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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
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
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.
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
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 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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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 - - - - + + - +
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
5
Review of Literature
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:
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
6
Review of Literature
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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
7
Review of Literature
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
8
Review of Literature
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
9
Review of Literature
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.
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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.
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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 - - - - - + - -
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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 ]
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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
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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.
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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
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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.
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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.
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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.
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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.
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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:
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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:
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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.
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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.
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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.
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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
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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
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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
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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
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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:
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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,
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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
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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,
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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.
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• 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.
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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
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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.
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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.
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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
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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
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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.
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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
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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.
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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 - - - + - +
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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.
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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.
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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.
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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
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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
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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.
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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.
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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
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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.
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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.
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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
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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.
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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
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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
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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.
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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.
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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
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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.
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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
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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.
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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.
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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
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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.
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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
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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.
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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.
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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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
95
Review of Literature
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
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Review of Literature
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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
Review of Literature
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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
Review of Literature
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Review of Literature
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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
Review of Literature
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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
Review of Literature
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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
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.
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Pharmaceutical Study
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.
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Pharmaceutical Study
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.
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Pharmaceutical Study
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.
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Pharmaceutical Study
• 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.
Date of Commencement : 1-9-2005
Date of Completion : 21-9-2005
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.
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Pharmaceutical Study
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
Date of Commencement : 26-9-2005
Date of Completion : 29-9-2005
Reference : Rasatarangini 5/38,39
Materials : Hingula chakrika – 240gms
Method : Urdwapatana
Equipment : Damaruyantra, Gas stove, cloth
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Pharmaceutical Study
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.
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Pharmaceutical Study
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.
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Pharmaceutical Study
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.
Date of Commencement : 8-10-2005
Date of Completion : 12-10-2005
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 Commencement : 15-11-2005
Date of completion : 17-11-2005
Materials : Hingulotha Parada : 250gms
Haridra: 16 gms
Method : Mardana
Equipment : Khalvayantra
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Pharmaceutical Study
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
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Pharmaceutical Study
Practical No. 6
Name of the practical : Kasisa Shodhana
Reference : BrihatRasaRajaSundara (28.chpt)
Date of commencement : 21-11-2005
Date of completion : 22-11-2005
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.
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Pharmaceutical Study
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
Date of commencement : 5-12-2005
Date of completion : 19-12-2005
Materials : Amber colored glass bottle, cloth,
Multani mud, Scissors.
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Pharmaceutical Study
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.
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Pharmaceutical Study
Practical No. 8
Name of the practical : Preparation of Prematerial
Reference : RT 6/29
Date of commencement : 9-1-2006
Date of completion : 14-1-2006
Materials : Hingulotha Parada : 240gms
Shodhita Kasisa : 240gms
Saindhava lavana : 240gms
Method : Mardana
Equipment : Khalvayantra
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)
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Pharmaceutical Study
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
Date of commencement : 7-2-2006
Date of completion : 9-2-2006
Materials : Prematerial 100gms
Method : Kupipakwa Method (Bahirdhuma
Vidhi)
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Pharmaceutical Study
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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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Pharmaceutical Study
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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Pharmaceutical Study
• 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 -
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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Pharmaceutical Study
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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Pharmaceutical Study
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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Pharmaceutical Study
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
Name of the practical : Preparation of Rasapushpa
Date of commencement : 20-2-2006
Date of completion : 22-2-2006
Same as practical No.9
Results
Weight of the Prematerial : 100gms
Weight of Rasapushpa obtained : 15gms
Weight of Residue obtained : 65gms
Practical No. 11
Name of the practical : Preparation of Rasapushpa
Date of commencement : 6-3-2006
Date of completion : 8-3-2006
Same as practical No.9
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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Pharmaceutical Study
Results
Weight of the Prematerial : 100gms
Weight of Rasapushpa obtained : 18gms
Weight of Residue obtained : 56gms
Practical No. 12
Name of the practical : Preparation of Rasapushpa
Date of commencement : 19-4-2006
Date of completion : 21-4-2006
Same as practical No.9
Results
Weight of the Prematerial : 100gms
Weight of Rasapushpa obtained : 14gms
Weight of Residue obtained : 60gms
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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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
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Analytical Study
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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%
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Analytical Study
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
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131
Antimicrobial Activity
Antimicrobial Activity
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
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Antimicrobial Activity
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.
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Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
134
Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
135
Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
136
Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
137
Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its 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
Review of Literature
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.
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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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Pharmaceutical study
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.
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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%
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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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:
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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.
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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.
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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
100 gms 18 gms 56 gms
100 gms 14 gms 60 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.
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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.
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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.
Antimicrobial Activity
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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
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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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Pharmaceutical study
• 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.
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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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Pharmaceutical 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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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.
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
160
Biblography
BIBLOGRAPHY
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Edition, Edition, Vanranasi, Chowkambha Sanskrit Bhawan 2000, 2nd Chapter Shloka-69, pp 272.
3. Acharya Yadavaji Trikramji, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Vanranasi, Chowkambha Sanskrit Prakashana 2003, 1st Chapter pp 26. 4. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition
New Delhi, Motilal Banarasidas publications., 1979, 9th Taranga, Shloka 1-2, pp 198.
5. Acharya Yadavaji Trikramji, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Vanranasi, Chowkambha Sanskrit Bhawan 2003, 1st Chapter pp 26. 6. Shri Siddanandana Mishra, Ayurevda Rasashastra, 5th Edition, Varanasi,
Chowkambha Orientalia, 1994, Sadharanarasavarga, pp 484. 7. Acharya Yadavaji Trikramji, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Vanranasi, Chowkambha Sanskrit Prakashana 2003, 1st chapter pp 26. 8. Shri Gopal Krishana, Rasendra Sara Sangraha, translated by Dr. Ashok D.
Stapute, 1st Edition Vanranasi, Chowkambha Krishnadas Academy, 2003, 1st Chapter, pp 148.
9. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition
New Delhi, Motilal Banarasidas publications., 1979, 19th Taranga, Shloka 41, pp 301.
10. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd
Edition, Varanasi, Chowkambha Bharati Academy, 1999, 2nd Chapter, Shloka-78-82, pp 275-276.
11. Srimad Govind Bhagvatpada, Rasahridaya Tantra, Edited by Acharya Daulatram
Rasashastri, 2nd Edition, Varanasi, Chowkambha Publications, 2001, Shloka 4, pp 26. 12. Shri Indradev Tripathi, Rsarnava, 4th Edition Varanasi, Chowkambha Sanskrit
Series, 2001, 7th Patala, Shloka 2, pp 86. 13. Shri Gopal Krishana, Rasendra Sara Sangraha, translated by Dr. Ashok D.
Stapute, 1st Edition Varanasi, Chowkambha Krishnadas Academy, 2003, 1st Chapter, Shloka 114-115, pp 58.
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14. Shri Dattaram Chowbe, Brihatrasaraja Sundara, 3rd Edition, Varanasi, Chowkambha Orintalia, 2000, pp 164.
15. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd Edition, Varanasi, Chowkambha Bharati Academy, 2000, 2nd Chapter Shloka1,
pp 252. 16. Shri Bhudeb Mookerjee, Rasajalanidhi, 2nd Edition, Varanasi, Shri Gokul
Mudranalaya, 1984, II-Volume, 1st Chapter, pp 2. 17. Acharya Somadeva, Rasendra Chintamani, 1st Edition, Varanasi, Chowkamha
Orientalia, 1984, 11th Chapter, Shloka 60-61, pp 190. 18. Shri Vagbhatacharya, Rasa Ratna Samucchaya, Edited by Indradev Tripathi, 2nd
Edition, Varanasi, Chowkambha Sanskrit Bhavan, 2003, 3rd Chpter, Shloka 126-127, pp 37.
19. Acharya Yashodhara, Rasaprakasha Sudhakara, Siddiprada commentary by
Siddanandana Mishra, 1st Edition, Varanasi, Chowkamha Orientalia, 1983, 6th Chapter, pp 127.
20. Acharya Yadavaji Trikramji, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Vanranasi, Chowkambha Sanskrit Bhavan, 1998, 4th Chapter pp 26. 21. Acharya Somadeva, Rasendra Chintamani, 1st Edition, Varanasi, Chowkamha
Orientalia, 1984, 11th Chapter, Shloka 107, pp 195. 22. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd
Edition, Varanasi, Chowkambha Bharati Academy, 1999, 2nd chapter Shloka-69-71, pp 272-273.
23. Shri Vagbhatacharya, Rasa Ratna Samucchaya, Edited by Dr. Indradev Tripathi,
2nd Edition, Varanasi, Chowkambha Sanskrit Bhawan, 2003, 3rd Chapter, Shloka 147, pp 41.
24. Acharya Yashodhara, Rasaprakasha Sudhakara, Siddiprada commentary by
Siddanandana Mishra, 2nd Edition, Varanasi, Chowkamha Orientalia, 1999, 6th Chapter, Shloka 85, pp 130.
25. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition,
Varanasi, Motilal Banarasidas, 2000, 9th Taranga, Shloka 4, pp 199. 26. Yadavaji Trikramji Acharya, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Varanasi, Chowkambha Sanskrit Bhawan, 2003, 6th chapter Shloka 85, pp 130.
27. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition,
Varanasi, Motilal Banarasidas, 1979, 9th Taranga, Shloka 3, pp 1999. 28. Ibid, 9th Taranga, Shloka 11, pp 200.
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29. Shri Dattaram Chowbe, Brihatrasaraja Sundara, 3rd Edition, Varanasi, Chowkambha Orintalia, 2000, uparasa Prakarana, pp 168.
30. Shri Indradev Tripathi, Rsasrnava, 4th Edition, Varanasi, Chowkambha Sanskrit
Series, 2001, 7th Patala, Shloka 236, pp 60. 31. Acharya Yashodhara, Rasaprakasha Sudhakara, Siddiprada commentary by
Siddanandana Mishra, 2nd Edition, Varanasi, Chowkamha Orientalia, 1999, 6th Chapter, Shloka 86, pp 130.
32. Shri Vagbhatacharya, Rasa Ratna Samucchaya, Edited by Dr. Indradev Tripathi,
2nd Edition, Varanasi, Chowkambha Sanskrit Bhawan, 2003, 3rd Chapter, Shloka 152, pp 41.
33. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition,
Varanasi, Motilal Banarasidas, 1979, 9th Taranga, Shloka 14-15, pp 202. 34. Ibid, 9th Taranga, Shloka 16-17, pp 202. 35. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd
Edition, Varanasi, Chowkambha Bharati Academy, 1999, 2nd Chapter Shloka-77, pp 275.
36. Acharya Yashodhara, Rasaprakasha Sudhakara, Siddiprada commentary by
Siddanandana Mishra, 2nd Edition Varanasi, Chowkamha Orientalia, 1983, 6th Chapter, Shloka 87, pp 130.
37. Shri Vagbhatacharya, Rasa Ratna Samucchaya, Edited by Dr. Indradev Tripathi,
2nd Edition Varanasi, Chowkambha Sanskrit Bhawan, 2003, 3rd Chapter, Shloka 150, pp 41.
38. Acharya Somadeva, Rasendra Chintamani, 1st Edition, Varanasi, Chowkamha
Orientalia, 1984, 11th Chapter, Shloka 108, pp 165. 39. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd
Edition, Vanranasi, Chowkambha Bharati Academy, 1999, 2nd Chapter Shloka-72, pp 275.
40. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition
Varanasi, Motilal Banarasidas, 1979, 9th Taranga, Shloka 18, pp 202. 41. Acharya Yadavaji Trikramji, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Varanasi, Chowkambha Sanskrit Bhawan, 2003, 4th Chapter shloka 52, pp 26.
42. Acharya Yashodhara, Rasaprakasha Sudhakara, Siddiprada commentary by
Siddanandana Mishra, 2nd Edition Varanasi, Chowkamha Orientalia, 1999, 6th Chapter, Shloka 108-109, pp 195.
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43. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition Varanasi, Motilal Banarasidas, 1979, 9th Taranga, Shloka 18-19, pp 202.
44. Ibid, 9th Taranga, Shloka 38, pp 82. 45. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd
Edition, Varanasi, Chowkambha Bharati Academy, 1999, 2nd Chapter Shloka-83-85, pp 277.
46. Shri Vagbhatacharya, Rasa Ratna Samucchaya, Edited by Dr. Indradev Tripathi,
2nd Edition Varanasi, Chowkambha Sanskrit Bhavan, 2003, 3rd Chapter, Shloka 154, pp 41.
47. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd
Edition, Varanasi, Chowkambha Bharati Academy, 1999, 2nd Chapter Shloka-76, pp 275.
48. Shri Dattaram Chowbe, Brihatrasaraja Sundara, 3rd Edition, Varanasi,
Chowkambha Orientalia, 2000, uparasa prakarana pp 165-166. 49. Shri Govindadas, Bhaishajya Ratnavali, Edited by Kaviraj Ambikadatta Shastri,
17th edition, Varanasi Chowkambha Sansthana, 2002, 5th Chapter, Shloka 473-482, pp 82.
50. Ibid, 5th Taranga, Shloka 483, pp 82. 51. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition,
Varanasi, Motilal Banarasidas, 1979, 24th Taranga, Shloka 106-109, pp 667. 52. Ibid, 9th Taranga, Shloka 31-33, pp 204. 53. Shri Damodar Joshi, Rasashastra, edited by Dr. K.P. Saikumari Amma, Trivandurum Publication division, Ayurveda college, pp 174-175. 54. htpp =1/mineral galleries. Com/mineral data/sulfide/cinnabar/cinnabar. 55. Sri Vidya vasudeva Malashankara Dwivedi, Parada Vijnaneeyam, 3rd edition, Sri
Sharma Ayurveda Mandira, 1997, 2nd Chapter pp 22-23. 56. Acharya Madhava, Ayurevda Prakash, Edited by Gulraj Sharma Mishra, 2nd
Edition, Varanasi, Chowkambha Bharati Academy, 2000, 2nd Chapter Shloka-83-85, pp 277.
57. Shri Vagbhatacharya, Rasa Ratna Samucchaya, Edited by Dr. Indradev Tripathi,
2nd Edition, Varanasi, Chowkambha Sanskrit Bhawan, 2003, 3rd Chapter, Shloka 154, pp 41.
58. Acharya Yadavaji Trikramji, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Varanasi, Chowkambha Sanskrit Bhawan, 2003, 4th Chapter Shloka 16, pp 11-12.
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59. Shri Siddanandana Mishra, Ayurvediya Rasashastra, 5th Edition, Varanasi,
Chaukambha Orientalia, 1994, Parada prakarana, 3rd chapter, PP 109. 60. Shri Sadananda Sharma, Rasatarangini, Edited by Kashinath Shartri, 11th Edition
Varanasi, Motilal Banarasidas, 1979, 5th Taranga, Shloka 38, pp 82. 61. Shri Gopal Krishana, Rasendra Sara Sangraha, translated by Dr. Ashok D.
Stapute, 1st Edition, Varanasi, Chowkambha Krishnadas Academy, 2003, 1st Chapter, pp 6.
62. Acharya Yadavaji Trikramji, Rasamritam, translated by Sri Damodar Joshi, 2nd
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Sharma Ayurveda Mandira, 1997, 1st Chapter pp 2. 64. Acharya pandit Vishwanath Dwivedi, 2nd edition, Varanasi, Nagpur, Sharma
Ayurveda Mandira, 5th Chapter, pp 154. 65. Acharya Trikramji Acharya, Rasamritam, translated by Sri Damodar Joshi, 2nd
Edition, Varanasi, Chowkambha Sanskrit Bhawan, 2003, 1st Chapter pp 3. 66. Dr. Nadakarni, Indian Materia Media, 2nd edition, Mumbai, Popular Prakashana,
1976, volume 2, pp 67. 67. Sri Vidya vasudeva Mulashankara Dwivedi, Parada Vijnaneeyam, 3rd edition, Sri
Sharma Ayurveda Mandira, 1997, 1st Chapter pp 2. 68. Agniversha, Charaka Samita, 7th Chapter, Chikitsasthana, Shloka 70-71,
Kashinath Shartri, 5th edition, Varanasi, Chowkambha Sanskrit Samsthana, 1997, p 260.
69. Shushruta Acharya, Sushruta Samhita, 25th Chapter, Chikitsasthana, Shloka 39,
Kaviraj Ambikadatta Shastri, 8th edition, Varanasi Chowkambha Sanskrit Samsthana, 2000.
70. Shri Vagbhatacharya, Asthanga Sangraha, 32nd Chapter, uttaratantra, Shloka 31-
32, K.R. Srikantha Murthy, 1st Edition, Varanasi Chowkambha Orintalia, 1996, pp 503.
71 Acharya Vagbhata, Rasa Ratna samucchaya,_Edited by Ambikadatta Shastri 9th
Edition, Varanasi, Chaukambha Amarabharati Prakashana, 1998, 1st Chapter, Shloka 88; pp 10.
72 Shri Gopalkrishna, Rasandra Sara Sangraha, translated by Dr. Ashok, 1st Edition,
Varanasai, Chaukambha krishnadas Academy, 2003, 1st Chapter, pp 8
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73 Acharya yashodhar, Rasaprakash Sudhakara, Siddiprada commentary by S Siddanandana Mishra, 1st Edition, Varanasi, Chaukambha Orientalia, 1983, 6th chapter, shloka 87, pp 130.
74 Acharya Indradev Tripathi, Rasarnava, 4th Editon, Varanasi, Chaukambha
Sansrkrit Series, 2001, 10th Patala, shloka 4, pp 130 75 Acharya Madhava, Ayurveda Prakash, Edited by Shri Gulraj Sharma Mishra, 2nd
Edition, Varanasi, Chaukambha Bharati Academy, 1999, 1st Chapter, pp 18. 76 Shri Dattaram choube, Brihat Rasa Rajasundara, 3rd Edition, Varanasi,
Chaukambha Orientalia, 2000, 1st Chapter, Shloka pp 5 77 Shri Vagbhatacharya, Rasa Ratna Samucchaya, Edited by Dr. Indradev Tripathi,
2nd Edition, Varanasi, Chaukambha Orientalia, 2000, 1st Chapter, Shloka 67, pp 7. 78 Ibid, 1st Chapter, Shloka 67, pp 7. 79 Acharya Madhava, Ayurveda prakash, Edited by Gulraja sharma Mishra, 2nd
Edition, Varanasi, Chaukambha, Bharati Academy, 1999, 1st Chapter, Shloka 16-18, pp 22.
80 Shri Bhudeb Mookarjee, Rasajala nidhi, Vol I, Edited by Siddinandana Mishra
2nd Edition,Varanasi, Chaukambha Sanskrit Bharti, 1998, 4th Chapter, pp 37. 81 Dr. Chandrabhushan Jha, Ayurveda Rasashastra, 2nd Edition, Varanasi,
Chukambha Surabharati,Prakashana, 1998, 9th Chapter, pp 127. 82 Acharya Madhava, Ayurveda prakash, Edited by Gulraj Sharma, 2nd Edition
Varanasi, 1st chapter, shloka 105, pp 92. 83 Shri Sadananda Sharma, Rasastarangini, Edited by Kashinatha Shastri, 11th
Edition, Varanasi, Motilal Banarasidas, 2000, 5th, Taranga, Shloka 31, pp 80. 84 Ibid, 5th Taranga, Shloka 35, pp 81. 85 Acharya Madhava, Ayurveda Prakash, Edited by Gulraj Sharma, 2nd Edition,
Varansi, Chaukambha Bharati Academy, 1st Chapter, Shloka 36, pp 32. 86 Acharya Yadavji Trikramji, translated by Dr.Damodar Joshi, 2nd Edition, Varansi,
Chaukambha Snskrit Bhawan, 2003, 1st Chapter, pp 4. 87 Shri Sadananda Sharma,Rasatarangini,Edited by Kashinath Shastri Varanasi,
Motilal Banarasidas,7th Taranga, Shloka 6, pp 5.
88 Ibid,7th Taranga,Shoka 87-88,pp 169. 89 Acharya Madhava, Ayurveda Pakash,Edited by Gulraj Sharma Mishra ,2nd
Eddition,Varanasi,Chaukambha Bharati Academy, 1t Chapter, Shloka 375 pp 193.
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90 Dr. K.N.Nadakarni, Indian Materia Medica 2nd Editon, Mumbi, Popular
Prakashana, 1976, Volume II, pp 35. 91 Achary Yadvaji Trikramji Rasamritam, translated by Damodar Joshi, 2nd Edition
Varanasi, Vhaukambha Sanskrit Bhawan, 2003, 1st chapter, Shloka 22-26, pp ;15. 92 Acharya Yashodhara, Rasaprakasha sudhakara, Siddiprada commentary by sidda
nandana Mishra, 1st Edition, Varanasi, Vhaukambha Vrientalia, 1983, 8th Chapter, Shloka 72-73, pp 159.
93 The pharmacological basis of therapeutics, Edited by Louis Goodman and Alfred
Gilman, 5th Edition, New york, Macmillan Publishing co., INC, 1976, 11th section, 46th Chapter, pp 435-937.
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
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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.
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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
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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
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170
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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
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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,
Preparation, Physico Chemical analysis of Rasapushpa and its Antimicrobial Activity
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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.
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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)