. In the following list, the first six are

inflammatory in origin whereas the last 2 are toxin mediated.

1. Skin infections include impetigo, furuncles, cellulites, surgical wound

infections and post partum breast infections.

2. Bacteremia from any localized lesion, especially wound infection or as a result

of intravenous drug abuse (Bacteremia may lead to endocarditis).

3. Endocarditis

4. Osteomyelitis

5. Pneumonia in postoperative patient or following viral respiratory infection

especially influenza (Staphylococci often leads to emphyema).

6. Abscess

7. Food poisoning (characterized mainly by its vomiting) due to ingestion of

enterotoxin that is preformed in foods and hence has a short incubation period.

8. Toxic shock syndrome which includes fever, hypotension.


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Laboratory Findings

Smears from local lesions or pus reveal G(+)ve cocci in grape like clusters.

Culture yields white or golden yellow colonies.

Treatment

90% of Staph aureus strains are resistant to penicillin which should be used

only if the organism is shown to be sensitive Staph aureus strains resistant to all

antibiotics except vanomycin. Some strains of Staphylococci exhibit tolerance i.e.

they can be inhibited by antibiotics but are not killed. Tolerance may be due to failure

of drugs to inactivate inhibitors of autolytic enzymes that degrade the organism.

Tolerated organisms should be treated with drug combinations.

Prevention

There is no effective immunization. Cleanliness, frequent hand washing and

aseptic management of lesions help to control spread of Staph aureus. Dissemination

from the nose or skin of carriers can be reduced by topical application of

antimicrobial agents (or by systemic treatment), but it is difficult to arrest altogether.

Shedders may have to be removed from high-risk areas, e.g.: operating rooms and

newborn nurseries.

Streptococcus Pyogenes189, 190

Streptococci are spherical cocci usually arranged in chains or pairs. Although

the genus is usually considered aero tolerant, some strains can tolerate only less levels

of oxygen and some are aerobic and some are anaerobic. Nutritional requirements are

complex including several amino acids and vitamins.


83


Most Streptococci are parasites of humans and animals and several species are

pathogenic. There are many species of Streptococci such as Strepto pyogenes, Strepto

mutans, Strepto fucecabs and Strepto pneumoniae.

Clinical manifestations

Streptococci produce a wide variety of infections. All species can cause

septicemia. Strepto pyogenes is the most common bacteria that cause sore throat,

tonsillitis, scarlet fever, glomerulonephritis, rheumatic fever, purpureal sepsis, skin

and soft tissue infections, bone and joint infections.

Laboratory Diagnosis

All smears are stained. Smears from skin lesion or wounds that reveal

streptococci are diagnostic.

Treatment and prevention

Strepto pyogenes are susceptible to penicillin but neither rheumatic nor acute

glomerulonephritis patients benefit from penicillin after onset. Hence most of

Streptococci infection has to be treated by drug combinations.

Prevention

Prevention of Rheumatic fever involves prompt treatment of streptococcal

pharyngitis with penicillin. There are no vaccines available against streptococcal

infection.

Pseudomonas Aeusrginosa191

Pseudomonas are G (-) ve bacterias resemble the members of the

enterobacteriacea but differ in that they are strict aerobes i.e. they derive their energy

only by oxidation of sugars rather than by fermentation because they do not ferment

glucose, they are called non fermenters in contrast to the enterobacteriacea which do

ferment glucose.


84


These bacterias are widely distributed in soil and water. Several species are

pathogenic for humans or animals. Some cause spoilage of meats and other foods.

Pseudomonas is able to grow in water containing only traces of nutrients, e.g.: tap

water and this favors their existence in hospital environment. These bacteria contain 5

genetically distinct groups. Among them Pseudomonas aeurginosa and papacia have

a remarkable ability to withstand disinfectants. This accounts in part of their role in

hospital-acquired infections.

Pseudomonas aeurginosa produces two pigments useful in clinical and laboratory

diagnosis

1) Pyocyanin which can colour the pus in a wound blue

2) Pyoverdin (fluorescin) a yellow –green pigment that fluoresces under

ultraviolet light, properties that can be used in the early detection of skin

infection in burn patients.

Strains of Pseudomonas aeurginosa isolated from cystic fibrosis. Patients have

a prominent slime layer (glycocalyx), which gives their colonies a very mucoid

appearance. The slime layer mediates adherence of the organisms to mucus

membranes of the respiratory tract and prevents antibody from binding to the

organism.

Pathogenesis and Epidemiology

Pseudomonas aeurginosa is found chiefly in soil land water, although approximately

10% of people carry it in the normal flora of colon. It is found on the skin in moist

areas and colonize the upper respiratory tract of hospitalized patients.

Pseudomonas aeurginosa is primarily an opportunistic pathogen that causes

infections in hospitalized patients, e.g. those with extensive burns, in whom the skin

host defenses are destroyed, those with chronic respiratory disease (e.g.: cystic


85


fibrosis) in whom the normal clearance mechanisms are impaired, those who are

immune suppressed, these with neutrophil counts of less than 500/ μl and those with

indwelling catheters. It causes 10-20% of hospital-acquired infections.

Clinical findings

Pseudomonas aeurginosa a can cause infections virtually any where in the body, but

urinary tract infections, pneumonia and wound infections especially burns

predominate. From these sites, the organism can enter the blood, causing sepsis,

patients with Pseudomonas aeurginosa sepsis have a mortality rate of over 50%.

Sever external otitis and other skin lesions occur in users of swimming pool and hot

tubs in which the chlorination is inadequate.

Treatment

Because Pseudomonas aeurginosa is resistant to many antibiotics treatment

must be tailored to the sensitivity of each isolate and mentioned frequently resistant

strains can emerge during therapy. The treatment of choice is penicillin e.g. ticarcillin

or pipercillin and amino glycoside e.g. gentamycin or amikacin.

Prevention

Prevention of Pseudomonas aeurginosa infections involves keeping neutrophil

counts above 500% ml, removing indwelling catheters promptly, taking special care

of burns and taking other similar measures to limit infection in patients with reduced

host defenses.


86


Escherchia Coli 192,193

E-coli is the G (-) ve rod species. It is most abundant facultative anaerobe in

the colon and feces. E-coli occurs in the lower portion of the intestines of humans and

warm-blooded animals, where it is part of the normal flora. Some strains can cause

gastroenteritis and others can cause urinary tract infections.

Pathogenesis

E-coli has several clearly identified components that contribute to its ability to

cause disease viz pili, capsule endotoxin and 2 hexotoxins (enterotoxin)

Intestinal tract infection

First step is the adherence of organism to the cells of jejunum and ileum by

means of pili that protrude from the bacterial surface. Once attached, the bacteria

synthesize enterotoxin (exotoxins that act in the enteric tract), which act on the cells

of the jejunum and ileum to cause diarrhea. The enterotoxin-producing strains do not

invade the intestinal mucosa but some strains of E-coli are entero pathogenic and

cause diarrhea by invasion of the epithelium of large intestine causing bloody

diarrhea.

Systemic infection

The other two structural components, the capsule and the endotoxin, lay a

more prominent role in the pathogenesis of systemic rather than intestinal tract,

diseases. The capsular polysaccharide interferes with phagocytosis, enhancing the

organism’s ability to cause infections.

Clinical findings

E-coli causes a variety of disease both within and outside the intestinal tract. It

is the leading cause of community acquired urinary tract infections. Ascending


87


infection into the bladder is common in women. On the whole gastroenteritis, cystitis,

pyelonephritis, neonatal meningitis, hospital acquired sepsis are most common

manifestations of E-Coli.

Treatment

Treatment of E- coli infections depends on the site of disease and the

resistance pattern of specific isolate.

Lower urinary tract infection can be treated for just 1-3 days with and oral

sulfonamide or oral penicillin e.g.: ampicillin. E- coli sepsis requires treatment with

parenteral antibiotics (e.g.: third generation cephalosporin such as cefotaxime)

In neonatal meningitis combination of ampicillin and cefotaxime is given. Antibiotic

therapy is not indicated in E-coli, as diarrheal diseases are self-limiting.

Prevention

There is no specific prevention for E-coli infections, such as active or passive

immunization. However various general measures can be taken to prevent certain

infections caused by E-coli, for ex: the incidence of urinary tract infections can be

lowered by the judicious use and prompt withdrawal of catheters. Some cases of

sepsis can be prevented by prompt removal of switching the site of intravenous lines,

caution regarding uncooked foods and unpurified water, while traveling in certain

countries is also advisable.

Fungi 194

Fungi are large diverse group of heterotrophic organisms that exist as

saprophytes parasites or commensals most of them are found over decaying organic

material and in the soil202. Fifty thousand to one lakh species are known, though all

are not human pathogens.


88


Fungi lack chlorophyll and are eucaryotic organisms. They have a diversity of

morphological appearances depending on the species. They reproduce both sexually

and asexually. Fungi comprise molds and yeasts. Yeasts grow as single cells that

reproduce by asexual budding. Molds grow as long filaments (hyphae) and form mat

(mycelium).

Several important fungi are thermal dimorphic i.e. they form different

structures at different temperatures. They exist as mold in saprophytic free-living state

at ambient temperature and as yeast in host tissues at body temperature.

Most fungi are obligate aerobes, some are facultative anaerobes but none are

obligate anaerobes. All fungi require a preformed organic source of carbon, hence

their frequent association with decaying matter. The natural habitat of most fungi is

therefore the environment. An important exception is Candida albicans that is a part

of normal flora.

Candida Albicans195, 196

Candida albicans is a oval yeast with single bud. It is part of the normal flora

of mucus membranes of the upper respiratory gastro intestinal, and female genital

tracts. In tissue it may appear as budding yeasts or as elongated budding

“pseudohyphae” It is not usually found in non-living habitat. It measures 2-6 mm×3-

9mm in size.

Apart from Candida albicans, the genus Candida includes over 100 species,

most of which are neither commensals nor parasites in man.

Transmission

As a member of the normal flora, it is not transmitted.


89


Pathogenesis and Clinical findings

When local or systemic host defenses are impaired, disease may result

overgrowth of Candida albicans in the mouth produces white patches (thrush), vulva

vaginitis with itching and discharge is favored by high pH, diabetes or use of

antibiotics. Skin invasion occurs in moist areas, which become red and wheeping.

Fingers and nails become involved when repeatedly immersed in water; persons

employed as dish washers in restaurant and institutions are commonly affected.

Thickening or loss of the nail can occur. It seems that predisposition factors such as

other diseases, physiological disorders, obesity, alcoholism, prolonged use of broad-

spectrum antibiotics and steroids can create conditions in which Candida albicans can

cause disease. This makes the fungus an opportunistic pathogen.

80% of normal individuals may show Candida albicans as a saprophyte with

colonization of Oropharynx, GIT tract and Vagina.

Laboratory Diagnosis

In exudates of tissues, budding yeasts and pseudohyphae are seen

microscopically. Such specimens grow typical yeasts when cultured. Germ tubes form

in serum at 370C, which serves to distinguish Candida albicans from most other

Candida species.

Treatment and prevention

Treatment of local infections e.g.: thrush, consists of oral or topical antifungal

drugs, e.g.: clotrimazole or nystatin.

The mucocutaneous candidiasis can be controlled by ketaconazole. Treatment

of disseminated candidiasis consists of either amphotericin B, with or without

flucytosine or ketaconazole. Treatment of candidial infections with antifungal drugs


90


should be supplemented by reduction of predisposing factors. Certain candidal

infections such as thrush can be prevented by oral clotriamazole or nystatin.

There is no vaccine for prevention of candidal infections.

Aspergillus Flavus 197

Aspergilli are widespread in nature, being found on fruits, vegetables and

other substances that may provide nutriment. There are about 900 species in genus;

only a few species are constantly associated with human disease some species are

involved in food spoilage. Aspergillus species exist only as molds, they are not

dimorphic.

Transmission

These molds are ubiquitous and can be isolated from all environments. They

grow on decaying vegetation.

Pathogenesis and clinical findings

Aspergilli grow in high concentrations of sugar and salt, indicating that they

can exact water required for the growth from relatively dry substances.

Aspergillus flavus growing on cereals or nuts produce aflatoxins that may be

carcinogenic or acutely toxic. Aflatoxins are coumarine derivatives are identified as 6

kinds B1, G1, B2, G2, B2a and G2a in decreasing order of toxicity.

Aspergillus flavus that cause liver damage and tumors in animals and are

suspected of causing hepatic carcinoma in humans. Alfatoxins are ingested with

spoiled grains and peanuts and are metabolized by liver to the epoxide, a potent

carcinogen.

There is no specific means of prevention.


91


Antimicrobial Activity

Microorganisms are not visible to naked eye but they are present in soil, air,

water and food. Among them some microorganisms are ubiquitous in distribution.

Some are harmless and useful, some other are harmful to mankind198. It is necessary

to control virulent microorganisms from our living environment, as they may cause

diseases. Some agents destroy the microbes, where as others only inhibit their growth.

In general these agents are called antimicrobial agents. The antimicrobial agents may

be naturally occurring or may be synthesized. Selection of these agents from the

literature were made on the basis of their use in the treatment of infectious diseases

such as diarrhea, dysentery, skin diseases etc.

To evaluate the efficiency of these agents for their antimicrobial activity

different scientific procedures are established. Antimicrobial activity is a process by

which response of an organism to a drug or a crude extract can be evaluated.

Screening Methods for Antimicrobial Agents

The inhibition of microbial growth under standardized conditions may be

utilized for demonstrating the therapeutic efficacy of different medicinal drugs. Any

change in the antibiotic molecule, which may not be detected by chemical methods,

will be revealed by a reduction in the antimicrobial activity and hence microbiological

assays are very useful for resolving doubts regarding possible change of potency of

antibiotics and their preparations. The microbiological assay is based upon a

comparison of inhibition of growth of bacteria by measured concentrations of

antibiotics to be examined with that produced by known concentrations of a standard

preparation of the antibiotic having a known toxicity. Many methods are employed


92


for the evaluation of antimicrobial (antibacterial and antifungal) activity, which are

below mentioned.

Techniques199

Cup plate method.

Serial dilution method.

Solid dilution method.

Ditch plate technique.

Gradient plate technique.

Disc diffusion method.

1. Cup plate method

In this technique the test solution is placed in contact with agar which is

already inoculated with test organism. After incubation zone of inhibition are

observed. The test solution may be placed in a small cup sealed to agar surface with a

sterile cork borer of 8mm diameter.

2. Serial dilution method

In the serial dilution technique, the graded doses of test substances are

incorporated into broth and the tubes inoculated with test organism are incubated. The

lowest concentration at which no growth occurs is taken as minimum inhibitory

concentration.

3. Solid dilution method

In this method the dilutions of the substance under test are made in agar

instead of broth. The agar containing the substance under test subsequently poured

into a petri dish, then incubated and observed for any failure of growth. It has the

advantage for any one concentration of the test substance, several organisms may be

tested.


93


4. Ditch plate technique

This test allows a single compound or formulation to be tested against a range

of organisms. A solution of the compound or compound mixed with agar or a

semisolid formulation is placed in ditch cut in a nutrient agar plate. Various test

organisms are then streaked across the agar at right angles to the ditch. After the

suitable period of incubation at relevant temperature, the extent of inhibition is noted.

The ditch plat test allows spectrum of a compound to be obtained comparatively

quickly. Its main use similar to that of agar cup test for semisolid formulations, e.g.

Creams, ointments and powders.

5. Gradient plate technique

In this technique the concentration of drug in an agar plate may be varied

infinitely between zero to a given maximum. It consists of an agar plate with two

layers of agar. The nutrient agar is melted and mixed with the test solution and the

mixture poured into a sterile Petridish and allowed to set in the form of wedge. A

second amount agar is poured on to the wedge and allowed to set with the Petridish

flat on the bench. The plates are incubated overnight to allow diffusion of drug and to

dry the surface. The test organisms must be streaked in a direction running from the

highest to the lowest concentration. In this way up to six organisms may be tested.

6. Disc diffusion method

This is very simple and effective method to evaluate antimicrobial activity of a

drug. The principle of it is to allow the drug to diffuse through a solid medium. The

concentrations of drug being highest near the site of application of drug and

decreasing with distance. Whatmann filter paper discs of 6mm in diameter are

charged with required concentration of drugs and are stored dry in the cold. They may

be prepared in the laboratory or purchased commercially.


94


Pure bacterial and fungal cultures are inoculated into nutrient agar individually

and poured into sterile petriplates. After proper solidification prepared disc are

applied with sterile forces on agar media. Then plates are incubated for specified time

at particular temperature. Then the degree of sensitivity is determined by measuring

the zone of inhibition. Growth of microbes will be inhibited around discs containing

antibiotics to which bacterium is susceptible zone of inhibition are measured to

nearest whole millimeter using sliding calipers, ruler.

Culture Media200

Culture media gives artificial environment stimulating natural conditions

necessary for growth of bacteria. By appropriate procedures they have to be grown

separately on culture media and obtained as pure cultures for study.

Requirement of Culture Media

For the microbial growth certain consumable and suitable environmental

factors are required. The consumables represent the essential food or nutritional

requirements. They include sugars, starches, protein, vitamins, trace elements,

oxygen, carbon dioxide and nitrogen. The main environmental determinants of

microbial growth are PH and temperature.

In short the requirements for the microbial growth are listed as below:

Energy source

Carbon source

Nitrogen source

Salts like sulphates, phosphates, chlorides and carbonates of sodium,

potassium, magnesium, ferric, calcium, and trace elements like copper etc.

Satisfactory pH, 7.2 to 7.6

Adequate oxidation


95


Characteristics of ideal culture medium

♦ Must give a satisfactory growth from single inoculum.

♦ Should give rapid growth.

♦ Should be easy to grow.

♦ Should be reasonably cheap.

♦ Should be easily reproducible.

Classification of Media201

For the culture of microbes many culture media have been devised. They are

represented below in three sets:

I Set

Solid media

Liquid media

Semisolid media

II Set

Simple media

Synthetic media or defined media

Complex media

Semi defined media

Special media

Special medias are further divided as under

♦ Enriched media

♦ Enrichment media

♦ Selective media

♦ Indicator and differential media

♦ Sugar media

♦ Transport media

III. Aerobic media and Anaerobic media


96



97

Choice of Culture Medium

Each kind of microorganism has specific growth requirements. Many

microorganisms can be grown on a culture medium in the laboratory. Some microbes

can grow in a medium containing only inorganic compounds where as others require a

medium containing organic compounds (amino acids, sugars, vitamins) and other

erves as a very complex

anisms.

contain interfering or competing materials.

eat extract, sodium chloride and water. Nutrient broth is used

olidif

o, has not been replaced by any other agent. Agar is as important now as

it was t

require complex natural substances. (Peptone, yeast, blood cells or blood serum).

Some organisms cannot be grown in an artificial laboratory medium and can

be propagated only in a living host or cells. The host s

medium for such nutritionally demanding microorg

In general, it is ideal to use simple media because

1) They tend to have less batch-to-batch variation.

2) Less likely to

Simple Media

It is also called basal media. An ideal example is nutrient broth.

It consists of peptone, m

to culture the bacterias.

S ication Agent202

Agar is introduced, as a solidifying agent in microbiological media just about

100 years ag

hen.

The earliest solid medium was cooked cut potato used by Robert koch. This

proved unsatisfactory for variety of reasons. He tried gelatin as a solidifying agent,

but it was not suitable as gelatin is liquefied at 240C and also by many proteolytic

bacteria. A German housewife, “Faraultesse”, provided the solution to this problem in



98

1883. H r hus

f

elts at 980C and, usually sets at 420C

ow more slowly than bacteria. But to avoid the possible bacterial

ontam

xtrose agar media is suitable for fungal culture as it has got low pH

rias.

potato, dextrose, Agar and distilled water. pH maintained at

be expressed

in following points.

• prepared and incubated.

e band was one of the investigators in Koch’s laboratory. She suggested

her husband an use of agar, who had seen her mother using it for making jellies.

From then agar is universally used for solidification. Agar is obtained

from some types of seaweeds. Its chief constituent is a long chain polysaccharide. It

also contains verifying amounts of inorganic salts and small quantities of a protein

like substance. It has virtually no nutritive value and is not affected by growth o

bacteria. Its unique property is that it m

depending on agar concentration. Approximately 2% agar is applied for solid media.

Media used for Fungal Cultures

For the cultivation of fungi a medium bearing acidic pH has to be selected. It

facilitates the growth of the fungi but is not optimal for the growth of bacteria.

Fungi can be cultivated by the same general culture methods used for bacteria.

Most of them gr

c ination, it is good practice to use specific media for fungal culture viz “ Potato

Dextrose Agar”

Potato de

and a relatively high concentration of sugar tolerated by fungi but is inhibitory to

many bacte

It consists of

5.6 to 5.8

Procedure activity

To summarize, the whole procedure of antimicrobial activity can

Fresh cultures of selected organisms are

Bacteria : 37 C for 18 to 24 hrs 0



99

Fungi : 270C for 42 to 72 hrs.

• Discs of standard and trial drug are prepared.

• The nutrient agar medium prepared and sterilized.

• The sterile medium was cooled to 450C and mixed with 20% of bacterial and

ia was shaken well and poured to respectively labeled sterile petri

fication; standard and trial discs prepared are applied over agar

were kept in a refrigerator for 2hrs for the diffusion of the drug into

hrs .

• Fungi are kept at room temperature for 42-72 hrs.

• The plates were observed e of inhibition and measured in

mm.

comes under Cephalosporin

nd their chronological development. The

fungal culture individually (i.e. 80ml media and 20ml culture).

• The Med

plates.

• After solidi

surface.

• The plates

the media.

• After 2 hrs, plates of bacterial culture were kept in an incubator at 370C for 18

to 24

for appearance of zon

Cefotaxime203, 204.

Antibiotic is a chemical substance derived from or produced by a living

organism, which is capable in small concentrations of inhibiting the life processes of

microorganisms. Cefotaxime is an antibiotic, which

group. Cephalosporins are group of semi synthetic antibiotics which possess a wide

range of activity against G(+)ve and G(-)ve bacterias.

These are safe antibiotics derived from Cephalosporin Cremonium that is

obtained from marine fungus. The nucleus of Cephalosporins resembles that of

penicillin. Cephalosporins have been conventionally divided into 4 generations based

on their potency, antibacterial spectrum a

first generation compounds begat the second generation compounds which begat the

third generation & so on.



100

Cefotaxime is a third generation compound and is less active against Gram

(+)ve bacterias than those of the second generation but more active against G(-)ve

ents protein synthesis in bacterial cell wall.

enal excretion is remarkably reduced in renal insufficiency.

halosporins, Cefotaxime is partly metabolized by the liver before its

uscular injections are painful, irritation may be present at the site of injection.

us injections rarely may cause thromboplebits, allergy, fever, chills, and

rash.

Total dose should not exceed 4gms and it is available as tablet and injection

nging from 125 mg to 1gm.

bacterias. Bactericidal in action and it prev

It has got variable susceptibility against Pseudomonas.

Absorption, Distribution and Excretion

Cephalosporins are administered either orally or intravenously or through intra

muscular route. Intramuscular administration is painful. Their distribution in the body

is similar to that of penicillin but the concentration in the eye and CSF is poor.

Eliminated mainly through renal excretion and high concentrations are achieved in

urinary tract. Probenecid administered concomitantly by oral route does not have any

adverse effects. The r

Unlike other Cep

excretion by kidneys.

Adverse effects

In general Cephalosporins are well tolerated.

Local reactions

Intram

Intraveno

Dosage

ra



101

l use and infusion for intravenous use.

ectrum antifungal activity.

and body. The drug has longer half-life thus

rmitt

proximately 10 times higher in

concentration after a single dose.

rrent cryptococcal meningitis

nd mucocutaneous candidiasis in patients with AIDS.

Fluconazole205

Fluconazole is a triazole derivative, which is effective both orally and through

intravenous route. It penetrates readily into the CSF. It is used in the treatment of

local and systemic candidiasis, cryptococcal infections including meningitis. The drug

may cause nausea, gastrointestinal disturbances and abnormalities of liver enzymes. It

is water soluble and available as capsule for ora

It possess a broad sp

Pharmacokinetics

The drug is highly water-soluble and is well absorbed through oral route. The

drug penetrates well into tissues

pe ing its single dose regimen.

Fluconazole shows a low affinity for protein binding and 80% is excreted

unchanged in the feces and urine. It rapidly penetrates CSF. Pharmacokinetic studies

demonstrate that the Fluconazole concentration is ap

CSF than the serum

Therapeutic uses

It is very effective in dermatophytic infections and in cutaneous and

oropharyangeal candidiasis. It is also effective in deep mycoses. In addition it

prevents the development of fungal infections in individuals predisposed to such

conditions. Fluconazole is the drug of choice for recu

a



102

Dose regimen

A dose of 150mg, once in week for 4-6 weeks is quite effective in most of the

dermatophytic infections and in cutaneous candidiasis, vaginal candidiasis and

osthitis. Single dose of 150mg is effective.

nts with renal impairment.

onazole resistance has also been reported.

80% reaches the CSF). It has a longer half-life

No. erties of

No istics

candidial balanop

Adverse effects

The common side effects are nausea, headache, pain in abdomen and diarrhea.

Fluconazole is contraindicated in pregnancy and recent studies shows that even a

single dose taken by a pregnant women can lead to birth defects in infants. The dose

has to be regulated and should be decreased in patie

Recently, Fluc

Advantages

The advantages of Fluconazole over other antifungals are that it has higher

water solubility, good oral absorption, approximately 90% bioavailability and crosses

the blood brain barrier readily (

permitting a single dose regimen.

Table 29 Showing prop Fluconazole

Sl. Character Fluconazole 1 Antifungal spectrum Broad 2 Solubility Readily soluble in water 3 Absorption Effective absorption 4 IV Administration ible and available Poss5 Nephrotoxicity incidence Nil 6 Anaemia Nil 7 Leucopenia Nil 8 Gastro intestinal upset Mild 9 Overall toxicity Low

Pharmaceutical Study

Methodology

Methodology adopted in the present study includes following headings.

1) Pharmaceutical study

2) Analytical study

3) Experimental study


Pharmaceutical study means the practical experience of preparing medicines

from raw drugs. Practical experience is most essential for vaidya as described in

Rasaratna Samucchaya (RRS 6/4) that Rasa shastrajna must have the quality of

Kushala Rasa Karamani.

Rasashastra is a science, which mainly deals with minerals and metals. These

minerals cause some toxic and untoward effects if are not properly processed. Hence

preparation of mineral drugs requires more skill and only way of obtaining skill is

through repeated practicals and careful observations during the process.

This section deals with identification, selection and processing of raw drugs

and preparation of Rasapushpa, which is Nirgandha Kupipakwa preparation explained

in Rasatarangini (6/29-31).

Study design

A detailed and clear description of steps taken to prepare the trial drug

Rasapushpa is being put under following headings:

Step 1 : Identification and collection of raw drugs

Step 2 : Shodhana of raw drugs

Step 3 : Preparation of Rasapushpa.


103


Method

Step 1 : Identification and collection of raw drugs

Proper identification and collection of raw drugs are need of the hour for the

Ayurvedic formulations by which quality of the medicine can be assured. Rasapushpa

comprises following ingredients:

♦ Hingulotha Parada

♦ Kasisa

♦ Saindhava Lavana

Special request was made to the local herbomineral drugs shop dealer to get

the particular quality raw drugs and those were screened for classical grahya and

agrahya laxanas and were certified by concerned departments.

Step 2: Shodhana of raw drugs

Shodhana is the process, which makes metal and minerals fit for therapeutic

use by eliminating toxic substances present in the drug. Shodhana is done by many

methods viz Mardana, Bhavana, Swedana, Nirvapa etc with particular vanaspati

dravya swarasa or kwatha etc. it is necessary to increase therapeutic efficiency of

drug, hence proper shodhana of ingredients used in Rasapushpa preparation was

conducted.

Practical No 1

Name of the practical : Hingula Shodhana

Reference : Rasatarangini 9/16-17

Date of Commencement : 5-8-2005

Date of Completion : 19-8-2005

Materials : Hingula - 500gms, Nimbu swarasa -QS

Method : Bhavana

Equipment : Khalavayantra, Juice extractor.


104


Procedure

• Hingula shodhana was done by giving 7 bhavanas with Nimbu swarasa.

• 500 gm of Hingula was taken and finely powdered in Khalva yantra.

• Required quantity of Nimbu swarasa was extracted from lemons with the help of

juice extractor.

• For the first Bhavana, 170ml of Nimbu swarasa, the quantity sufficient to

immerse Hingula was added.

• It was subjected for continual and cautious mardana till powder completely

absorbs the swarasa. Mardana has to be continued till Hingula dries up

completely & becomes powder again. This completes one bhavana.

• Like this bhavana was repeated for another six times taking fresh swarasa each

time.

Observations

• For first bhavana the quantity of Nimbu swarasa required was quite more than

the subsequent bhavanas.

• The Hingula was solid in form, and red in color with glistening

white/mercurial lines.

• It took 30 minutes to powder the block of Hingula and it possessed glistening

particles at the initial stage of mardana.

• Glistening particles disappeared at the end of 30 minutes and Hingula was

finely powdered.

• The clump of Hingula was comparatively dull in color but brilliant red color

could be appreciated only after it was finely powdered.


105


Table No. 30 Observations made during Hingula shodhana.

No. of Shodhana Day Quantity of swarasa in ml Time taken

01 1 175 ml 8

02 3 170 ml 8

03 5 165 ml 7.45

04 7 165 ml 8

05 9 165 ml 8

06 11 165 ml 8.30

07 13 165 ml 9

• After completion of 7 bhavanas, Hingula was taken out from the khalva and

washed in a steel vessel with water thoroughly and allowed to settle.

• Washing of Hingula was done for another 2 times

• Settling of Hingula at the bottom took 6 hours after which the water was

decanted.

• Totally it took 17 days for Hingula shodhana.

Precautions

• Khalva yantra was clean and dry before carrying out the procedure.

• Hingula was finely powdered before adding Nimbu swarasa.

• Bhavana dravya i.e Nimbu swarasa was just sufficient to immerse the powder

of Hingula so as to avoid more liquidity resulting in spilling of the drug from

Khalva.

• Mardana was carried out cautiously allowing peshani to move entire length of

Khalva yantra.

• At the end of each bhavana, mardana was done slowly as the material becomes

stickier.


106


• When the material was completely dried up by mardana then only it was

considered as the completion of one bhavana and fresh swarasa was added for

next bhavana.

• After the completion of 7 bhavanas, Hingula was washed with water until it

loses its snigdhata and amlatva and attain ujjwala varna.

Result

Table No.31 Showing the results of Hingula shodhana

Initial weight of Hingula 500 gms

Final weight of Shodhita Hingula 550 gms

Weight after prakshalana 530 gms

Total weight gain 30 gms

Causes of weight gain

Due to the addition of solid contents present in Nimbu swarasa.

Practical No. 2

Name of the practical : Preparation of Hingula Chakrikas.



Materials : Shodhita Hingula – 500gms,

Nimbu swarasa - 130ml

Method : Bhavana

Equipment : Khalvayantra

Procedure

• 500 gms of Shuddha Hingula taken and finely powdered in khalva yantra.

• Then 130ml of Nimbu swarasa added and mardana was done.

• Mardana was continued till the proper consistency was obtained.

• Then chakrikas were prepared and dried in shade.


107


Observations

• It took 5 hrs of mardana to get the proper consistency.

• Chakrikas dried completely in 20 days.

• The average measurement of chakrikas

Thickness – 3.5 mm

Circumference – 3-31/2cm

Precautions

• Clean and dry Khalva yantra was taken.

• Mardana was done carefully to avoid the spillage of content from khalva

yantra.

Results

Initial weight of Hingula : 500gms

Weight of Chakrikas : 480gms

Cause of weight loss

• Due to the adherence of Hingula to Khalva yantra.

• While preparing chakrikas.

Practical No. 3

Name of the practical : Hingula Satwapatana



Reference : Rasatarangini 5/38,39

Materials : Hingula chakrika – 240gms

Method : Urdwapatana

Equipment : Damaruyantra, Gas stove, cloth


108


Procedure

Whole procedure was divided into 3 stages.

1. Preparation of Chakrikas.

2. Preparation of Urdwapatana yantra

3. Parada Nishkasana.

Preparations of Chakrikas:

Same as Practical No. 2

Preparations of Damaruyantra

Materials required – Two earthen pots of equal size

Cloth – 4 x 60cms, Multani mrittika.

Method

Mouth surface of two pots was rubbed on a smooth stone with sand to make

the facing surfaces of mouth even.

Dried chakrikas of shodhita Hingula was weighed as 240gms and were kept in

a lower pot.

This pot was covered with another pot of same size.

The gap left at the union of two mouths of pot was with multani mrittika

smeared thread.

Then sandhi bandhana was done with cotton cloth strip smeared with multani

mrittika and it was allowed to dry for a day.

Parada Nishkasana

When sandhi bandhana dried i.e. on next day Damaruyantra was kept on gas

stove and heat was given continuously for 8 hours.

While heating cold pad was maintained on the upper pot for condensation of

sublimed parada.


109


Temperature was maintained at 350o –400oC

After swangasheeta, Damaruyantra, sandibandhana was carefully removed.

Parada, which was sublimated on upper pot was collected by doing

prakshalana with hot water.

After prakshalana, parada was filtered through a clean cloth and was collected

in a clean and dry glass jar.

Observations

Table No. 32 Temperature recorded during the procedure.

Time Temperature

0 Hours 300C

1st Hour 1500C

3 Hours 2800C

5 Hours 3600C

7 Hours 3800C

8 Hours 4050C

• Temperature was recorded with the help of pyrometer at regular

intervals.

• After one hour of agni smell of Gandhaka was noticed.

• After opening of Damaruyantra, globules of mercury were seen

adhered to the upper part of the pot.

• Chakrikas of Hingula in the lower part was completely burnt.

• Mercury obtained was very much shining.

Precautions

o The upper pot was maintained with cold pad to ensure proper condensation of

Parada.


110


o Sandibandhana was properly done without leaving a gap at the junction of two

pots.

o The water has not wet the sandibandhita area.

Results

Total time taken : 3days

Weight of Chakrikas : 240 gms

Weight of Parada extracted : 135 gms

Note: The same extraction procedure was followed for another sample.

Practical No. 4

Name of the practical : Hingula Satwapatana.



Same as practical No. 03

Results

Total time taken : 2 days

Weight of chakrikas of Hingula : 240

Weight of parada obtained : 130

Practical No. 5

Name of the Practical : Parada shodhana

Reference : Rasatarangini 5/40,41


Date of completion : 17-11-2005

Materials : Hingulotha Parada : 250gms

Haridra: 16 gms

Method : Mardana



111


Procedure

16 gms of Haridra taken and finely powdered in Khalva yantra.

250gm of Parada was added and subjected for continual and cautious

mardana.

After 30 minutes of mardana, little quantity of Parda split up into small

particles.

After 1 hour of mardana yellow color of Haridra turned into pale

brown color.

As mardana continued pale brown color gradually depended.

Mardana was done for 2 days. Each day 8 hours mardana is conducted.

Then on 3rd day Parada is squeezed out with the help of cloth.

Observations

• A little quantities of Parada disintegrated into small particles.

• After one hour of mardana yellow color of Haridra changed to brown

colour which become more deepened with mardana.

• Haridra was shiny in appearance.

• Parada secured after 2 days of mardana

• It was very bright in appearance.

Precautions

o Khalva yantra was clean and dry before carrying out the procedure.

o Haridra was finely powdered before adding parada.

o Mardana was done cautiously so as to avoid spilling of Parada.

Result

Initial weight of Parada : 250 gms

Final weight : 245 gms


112


Practical No. 6

Name of the practical : Kasisa Shodhana

Reference : BrihatRasaRajaSundara (28.chpt)

Date of commencement : 21-11-2005


Materials : Kasisa: 300 gms

Nimbu swarasa: 100 ml

Method : Bhavana.

Apparatus : Khalva yantra.

Procedure

Kasisa was taken in Khalva yantra and finely powdered.

100ml of Nimbu swarasa was taken in measuring flask and was poured slowly

into Khalva yantra containing Kasisa churna.

Kasisa churna was completely immersed in Nimbu swarasa.

Then mardana was done continuously and cautiously till Kasisa completely

absorbs the swarasa and becomes dried.

Observations

• Ashodhita Kasisa was bluish green in colour, lustrous, and crystalline in nature.

• After powdering Kasisa became lusterless.

• It took 25 minutes to powder the kasisa finely.

• On After 2 hours of mardana Kasisa turned into viscous mixture.

• Mardana was done for 10 hours.

• After shodhana Kasisa become whitish green in colour.


113


Precautions

o Before adding Nimbu swarasa, Kasisa was finely powdered.

o Mardana was done slowly to avoid loss of Kasisa by spilling out.

o After shodhana, Kasisa dried well and weighed.

Table No. 33 Physical examination of Ashodhita and Shodhita Kasisa

Observation Ashodhita Shodhita

Consistency Shining crystals Fine powder lusterless

Color Bluish green Pale green

Touch Hard, solid, rough Soft, fine

Smell Slightly metallic Smell of Nimbu swarasa

Result

Quantity of Kasisa : 300 gms

Quantity of Kasisa obtained : 275 gms

Loss of weight : 25 gms

Causes of weight loss

Some amount of Kasisa adhered to Khalva yantra.

Some amount of Kasisa spilled out during mardana.

Practical No. 7

Name of the practical : Preparation of Kupi



Materials : Amber colored glass bottle, cloth,

Multani mud, Scissors.


114


Method

A clean and dry bottle was taken and paste of multani mud was applied to the

base so as to level the concavity of the bottom.

A cloth smeared with paste of multani mud measuring 12 cms in width and 64

cms in length was applied over the kupi in such a way that both ends of cloth

come at opposite side of the neck of the kupi.

Such a type of smearing with the mud-cloth and at a stretch covering whole

kupi helps in uniformity of mrutkapata and was allowed to dry completely.

In this way all the 7 layers were applied.

Observation

• Fine powder of multani mud helps in preparing uniform smooth surface of

Kupi.

• The color of cloth was entirely changed to cream colour after it was smeared

with mud.

• The drying of each covering took one day.

• Measurement of cloth was remained same for all the 7 layers.

• In initial covering, the bottle was thin, as the layers were applied it became

thick.

Precautions

o The selected bottle was clean and dry.

o Finely powdered multani mud was used.

o Paste of the mud was prepared by adding little by little water as per

requirement.

o The bottle was completely allowed to dry after each covering.


115


Practical No. 8

Name of the practical : Preparation of Prematerial

Reference : RT 6/29



Materials : Hingulotha Parada : 240gms

Shodhita Kasisa : 240gms

Saindhava lavana : 240gms

Method : Mardana


Procedure

Hingulotha Parada, shodhita Kasisa and Saindhava lavana were taken in equal

quantity and mardana was done in Khalva yantra for 48 hours.

As mardana continued, Parada was split up into smaller and smaller particles.

Daily 8 hours of mardana was done and it took 6 days for completion of this

procedure.

Observations

Laxanas of Prematerial were observed only after 48 hours of mardana.

At the end of 5 hours whole mixture became very fine powder.

The consistency of Prematerial was kajjalabha i.e. very soft and smooth except

the kajjala varna.

When Prematerial was rubbed between the fingers it filled the furrows of the

finger (rekhapurnatwa)


116


For confirmation of nischandrata, a little pinch of Prematerial was added to a

drop of water on the palm and rubbed gently and observed in sunlight. No free

particles of mercury were traced in Prematerial.

Precautions

o Clean and dry Khalva yantra was used.

o Uniformity of mardana was maintained through out the procedure.

o Mardana was done carefully to avoid spillage of fine powder of

Prematerial.

Results

o Initial weight of ingredients : 720gms

o Weight of Prematerial : 685gms

o Loss of weight : 35gms

Causes of weight loss

• Spilling of Prematerial during mardana.

• Some particles of Prematerial adhere to Khalva yantra which becomes

difficult to collect.

Practical No. 9

Name of the practical : Preparation of Rasapushpa

Reference : RT 6/29-31



Materials : Prematerial 100gms

Method : Kupipakwa Method (Bahirdhuma

Vidhi)


117


Equipment : Mritlepita kachakupi, Valuka yantra,

Loha shalaka, Cork, Copper foil,

Khalva yantra, Pyrometer with thermocouple.

Procedure

Whole procedure was divided into 3 phases.

1. Poorva karma

2. Pradhana karma

3. Paschat karma

1) Poorva karma

a) Preparation of Kachakupi

b) Preparation of Prematerial

c) Filling of Prematerial into Kachakupi

d) Placing of Kachakupi in Valuka yantra.

2) Pradhana karma

a) Heating schedule

b) Observation and Recording of Temperature

c) Corking, sealing of Kupi and self-cooling of Valuka yantra.

3) Paschat karma

a) Removal of Kupi from Valuka yantra

b) Breaking of kupi

c) Collection of Rasapushpa

1. Poorva karma

Preparation of kachakupi : same as practical No. 7

Preparation of Prematerial: same as practical No. 8


118


Filling of Prematerial and placing of kupi in valuka yantra

• 100gms of Prematerial taken and was cautiously filled into the kachakupi.

• Then valuka yantra was placed exactly at the center of gas.

• About 3 angulis of sand was spread uniformly within the valuka yantra.

Now kupi filled with Prematerial is kept over sand at the center with sand.

Then remaining part of yantra was filled with sand up to the neck of the

kupi.

• Care was taken while putting the sand as it may contaminate the ingredients

inside kupi for which it was duly corcked.

• The cork was prepared by ishtika exactly fitting to the mouth of kupi.

double layers of cloth smeared with multani mrittika was applied over cork

and dried.

Pradhana karma

• The Pooja was performed and Aghora mantra was chanted, after valuka yantra

was subjected to agni.

• The kupi in valuka yantra was heated for 6 hours in three stages of graded

heating i.e. mrudu agni, madhyamagni and teevragni.

• The temperature was recorded with the help of digital pyrometer by inserting

the thermocouple in valuka yantra; the tip was kept nearer to the bottom of

kupi.

• Heat was increased gradually and regulated as per requirement. Temperature

was recorded at every hour.

• During the procedure water vapours along with fumes were expelled out from

the kupi initially and as the time passed amount of vapours and fume

decreased.


119


• The temperature was maintained at 3 stages ranging from mrudu agni 100-

150oC for 2hours, madhyamagni – 150-250oC for 2 hours, tikshnagni 250oC –

350oC for 2 hours.

• After paka laxanas the corcking was done with the help of ishtika cork, and

was sealed with cloth smeared with multani mitti.

• The kupi paka laxanas noticed were:

o Complete cessation of water vapours

o The sticking of materials to cold shalaka was whitish.

• After crocking, the sand layer of 4 angulis surrounding the kupikantha was

moved aside.

• Later heat was given for 2 hours and temperature was maintained at 250o –

350oC.

• Vauklayantra with kupi was allowed to become swangasheeta.

Observations

The valuka yantra was kept on agni at 10am of 8th of February 2006 and room

temperature was 30oC.

The kupi was subjected to kramagni for 6 hours and specific observations

made mean while are represented in Table No. 34:

Table No. 34 Showing the observations made during the procedure

Time in hours Temp in C0 Specific observations 0 Hour 300C - 1st Hour 1100C Prematerial was moistened

Water vapours present Mild fumes present

2nd Hour 1600C Water vapours and fumes increased

3rd Hour 2200C Fumes decreased 4th Hour 2650C Water vapours absent 5th Hour 3100C - 6th Hour 3600C -


120


After one hour Prematerial was found moistened and was tested by sheeta

shalaka. Water vapours started, which gradually increased and was ascertained

by tamra patra (Copper coin)

Fumes also started in the same hour, which increased with time.

After four hours water vapours disappeared which was confirmed by keeping

Tamra patra at mouth of kupi.

Then sheeta shalaka was introduced into the kupi, which had the white

granular coating of compound on it.

The maximum temp, attained throughout the procedure was 360oC.

After 6 hours, agni was stopped and valuka was allowed to cool by itself.

Precautions

o The Prematerial was again done mardana for half an hour before filling into

the kupi.

o The kupi was kept exactly at the center of valuka yantra so that homogenous

temperature would be obtained.

o The maintenance of temperature was done carefully with the help of the

pyrometer.

o Steady rise in temperature was maintained throughout the procedure which

was in proportion with respect to kramagni.

o Care was taken while inserting shalaka so that it does not touch the bottom of

kupi.

o Corcking was done after complete cessation of water vapours and sheeta

shalaka test.

o After corcking sand at the neck portion was removed.


121


Paschat karma

♦ After complete cooling, kupi was taken out from valukayantra carefully by

removing the sand surrounding kupi.

♦ The layer of kapadamitti was removed by scraping with knife and the external

surface of kupi was cleaned with wet cloth.

♦ Jute dipped in kerosene was tied to the kupi 2-3 cm below the level of

sublimated product and ignited. When the whole thread was burnt. It was

wrapped by wet cloth.

♦ Kupi was broken into 2 halves with a breaking sound.

♦ From the neck region, Rasapushpa was collected and was stored in clean

airtight container.

Observations

The lower portion of outer layer of kupi was black in color.

The sublimated compound was whitish and shining when observed in sunlight.

No glass piece was seen along with medicine.

In the bottom residue, which was red in colour was found.

Precautions

o Scraping of layers of mrutkapata over the kupi was done carefully.

o Thread dipped in kerosene was tied in only one circle.

o No force was applied to break the kupi to avoid mixing of glass pieces with

the final product.

Result

Amount of Prematerial taken : 100 gms

Amount of Rasapushpa obtained : 10 gms

Amount of Residue obtained : 60 gms.


122


Note

o Total duration of Kupipakwa was 6 hours.

o Water vapours observed totally for 4 hours.

o Cold shalaka was inserted to access the state of Prematerial at different stages

of agni.

o For corking the mouth ishtika cork was used.

o Heat was increased in three successive stages.

Rasapushpa preparation was conducted for another three times with same

proportion of Prematerial.

Practical No. 10




Same as practical No.9

Results

Weight of the Prematerial : 100gms

Weight of Rasapushpa obtained : 15gms

Weight of Residue obtained : 65gms

Practical No. 11






123


Results




Practical No. 12





Results




Table No. 35 Showing the yield of Rasapushpa in different practicals

Practical No. Weight of

Prematerial

Weight of

Rasapushpa

Residiue left

9 100gms 10 gms 60 gms

10 100 gms 15 gms 65 gms

11 100 gms 18 gms 56 gms

12 100 gms 14 gms 60 gms


124

Graph Showing the Temperature and Time during Valuka Yantra Pachana of Rasapushpa

Temperature Hour Chart

30

110

160

220

265

310

360

200

100

5025

00

50

100

150

200

250

300

350

400

0 1 2 3 4 5 6 8 9 10 11 12

Time in Hours

Tem

pera

ture


125

Analytical Study

Analytical Study

The Rasoushadhies mentioned in Ayurvedic Pharmacopoeia should be

analyzed for physical and chemical properties to confirm the genuinity and safety

before administration to the patients. Hence it is essential to adopt modern analytical

methodology for better understanding and interpretation of physico-chemical changes

occurred during the process.

In the present study, Rasapushpa prepared according to Kupipakwa method is

collected and subjected to modern analytical methods in Bangalore test house at

Bangalore.

Some physical analysis is done at J.T. Pharmacy College, Gadag. The test

performed are Fineness of particle test, Flow rate of Rasapushpa, Acid insoluble ash,

Water soluble ash content and Organoleptic characters of Rasapushpa. Rasapushpa is

also accessed according to the Ayurvedic parameters, which is below mentioned.

Test: To test the genuinity of Rasapushpa, clean shiny iron pan is taken and a drop of

water put on it. Then pinch of Rasapushpa added on that drop. After 1 or 2 minutes

water drop is removed, there is no blackening at that site indicating that Rasapushpa is

pure one.

Analysis of Hingula

Before shodhita Hingula was subjected for Satwapatana process, it was

analyzed for mercury content present in it. It is also tested for organoleptic characters.

Organoleptic Characters

Colour : Red

Smell : Odourless

Touch : Fine


126

Analytical Study


127

Determination of Sulphur:

Eschka Mixture:

Mix two parts by weight calcined magnesia with one part by weight of

anhydrous sodium carbonate.

Procedure:

Cover the bottom of a 50ml crucible uniformly with 0.5g of Eschka’ s

mixture. Weigh accurately the appropriate quantity of the sample material and mix it

intimately with 2gms of Eschka’ s mixture and put evenly on the previously weighed

Eschka mixture. Level the contents by tapping gently on a bench. Cover this

uniformly with 0.5 gm of Eschka’ s mixture. Place the crucible into the cold muffle

furnace. Raise the temperature from room temperature to 8000C ± 250C in about one

hour and then heat for a 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

50ml of hot distilled water; add the washings to the contents of the beaker.

Add carefully sufficient quantity of concentrated hydrochloric acid to dissolve

the solid matter, warming the contents of the beaker to effect solution. Boil for 5

minutes to expel carbon dioxide. Add drop wise from a pipette; warm 5% of Barium

chloride solution. Stir the solution constantly during the addition. Allow the

precipitate to settle for a minute or two.

Then test the supernant liquid for complete precipitation by adding a few

drops of Barium chloride solution. If a precipitate is formed, add slowly a further 3ml

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

precipitating agent has been added, keep the covered solution hot, but not boiling for

Analytical Study

an hour (steam bath) in order to allow time for complete precipitation. The

precipitation should settle and a clear supernant liquid should be obtained. Test the

later 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 ( Whatman 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

ensure. 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 weigh is attained.

A blank is necessary. Calculate the percentage of sulphur converting Barium

sulphate X 0.1374.

Sulphur content present in shuddha Hingula :10.15%. w/w

Analysis of Rasapushpa

1) Organoleptic characters

Rasapushpa is white in colour, odorless and shiny fine powder.

2) Loss on drying at 1100C

2gms of Rasapushpa weighed accurately in a silica crucible and dried in a hot

air oven at 1100C till a constant weight is obtained. The difference in weight was

calculated and the result is attached.

Result: 0.34%


128

Analytical Study


129

3) Determination of Total Ash

Take about 2gm accurately weighed, ground drug in a previously tared 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 weigh. Calculate the percentage of

ash with reference to the air-dried drug.

Result: 0.4%

4) Acid Insoluble Ash

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

ash less 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: 0.23%

5) The fineness of particle test

It can be possible to use the ordinary microscope for particle size measuring in

the range of 0.2 micrometers to about 100 micrometers. According to microscope

method, the fine powder was sprinkled on the slide covered with covering slip and

placed on a mechanical stage. Initially standardization of micrometer was carried out

by coinciding with the lines of both ocular micrometer and 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. The particles are measured along the

Analytical Study


130

arbitrarily chosen fixed lines covered by the particles using the ocular micrometer.

The size of the particle was calculated using the standard value.

Result : Arithmetic Mean : 9.34 Micrometer.

6. Flow Property:

Rasapushpa, which is a very fine powder is subjected to flow property test i.e

“Angle of response” by which we can analyze goodness of flow property.

Angle of repose: It is the maximum angle that can be obtained between the

freestanding surface of a powder heap and the horizontal plane i.e. tan θ = 2h/D.

Where D is the diameter of the circle and “h” is the height of the powder heap.

This test involves the hollow cylinder, half is filled by Rasapushpa 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 : Angle of repose – 40.140.

7. Flow rate:

A simple indication of the ease with which a material can be induced to flow

is given by application of a compressibility index “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.

In this procedure, one measuring cylinder is taken and is filled with

Rasapushpa. The level of the Rasapushpa should be noted. Then at a height of 2 cm

continuous 10 tapping should be done after that the level of the Rasapushpa in the

Analytical Study

cylinder is once again noted and value “I” is calculated with respect to the V0 and V

value. If the value of “I” is below 15% usually having good flow rate.

Result : I = 30%

8. Determination of Mercury:

Procedure

Dissolve about 0.3gms of sample in 5ml of aquaragia and add 100ml of water.

Add 40ml of 0.05 NEDTA, 5ml of Solo chrome black Indicator. Titrate the solution

with 0.05 M zinc sulphate until the blue color changes to purple (do not overshoot the

end point). Add 3gm of potassium Iodide, swirl to dissolve. Allow standing for two

minutes, and then continuing the titration with zinc sulphate solution to the same end

point as before. Each ml of zinc sulphate solution required after addition of potassium

Iodide = 0.6103 Hg

Result: Percentage of Mercury: 84.3% in Rasapushpa.

Percentage of Mercury: 71.2% in Shodhita Hingula

9. Estimation of Chlorides

Weigh accurately appropriate quantity of sample and dissolve in sufficient

distilled water. Add 0.1 ml of potassium chromate and titrate the solution with 0.1 M

silver Nitrate until the color changes to reddish brown.

Calculate the percentage of chlorine in the sample. The factor for converting

silver chloride to chlorine is 0.03545.

Result: Percentage of Chloride: 13.6%

Solubility tests

Water : Practically insoluble

Chloroform : Sparingly soluble

Alcohol : Sparingly soluble.


131



There may be wide variations in the susceptibility of different strains of the

same bacterial species to antibiotics. Several technologies are now available for

determination of microbial sensitivity to antimicrobial agents. Some of them are cup-

plate method, disc-diffusion method, serial dilution method and gradient plate

method. In present context, disc-diffusion method is adopted to assay the

antimicrobial activity of Rasapushpa.

Antimicrobial activity was carried out at Pharmaceutical, Microbiology and

Biotechnology, Department, S.C.S. College of Pharmacy, Harapanahalli dated 08-05-

2006 to 15-05-2006.

Materials and methods

Materials

Drugs : Rasapushpa (Trial drug)

Cefataxime, Fluconazole (Standard drugs)

Test strains : Staphylococcus aureus, Steptococcus pyogenes,

E-coli, Pseudomonas aeurginosa,

Candida albicans, Aspergillus flavus

Adhesive Agent : Gum acacia.

Chemicals : Nutrient broth, Potato dextrose agar, Nutrient agar,

Equipment : Autoclave, Incubator, Conical flask, Petri plates, Test

tubes, Measuring cylinder, Syringe, Cotton,

Vernier calipers.

Method : Disc diffusion method


132


Disc Diffusion method:

This technique is simple to perform and relatively inexpensive. This test is

performed with the help of 8mm disc prepared of Whatman filter paper No.1. The

discs are impregnated with specific quantities of drug and applied on the surface of

agar plates, which is already inoculated with test organisms. After proper incubation,

zone of inhibition around the disc was determined.

Procedure:

All the chemicals used were of Highmedia Company, USA, and the

glasswares used for study were sterilized by following standard procedure.

Complete procedure was carried out in 3 stages for antibacterial as well as antifungal

activity of standard and trail drugs.

Stage I

♦ Preparation of inoculum

♦ Preparation of discs of different drug concentration (standard and trial)

Stage II

♦ Preparation of agar media.

♦ Inoculation of test organisms

♦ Application of discs.

♦ Incubation

Stage III

♦ Reading of zone of inhibition

♦ Interpretation of Results.


133


Preparation of Inoculum

Two G(+)ve and two G(-)ve bacterias were chosen for the study.

Bacterias selected for activity are as mentioned below:

Gram Positive bacterias

1) Staphylococcus aureus MTCC 87

2) Streptococcus pyogenes MTCC 32

Gram Negative Bacterias

1) E- coli MTCC 46

2) Pseudomonas aeurginosa MTCC 442

Fresh bacterial cultures were prepared by using sterile nutrient broth.

Composition of Nutrient Broth (HIMEDIA)

Table No.36 showing the composition of nutrient broth

PH of the media: 7.2 to 7.4 Sl.No Ingredients Gms/lit 1 Peptone 5gms 2 Beef extract 3 gms 3 Sodium chloride 5 gms 4 Distilled water 1000ml

Required quantity of nutrient broth was prepared as per the standard ratio and its

pH was checked as 7.4.

Required quantity nutrient broth was taken in 4 conical flask and are sealed with

aluminum files. Then they are sterilized by autoclaving at 15lbs/sq.inch for

15minutes.

After complete cooling, 0.1 ml of selected bacterial cultures were inoculated and

shaken well and incubated at 37 ± 20C for 24 hrs.


134


Fungal Culture

Fungal organisms selected for antifungal activity are mentioned below:

1. Candida Albicans MTCC 35

2. Aspergillus flavus MTCC 62

Composition of Potato dextrose Agar (HIMEDIA m096)

Table No.37 showing the composition of Potato dextrose agar

PH of the media: 5.6 to 5.8 Sl.No Ingredients Gms/lit 1 Potato 200gms 2 Dextrose 20 gms 3 Agar 15 gms 4 Distilled water 1000ml

Required quantity of PDA was prepared as per the standard ratio.

The solution was sterilized in an autoclave for 15minutes at 15lbs/sq.inch

After cooling, individual fungal cultures are inoculated into separate conical flask

by adding 0.1ml of stock culture and are shaken well to assure proper mixing.

They are incubated at 270 ± 20C temperature for 48 hours.

Preparation of discs of Trial and Standard drugs

Preparation of disc of Rasapushpa

Disc of different concentration of Rasapushpa were prepared manually because till

date no ready-made disc of it is available in the market. Different concentration of

drugs was prepared making a suspension in 5% gum acacia, as Rasapushpa is

insoluble in water.

Disc of approximately 8mm diameter were prepared from Whatman filter paper

No.1 using stationary paper punch. They were sterilized in hot air oven at 1600C

for 1 hour.


135


The 50μg/ml and 100μg/ml concentration of discs were prepared by dipping the

100 sterile discs of 8 mm diameter in 1ml of Rasapushpa Suspension containing

respective concentrations.

They are labeled as “T1 and T2” i.e. T1: 50μg/ml T2: 100μg/ml.

Preparation of discs of standard drugs

• Cefotaxime is selected as standard drug for antibacterial activity and

Fluconazole is selected as standard drug for antifungal activity. Discs of both

standard drugs were prepared at two different concentrations viz 50μg/ml and

100μg/ml.

• The solutions of standard drugs were prepared in distilled water.

• The discs of both concentrations were prepared in same way as Rasapushpa.

• The discs prepared were labeled as S1 and S2 respectively i.e S1 50μg/ml and

S2100μg/ml.

Preparation of Nutrient Agar media

Agar media is common for both antibacterial and antifungal activity. It is

essential for solidification.

Composition of Nutrient Agar Media (Highmedia REF 1001)

Table No. 38 showing the composition of nutrient agar

PH of the media: 7.4 ± 0.2 Sl.No Ingredients Gms/lit 1 Peptone 5gms 2 Beef Extract 3 gms 3 Sodium chloride 5 gms 4 Agar 15gms 5 Distilled water 1000ml

Required quantity of agar solution was prepared according to the standard

ratio. pH of the solution was observed as 7.2.


136


The prepared media was sterilized by autoclaving at 15lbs/sq.inch for 15

minutes.

Inoculum is mixed in media before pouring into Petri plates.

Preparation of Inoculum

Bacterial culture

The sterile Nutrient Agar medium was cooled to 450C and mixed with 20% of

respective bacterial culture individually in a

[i.e. 80ml medial and 20ml culture]

Fungal Culture

Here also Agar media and fungal cultures were mixed in 80:20ml ratio

individually.

Application of Disc [trial and standard]

The inoculation prepared was transferred to petriplates immediately. After

proper solidification of media, disc of different concentration of trial and

standard drug [50μg/ml and 100μg/ml] were applied with sterile forceps over

the dried inoculum plate.

Discs were placed at equidistant. They were pressed gently to ensure even

contact with medium. For the identification, on the petriplates names of test

strains was specified and T1, T2, and S1, S2 was marked with marker.

Inoculated media is poured into the petriplates to a depth of 3 to 4 mm. It was

ensured that the layer of media was uniform in thickness

Incubation

After introduction of test and standard drugs, the plates were placed in

refrigerator at 80-100C for diffusion of drugs into the media.


137


After 2 hrs of cool incubation the petri plates were transferred to incubator at

37 ± 20C for 24 hrs. for bacteria.

Fungal cultures were incubated at room temperature for 48 hrs.

After the incubation period the petri plates were observed for zone of

inhibition, which was measured using vernier calipers, and represented in

Table No. 39 and 40.

Observations

♦ Bacterial culture was incubated at 37 ± 20C and fungi at 27 ± 20C.

♦ The growth of Bacterial and fungal cultures were indicated by the turbidity of

the media.

♦ Inoculated media was poured on petri plates to get uniform spreading of the

media.

♦ Agar media was observed for its uniform thickness.

♦ While mixing bacterial or fungal cultures, the temperature of agar was

maintained above 450C.

♦ Zone of inhibition was measured by Vernier calipers.

Precautions

♦ pH of all the media were accurately maintained.

♦ Petridish, conical flask etc, were properly sterilized by autoclaving at

15lbs/sq.inch for 15 minutes.

♦ Activity was conducted by wearing gloves and mask and the activity was

carried in the Laminar airflow.

♦ Zone of inhibition was recorded by placing the petri plates on colony counter.


138

Results

Results

The results of the study are interpreted on the basis of readings of zone of

inhibition of particular organisms. The readings were measured by using Vernier

Calipers.

In present study two G (+) ve organisms viz staphylococcus aureus MTCC87,

streptococcus pyogenes MTCC 32 and two G (-)ve organisms viz E-coli MTCC 46,

pseudomonas aeurginosa MTCC 442 and 2 fungal organisms Candida albicans

MTCC35, Aspergillus flavus MTCC 62 were used to evaluate antimicrobial activity

of Rasapushpa.

Antibacterial and antifungal activity of respective standard and trial drugs was

done in the two different concentrations (50μg/ml and 100μg/ml). The antimicrobial

activity was carried by using six petri plates for each organism. The mean of zone of

inhibition was obtained by the six values of each concentration of standard and trial

drugs. The mean value calculated represents accurate zone of inhibition for standard

and trial drugs. Mean values of both (standard and trial) are shown in table No. 39 and

40.

Table No. 39 Showing the efficacy of standard and trial drugs against Gram positive and Gram-negative organisms

Sl.No Organisms Standard Trial

50μg/ml 100μg/ml 50μg/ml 100μg/ml 1 Streptococcus

aureus 32.00mm 34.66mm 18.83mm 20.33mm

2 Staphylococcus aureus

30.66mm 33.16mm 22.83mm 23.33mm

3 Pseudomonas aeurginosa

32.00mm 33.83mm 24.83mm 27.03mm

4 E- coli 29.05mm 34.00mm 23.83mm 24.83mm

Results are mean of six readings of zone of inhibition.


139

Results

The results reveals that organisms have shown varied response to Rasapushpa.

Both G (+)ve and G (-)ve organisms have shown appreciable zone of inhibition for

Rasapushpa supporting its antibacterial activity. However compared to standard drug

cefotaxime, Rasapushpa is having less zone of inhibition.

Antifungal activity

Table No. 40 Showing efficacy of standard and treated drug against fungus

Sl.No Organisms Standard Drug Trial Drug

50μg/ml 100μg/ml 50μg/ml 100μg/ml

1 Candida albicans 14.18mm 14.83mm 19.33mm 19.83mm

2 Aspergillus flavus 10.50mm 13.66mm 16.83mm 19.96mm

Results are mean of six readings of zone of inhibition

In contrast to antibacterial activity, Rasapushpa has shown significant

antifungal activity. Rasapushpa has shown better zone of inhibition than standard drug

Fluconazole at 50μg/ml and 100μg/ml concentrations.

Thus by observing the readings, it can be said that Rasapushpa has got good

antimicrobial activity.


140

Discussion

DISCUSSION

This chapter deals with analysis of apparent reasons and interpretations of

observations, and results of the study. It can be studied under following headings:

1) Review of Literature

2) Pharmaceutical study

3) Analytical study

4) Experimental study


Kupipakwa Rasayanas are one of the important therapeutic modes of

presentation of Rasa oushadhies. We get the references of kachakupi after 10th century

AD. But preparation of Kupipakwa rasayana came into existence from 13th century

AD onwards. Its preparation is first mentioned by Acharya Yashodhar. Gandhaka

Jarana procedure mentioned in Rasahridaya tantra developed and came into light as

Kupipakwa rasayana.

Kramagni pattern of heating to prepare the compounds, use of valuka yantra,

time of corcking and phenomena of swangasheeta make this process more potent.

The kramagni pattern of Kupipakwa differs from one yoga to other. Nirgandha

preparations need relatively less intensity of agni. Stage of boiling of kajjali,

appearance of profuse fumes, and blocking of Kupi are not obtained in them. Because

there is no such ingredient present which on heating can boil and which may be

inflammable as Gandhaka.

As Kupipakwa Rasayanas are subjected to agni for longer duration, the

therapeutic potency of compound prepared by them is good. Hence only less dosage is

sufficient to achieve desired results.


141

Discussion

Because of these specific features, Kupipakwa method has been selected for

Rasapushpa preparation.

Rasapushpa

Rasapushpa is nirgandha Kupipakwa kalpana. It is described only by Acharya

Sadananda Sharma in his text Rasatarangini.

The references of nirgandha kupi preparations are found from 16th century

onwards. Rasendra Sara Sangraha was first to describe Rasakarpoora by kupi method.

Nirgandha kalpanas are most neglected part in Rasashastra as they are more prone to

cause Parada vikaras, but risk exists only if, they are not properly processed and

continued even after the cure of the disease.

The ingredients of Rasapushpa are Hingulotha Parada, Kasisa and Saindhava

lavana. In case of Rasapushpa preparation by damaruyanatra Sphatika is used as an

additional ingredient. Chemically Rasapushpa is considered as mercurous chloride

and it is obtained by the process of sublimation.

It is important to mention here that, the chemical nature and composition of

Rasapushpa and Rasakarpoora are very much doubtful.

Rasakarpoora is a mercuric chloride. But according to V.M. Dwivedi,

Rasakarpoora prepared by ancient method is mercurous chloride while recent method

is mercuric chloride.

Dr. Nadakarni described it as calomel. P.C. Ray, opined that Rasapushpa and

karpoora preparatory methods are of mix compounds.

It is Rasatarangini who first differentiated it and called Rasapushpa as

mercurous chloride and Rasakarpoora as mercuric chloride.


142

Discussion

Hingulotha Parada

Hingulotha Parada is considered to be having qualities equivalent to ashta samskarita

Parada and samaguna balijarita Parada. Because during the formation of Hingula,

either in natural form or synthetic the process of heat on the proportionate

combination of Parada and Gandhaka is of vital importance. This probably might help

in eliminating the impurities present in the Parada. Ashtasamarkar is a difficult and

time taking procedure. Hence Hingulotha Parada is taken for preparation of

Rasapushpa without compromising the quality.

Saindhava Lavana

Saindhava lavana is a mineral salt, which forms essential constituent of blood

serum. In the preparation of Rasapushpa compound Saindhava lavana is an essential

ingredient. Saindhava lavana is considered best among panchalavana. Because it is

easily available and satmya to the body. Unlike other lavanas it possess madhura rasa,

tridoshaghna, and sukshmagamitwa properties. Hence whenever lavana is told

without any prefix, Saindhava lavana has to be considered.


The pharmaceutical study was carried out in 3 stages.

1) Shodhana of constituent drugs.

2) Hingula Satwapatana.

3) Rasapushpa Nirmana.

For present study, Rasapushpa was prepared according to Kupipakwa method as

quoted in Rasatarangini 6/29.

Shodhana of constituent drugs

All the ingredients of trial drugs were selected strictly according to classical

reference and were purified.


143

Discussion

Hingula shodhana

The practical No. 01 deals with shodhana of Hingula. It was carried out by

bhavana with Nimbu swarasa for 7 times. Slight reduction in quantity of

Nimbuswarasa was noticed in each succeeding bhavana. On an average 170ml of

Nimbu swarasa is essential in each bhavana for 500gm of Hingula.

Nimbu swarasa might help in detoxification of Hingula due to its amla rasa.

Nimbuswarasa is rich in complex of organic acids such as citric acid, mallic acid,

which may react with the unwanted materials in Hingula and form a complex, which

is soluble in water.

Then Hingula was washed with water thoroughly so that it may help in

separation of water-soluble complex of impurities. Prakshalana continued till Hingula

attained ujjwala varna and loses amlatva of bhavana dravya.

Hingula gained weight after shodhana; may be due to the addition of solid

contents present in Nimbuswarasa.

Hingula Satwapatana

Hingula Satwapatana was done by urdwapatana method using damaruyantra.

Total duration of heat given was 8 hours. Throughout the procedure temperature was

maintained at 3500-4000C. Cold pad was maintained on upper pot for proper

condensation of Parada.

Hingula Satwapatana was done for 2 times under the heading of practical no.3

and 4.

In practical no3, for 240gms of Hingula, 135gms of Parada was secured and

practical no.4, for 240gms of Hingula 127gms of Parada was collected.

Hence and average yield of Satwapatana here is taken as around 54%


144

Discussion

♦ The percentage of mercury content present in the Hingula sample taken for

present study is 71%.

♦ The loss may be due to following reasons.

Some percentage of loss is evident in dhoomgati during heating

procedure.

Some percentage of loss during Prakshalana through jalagati and

malagati.

Hingula shodhana was done before it is being subjected to Satwapatana

process. By bhavana process, Hingula converts into finest state of sub division. In this

state maximum amount of Parada can be extracted.

As regards to modern chemistry, it is a auto-reduction process. Mercury is

extracted from ore cinnabar by heating in current of air. The reaction-taking place

during the procedure is.

Hgs + 02 Hg + So2

Parada shodhana

Many Rasa acharyas believe that shodhana is not essential for Hingulotha

Parada as it is devoid of naga, vanga and saptakanchukadi doshas. But for the present

study, procedure mentioned in Rasatarangini (5/38-41) was adopted. Hingulotha

Parada was mixed with 1/16th part of Haridra and mardana was done for 2 days. Then

Parada was filtered through cloth and weighed. 5 gms loss was noticed during the

procedure, which may be due to malagati of Parada.

The mixture was yellow initially, which gradually turned into brown colour.

Haridra is very sensitive to the chemical changes and may be impurities in Parada

have colour changing activity on Haridra.


145

Discussion

Haridra possess shothaghna, vishagana, krimighna and many other important

properties. It is also known antioxidant. By the process of mardana, some qualities of

Haridra may get inherited in Parada due to samskara effects.

Kasisa Shodhana

Kasisa shodhana was done by giving bhavana with Nimbuswarasa. Different

methods of shodhana of kasisa were described by various acharyas, but this method

was being easiest and commonest in routine practicals. All instructions mentioned for

bhavana, in classics are followed. During the procedure 25gms of Parada was lost

which might be because of adherence of kasisa to equipments and spilling out of

Khalva yantra.

Chemically kasisa is ferrous sulphate, which is soluble in water, hence the

purity of kasisa is related to its water insoluble content. By bhavana with

Nimbuswarasa, water insoluble content of kasisa might reduce and thus help in

shodhana of kasisa.

Rasapushpa preparation

For the present study, Rasapushpa nirmana by Kupipakwa method as

mentioned in Rasatarangini (6/29-31) was adopted.

Preparation of Prematerial

For the preparation of Rasapushpa, equal quantities of Hingulotha Parada, shodhita

Kasisa and Saindhava lavana was taken and mixed in Khalva yantra uniformly.

Before preparing Prematerial, shodhita Kasisa, and Saindhava lavana were powdered

separately. This was done in order to ensure the proper mixing of ingredients.

Preparation of kupi.

Amber coloured bottles of 650ml capacity were selected for procedure. The amber

colour bottles are more pyrosensitive, hence selected for preparation of kupi. Kupi


146

Discussion

was prepared by wrapping mud-smeared cloth up to the mouth of kupi in seven

consecutive layers. After drying of previous layer, next layer is applied. This avoids

presence of any air bubbles inside mrutkapata. Air space between the layers may lead

to breakage of bottle during heating.

The application of mrutkapata may help in regulation of temperature inside the kupi

and it might also make kupi resistant to heat so that it can withstand high and

continuous temperature given during the procedure.

Filling of Prematerial into Kupi

The kupi was filled with 100 gms of Prematerial that acquired lower 1/5th

space, as large quantity may hinder the sublimation of final product.

Placing of Kupi in Valuka yantra

Initially three angulas of sand was put in valuka yantra to avoid the

direct contact of yantra with kupi. Then kupi was placed firmly in the center, then

carefully sand was put in remaining portion of yantra. While filling the sand, kupi was

duly corcked to avoid contamination of Prematerial with dust.

For preparation of valuka yantra, mud pot is taken and 2 layers of mud

smeared cloth is wrapped from the base to 3/4th portion of pot. After it is completely

dried, used for preparation of Rasapushpa.

Totally 10kg of coarse sand was filled. It renders resistance to the apparatus

from atmospheric temperature variations.

Inferences made during procedure

Total duration of heat given during the procedure was 6 hours. It was

classified into 3 stages as mrudu, madhyama and teevragni. The temperature

maintained at these stages as mentioned as follows:


147

Discussion

Mrudu agni : 100-1500C

Madhyama : 100-2500C

Teevragni : 250-3500C

• Regular temperature monitoring was done by digital pyrometer. The

thermocouple was inserted into the valuka yantra up to level of base of kupi to

measure the temperature.

• The duration of heat given in each stage was 2 hours because it was usually

noticed that in the preparation of kupi rasayana, initial 3/4th part would be

supplied with mrudu and madhyamagni and last 1/4th portion with teevragni.

Here, for first 4 hours mrudu and madhyamagni was maintained and last 2

hours teevragni was applied to get the compound.

Shalaka sanchalana

Sheeta shalaka was used in the procedure to note the state of Prematerial whether it is

in powder form or moistened or in sublimating compound state.

Sheeta shalaka was used to confirm the formation of compound and thus helps in

deciding corcking time of the kupi.

Observations of Fumes and Water vapours

This forms the important factor while preparing Rasapushpa. Fumes were not so

profuse as we notice in case of any sagandha preparations. It was slightly irritating.

Fumes did not cause blocking of kupikantha, as it was not thick. So tapta shalaka was

not used during the procedure the presence of water vapours was tested by means of

tamra patra and was ascertained by collection of water droplets on the surface of

tamra patra. Tamra patra test was done to detect the evaporation of Parada but no

discolouration of tamra patra or no crystals of Parada was found over it.


148

Discussion

Corcking

Time of corcking is very important. After complete evaporation of water

vapours and sheeta shalaka test, corcking was done. It is the land mark for teevragni

stage of kupi rasayana. Delayed and improper corcking may lead to loss of finished

product. Corcking is also essential to obtain kanthastha rasayanas.

Corck was prepared by ishtika, which was wrapped with two layers of mud-smeared

cloth. It should exactly fit into the mouth of kupi otherwise may result in loss of final

product.

Removal of sand

After crocking, 4 angulis of sand surrounding the neck of kupi was removed.

This was done to ensure proper sublimation and condensation of compound.

Swangasheeta

It is a good concept established in Ayurveda. After 6 hours of heating, agni

was stopped and valuka yantra was left for self cooling. This helps in sublimation and

condensation of compound.

Kupi bhedana

After swangasheeta, kupi-bhedhana was done. Thread was tied 3 inches below

the sublimated compound in a single circle. This ensures proper breaking.

Collection of compound

Rasapushpa was collected carefully and weighed. 4 practicals of Rasapushpa

nirmana was done. Each time 100gms of Prematerial was taken, the results of which

are mentioned below.


149

Discussion

Table No. 41 showing the yield of Rasapushpa

Weight of Prematerial Weight of Rasapushpa Residiue left

100gms 10 gms 60 gms

100 gms 15 gms 65 gms



The result reveals that on an average 14% yield of Rasapushpa was found.

Rasapushpa was also prepared according to antardhooma method for two

times i.e kupi was subjected to agni after corcking. On an average 18% yield was

found which is more compared to bahirdhooma procedure. Duration of heating and

temperature was maintained as according to bahirdhooma procedure.

Rasapushpa was also prepared twice following damaruyantra method. In

Rasatarangini, it is stated to prepare Rasapushpa on antimandanala. This term

suggests requirement of relatively less intensity of agni compared to Kupipakwa

method. After discussing with guide and experts. For the first hour 1100C of heat was

given which was raised to 2500C gradually and same temperature maintained

throughout the procedure. On an average 27% yield was obtained.

Analytical part

Analysis of any drug should be known before experimental and clinical trials.

In present study Rasapushpa was sent for physical and chemical analysis at Bangalore

test house, Bangalore.

Analysis of Hingula

Shodhita Hingula was analyzed for organoleptic characters and total mercury and

sulphur content.


150

Discussion

An organoleptic character of Shodhita Hingula reveals that it is red coloured,

odourless, fine powder.

These reports are similar to grahya Hingula laxanas given in literature i.e. Japakusuma

nibha which means red colour.

Total mercury content estimated in Hingula is 71.2% w/w and sulphur content is

10.15% w/w

According to British pharmacopoeia Cinnabar should contain 84% w/w of total

mercury as Hg. Total mercury content in shodhita Hingula was found to be slightly

less.

Rasapushpa

Organoleptic Characters reveals that Rasapushpa is practically insoluble in water,

cold and dilute acids and sparingly soluble in alchol and chloroform.

The insolubility is bar to toxicity of Rasapushpa. Hence it can be administered in

proper dose without any risk.

Loss on drying is 0.34%, which suggests the presence of negligible amount of

moisture in Rasapushpa. Hence product will not be affected from microorganisms.

Total ash is 0.4%, which indicates presence of organic matter in the final product,

which may be inherited during shodhana procedure.

Acid insoluble ash is 0.23% indicates that negligible amount is soluble in acid.

Water-soluble content is nil. Rasapushpa is chemically mercurous chloride and

insoluble in water. Absence of water-soluble content indicates that Rasapushpa

sample does not contain any impurities. The probable impurity present in Rasapushpa

is mercuric chloride, which is soluble in water.


151

Discussion

Flow property: Since the drug is in powder state, it was tested for its flow property.

This investigation suggests necessity of any adjuncts for proper flow of drug during

capsule or tablet preparation. Flow property was identified by angle of Repose (Tan θ)

and flow rate by compressibility index (I)

Angle of Repose (Tan θ) is 40.14 degree, which suggests that its flow property

is good. Its compressibility index is 30%, which suggests that flow rate is moderate

and need adjuncts in tablet or capsule preparation.

Particle size: Particle size of drug was viewed under microscope and calculated

according to procedure. Arithmetic mean was calculated.

Arithmetic mean of Rasapushpa is 9.34 micrometer. It signifies the fineness of

particle size. The test shows that smaller the particles, better the absorption capacity

of compounds.

The percentage mercury and chloride

The total content of mercury in Rasapushpa is 84.3% w/w. according to

British pharmacopoeia mercury content in mercurous chloride should be 84.09% w/w.

Percentage of Hg in Rasapushpa sample is very near to the value mentioned in B.P.

The percentage of chloride in Rasapushpa is 13.6% w/w. This value is nearer

to the limit prescribed in pharmacopoeial Standards of Ayurvedic formulations i.e.

15-20% w/w.


There are wide variations in susceptibility of different strains of same species

of organisms to the drug. Antimicrobial activity is a technique in which response of

an organism to a particular antimicrobial agent can be established. Many methods are

employed for evaluation of antimicrobial activity of a drug. For the present study


152

Discussion

disc-diffusion method was adopted. This technique is simple and relatively

inexpensive which makes it still the method of choice for the average laboratory.

Two G(+)ve (Staphylococcus aeures, Streptococcus pyogenes) and two

G(-)ve bacterias (E-coli, Pseudomonas aeurginosa) and two fungi (Candida albicans,

Aspergillus flavus) are taken the study. As these microorganisms are responsible for

manifestation of majority of pathological conditions such as gastroenteritis, pyogenic

infections, urinary tract infections, wound infections etc.

Nutrient broth is used to culture the bacteria and potato dextrose agar is used

to culture fungi. Both are simple culture media, which are usually preferred because

most of the bacterias are easily grown in this media. They tend to have less batch-to-

batch variations.

Potato dextrose agar media is suitable for fungal culture as it has got low pH

and a relatively high concentration of sugar tolerated by fungi but are inhibitory to

many bacterias.

Growth of organisms was confirmed by turbidity of the media.

Cefataxime and Fluconazole are the standard drugs selected for antibacterial

and antifungal activities respectively.

As readymade discs were not available in the market, discs of Rasapushpa

Cefataxime and Fluconazole of two different concentrations (50mg/ml and

100mg/ml) were prepared in the laboratory.

Rasapushpa is insoluble in water and soluble only in Con HNo3 on heating.

But Con HNo3 changes the chemical nature of Rasapushpa and it possess good

antimicrobial activity. Hence gum acacia was selected to make the suspension of

Rasapushpa. Gum acacia was tested for its antimicrobial activity. Discs were prepared


153

Discussion

only by 5% gum acacia and tested and we found no zone of inhibition. This indicates

that it has got no activity against microorganisms.

Agar was used as solidifying agent in microbiological media. It is common for

both bacterias and fungi. It is suitable because, it has virtually no nutritive value and

is not affected by growth of bacteria.

Then prepared discs were placed on petriplates containing inoculum and then

kept in refrigerator for 2 hours for proper diffusion of drugs. Then bacterias were

incubated at 37± 20c for 24 hours and fungi incubated at 27± 20c for 48 hours.

Aseptic care was taken throughout the procedure. Results were studied on the

basis of zone of inhibition. It revealed that measured zone of inhibition of Rasapushpa

were less for bacteria compared to standard drug. In contrast, for fungi Rasapushpa

has shown significant zone of inhibition. Antimicrobial activity depends upon rate of

diffusion of drug through an agar surface. Water-soluble drugs readily diffuse where

as water insoluble compounds take relatively longer duration to diffuse through the

surface. May be because of this reason, Rasapushpa has shown less zone of inhibition

for bacteria. It does not mean that they don’t possess antimicrobial activity.

Probable mode of action

Parada is a main ingredient of Rasapushpa. It is having properties such as Krimighna,

Tridoshaghna, Rasayana, Vrana Ropana, Yogavahi etc. Rasapushpa also shows

bhutavishapaha, Krimighna, Twakdoshara, properties. By this it is evident that the

properties of parada exists in Rasapushpa.

Rasapushpa is chemically identified as mercurous chloride. It is said that

mercurial salts in small proportions possess bactericidal action against G (+) ve and

G (-) ve organisms.


154

Discussion

Rasapushpa in its finest state of subdivision slowly absorbs into the gut and

produces systemic action. It probably exhibits bacteriostatic action by inhibiting the

sulphahydral enzymes of bacterial cells. Thus it interferes with cell metabolism may

prove bacteriostatic.

It has got an astringent property and soothing action on skin by which it acts

as an effective antiseptic agent.


155

Introduction

Conclusion

The conclusions are drawn on the basis of observations made during present study.

• Rasapushpa is a Kupipakwa rasayana and is Sagni, Nirgandha, and

Bahirdhuma moorchana of Parada.


• Shodhana of Hingula, Kasisa with Nimbu swarasa bhavana and Parada with

Haridra, certainly have a role in detoxifying and potencifying the drugs.

• The yield of Parada varies inversely to the residue left at the end of the

procedure.

• Urdwapatana is simple method to obtain Hingulotha Parada.

• On the basis of observations made during the preparation, the heating pattern

for Rasapushpa may be concluded as:

Mrudu agni : 100-1500C

Madhyamagni : 150-2500C

Teevragni :250-3500C

Analytical study

• Rasapushpa is white in colour, odourless, insoluble in water, and spairingly

soluble in chloroform and alcohol.

• Loss on drying is 0.34% revealing the presence of negligible amount of

moisture content.

• Total ash is 0.4%, which suggests that Rasapushpa has got low organic

content.

• Particle size is 9.34 micrometer, which signifies the fineness of particles.

• The total mercury (84.3%) and chloride (13.6%) content present in

Rasapushpa is near to the standards specified.


156

Introduction

Antimicrobial study

• The bacterias and fungi selected for present study are responsible for

manifestation of majority of pathological conditions.

• Disc-diffusion is easy, economical, and suitable method for evaluation of

antimicrobial activity of Rasapushpa.

• The result reveals that, Rasapushpa has got less activity against bacterias

compared to standard drug.

• Rasapushpa has shown better zone of inhibition for fungi compared to

standard drug.

• Thus it can be concluded that Rasapushpa possess significant antimicrobial

activity.

Limitations

1. The duration of the study was precise.

2. Minimum instrumental and investigatory facilities.

3. The study was conducted on selected organisms.

Further scope

1. Rasapushpa preparation can be done with varied proportions of ingredients,

procedures along with their toxicity study.

2. Study of specific chemical reactions going on inside the kupi may be studied

making use of modern techniques.

3. Actual temperature inside the kupi may be studied with the aid of optical

pyrometer.

4. Use of vertical muffle furnace in the preparation of Rasapushpa may be

carried out and compared with present sample in all aspects.

5. A clinical study is essential to evaluate the therapeutic effect of Rasapushpa.


157

Summary

Summary

The present dissertation work is entitled as “preparation, physico chemical

analysis of Rasapushpa and its antimicrobial activity”. In this study an attempt was

made to prepare Rasapushpa following classical procedures. Its analysis and

antimicrobial activity was accessed following disc-diffusion method.

The study includes topics such as Introduction, Objectives, Review of

Literature, Methodology, Observation and results, Discussion and Conclusion.

Introduction

It depicts the importance of Ayurveda and Rasashastra. The significance of

Kupipakwa Rasayana and therapeutic values of Rasapushpa are mentioned. The

virulence of microorganisms and their role in causing epidemics is described. The

concept of krimis according to our classics is mentioned. Finally the importance and

need for the present study is discussed.

Objectives

Aim and objectives of the study is stated.

Review of Literature: It includes drug review, procedure review, review of krimis

and anti microbial activity.

It includes description of Hingula according to both Ayurvedic and Modern

concepts. Different methods for Hingula Satwapatana are explained. Then ingredients

of Rasapushpa viz Parada, Kasisa and Saindhava lavana are described. Different

opinions of doctrines of Rasashastra regarding their paryaya, bheda, shodhana, and

pharmacological properties are included. Then Haridra and Nimbuswarasa were

described.


158

Summary

The procedure of Kupipakwa method was dealt in detail. Then different

pharmaceutical procedures of Rasapushpa along with its paryaya, matra and guna are

included.

It includes description of krimis as per Vedic and Ayurvedic science. Then

special features and virulence caused by bacteria and fungi selected for the study are

described.

It includes historical background of the evolution of antibacterial activity and

different methods to evaluate the sensitivity of drug. Different culture media essential

for growth are described.

Methodology

It involves, pharmaceutical, analytical and experimental study.


It deals with practical works performed during present study. Totally 12

practicals were conducted. They are shodhana of Hingula, Hingula Satwapatana,

Parada and Kasisa shodhana. Practicals done for preparation of Rasapushpa are

mentioned. For each practicals. Observations and precautions taken are mentioned. At

the end of the practical, quantity of initially taken material and end product was

narrated.

Analytical study

It deals with physico-chemical analysis of Hingula and Rasapushpa.

Hingula was analysed for organoleptic characters and for the percentage of mercury

and sulphur content.

Rasapushpa analyzed to know the proportions of mercury and chloride. Other

analytical procedures such as loss on drying, total ash, acid insoluble ash, fineness of

particles and flow property were also carried out.


159

Summary

Experimental study: It deals with the antimicrobial study.

Antimicrobial study

The study was carried out by disc diffusion method. Cefataxime and

Fluconazole are selected as standard drugs for antibacterial and antifungal activities

respectively. Two gram positive, and gram negative bacterias and 2 fungi are taken

for the study.

Antimicrobial activity of Rasapushpa and standard drugs were studied and

compared at two different concentrations (50 μg/ml, 100 μg/ml). All necessary aseptic

precautions were taken during the study.

Results

Results of the study were interpreted on the basis of observed zone of

inhibition. Zone of inhibition was calculated by Vernier Calipers. Results reveals that

Rasapushpa has got less antibacterial activity compared to standard drug Cefataxime

where as antifungal activity of Rasapushpa is significant.

Discussion

The whole work was elaborately discussed along with reasoning. The concepts

regarding the shodhana of Hingula, Parada, Kasisa and Hingula Satwapatana are

discussed. Few rationales are put forward in the discussion of Rasapushpa

preparation, analytical study and antimicrobial study.

Conclusion

Conclusion has been reported based on the observation and interpretations

made during whole study.


160

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94 R. Ghosh’s, Pharmacology Materia Medica and Therapeutics, Edited by

Sadhanda Sekharsen Gupta, 23rd Edition, Calcutta, Hilton and co, 1947, Group X, pp. 553

95 Dr. Vilas A.Dole, Text book of Rasashastra, Ist Edition, Chaukambha surabharati prakashana, 2004, 6th Chapter, pp. 78-79. 96 Acharya Gopal Krishna, Rasendra Sara Sangraha, translated by Dr. Ashok. D. Satpute 1st Edition, 1st Chapter, pp 129 97 Acharya Yashodhara, Rasaprakasha Sudhakara, Siddiprada commentary by sidda

nandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 3rd Chapter, Shloka 72-73, pp 98.

98 Acharya Gopal Krishna, Rasandra Sara Sangraha, translated by Dr Ashok D. Satpute, 1st Edition, Varanasi Chaukambha Krishnadas Academy, 2003 ,

1st Chapter, pp 129

99 Acharya Madhava, Ayurveda Prakasha, Edited by Gulraj Sharma Mishra, 2nd Edition, Varanai, Chaukambha Bharati Academy, 1999, 2nd Chapter, 273 Shloka, pp .325 100 Acharya Yashodhara, Rasaprakasha Sudhakara, Siddi prada commentary by

Sidda nandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 3rd Chapter, pp 98.

101 Shri Sadananda Sharma, Rasastarangini Edited by Kashinatha Shastri, 11th

Edition, Varanasi, Motilal Banarasidas, 2000 5th Taranga, 21st Chapter, Shloka 227, pp 562

102 Acharya yashodhara, Rasaprakasha Sudhakara, Siddi prada commentary by

Siddanandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 3rd Chapter, pp 99.

103 Acharya vagbhata, RasaRatna Samucchaya, Edited by Dr. Indradev Tripathi, 2nd


167

Biblography

Edition, Varanasi, Chaukambha Orientalia, 2000, 1st Chapter, Shloka 67 pp 7. 104 Shri Sadananda sharma, Rasastarangini Edited by Kashinatha Shastri, 11th

Edition, varanasi, Motilal Banarasidas, 2000, 5th Taranga, 21st Chapter , pp 562 105 Acharya Yashodhara, Rasaprakasha sudhakara, Siddi prada commentary by

Siddanandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 3rd Chapter, pp 98.

106 Acharya Gopal Krishna, Rasandra Sara Sangraha, translated by Dr. Ashok. D.

Satpute, 1st Edition, 2st Chapter, pp 129-130. 107 Shri Sadananda Sharma, Rasastarangini, Edited by Kashinatha Shastri, 11th

Edition, Varanasi, Motilal Banarasidas, 2000 5th Taranga, 21st Chapter Shloka 225, pp 563.

108 Achary Madhava, Ayurveda Prakash , Edited by Gulraj Sharma Mishra, 2nd

Edition,Varanasi, Chaukambha Bharati Academy, 1999, 2nd Chapter, Shloka 273 pp.325.

109 Acharya Vagbhata, RasaRatna Samucchaya, Edited by Dr. Indradev Tripathi.

2nd Edition, Varanasi, Chaukambha Orientalia, 2000, 3rd Chapter, Shloka 52, pp 31.

110 Acharya Somadeva, Rasendra Chudamani, 1st Edition, Varanasi, Chaukambha

Orientalia, 1984, 11th Chapter, Shloka 77, pp 187. 111 Acharya Yashodhar, Rasaprakash Sudhakara, Siddiprada Commentary by

Siddanandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 6th

Chapter, Shloka 63, pp 125 112 Acharya Indradev Tripathi, Rasarnava, 4th Edition, Varanasi, Chaukambha

Sanskrit Series 2001, 10th Patala, Shloka 81, pp 91. 113 Acharya Pandit Ramprasad, Rasendra Purana, Khemaraj Shri Krishnadas,

Mumbai, 2000, 9th chapter, Shloka 22, pp 223.

114 Acharya Madhava, Ayurveda Prakash, Edited by Shri Gulraj Sharma Mishra, 2nd Edition, Varanasi, Chaukambha Bharati Academy, 1999, 28th Chapter, pp 162

115 Shri Bhudeb Mookarjee, Rasajalanidhi, Vol I, Edited by Siddinandana Mishra.

2nd Edition, Varanasi, Chaukambha Sanskrit Bhawan, 1998, 2nd Chapter, pp 149. 116 Ibid, 2nd Chapter, pp 150. 117 Shri Dattaram Choube, Brihat Rasa Raja Sundara , 3rd Edition, Varanasi,

Chaukambha Orientalia, 2000, 28th Chapter, pp 162. 118 Ibid, 28th Chapter, pp 161.


168

Biblography

119 Shri Sadananda Sharma, Rasatarangini, Edited by Kashinatha Shastri, 11th

Edition, Varanasi, Motila Banarasidas, 2000 21st Taranga, Shloka 230, pp 564. 120 Acharya Yashodhar, Rasaprakash Sudhakara, Siddiprada Commentary by

Siddanandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 6th

Chapter, Shloka 65, pp 125. 121 Acharya Madhava, Ayurveda prakash, Edited by Gulraj Shrma Mishra, 2nd

Edition,Varanasi, Chaukambha Bharati Academy 1999, 2nd Chapter, Shloka 274, pp 325.

122 Acharya yashodhara, Rasaprakasha sudhakara, siddiprada commentary by Sidda

nandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 6th Chapter, Shloka 64, pp 125.

123 Acharya Madhava, Ayurveda Prakash, Edited by Gulraj Shrma Mishra, 2nd

Edition, Varanasi, Chaukambha Bharati Academy 1999, 2nd Chapter, Shloka 274, pp 325.

124 Ibid, 2nd Chapter, Shloka 275, pp 325. 125 Shri Sadananda Sharma, Rasatarangini, Edited by Kashinatha Shastri, 11th

Edition,Varanasi, Motilal Banarasidas, 2000 21st Taranga, Shloka 232-33, pp 564.

126 Ibid, 21st Taranga, Shloka 255- 258, pp 568, 127 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd

Edition, Varanasi, Chaukambha Sanskrit Bhawan 2003, 3rd Chapter, Shloka 59-60, pp 99-100.

128 Shri Sadananda Sharma, Rasatarangini, Edited by Kashinatha Shastri, 11th

Edition, Varanasi, Motila Banarasidas, 2000, 21st Taranga, Shloka 260, pp 569. 129 Ibid, 21st Taranga, Shloka 242, pp 566. 130 Shri Dattaram Choube, Brihat Rasa Raja Sundara , 3rd Edition, Varanasi,

Chaukambha Orientalia, 2000, 28th Chapter, pp 163. 131 The Pharmacological Basis of therapeutics, Edited by Louis Goodman and

Alfred Gilman, 5th Edition, Newyork,Macmillan Publishing Co,INC, 1976, XVI Section, 63rd Chapter, pp 1315. 132 R. Ghosh’s, Pharmacology Materia Medica and Therapeutics, Edited by

Sudhendu Shekharsen Gupta, 23rd Edition, Calcutta, Hilton and co, 1947, Group IV, pp 367

133 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd


169

Biblography

Edition, Varanasi, Chaumabha Sanskrit Bhawan 2003, 7th chapter, pp 128. 134 Dr.K.N Nadakarni, Indian Materia Medica, 2nd Edition, Mumbi, Popular

Prakashana, 1976, Volume II, pp 98. 135 Shri Sadananda Sharma, Rasastarangini, Edited by Kashinatha Shastri, 11th

Edition, Varanasi, Motilal Banarasidas, 2000, 14th Taranga, Shloka 117-118 pp 347. 136 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd Edition, Varanasi, Chaukambha Sanskrit Bhawan ,2003, 7th chapter, pp 128. 137 Pandit Narahari, Rajanighantu, Edited by Indradev Tripathi, 3rd edition,

Varanasi, Krishnadas Academy, 2003, Pippalyadi varga, Shloka 88, pp 152. 138 Acharya Kaidev, Kaidev Nighantu, Edited by P.V. Sharma, dhatuvarga, Shloka 96-97, pp 290. 139 Dhanvantari Nighantu, Edited by P.V. Sharma, Varanasi, Chaukambha

Orientalia, 2000, Shatapushpadi varga, Shloka 25, pp 74. 140 Shri Madanapala, Madanapala Nighantu,Shuntyadi Varga, Edited by Pandit Ramaprasad Vaidya, Mumbi, Khemaraj Shri Krishnadas Prakashana, 1998, pp173. 141 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd Edition, Varanasi, Chaumabha Sanskrit Bhawan, 2003, 7th Chapter, pp 128. 142 Ibid, 7th Chapter, pp 128 143 Dr.K.N. Nadakarni, Indian Materia Medica, 2nd Edition, Mumbi, Popular prakashana, 1976, volume II, pp 108. 144 Ibid, Volume II, pp 108. 145 Shri Sadananda Sharma, Rasastarangini Edited by Kashinatha Shastri, 11th Edition, varanasi, Motilal Banarasidas, 2000, 14th Taranga, Shloka 119-120, pp 347. 146 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd Edition, Varanasi, Chaukambha Sanskrit Bhawan 2003, 7th Chapter, pp 129 147 Shri Madanapala, Madanapala Nighantu,Shuntyadi Varga, Edited by Pandit Ramaprasad Vaidya, Mumbi, Khemaraj Shri Krishnadas Prakashana, 1998, pp173. 148 Dhanvantari Nighantu, Edited by P.V. Sharma, Varanasi, Chaukambha Orientalia, 2000, Shatapushpadi varga, pp 74. 149 Acharya Kaidev, Kaidev Nighantu, Edited by P.V. Sharma, dhatuvarga, Shloka


170

Biblography

96-97, pp 290. 150 Pandit Narahari, Rajanighantu, Edited by Indradev Tripathi, 3rd edition, Varanasi,Krishnadas Academy, 2003, Pippalyadi varga, pp 152. 151 Shri Sadananda sharma, Rasastarangini Edited by Kashinatha Shastri, 11th Edition, Varanasi, Motilal Banarasidas, 2000, 14th Taranga, Shloka 119-120, pp 347. 152 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd Edition, Varanasi, Chaumabha Sanskrit Bhawan, 2003, 7th Chapter, pp 129 153 Pandit Narahari, Rajanighantu, Edited by Indradev Tripathi, 3rd edition, Varanasi,Krishnadas Academy, 2003, Pippalyadi varga,Sloka 89-90 pp 152.-153 154 Dhanvantari Nighantu, Edited by P.V. Sharma, Varanasi, Chaukambha Orientalia, 2000, Shatapushpadi varga,Sloka 89-90 pp 152-153. 155 R. Ghosh’s, Pharmacology Materia Medica and Therapeutics, Edited by Sudhendu Shekharsen Gupta, 23rd Edition, Calcutta, Hilton and Co, 1947, pp 922. 156 Vaidya U.M.Gogte, Ayurvedic Pharmacognosy and Therapeutic uses of

Medicinal Plants, 1st Edition Mumbi, Bharateeya Vidya Bhavan, 2000, Part II, pp 412.

157 Kirtikar and Basu, Indian Medicinal plants, 2nd Edition, 1981, International

Book Distributors. Delhi, No. IV, pp 2423. 158 I bid, vol I, pp 489. 159 Vaidya U.M.Gogte, Ayurvedic Pharmacognosy and Therapeutic uses of

Medicinal Plants, 1st Edition Mumbi, Bharateeya Vidya Bhavan, 2000, Part II, pp 412

160 Dr Chandrabhushan Jha,Ayurvediya Rasashastra, 1st Edition, Varanasi Chaukambha Surabharati Prakashana, 2004, pp 173-174. 161 Ibid, 5th Chapter, pp 175-176. 162 Shri Harisharnanda Sharma, Kupipakwa Rasanirmana Vijnana, 1st Edition, Amritsar, 1941, pp 12-34. 163 Acharya Vagbhata, RasaRatna Samucchaya, Edited by Dr. Indradev Tripathi. 2nd Edition, Varanasi, Chaukambha Orientalia, 2000, 9th Chapter, Shloka 36, pp103 164 Dundukanath, Rasendra Chintamani, Edited by Prof. Siddanandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 2000, 2nd Chapter Shloka 18, pp 14 165 Acharya Madhava, Ayurveda Prakash, Edited by Gulraj Sharma Mishra, 2nd


171

Biblography

Edition, Varanasi, Chaukambha Bharati Academy 1999, 1st Chapter, Shloka 199, pp 104. 166 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd Edition, Varanasi, Chaumabha Sanskrit Bhawan 2003, 1st chapter, pp 20-21 167 Shri Harisharnanda Sharma, Kupipakwa Rasanirmana Vijnana, 1st Edition, Amritsar, 1941,Shloka 399-400, pp 195 168 Viadya Vasudeva Moola Shankara Dwivedi, Parada Vijnaneeyam, 3rd Edition, Shri Sharma Ayurveda Mandira, 1997, pp 245. 169 Ibid, pp 246 170 Ibid, pp 246 171 Dr Chandrabhushan Jha,Ayurvediya Rasashastra, 1st Edition, Varanasi Chaukambha Surabharati Prakashana, 2004, pp 177. 172 Acharya Yadavaji Trikramji, Rasamritam, translated by Damodar Joshi, 2nd Edition, Varanasi, Chaumabha Sanskrit Bhawan 2003, 5th chapter, pp 177 173 Shri Sadananda Sharma, Rasastarangini Edited by Kashinatha Shastri, 11th Edition, Varanasi, Motilal Banarasidas, 2000, 6th Taranga, shloka 22-31 pp 107-114 174 Pandit Vishwanath Dwivedi, Bharatiya Rasashastra ,2nd Edition .Nagapur, Shri Sharma Ayurveda Mandira, 2000, pp 232-233. 175 P.L.Soni, Text Book of Inorganic Chemistry, 18th Edition, 3rd Chapter, pp 323. 176 Shabda Kalpa Drauma, Part II, 3rd Edition, Varanasi, Chaukambha Sanskrit Series, 1967, pp 178. 177 Atharvaveda, Samhitapart I, 7th Edition, Delhi, Parimala Publications, 2nd Kanda, 31st Sukta, Shloka 2-4 pp 25-26. 178 Acharya Agnivesha, Charaka Samhita, Commentary by Kashinath Shastri, Edited by Gangashaya Pandya, 5th Edition, Varanasi, Chaukambha Sanskrit Samsthana, 7th Chapter, Vimanasthana, Shloka 11,pp 726. 179 Acharya Sushruta, Sushruta Samhita, Commentary by Kaviraja, Dr.Ambikadatta Shastri, Varanasi, Chaukambha Sanskrit Samsthana. Reprint 2005, Nidanasthana, 5th Chapter, shloka 32-33, pp 257. 180 Acharya Vaghata, Ashtanga Hridaya, 3rd Edition, Varanasi, Krishnadas Academy Publications, Sutrasthana, 29th chapter, Shloka 24, pp 33. 181 Acharya Agnivesha, Charaka Samhita, English Translation by Bhagawan dash,


172

Biblography

Ramkaran Sharma, 5th Edition, Chaukambha Sanskrit series, 1998, Vimanasthana, 7th Chapter, Shloka 7, pp 199. 182 Ibid,7th Chapter, Shloka 10-13, pp 200-202. 183 The Short Textbook of Microbiology,8th Edition, Jaypee Brothers Medical

Publishers, 1st chapter, pp 12-13. 184 W.B. Hugo and A. D. Rusell, Pharmaceutical Microbiology, Edited by

W.B.Hugo, 2nd Edition, London, Blackwell Scientific Publications,1980, Part I, 1St Chapter, pp 3-4.

185 Warren E. Levinson, Medical Microbiology and Immunology, 3rd Edition, 1992

Prentice-Hall, International Inc, Part I ,1st Chapter, pp 5-6. 186 W.B. Hugo and A. D. Rusell, Pharmaceutical Microbiology, Edited by

W.B.Hugo, 2nd Edition, London, Blackwell Scientific Publications,1980, Part I, 1th Chapter, pp19

187 Warren E. Levinson, Medical Microbiology and Immunology, 3rd Edition, 1992 ,Prentice-Hall, International Inc, Part II ,15th Chapter, pp 68-70. 188 The pharmacological basis of therapeutics Edited by Louis Goodman and Alfred

Gilman 5th Edition, New york, Macmillan publishing co., INC 1976, section XIV, 55th chapter, pp 1097.st Chapter, pp22.

189 I bid, 55th chapter, p 1097 190 Warren E. Levinson, Medical Microbiology and Immunology, 3rd Edition, 1992 Prentice-Hall, International Inc, Part II ,15th Chapter, pp 70-72. 191 Warren E. Levinson, Medical Microbiology and Immunology, 3rd Edition, 1992, Prentice-Hall, International Inc, Part I ,18th Chapter, pp 102-103 192 WB. Hugo and A. D. Rusell, Pharmaceutical Microbiology, Edited by

W.B.Hugo, 2nd Edition, London, Blackwell Scientific Publications,1980, Part I, 2nd Chapter, pp 22.

193 Warren E. Levinson, Medical Microbiology and Immunology, 3rd Edition, 1992

Prentice-Hall, International Inc, Part I ,18th Chapter, pp 91-93 194 I bid, 47th Chapter, pp 235. 195 W. B. Hugo and A. D. Rusell, Pharmaceutical Microbiology, Edited by

W.B.Hugo, 2nd Edition, London, Blackwell Scientific Publications,1980, Part I, 2nd Chapter, pp 42.

196 Warren E. Levinson, Medical Microbiology and Immunology, 3rd Edition, 1992 ,Prentice-Hall, International Inc, Part II ,50th Chapter, pp 245.


173

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197 I bid, 50th Chapter, pp 247. 198 Harsha Mohan, Text Book of Pathology, 4th Edition New delhi, Jayapee

Brothers Medical Publishers, Limited, 2002, Chapter 6, PP 161 199 W.B. Hugo and A. D. Rusell, Pharmaceutical Microbiology, Edited by

W.B.Hugo, 2nd Edition, London, Blackwell Scientific Publications,1980, Part II, 10th Chapter, pp 189-191.

200 Satish Gupta, The short text Book of Medical Microbiology, 8th Edition,

Jayapee brothers, New Delhi, 6th Chapter, pp 43. 201 Ibid, 6th Chapter, pp 43-44 202 Michael pelczar, Text book of Microbiology, 2nd edition, Part I, 1st Chapter

pp 2. 203 Warren E. Levinson, Medical Microbiology and Immunology, 3rd Edition,

1992 Prentice-Hall, International Inc, Part I ,10th Chapter, pp 45 204 Acharya Sushruta, Sushruta Samhita, Commentary by Kaviraja,Dr.Ambikadatta Shastri, Varanasi, Chaukambha Sanskrit Samsthana. Reprint 2005, Part II, 4th Chapter, pp 74. 205 R.S.Satoskar, Pharmacology and Pharmaco therapeutics, 16th Edition, 1999,

Popular Prakashana, Mumbi, 45th Chapter. pp 682.


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K.L.E.Society’s COLLEGE OF PHARMACY J.T.COLLEGE CAMPUS, P.O.Box No.23

GADAG 582101. (Karnataka) Phone: 08372-230042 (O), 08372-231887(R), Tel Fax: 08372 – 221443 To, Dr. ANITHA. H P.G. Scholar, Dept of Rasashastra, D.G.M.A.M.C, GADAG. Sub: Evaluation report of Ayurvedic sample (Rasapushpa) Sir, The evaluation results of the above-referred sample are given below -

1. Orgenoleptic Character:

1 Physical State Solid 2 Colour White 3 Taste - 4 Odour Odourless 5 Texture Smooth Powder 6 Appearance Shiny

2. Size of Particle:

Sample Arithmetic Mean Rasapushpa 9.34μmeter

3. Flow Rate and Property:

Sample Angle of repose Compressibility Index (I)

Rasapushpa 40.14 0 30% 4. Ash Content:

Sample (Rasapushpa) Total Ash 1.47% w/wAcid Insoluble Ash 0.98% w/wWater soluble Ash Nil

5. Solubility:

SL No Test Results

1 Solubility Soluble in Con HNO3 on Heating

V.K.Chandur. Suresh.H.M. (Dept of Pharmaceutics) (Dept of Pharmacognosy)

Rasapushpa antimicrobial rs014-gdg

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