butea monosperma#dg03 blg

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“ SCREENING OF ANTIMICROBIAL ACTIVITY OF

BUTEA MONOSPERMA (LAM) SEED ”

By Dr. Manjiri Kulkarni

B.A.M.S.

Dissertation Submitted to The Rajiv Gandhi University of Health Sciences,

Karnataka, Bangalore.

In Partial Fulfillment of the requirements for the Degree of

AYURVEDA VACHASPATHI in

DRAVYAGUNA

under the guidance of Dr. S. K. Hiremath

M. D. (Jamnagar)

POST GRADUATE DEPARTMENT OF DRAVYAGUNA

K. L. E.’s Shri. B. M. Kankanawadi Ayurved Mahavidyalaya, Post - Graduate Studies Cum Research Center,

Shahapur, BELGAUM. ____________________________________________________________________

2007-08.

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“SREENING OF ANTIMICROBIAL ACTIVITY OF ‘PALASH SEED’ (BUTEA MONOSPERMA LIM.)”

0

Ayurmitra

TAyComprehended

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The Rajiv Gandhi University of Health Sciences, Karnataka, Bangalore.

DECLARATION BY THE CANDIDATE

I here by declare that this dissertation / thesis entitled ‘SCREENING OF

ANTIMICROBIAL ACTIVITY OF BUTEA MONOSPERMA (LAM.)’. Is a bonafide and

genuine research work carried out by me under the guidance of Dr . S. K. Hiremath

M. D. Professor.

Date : Signature of the Candidate Place : Belgaum. Dr. Manjiri Kulkarni P. G. Scholar

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CERTIFICATE BY THE GUIDE

This is to certify that this dissertation entitled ‘SCREENING OF

ANTIMICROBIAL ACTIVITY OF BUTEA MONOSPERMA (LAM.)’ is a bonafide

research work done by Dr. Manjiri Kulkarni in partial fulfillment of the requirement for the

degree of AYURVEDA VACHASPATHI.

Date : Signature

Place : Belgaum. Dr. S. K. Hiremath, M. D.

Professor P. G. DEPARTMENT OF DRAVYAGUNA K. L. E.'s Shri. B. M. Kankanawadi Ayurved Mahavidyalaya, Post - Graduate Studies Cum Research Centre, Shahapur, BELGAUM.

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ENDORSEMENT BY THE HOD, PRINCIPAL OF THE

INSTITUTION

This is to certify that this dissertation entitled ‘ SCREENING OF

ANTIMICROBIAL ACTIVITY OF BUTEA MONOSPERMA (LAM.) ’ is a bonafide

research work done by Dr. Manjiri Kulkarni under the guidance of Dr. S. K. Hiremath

M. D. Professor.

Dr. S. K. Hiremath, (M. D. Jamnagar) Dr. B. S. Prasad M. D. Ph. D.

Professor Principal,

P. G. DEPARTMENT OF DRAVYAGUNA K. L. E.'s Society.

K. L. E.'s Shri. B. M. Kankanawadi Belgaum.

Ayurved Mahavidyalaya,

Post - Graduate Studies Cum Research Cemtre,

Shahapur, BELGAUM.

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COPYRIGHT

DECLARATION BY THE CANDIDATE

I hereby declare that the Rajiv Gandhi University of Health Sciences, Karnataka

shall have the right to preserve, use and disseminate this dissertation / thesis in print or

electronic format for academic / research purpose.

Date : Signature of the Candidate

Place : Belgaum. Dr. Manjiri Kulkarni P. G. Scholar

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CONTENTS

TABLE OF CONTENTS

Sl. No. Contents Page No.

1 Introduction 1-2 2 Objectives 3 3 Review of Literature a) Drug review 4-27 b) Microbiology review 28-41 c) Extraction Procedure review 41-45 4 Methodology 4.1) Collection 46 4.2) Authenification 46 4.3) Collection of Test Drugs and storage 46 4.4) Pharmacognostic Study 47 4.5) Physico – Chemical Study 48-53 4.6) Schematic Chart of extraction 54 4.7) Ethanol extraction 55-57 4.8) Aqueous extraction 57-58 4.9) Preliminary phytochemical screening 59-62 4.10) Qualitative confirmation (T.L.C., H.P.T.L.C.) 63-72 4.11) Evaluation of Antimicrobial activity 72-78 5 Results 79-86 6 Discussion 87-90 7 Conclusion 91 8 Summary 92 9 Biblography references 93-98 10 Annexure I-V

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LIST OF TABLE

Sl. No. Tables Page No.

1 Showing synonyms according to different authors 13 2 Rasa, Guna, Veerya, Vipak, Doshghanata 16 3 Prayojyange according to different authors 16 4 Karma of Palash 17 5 Prayoga (uses) of Palash 18 6 Doshaghnata according to different authors 18 7 Prayojyanya according to different authors 26 8 Organoleptic characters 46 9 Preliminary phyto-chemical screening 81-82 10 Anti microgial results 84 11 E-coli 85 12 Staphylococcus Aureus 85 13 Bacillus subtillis 85 14 Candida albicans 86

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LIST OF PHOTOS AND FIGURES

Sl. No. Plates Contents Page No.

1 Plates No. 1 Palash plant with flowers, pods and seeds 1

2 Plates No. 2 Distillation apparatus 1

3 Plates No. 3 Soxhlet apparatus 1

4 Plates No. 4 Muffle furnace 1

5 Plates No. 5 Incubator 1

6 Plates No. 6 Ash of Palash seed 2

7 Plates No. 7 Water soluble extractive 2

8 Plates No. 8 Alcohol soluble extractive 2

9 Plates No. 9 Cold water extract of Palash Beeja 2

10 Plates No. 10 Culture medium 3

11 Plates No. 11 Micro organisum 3

12 Plates No. 12 Micro Pippete 3

13 Plates No. 13 Micro scopic view of Palash seed 3

14 Plates No. 14 Bacillus subtillus 4

15 Plates No. 15 Candida albicans 4

16 Plates No. 16 E-coli 4

17 Plates No. 17 Staphylococcus 4

18 Plates No. 18 Water sol. extractive 5

19 Plates No. 19 Alcohol soluble extractive 5

20 Plates No. 20, 21 Water bath evaporation of Alc.and water sol. ext. 5

21 Plates No. 22 Evaporate extractive 5

22 Plates No. 23 Water and alcohol extract of Palash seed 5

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ACKNOWLEDGEMENT

A dissertation is an aim for every student. It is very tough and a field which is a very

ailien for any student. Therefore during the compilation of this entire project. I have been

blessed with a lot of senior faculty members who have gone to extreme limits in helping me

whole heartedly to complete my research work project. I owe a lot of gratitude to them. I

have strived to do my best of my knowledge. I hope I have achieved my success because of

everybody's keen participation.

It is my privilege for having worked under the guidance of Dr.Shivamurti

K.Hiremath M.D. (Jamanagar) (Professor) Head of Department, postgraduate department,

DRAVYAGUNA for his valuable support and guidance. I express my deep sense of

gratitude for suggesting this study and for his constant encouragement with allmost patience

throughout the course of my research work project.

I express my sincere thanks to our Principal Dr B. S. Prasad M.D PhD, for

provided me valuable, timely given suggestions and proper guidance and

encouragement for my research work . I thank Dr. Yogini. R. Kulkarni M.D. PhD,

Dr.S.R.Kulkarni M.D. and Dr Arun Chougale M.D. for their valuable guidance throughout my

studies. I am very much thankful to Dr. R.V. Savadi and Dr Kalpana Patil M Phram PhD, of

K.L.E's Pharmacy College, Belgaum for their valuable support during Analytical studies.

I am also thankful to Dr. S. D. Kolkute, Mr. Shripad Bhat and Dr Harsha Hegde,

Research officer of RCMR Belgaum. I also thank Mr. Haneef of RCMR for carrying out

Antimicrobial studies.

I also thank Dr Anand. S. Ammanagi M.D.DNB, MNAMS for their valuable guidance in

carrying out antimicrobial studies.

I thank Mr. Prakash Kokate NAFARI Pune. For carrying out TLC and HPTLC

analysis.

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I express my sincere sense of gratitude to the management committee and Principal

Dr Gangadharan. (Ayurved Medical College, NIPANI) who gave me full moral support,

encouragement and kind co-operation.

I also would like to thank always cherish memories of my senior and junior

colleagues Dr. Ramesh Konakeri, Dr. Deepak Mummigatti, Dr. Mahadev Gundakalle, Dr.

Manisha, Dr. Poornima B, Dr. Poornima Undi, Dr Ajeet Herwade, Dr. Nayana.Patil who

helped whenever needed at the studies.

I express my sincere heartfelt thanks to all the teaching staff members of other

departments for their valuable suggestion and support during my postgraduate studies.

I Thank our college librarian Mrs. G.C. Gulla who provided me necessary books and

journals for my studies.

I am very much thankful to my family friends Shailaja Katti and Mr. Anand

Deshpande for their moral support and kind co-operation.

I indebeted to my beloved parents and family members, family friends Shailaja Katti

and Mr. Anand Deshpande for their moral support and kind co-opration.

Lastly Vighnaharta Graphics D.T.P. Center, Nipani.

For bringing out the dissertation in the present form.

Date : / / Signature of Candidate

Place : Belgaum (Dr. Manjiri Kulkarni)

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LIST OF ABBREVIATIONS

1. A. H. - Ashtanga Hrudaya 2. A. H. Ni - Ashtanga Hrudaya Nidana Sthana 3. A. H. Su - Ashtanga Hrudaya Stura sthan 4. A. K. - Amarakosha 5. A. R. - Abhidana Ratnamala 6. A. S. - Ashtanga Sangraha 7. B. A. - Bruhat Dravyaguna Adarsha 8. B. P. N / B. P. N - Bhavaprakash Nighantu 9. B. R. - Bhaishajya Ratnavali 10. Cha - Charaka 11. Cha. Chi - Charaka Chikitsa 12. Cha. Su - Charaka Chikitsa Sutra Sthana 13. Cha. S. N. - Charaka Chikitsa Samhita Nidhana Sthana 14. Cha. S. Vi - Charaka Chikitsa Vimana Sthana 15. C. da/c. d. - Chakradata 16. D. N. - Dhanvantri Nighantu 17. D. G. H. - Dravya Guna Hastamalaka 18. D. G. (V. M. G.) - Dravya Guna Hastamalaka Vijanana by V. M. Gogte 19. H. S. - Haritha Samhita 20. K. N. - Kaiyadeva Nighantu 21. L. S. - Longitudinal Section 22. M. N. - Madhanapala Nighantu 23. Mau. N. - Mahaushadha Nighantu 24. MIC. - Minimal inhibitory concentration 25. N. A. - Nighantu Adarsha 26. Sha - s - Sharangadhara Samhita 27. S/k - Shloka 28. Su - Sushruta 29. Su. S. N. - Sushruta Samhita Nidan Sthana 30. T. S. - Transverse Section 31. T. L. C. - Thin layer chromatography 32. H. P. T. L. C. - High performance Thin layer chromatography 33. Y. R. - Yoga Ratnakara

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ABSTRACT

A. Background and Objectives :

In modern medical acrence various diseases are treated with antimicrobials, which

plays vital role in chemotherapy. It is necessary to develop such drugs from natural sources.

"Palash" is described as 'Krimighna' therefore this work is under taken to evaluate

antimicrobial effect of ' Palash' seed.

Objectives :

1. Pharmacognostical study, preliminary phytochemical screening of 'Palash' seed.

2. Antimicrobial activity of ‘Palash’ seed extracts by different solvents.

B. Methods :

I. Organoleptic character such as colour, taste etc. are studied. Total ash value acid

insoluble ash value, water and alcohol extractive values were determined.

II. Extraction of Palash' seed was done by wring ethinol and water.

III. Preliminary phytochemical screening and TLC and HPTCL of extracts were also

carried out.

Antibacterial and antifungal studies were carried out by cup diffusion method at three

concentrations.

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C. Results :

1. Total Ash value of ‘Palash’ seed - 7.685 %

2. Acid insoluble ash value - 0.03 gms.

3. Water soluble extractive value - 38.72 %

4. Alcohol soluble extractive value - 20.72 %

5. Phytoconstituentes present in extract are - Glycosides, Carbohydrates,

Terpenoids, Tannins present in both

extract, Alkoloid present in aq. extract,

steroils present in alcohol extract.

6. T.L.C. and H.P.T.L.C. report of the samples showed different peaks for different Rf.

Values and ethnol extract values are double than water extract.

7. The antimicrobial tests were done by Mueller – Hinton agar cup diffusion method,

which shows negetive result with water extract for all microbes and ethnol extract

shows also registrant for Candida albicans, but sensitive for remaining microbes.

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1. INTRODUCTION

Potent substances are present in version plants and herbs. Modern medicines are not free of adverse effects, so the use of herbal formulation is ever increasing in western world. People in large scale wanted potent drugs. But at the same time they want drugs to be safe. Because of this reason traditional system of medicine is regaining its past glory once again. According to Ayurvedic literature 'PALASH' - ("Butea monosperma") is used to treat various diseases. It is widely and abundantly available. Different parts of Palash are used in various diseases. According to the classical reference having a prime property of "Krimighna"(1). It can be used to treat various skin diseases. An antimicrobial agent is one that inhibits or kills microbes but causes minimum harm to normal tissues of human being. As many micro organisms have become resistant to the newest antimicrobial agent. So now we have taken furthers step towards inventing new antimicrobial agents from the field of Ayurveda without isolating active constituents. Thus we have conducted preliminary study of “Palash” seed by phytochemical analysis and antimicrobial activity. (A) Botanical authentification of the test drug from the respective taxonomist export. (B) Pharmacogonostic study i.e. macroscopic study, microscopic study, ash value, water

soluble extractive values should be determined. (C) Preliminary phytochemical screening extracts of “Palash beeja” (seeds). (D) Antimicrobial activity of the alcohol and water extract of 'Palash' seed by Agar plate

method. As we are not isolating active principle from plant drug. So dose tends to be little higher, as compared to the synthetic preparation. Advantages are more than disadvantages. We are administering drug in natural form which is widely and easily accepted by the patient. We will be are giving these drugs without harming the patient and would be more effective without compromising with our basic principles.

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A) Review of Liturature :- Drug review, Microbiology review, extraction procedure review,

B) Methodology :- Collection of the drug, Authentification, Storage, Pharmacognostic study, Phytochemical screening, TCL, Antimicrobial activity, C) Result :- Aqueous extract showed resistant activity to bacteria and

fungi and Ethenol extract showed resitant activity to fungi and sensitive to bacteries.

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2. OBJECTIVES

(2.1) Pharmacognostic and physicochemical study of 'Palash' seed.

To study regarding phyto constituents of the drugs so that it maybe helpful to

interpret regarding its pharmaco kinetic in further studies.

(2.2) Preliminary phyto chemical screening of 'Palash' seed.

To know presence of phytochemicals which also may help in studying the pharmaco

kinetic of the drug.

(2.3) Anti microbial activity of 'Palash' seed.

To know the antimicrobial effect, so that we can interpret on the 'Krimighna' effect

which is stated in Ayurveda texts and then a step wise and complete antimicrobial

study can be done for better therapeutic effects.

Review of Literature ==================================================================

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(A) Drug Review

(1) Vedic Review (2), (3) :

We have got the references of use of ‘Palash’ since vedic period. In this kala single

drug thearpy was present and since then single drug therapy is being used

The use of Palash was common in vedic period not only to treat the ailments but also

in routine life and in holy rituals. In vedic era leaves, stem and flowers were more

used but there are references regarding use of the seed. In vedic kala Palash tree was

known as Shant Vruskha and 'Bramha Varchass'. Samidha of this plant were used at

the time of different Homa and Yadgnyas.

According to 'Koushika sutra', Palash is ' Medhajanan' and lepa of Palash these

leaves was applied in 'Jalodar' (Ascitis). Keshav told it was 'Sarvaroga Bheshaja' and

also it was used in Krimi Roga ( Ke.P. 9. 4/25/20 ).

In Rigveda ' Kinshuk' was the synonym given for Palash. 'Kinshuk' means who

shines brightly. This synonym is given because of its bright attractive colour of the

flower. In Rigveda kala, Palash leaves were used with Ashwath and we get the

referenes of uses of Palash leaves with Nyagrodha in Atharvaveda. In Upanayan

samskar 'Dand' (Stick) which is used by 'Brahmachari' was also made from Palash

wood. Palash was used frequently because it had a power to destroy the 'Rakshasas'

so also its stem was being used in Yadgnyas and patras were used to prepare

'Abhishek paatra. 'In Atharaveda “Parnamani” of Palash patra was used to gain Bala,

Aayu, Samruddhi and fame. According to 'Shrout sutra' 'patra valkala' of Palash was

used in preparation of curds.

The tree is considered sacred both by Hindus and Buddhist. Hindus consider it, as

holy because of the trifoliate formation of leaves which represents the Holy trinity of

Brahma (the creator) on the left, Vishnu (the preserver) in the middle and Shiva (the

destroyer) on the right. The flower of this plant are offered especially to Goddess

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Kali. In Krishnasthami Vratam, wood of Palash is also being used. Because of curse

of Goddess Parvati, Bramha was converted into the tree of Palash. In Navagraha

Stotra written by Vyasa the character of Ketu has been compared with Palash flower.

1) Samhita kala :

a) Charak Samhita : In charak samhita Palash is not included in any Gana but it is said

to be used in treating many diseases and in different ritual functions. We couldnot get

any synonym for Palash in this samhita. Leaf, flower, skin, seed and kshar are the

parts used in different yogas.

Palash is used in kasa, Grahani, Arsh, Udararoga, kushtha and in skin diseases. But

seed is mainly used in skin diseases Kushtha, Arsh and Udarroga (ChS.Chi 110/13)

(ChS.Chi 92/14)

b) Sushrut Samhita : As per Sushrutcharya Palash is included in Rodhradi,

Mustakadi, Ambavashtadi and Nyagrodhadi Gana. Kinshuk is used as synonym for

Palash in various explanation in Sushrut Samhita. Utility of Palash in kushtha,

Gulma, Udar Roga, Arsh, Bhagna, Netraroga and Raktaprasadan (Su. Chi 7/2, 9/7,

18/42, 19/49).

KASHYAP SAMHITA :

In this Samhita kwath of Palash is used to give Mukti from 'Sheetputana Grah' for

kids.

2) Sangraha Kala :

a) Ashtang Sangraha and Ashtang Hridaya : Vagbhata also includes Palash in

Rodharadi, Mustadi etc., (A.H.S 15/32, 15/38) Rogaghnata of Palash is given in

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Krimi gulma, Kushtha, Vaata vyadhi, Rasayan, Shwitra kushtha, Arsh (A.H. Chi

20/26, A.S. Chi 5/9).

b) Chakra Datta : He has told the utility of Palash same as Charak but he used

Kinshuk as synonym.(Ch.Chi.3/258)

c) Sharangdhar Samhita (4) : According to Sharangdhar, Palash seed is used in 'Loha

Rasayan' 'Yoni Sankochanarth lepa. Kshar of Palash is used for kesh Nirharnana lepa

and for Netra prasadan vidhi.

d) Bhavaprakash (5) : In Bhavaprakash, Palash includes in Vatadi Varg. Many

Synonyms given for Palash are found here. And it is mainly indicated in Krimi, Arsh,

Prameha, Vataja and Kaphaja Vikar, Kushtha, Gulma and Udara roga.

e) Bhaishajya Ratnavali : It is used in Krimi, Arsh and Vatrog many diseases. Seeds

are used in preparation of many yogas.

f) Yoga Ratnakar (6) : Palash beeja recommended in Krimi Roga, Arsh, Gulma etc.,

III) Nighantu kala : Many drugs have been described in detail by giving different

synonyms and their properties and uses. The drug Palash has been described in every

Nighantu because it is well known drug since Vedic Period.

1) Bhavaprakash Nighantu (7) : In this Nighantu Palash is mentioned in 'Vatadi Varg'

and given nine synonyms. Though it is used in many diseases mainly indicated in

Krumi Rog.

2) Shaligram Nighantu (8) : This nighantu includes Palash in 'Phala Varga' and gives

23 synonyms. Author stating that it is Krimihar and mainly seeds are used in Skin

diseases.

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3) Dhanwantari Nighantu (9) : In this Nighantu it is included in 'Amradi varg' and 14

synonyms are given for it. Krimihar property of seeds is mentioned.

4) Kaiyadeva Nighantu (10) : In this Nighantu it is included in Aushadhi varga and

twelve synonyms are given for it.

5) Madanpal Nighantu (11) : In this Nighantu it is included into the 'Vatadi Varga'.

Different Synonyms are given in this book. Types of flowers and its uses in different

Rogas are explained.

6) Raj Nighantu (12) : In this Nighantu it is included given in 'Karviryadi Varga' 11

synonyms are given for it. Nighantukar also mention that it is 'Krimihar' and its seeds

are used to treat various skin disorders. He has mentioned 4 types of Palash 1) Peeta

2) Shweta 3) Rakta and 4) Neel pushpa.Though all 4 are having same quality, in

those 'Shweta' is the best for treatment.

7) Shodhal Nighantu (13) : In this Nighantu seed is said to have Krimihar property.

8) Nighantu Adarsha (14) : Palash is classified in “Palashadi varga” so many references

of different authors regarding Palash are given in this Nighantu.

9) Priya Nighantu (15): He mentioned it in “Haritakyadi Varga”. Its seeds are Krimihar

in action and seven synonyms are given.

ADHUNIK KALA :

1) Indian Mateia Medica (16) : The author of this book has mentioned vernacular

names and chemical constituents of this drug.. Seeds are indicated in many skin

diseases. Internally for worms, externally for ringworm, boils, pimples, bubbles,

tumours and also in various other diseases.

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2) Data base on medicinal plants used in Ayurveda Vol 1 (17) : Detail explanation

regarding plant Palash has been given such as - Family, Classical names (Synonyms)

vernacular names, morphology, useful parts, uses and doses.Along with that

pharmacognosy, chemical constituents, pharmacological activities, toxicology and

therapeutic evaluation is explained.

3) Indian Herbal Pharmacopoeia (18) : Detail Information regarding Palash seed is

given i.e. macroscopic, microscopic examinations etc.,

4) Medicinal Plants Quality Standards of Indian (19) : Description regarding plant is

given and also all types of phytochemical tests are explained in detail.

5) Vanoushadi Nidarshika (20) : Containts description of seed and Im ¶e«dœ¦d «ddÎdd

is given.

6) Indian Medicinal Plants (21) : Detail description like names, vernacular names

distribution etc., are given. Also properties and uses of seeds are given.

7) Wild medicinal plants of India (22) : pharmacognasy and phytochemical description

and use of Palash seed in folk medicine

8) Indian medicinal plant by Kirtikar and Basu. (23) : Properties of seed in detail have

described that seeds are not dry digestible, antihelmentic, aperient used in urinary

discharge, piles, cure, skin diseases, tumours, abdominal trouble and also used in

scorpion stinge.

9) The Useful Plants of India (24) : Herpes in dhobie's itch seed are pounded with

lemon juice and used in skin diseases. Seed yield a fatty oil (18%).

ii) Synonyms and their Meanings and Interpretations : Morphology of the plant, prominent characters, utility all the things have been mentioned by way of synonyms. There is a class of Nighantus which describe the drugs by its synonyms. Acharya

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Narahari Pandit the Author of Raja Nighantu has tried to arrange the synonyms in a proper way and elaborated them into 7 categories.

The various synonyms of “Palash” as given in Nighantus as follows : DIFFERENT SYNONYMS :

1) nbme: (^m.) àeñVm{Z nbmemÝñ & Leaves are more succulent and Fleshy.

2) qH$ewH$: (^m.) "qH$ewH$mo @ `_²' ?> B{V ^«mpÝVOZH$: ewH$VwÊS>gÑe nwînËdmV² & The flowers resembles beak of parrot

3) H¥${_¿Z: (gmo.) H¥$_rZ² hpÝV, nbme~rOñ H¥${_amoJo à moJmV² & The seeds are used in krimirogahar

4) j mal oð>: (^m.) j mad¥j ofw l oð>: & Kshara prepared from this plant is superior.

5) I anU©: ({Z.) néf§ nU©_ñ & Leaves are rough.

6) {ÌnÌ : (gmo.) Ì r{U nÌH$mì`ñ nU} & Leaflets are present 3 in No.

7) nU©: (^m.) àeñVm{Z nUm©Ýñ & Plants having more leaves.

8) nwVÐþ: (Y.) nyV: n{dÌmo Ðw_: & This plant is used in Rituals.

9) ~rOñZoh: (gmo.) ~rO§ ñZoh_wŠV§ñ &

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Seeds are oily and snehayukta.

~«÷d¥j : (^m.) ~«÷Umo d¥j : d¡{XH$g§ñH$maofw à moÁ ËdmV² & 10)

Wood of the plant utilized in yajnya kunda as fuel.

11) `m{kH$: (^m.) `ko à wÁ _mZ: & It is used in yagnyas.

12) aŠVnwînH$: (^m.) aŠVm{Z nwînmÊñ & Flowers in red in colour.

13) dH«$nwînH$: (gmo.) dH«§$ nwîn_ñ & Flowers are curved.

14) dmVha: (^m.) dmVñ ha: em_H$:& dmVamoY: B{V Aï>m§J{ZKÊQ>m¡ "dmVnmoW:' B{V gmoT>bo nmR>: It is used in vata vikara.

15) dmZàñW: (n.) dZo àñWmo dmgmo @ ñ & We get these plants more in forest.

16) g{_Ûa: (^m.) `kofw à wŠVmZm§ g{_Ym§ l oîR>: & Wood is used in yadgnyakunda.

17) dmVnmoW: dmV§ nmoW{V B{V &

18) qH$ewH$ :- Its flowers resembele to nose of parrot in colour and shape.

19) j mal oð> :- Its alkali is best among alkaline materials.

20) nbme :- Its leaves are fleshy and beautiful or it develops the muscle.

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aŠVnwînH$ :- Its flowers are red in colour. 21)

22) da§ g{_ÜVo @ Z m B{V (ñd.) It mesmerize every one with its qualities and it improves appetite.

DESCRIPTION OF PLANT ACCORDING TO ITS SYNONYMS (25) :

Palash is sacred tree (Putadru) used in religious rituals and sacrifices (Yajnika,

brahmavruksha, Samidvara), It grows widely (Vanaprastha) has characteristic leaves (Palash,

parna) with three rough leaflets and curved (Kharaparna, triparna). Flowers are red (rakta

puspaka) and curved (Vakrapuspaka) typical of papilionate resembling parrot's beak

(Kimshuka). Seeds are oily (bijasneha) and make a potent antihelmintic drug (Krimighna). It

also pacify vata (vatahara). The plant is one of the best among the sources of alkalies

(Ksarasrestha).

SPECIFIC CHARACTERS :

1) Plant grows wild.

2) Leaves rough, trifoliate.

3) Flowers red, curved.

4) Seeds Oily.

5) Pacifies vata and one of the best sources of alkali, seeds antihemintic.

Synonmys according to different authors :

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11


a) {e. {Z. :- ÁdbÐŠVnwîn, {eå~r, {VŠV~rOH$, H¥$îUd¥ÝV.

b) A{ . _§. :- dH«$Hw$gy_, a§Jnwîn, g{_Ûa, n¥Wwe¥§Jr, dH«$s.

c) H¡$. {ZK§Qw> :- qH$ewH$, H$_m©, `mkH$, JUoéH$, dQw>nUu, ÛrOñZoh,

{Ìd¥Îm, aŠVnwîn, j mal oð>, dmVnmoW, ~«÷d¥Îm, g{_X²da.

d) _XZnmb :-nbme,qH$ewH$,{H${_©,`m{kH$,~«÷nmXn,j mal oð>,aŠVnwîn,{Ìd¥Îm,g{_V².{ZK§Qw>

e) AmXe©{ZKÊQw>§ :- nbme, qH$ewH$, ~«÷nmXn, j mal oð>, dmVnmoW, ~«÷d¥Îm, g{_X²da.

ABLE NO. 1

T

===============================================================================


12


SHOWING SYNONYMS ACC. TO DIFFERENT AUTHORS

Cha Su AS DN RN KN BP.N AS.H Sho.N

ynonym ShaS

+ + + + + + + + +

+ +

+ + + + + + + + +

+

+ + +

+ + + +

+ +

+ +

+ + + +

+ +

+ +

+ + +

+ + +

+ + + + +

+ + + + + +

+ + +

+

+ + + +

+ + +

+ + + + +

+

ERNACULAR NAMES:

V

===============================================================================


13


Sanskrit - Palash

Tesu, Palas, Chichra, Dhara, Faras Kankeri,

Chini agond

, Kamarkas

r Guj

rha.

ala, Paladula Modug Mooduga.

Midug chettu.

a, Muttagamara.

Hindi - Dhaka,

Beng - Palash, Ganch

Mar - Palasa.

Tree Guj - Khasathi Khakra.

Flowe - - Kemuda.

Seeds Guj - Palash Pap

Tam - Paras, Patsan, Cam

Tel - Modhung,

Kann - Muttuga, Muttal

Mal - Palashin Samatha, Camata, Pilacham, Muraklamar CV² - namew d - I mH$Q>r Jm - nbmg \y$b A§ - nDow y Branch buter.

Flame f the forest, parrot tree, Judas tree (Myths.and traditions) o

½dm{b`a - Beespak

n§Om~r - Tesh.

Amo[agm - Kinjuko Porasu.

CXw©

Bheda

Acc M p ta, Peeta, Shweta and Neel

( according to colour of the flowers)

- 1) Palash and 2) Valli Palash

ccord g to aj Nig ntu

Dos agnat :

- Palash Papra.

(Varieties) :

ording to adan al - Rak

In Abhidhanmanjiri

Shaligram - 1) Kinshulak 2) Hastikarnak.

A in R ha - Rakta, Peeta, Shweta and Neel.

Guna and h a

===============================================================================


14


PN - Kashay, Shaligram - Katu, Tikta, Kashay,

Katu Vipak

Ushnavirya.

ingdom - Plantae

phyta,

a

a

me nosperma

of Bu a Mo ) :

fter J tron of Botany

frondosa means leafy.

.

errya, Vipak, Doshaghnata

Vatakaphahar.

Taxanomical Classification :

K

Division - Magnolio

Class - Magnoliopsid

Order - Fabalea

Family - Fabaceae

Genus - Butea

Species - Monosperm

Latin Na - Butea mo

Meaning te nosperma (LAM

Butea = Named a ohn Ear of Butea, pa

Mono = One, sperma= seed.

Before Butea monosperma, Butea frondosa was the name,

Palash is famous for its leaves

Flesh, like flesh or blood

Or - Demon.

TABLE NO. 2 Rasa, Guna, V

===============================================================================


15


Rasa B.P.N MN RN DN NA KN Tikta + + + Katu + + + + Kashay + + + + Guna Laghu + + Rukha + + Snigdha + + Veerya Ushna + + + + + + Vipak Katu + + + + + Doshagnata a) Kaphaghna + + + + + + b) Vataghna + + + + + c) Pittakar + + +

TABLE NO. 3

rayojyanga according to different authors :

rayojyanga PN PV.S RN DG(VMG) DGH MN NA DN Sha.N

P

PBark + + + + + Flowers + + + + + + + + + Leaves + + + + + + + Seeds + + + + + + + + + Gum + + + + + + + + +

TABLE NO. K a Palash 4 : arm of “ ”

===============================================================================


16


Karma Bpn DN RW MN KN PN Deepan + + + + Vrushya + Krimihar + + + + + + Sangrahi + + Sarak + + + Vranahar + + + + Gulmahar + + + Grahani + + Arshya + Kusthahar + Rasayan +

Veeryavardhak +

USE OF "PALASH" SEED ACCORDING TO ORS :

DIFFERENT AUTH

nbmeíVw H$fmmoîU: H¥$_r Xmof {dZmeZ: & VX²~rO§ nm_mH$ÊS>y{VXXwËdXmof ZmeH¥$V² && (em{bJ«m_ {ZKÊQw>)

H$fm§ na§ J«m{h nwîn§ gwaå§ H¥${_¿Z§ M ~rO§ _V§ nn©Q>m_² && ([à {ZKÊQw>>)

~rO§ Vw H$Qw>H§$ pñZ½Y_wîU§ H¥${_~bmg{OV² & (YÝd§. {ZKÊQw>) \$b§ bKwîU _ohme©: H¥${_dmVH$\$mnh_² & {dnmHo$ H$Qw>H§$ éj § Hw$ð> Jwë_moXa àUwV² && (^m.à.)

VX²~rO§ H¥${_{dÜd§{g H$mÊS>mo agmZo {hV: & (gmo.{Z.)

\$b~rO§ M pñZ½YmoîU§ H$Qw> H¥${_H$\$mZ² O oV² && ({Z.a.)

VX²~rO§ H¥${_{dÜd§{g {hV: H$mÊS>mo agmZo & emoT>b. TABLE NO. 5

===============================================================================


17


XII) Prayoga (Uses) :

DN RN KN MN SN

Prayoga BPNPrameha + + + + Arsha + + + Krimi + + + + + + Kushtha + + + + Gulma + + + + Udar Roga + + Twakvikar + + + Kandu + + Pleeha + Shool + Vatarakta + + +

Raktapitta +

TABLE NO. 6

natha according to different authers :

BPN DN RN

XIII) Doshag

Doshaghnata ShaN KN PN Kaphavata Shamak + + + + + Pittavardhak + +

II) INTRODUCTION OF PLANT (GENUS AND SPECIES) (27) :

to family Fabaceae.

was found that this genus Butea consist of around 35 species. Out of which literature

(V

This plant was identified as Butea monosperma (Lam) belonging

It

regarding the phytochemical and pharmacological profiles was available on only few

species. Most of the reports available were on the Butea monosperma and very few reports

are available on Butea superba and Butea parviflora. But as per literature seeds are well

documented for various activities,

===============================================================================


18


BUTEA SPECIES :

pecies available in the Butea genus. Generally they are trees or

andent shrubs. The general characters of Butea species are as below :

aves

paniculate, or

.

incurved and acute, or

ma capitate or truncate.

ni l desc ption pecies of Butea are as follows :

its characters are

mentioned in the following paragraphs.

Trunk ft to right, attaining

0.6 0.9 m girth.

cm

ge scarlet, borne in great

There are 35 s

sc

Le - Pinnately, 3-foliolate, stipellate.

Flowers - Orange, purple, rose or white, densely fasciculate;

Fascicles recemose or fasciculate

ample paniculate.

Calyx - Campanulate, the upper two teeth or lobes connate

Petals - Subequal or unequal; keen

Straight and obtuse.

Stamens - Ten, Vexillar stamen free, the other connate; anthers

uniform.

Ovary - Sessible or stipitate, 2-4 (-7) ovulate; style incurved

not bearded; stig

Pod - Pod is one seeded.

Bota ca ri of the important s

1) Butea Superba : Butea superba is a gigantic climbing shrub and

- Trunk is climbing from le

Leaflets - The leaflets attain 30-45 cm and sometimes 50

in young plants.

Flowers - The flowers are larger than other species, 4.5-6.3

cm long, orange or oran

profusion along the leaflets branches on racemes

which are 30 cm long.

===============================================================================


19


P - Pod is about 12.5 15 cm long. od

istribution of Butea superba :

in the forest of Oudh and Bundelkhand, Chota Nagpur,

urma, Konkan, N. Kanara and Central. It is also found in South India.

istribution :

out India, in deciduous forests in areas up to 1,200 m elevation, also in open

reas.

ant :

-sized deciduous tree, with a somewhat crooked trunk,10-15' in height and

-6' in girth. The bark is bluish-grey or light-brown and yields a gum. Its bright orange-red

p to 4,000'),

xcept in very arid parts. Generally, it grows gregariously on open grasslands and scattered

mixe

.

wish-brown, but very liable to sap -stain

nd often turns greysh-brown or grey. It is a light wood and can be seasoned without much

D

Butea superba is distributed

B

D

Through

a

The Pl

A medium

5

flowers (1.5-2” long) bloom in great profusion at the beginning of the hot season before the

appearance of new leaves. The pod contains a single seed (1”x3/4”) at its apex.

Butea monosperma is common throughout India, Burma and Ceylon (u

e

in d forests along with Shal (Shorea robusta). It is frost-hardy and drought-resistant and

is a valuable species for reclaiming saline soils.

Parts used : Bark, leaves, flowers, seeds, gums

Butea monosperma bark is white or yello

a

difficulty, but it contracts considerable during seasoning. It is not strong or durable in

exposed situations, but is said to last much longer under water. It is easy to work either by

===============================================================================


20


hand or machine, and is used mainly for well-curbs, water-scoops and for fuel. It can also be

employed as a cheap board-wood .

The leaves are much used throughout the country for making plates, cups, etc. They

re eaten by buffaloes and elephants.

but very fugitive yellow colouring matter. This is

ontained in the sap and may be obtained in the form of a decoction or infusion from dried

ark furnishes a very important exudation which hardens into a red brittle resin

nown as butea-gum or Bengal kino or magugo, largely used “as a substitute for the 'Kino' in

ks and artificial incisions in the bark, and it

ardens into a vitreous, ruby-red gum, known as 'Butea gum or 'Bengal kino'. It is

ve long been valued as anthelmintic. Birdwood prescribes them for

und worms and tapeworms. According to Chopra freshly powdered new seeds give fairly

a

The flowers yield a brilliant

c

flowers. The addition of alum, lime or an alkali deepens the colour to orange and also makes

it less fugitive. The sap contains the chalcone, butein C15H12O5 (0.3%), orange yellow

needles, m.p., 2130c 2150c, and small quantities of butin, the colourless isomeric flavanone,

and its glucoside, butrin.

USES :

B

k

India and to a limited extent in Europe also”.

A red juice exudes from natural crac

h

distinguished from Malabar kino from Pterocarpus marsupium by its greater solubility in

water, and by the presence of corky particles. Butea gum contains a large proportion of

tannin and mucilaginous material. On dry distillation, it is reported to yield pyrocatechin. It

is a powerful astringent and is given in many forms of chronic diarrhoea. (Dymock, Warden

and Hooper, I, 454)

The seeds ha

ro

good results against Ascaris, but old worm-eaten ones, like those frequently found in the

market, shows little activity. The oil, powdered seeds, and an alcoholic extract of seeds,

provide quite ineffective against hookworms, etc. When pounded with lemon juice and

===============================================================================


21


applied, the seeds act as a powerful rubifacient and they have been successfully used in

curing a form of herpes, known as dhobie's itch. (Dymock et al., loc. Cit.) The seed is also

useful in skin diseases Dadru, Pama, Kandu, ringworm infections etc.

The seeds contain 18% of a yellow, tasteless oil (sp. Gr. 25o/25oC, 0.8983; sap. Val.,

78; iod. Val., 67.2, Katti and Manjunath, J. Indian Chem. Soc., 1929, 6, 639). Fresh seeds

e leaves are astringent, anti-inflammatory, anodyne and aphrodisiac, and are useful

pimples, boils, flatulence, colic, worm infestations, inflammations, arthralgia and

aemor

ers are astringent, sweet, cooling, constipating, aphrodisiac, haemostatic,

iuretic, febrifuge, depurative and tonic. They are useful in vitiated conditions of pitta and

ith other astringents and rock-salt is recommended by

HAKRADATTA, as an external application for “Pterygium” and opacities of the cornea.

c.

hey are useful in herpes, skin diseases, ringworm, ophthalmopathy, epilepsy, round worm,

PRAKASH recommends new seeds to be

iven in powder, 10 to 20 grains or as paste with honey added (“because the seeds are very

1

are reported to contain proteolytic and lypolytic enzymes. The former is a mixture of plant

proteinase and polypeptidase, and behaves like 'yeast trypsin' (Chatterjee, Ghosh and

Chopra).

Th

in

h rhoids.

The flow

d

kapha, diarrhoea, haemorrhoids, menorrhagia, strangury, fever, leprosy, skin diseases,

swellings, hyperdipsia, haemoptysis, arthritis, burning sensation, bone fractures, and are very

efficacious in birth control.

Gum combined w

C

The seeds are purgative, ophthalmic, anthelmintic, rubefacient, depurative and toni

T

arthritis, flatulence, constipation and diabetes.

As anthelmintic and aperient, BHAVA

g

unpleasant to take and often produce retching pain in the abdomen and occasionally

vomiting and giddiness”) thrice daily for three successive days (especially for ascaris round

worms) and followed on the fourth day by a dose of castor oil. For this, the seeds are soaked

===============================================================================


22


in water, shells removed and kernels powdered after being dried. “Some medical men

consider that the seeds can be advantageously substituted for “santonin” against round

worms”. Externally the powder is a remedy for ringworm; it may be applied better in the

form of a paste being pounded with lemon juice; also for herpes (Dhobis' itch).

“Moodooga oil is said to be practically inert and does not possess any anthelmintic

ctivity. Active principle of the nature of alkaloid, neutral principle or glucoside could not be

IFFERENT YOGAS :- According to Charak Samhita.

Palash beeja. Udar rog. Ch.chi (110/13.)

ja. .)

ruta. ,

DHYAYA : According to Sushrut Samhita.

YOGAS Indication References

han. Su.chi. (3/6)

a. .

hi.

CCORDING SHARANGDHAR SAMHITA :

a

isolated from the seeds.

USE OF PALASH IN D

Yogas Indication References

Palash bee Arsha Ch.chi (92/14

Choorna. Kasa. Ch.chi (77/18.)

P. Beeja + Gh Atisaara Ch.chi (27/19.)

P.Beeja. Visha Ch.chi (51/23.)

Palash Kshara Dwivruna Ch.chi (54/25.)

A

Palash Valkal Vruna band

Palash Choorna. Vaata Vyadhi Su.chi (4/32)

PalashKshara. Ashmari Su.chi (7/22)

Beeja lepa. Kushta. Su.chi (9/10)

Beeja choorna. Prameha. Su.chi (11/8)

Beeja choorn Udar rog Su.chi (14/13)

Beeja choorna. Maha vyad Su.chi (31/5)

A

===============================================================================


23


Palash beeja choorna Krumihar, 5/26.

hanartha lepa. 11/110

.

CCORDING TO ASHTANG SANGRAHA. (A.S) :

P. Vrunta Swaras. Kasa. A.S.Chi (3/65)

A.S.Chi (5/9)

)

CCORDING TO ASHTANG HRUDAYA. (A.H)

P. VruntaGhruta. Raktapitta. AH.Chi (2/44)

AH.Chi (3/101)

n

AND STORAGE OF DRUG :

were collected in the

onth of April and May near Belgaum forest area. The seeds were authenticated by

DULTRATION AND SUBSTITUTION (28) :

Palash beeja choorna Yoni sankoc

(Uttarakhanda)

Palash pushapa swaras Netrapasadan 13/78.

(Uttarakhanda)

A

P. Churna. Raj Yakshma.

P. Churna Arsha A.S.Chi (8/63)

P. Choorna Vaatshonita. A.S.Chi (22/45

A

P. Ghruta Arsha

P. Beeja. Krumi AH.Chi (20/26)

P.Kshara. Rasaya AH Kalpa Sthana

(39/37)

COLLECTION

In the present study the matured seeds of Butea monosperma

m

Taxonomist. Soon after authentification all the seeds were dried at room temp until they

were free from the moisture and subjected to physical evaluation with different parameters

like nature, colour, taste, odour, size, shape width, length etc. Nearly 1.5kg seeds were

collected and kept in air tight jars to avoid fungal infections. Veerya period is 1 year.

A

===============================================================================


24


Butea monasperma seeds are adulterated with the seeds of B. Superba (Roxb). These

an be identified on the basis of the size of seeds i.e the seed of B. monosperma are

ompar

I) CHEMICAL CONSTITUENTS REVIEW (29) :

asonin' isolated from seeds.

m seed oil.

ge

uantity of water soluble albuminoids.

ntimicrobial activity against fungi which are

athogenius and gram +ve bacteria strepto coccus pyogences, staphylo coccus aureus,

cillu

DHA :

oned in Siddha.

• Action and uses in Unani medicine.

d mare administered against piles, Eye

iseases and Enlargement of spleen (Spleenomegaly).

c

c atively larger in size. But in this case the seeds were collected by own. So there is no

chance of adulteration.

(X

A Nitrogenous acidic compound (I) along with 'Pal

α Amyrin, β Sitosterol, its glycoside and sucrase isolated from seeds

Glycerides of Palmitic stearic, Lignoceric, Oleic, and, linoleic acids fro

Seed contain 18% of fixed oil called moodooga oil small quantity of resin and lar

q

The compound (I) showed A

p

Ba s substilis gram ve bacteria ,E. Coli, Pseudomonas, maximum inhibitary effect was

shown by gram +ve bacteria,

ACTION AND USES IN SID

• Uses and Action are menti

The fruits and seeds are bitter and oily an

d

TABLE NO. 7

===============================================================================


25


Prayojyanga according to different authors:

Prayojyanga

S.H.N Names of the authors

D.N K.N N.A

Parts used

Or P.N BP.N R.N

Patra (Leaf) + + + + + + +

Stem + + + + +

Flower + + + + + + +

Seed + + + + + + +

Kshara + + + + + + +

OTE : Studies carried out on 'Palash' seed.

ls as:-

bial activity.

ty.

ity in female rats.

ioxidised activities in female rats.

ternet information –

) Kumari,N, Ehaum ,V, Mikosch, M, 2005 Seasonal photosynthetic performance and

adava R. N. Tiwali L.

N

Pharmacological activities and clinical trai

- Antihelminthic activity.

- Antifungal and Antimicro

- Antidiabetic activity.

- Antioestrogenic activi

- Ophthalmic disorder.

- Anti implantation activ

- Anti hepato toxic activity.

- Uterotropic and uterine per

- Antioxidant activity.

In

(1

nutrient realation Butea monosperma (Fabacea) in comparison to two other woody

species of a seasonal deciducos foresty in Nw India and planted tree in this area ,

Indian journal of forestry 2005 28:116-126.

Y

===============================================================================


26


(2) A potential antiwiral flavone glycoside from the seeds of Butea monosperma O.

excena, Ajit Kumar, Gupta

) A pharmacentical composition (extract of Butea monosperma flower) useful for the

. Khan, T. H. Prasad.

) Butea monosperma and chemomodulation protective role against thioacetamide

SES AND ACTIONS ARE NOT GIVEN IN SIDDHA :

Kuhne J-Acian-Nal-Pro-Res-2005 Apr. : 7(2) : 185-8.

S

(3

treatment of the hepatocellular caricinoma world intellectual property organization

2006?

A

(4

medicated hepatic alternation in wistar rates. Physiotherapy & Phytopharmacology

02/01/06.

U

===============================================================================


27


Actions and Uses in Unani (30) :

its and seeds are bitter and oily and are administered against piles, eye

) MICROBIOLOGY REVIEW :

Microbes (Krimi) In Vedas (Ayurveda) (31) :

From the vedic period itself acharyas have been aware of minute organisms of the

ABITAT :

The fru

diseases and enlargement of spleen. (Spleenomegaly).

B

I.

nature / environment. They have also described krimis, their habitats types, synonyms and

treatment. The acharyas had knowledge about jantus etc. that had harmful and harmless to

beings. under the heading of krimi's they have described about both Ñï> (drustha) AÑï>

H¥${_ (Adrushta krimis).

H

`o {H«$_ : nd©Vofw dZofw A¡²fYr~w newfw AßgdÝm: &`o Añ_mH§$ VÝd_m{d{dew gd© VX²Y{_ {H«$_rUm_² && (A. 2/31/5)

In atharvveda it is told that krimis are present in all factors of the nature. They are

inuteness of Krimis :

1)

present on parvath, vana aushadhi, pashu, etc.

M

gyú_ËdmÀM¡Ho$ ^dÝËÑí`m: &&( (C.S.VI 7/11)

(2) gm¡úåmV Ho${MXXe©Zm: && (vaghbhata)

===============================================================================


28


rimis are not vHere Acharyas says that K isible to our naked eyes.

1)

gm¡úåmV Ho${MXXe©Zm: &&(

Here Drushta and Adrush a types are specified.

terpretation by Acharyas :

)

t

In

H«$ì`o _oÜm{V B{V {H«$_r: && 1 One which survives on raw meat is krimi.

) 2 H$mVodm© One which has desires.

)

H«$_Vodm© ñ mV² gaUH$_©: & 3

One which crawls and moves.

AUSES OF INFECTION BY KRIMS :

C

1) CXHo$ ~hd: àmUm: n¥{Wì m_² M \$bofw M & gyú_ `moZr{Z ^yVm{Z VH©$ Jå{Z ^maV && nú_Umo Am{n {ZnmVoZ `ofm§ ñWmV² ñH$ÝY n ©: & o AÝZof {d{dÜmpÝV nmÌofw {n~Vmo OZmZ² & (` .d. 16/ 62) o

2) àg§JmV² JmÌg§ñnem©V² {Z:ídmgmV² gh moOZmV² & gheæmgZƒm{n dñÌ_më mZwbonZmV² && Hw$ð>§ ÁdaíM emofûM ZoÌm{ î §X Ed M & Am¡ng{J©H$ amoJmü g§H«$_pÝV ZamZ_² && (gw.g.{Z. 5/ 33-34) 3) ñne} H$mhma eæmXr godZmV² àm mo JXm: & gV} g§M[aZUmo ZoÌ ËdH²$ {dH$mam {deofV: & (A.h.{Z. 14/ 441)

===============================================================================


29


By dushita, vayu, jala annasevana, bhumi and by maithuna in cted rogi gatra

arsha of infected pt. infection by nishwasa etc. use of contiminated clothes, bed etc. of

fe

sp

infected patient / sahabhojana etc. krimis are said to be entering the body.

ORIGIN OF KRIMIS :

g§ŠboXmV² {H«${_í`mñ ^dpÝV CnhVmË_Z: & (M.g.gw. 17/ 38) Charaka says kleda in the body is one of main factors for the production of Krimi’s.

ROUTE OF ENTRY :

Am_o gwnŠVo e~i o {dnŠVo `mo _m {nemMmo AeZo XXå & VXmË_Zm àO {nemMm {d `m§V ÝVm_JXmoX_ñVw && (E 5/ 29)

Krimis enter the body through annapanadi and vruna mukhas.

`m¡ Aí`m¡ n[agn©Vmo `mo Zmgo n[agdm©{V & d§Vm `mo _Ü§ JÀN>{V V§ {H«$_r O `^m{g && (A. 5/ 23)

here they also enter through eyes, nostrils mouth to produce disea es.

rimis are given (1)

xternal (2) Internal * Internal krimis are further divided into 20 types eg. Kaphaja, Raktaja,

T s

In sharangdhar samhita, he mentioned main two types of k

e

Pureeshaga etc.

aŠVdm{h { amoñ mZm aŠVOm OÝVdmo AUd: & (A.h.gw. 14/ 42) Krimis presenting in blood are the minutest.

VYAKT KRIMI LAKSHANA'S :

===============================================================================


30


emo{UVOmZm§ Vw I bw Hw$îR>¡: g_mZ§ g_wñWmZ§ ñWmZ§ aŠVdm{hÝmo Xí` gwú_m¡ ËdmÀM¡Ho$

^dpÝV AÑí`m: d«UJVmZm§ Mhf© H$ÊSw> VmoX g§ñVn©Um{Z A{Vd¥X²YmZm§ M ËdHw$ {gamñZmw _m§gVéUmpñW `mgU{_Vr

Y_Ý: g§ñWmZ AUdmo d¥Vñnmnm

&& (M.g.d. 7/ 11)

gmÜdmZ{n Cnoú _mUm§ ËdHw$ _m§g eo{UV b{gH$m H$moW H$boX g§ñdoXOJ: {H«$_ moX² { _wÀN>©pÝV Vo ^ú pÝV ËdJmXrZ² & {H«$_ ñVw ËdJXríMVwa: & {eam: ñZmwíMmpñW M VéUmÝmXXVo && (M.g.{Z. 5/ 10)

Raktaja krimis that are minute and because of the size mynuteners they are

metimes visible or invisible when vranagata they cause kandu, to a etc. when excessively

rown they eat away twak, sira, snayu, mamsa tarunasthi.

In vedas explanation regarding sun as a krimighna is stated (Ri/191/18), (A 2/32/1)

aja are drushta krimis, while remaining 7 described as raktaja are invisible i.e.

ey are adrushta krimis, But acharya Charaka terms some krimis to be sukshma.

FERENCE :

This brief explanation suggests that there was tremendous knowledge regarding

m the vedic kala. Only the way of approach has been changed.

so d

g

Thus krimis minutest in the twak, mamsa lasika, sharira in kleda and sweda.

(R1/28/1).

Acharya Sushruta also clearly mentioned that 13 krimis are described as pureeshaja

and shleshm

th

IN

microbes right fro

===============================================================================


31


II) INTRODUCTION :

Antimicrobials (32) :-

The agents which have a capacity to kill the microbes but which causes minimum

tissue.

tory.

gents.

and fungi, become resistant to the

ewest antimicrobial agent.

) Has the most activity against the pathogen.

wards the patient.

) Has the least impact on the normal flora of the patient.

natural products but also micro organisms

se

on of natural compounds.

harm to the human body

It is not sufficient to report the name of microbe to the physician after isolating and

identifying clinically in the labora

Although physicians may have knowledge of antimicrobial agents and a general

pattern of susceptibility of certain antimicrobial a

Susceptibility of many bacteria, fungi, viruses to antimicrobial agents cann’t be

predicted that many micro organisms including bacteria

n

SELECTION OF DRUG :-

(1

(2) Has the least toxicity to

(3

Not only antimicrobial agents are made by

them lves.

Antimicrobial activity can improve by antibiotics which are synthetics, semisynthetic

and modificati

===============================================================================


32


Antibiotic which kills the organism is said to be “cidal” and which inhibit the growth

of organism know as “static”.

Antimicrobial agents have specific activity against bateria, fungi, and viruses.

The antibiotic which act on both gran + ve and -ve bacteria in known as broad

become disrupted in the intestine cauring diarrhoea. Vaginal flora can

e disrupted cauring fungal infections of genital tract.

ostate.

Fixation of dose based on two criteria which kills micro organisms but does’nt harm

) Has the least impact on the normal flora of the patient.

- Not completely destroying the intestinal flora. Not only antimicrobial agents are

spectrum antibiotic.

High dose of antibiotics leads to disturb the natural defense mechanism in human.

The normal flora can

b

To over come infection antimicrobial agent must attain effective level in infected

site. Some antibiotic enters easily in to C.S.F. Eye or pr

Some organisms cann’t be given through the mouth because of distruction by the

stomach.

Excretion of must of the druges are through kidney and some are broken down by the

liver.

to the patient. It is very difficult and tough to treat fungal diseases.

Synergic action can be seen by giving drugs in combination than they act

independently.

(3

made by natural products but also micro organisms them selves.

===============================================================================


33


One which kills the organism is said to be “cidal” and which inhibit the growth of

organism know as “static”.

Antimicrobial agents have specific activity against bacteria, fungi and viruses. The

antibiotic which act on both gram +ve and -ve bacteria is known as broad spectrum

ntibiotic. High dose of antibiotics leads to disturb the natural defense mechanism in

uman

Some organisms cannot be

iven through the mouth because of distractions by the stomach. Exception of must of the

ONCENTRATION (MIC) (33) :-

A test that determines the lowest concentration of the antimicrobial agent that inhibit

Sensitivity test, in which an organism is placed with antibiotics to determine which

ffectively kill the organism with smallest dose, also refers to the ability ? An

ntibiotic to inhibit growth of an organism.

e growth of organism.

a

h . The normal flora can become disrupted in the intestine curing diarrhoea. Vaginal

flora can be disrupted cauring fungal infections of genital tract.

To over come infection antimicrobial agent must attain effective level in infected

site, some antibiotic enters easily in to CSF. Eye or Pro state.

g

drugs are through kidney and some are broken down by the livers. Fixation of dose based on

two criteria which kills micro organisms but doesn’t harm to the patient. It is very difficult

and tough to treat fungal diseases. Suberic action can be seen by giving drugs in combination

then they act independently.

MINIMAL INHIBITORY C

the growth of an organism.

Sensitivity :-

antibiotic will e

a

Resistance :-

When an antibiotic dose not inhibit th

===============================================================================


34


Static :- Inhibiting growth of an organism.

Synergy :-

Two or more drugs in combinations having a great effect that the sum of the two

ndently.

I) BACTERIOLOGY (34) :-

) Scientific Classification :

e)

Domain - Bacteria

Kingdom - Bacteria

- Fermicutes

is a gram + ve cocci ich a rape like cluster when seen through

icroscope and has lagered, round golden yellow colonies, often withy B-hemolysis when

rown on blood agar plates.

acting indepe

II

1

(i) Staphylococcus (Gram+v

Phylum

Claso - Cocci

Urder - Bacillales

Family - Staphylococcaceae

Species - S. Aures.

S. Aureus wh ppears as g

m

g

===============================================================================


35


The bacteria’s name aureus means “golden” in Lactic. The most common cause of

staph infections is a special bacterium, frequently living on the skin or in the nose of healthy

person, that can cause minor skin infection as pimples boils, c, Celluloids and abscesses.

i) Bacillus subtilis

Kingdom - Bacteria

Phytum - Firmicutes

- Bacilli

Order - Bacillaceae

of genus bacillus. It is not

us, only 192 were shown to

e indi

S. Aureus Has about 2600 genues. The species has been subdivided in to two sub

species S. Aureus and S. Aureus anaerobes. S. Aureus may occur as a commensal on human

skin scalp, armpit, groin. It also seen in the nose, throat, and less commonly seen in urine.

S. Aureus can infect other tissues when normal barriers have been breached Eg. Skin

or mucosa lining this leads to biols and carbuncle I infected it can cause severe disease

staphylococcal scalded skin syndrome.

Infection of this can be spread through contact with pus from an infected wound, skin

to skin contact with an infected person and things. Which are used by this person.

(i

Class

Family - Bacillaceae

Species - Subtilis

It is a gram +ve, commonly found in soil. A member

considered as a human pathogen. It has approximately 4100 gen

b spensable.

===============================================================================


36


(i Escherichia Coli (Gram -ve) :

This genus

ii)

is named after Escherich who was the first to be deseribe the colon

acillus under the name bacteritim coloc commune (1885) based on minor differences in bio

hemical characteristics, colon bacilli were described under various names, but in view of

u ies in this group, they have all been included in the

oli which is further subdivided in to biotypes and serotypes. A few other species are E.

singly or in pairs.

is motile by peritricatate Hagelly, though some strains may be non motile capsules and

mbriae are found some strains, spores are not formed.

been recognized in E-coli namely

) Surface antigen (2) Toxins

of toxins (a) Haemolysines and (b) Entero toxins.

s.

t types of

-coli, enterotoxins has been identified (i) Heat liable toxin (ii) Heat stable toxin (iii) Vero

xin.

E-coli causes wound infection, abscesses and also septicemia.

b

c

the m tability of the bio chemical propert

C

forgusonii, E hermonii, E-vineries which are of less medical importance.

MORPHOLOGY :-

E-Coli is gram - ve, straight rod measuring 1-3x 0.7 mm arranged

It

fi

VIRULENCE FACTORS :

Theere are two types of virulence factors have

(1

E-coli produces 2 types

Heomolysines don't appear to be relevant in pathogenesis through they are produced

more commonly by virulent strain

Enterotoxins are very important in pathogenesis of diarrhoea. These distinc

E

to

===============================================================================


37


Because of it UTI. diarrhoea pathogenic infection like meningitis, peritonitis may

occur.

(IV) CANDIDA ALBICANS :-

ingdom - Fungi

Phylum - Ascomycota

Subphylum - Saccharo mycotina

Saccharaomycetoes

Order - Saccharo my cetales

les

m

sexual fungus ses (skin), oral and vaginal

ns.

These are many species of candida. The clinical spectrum of candida albicans range

om superficial infection of the skin to the life thereatening infection. They can be isolated

om h

The infection caused by organisms in the genus candida includes localised disease of

e skin and nails, diseases that affect the mucosal surfaces of oral cavity, vagina,

esophagus and brochealial trees.

K

Class -

Family - Saccaromyceta

Genus - Candida

Species - C-albicans.

Synoni - Candida stellatoidea

It is diploid a (a form of yeast. It cau

infection in huma

fr

fr ealthy mucosal surfaces of oral cavity, vagina, G.T. tract and malaria.

CLINICAL SYNDROMES :-

th

o

===============================================================================


38


Because of this fungal infection causes most areas or skin causing itchy lesions on

ant in pathogenesis of diarrhoea. The distinct types of E-

oil entero toxins has been identified .

stable toxin (c) Vero toxin

ningitis peritonitis may occur .

V) CANDIDA ALBICANS :-

Phylum - Ascomycota

Subphylum - Saccharo mycotina

Sacch aromycetes

Order - Saccharo mycetales

mycetaceae

ding subcutaeneous tissues

nd cau fections. It re s in m es.

most laboratory med od agar and sabouroud agar. It

ppears after 24 hours has tiny dots and by 48 hours the dots are soft, creamy colonies. It

mells

the perinium, including deeper rash in.

Enterotoxins are very import

c

(a) Heatliable toxin (b) Heat

E-coli causes wound infection , abscesses and also septicemia . Because of UTI,

diarrhoea, pathogenic infection like Me

(I

Kingdom - Fungi

Class -

Family - Sacchro

Genus - Candida

Species - C - albicans.

Scaling and ulceration in candida is also notorious for inva

a sing serious in side ost health care faciliti

It will grow in ia, including blo

a

s like baker's yeast.

===============================================================================


39


(V) SKIN DISEASES (35) :-

Severel infectional diseases are seen because of the various types of micro

rganisms. Palash seed is used in many skin diseases like Kustha, Pama, dadru and kandu.

nd also infection can be seen in.

So in skin diseases ring worm infection is also present. This infections are commanly

tion.

acilli, Pseudomonas sp.,

streptococci, Haemophilus influenzae.

s,

Gardnerell vaginallis, Trichomonas vaginalis, Candida albicans, B-Hemolytic

a pallidum, Chlamydia

4) A and B streptococci, Corynebacterium diptherise,

streptococcus pneumonicae, Haemophilus influenzae, Mycobacterium tuberculosis,

) Urinary Tract : Staphylococci, Diphtheroid Bacilli, Coliform bacilli, Enterococci,

Proteus sp., Lactobacilli and B-hemolytic streptococci, Bacillus sp., E.Coli.

o

a

found in male, female and in children also. According to our references palash seed is

usesful in skin manifestation. Fungal infections are difficult to cure fast. There is indirect

role of Bacillus subtilis in skin infe

(VI) Microorganisms Frequently Encountered In Body Sites :

1) Blood : Staphylococci, Coliform b

2) Genital Tract : Coliform bacilli, Enterococci, Bacteriodes sp., Lactobacillu

streptococci A,B and D. Neisseria gonorrohoeae, Treponem

tractomatis, Herpes simplex.

3) Wounds : Staphylococcus aureus, Streptococcus pyogenes, Coliform bacilli,

Bacteriodes sp., Pseudomonas sp., Clostridium sp., Anaerobic cocci.

Respiratory Tract : Group

Klebsiella pneumoniae, Pseudomonas aeruginosa.

5

===============================================================================


40


6) Eye, Ear : Staphylococcus aureus, Hemophilus sp., Streptococcus-pneumoniae,

Neisseria gonorrhoeae, and B-hemolytic streptococcus, Psedomonas species,

Corynebaterium diphtheriae.

7) Gastrointestinal Tract : Escherichia Coli, Clostridium sp., Salmonella sp., Shigella

sp., Yersinia sp., Campylobacter sp., Vibrio sp.

8) Cerebrospinal Fluid : Hemophilus influenzae, Neisseria meningitis, streptococcus

pneumonia.

FERENCE :

commonly encountered organisms in

arious sites of the body.

) EXTRACTION PROCEDURE REVIEW (36,37) :-

here are there types of extraction procedures are present.

type solid components extracted from solid substance. This

extraction always carried out before any further separation / processing.

cted with a liquid medium.

o liquids that aren't miscible

includes that substance to be extracted (solvent extraction).

lar size with

itable extraction medium. For extraction fresh or dried parts of plants are used. The parts

oid

hemical changes drying operation should be performed under controlled condition. Dry

IN

Here we can see that E.coli and staph.aureus are

v

C

T

(a) Solid-Solid in this

(b) Solid liquid extraction :- Where the solid drug is extra

(c) Liquid - Liquid extraction - In this process any of the tw

To get specific drug extract herbal drugs are extracting of certain particu

su

which are used for extraction must be dried before the procedure taking place. To av

c

===============================================================================


41


these parts as quick as possible. Drying procedure must be going on in room temperature and

place High temperature must be avoid for drying. This partin airy must be kept free from

fection.

the extent. When the tissue on repeated extraction is completely free of green

olour, it can be assumed that all the low molecular wt. compounds have been extracted

Selection of the solvent is very important aspect, not only for the yield of one more

incip

ing substance.

xhlation :-

The actual solvent proportions in the extracting chamber differ from that originally

sed in the collector.

in

The extraction depends on the texture and water content of the plant material and on

the type of substance that being isolated. Alcohol is good solvent for preliminary extraction.

After exhaustive extraction subsequently the material can be macerated in a blender

and filtered. After extraction of green tissues with alcohol chlorophyl is removed in to the

solvent at

c

when we extract continuously dried powdered plant material such as dried seeds, root, leaf,

heartwood in a soxhlet apparatus with a range of solvents we obtain organic constituents.

There are short cut methods in extracting procedures that one can learns with

practice.

SELECTIVITY OF SOLVENTS :

pr le substances but also for the qualitative and quantitative composition of the

acompany

(i) Hot continuous extraction - so

By the help of available soxhlet extractor one can prepare crude plant extract. The

procedure is done by the help of pure solvents.

u

===============================================================================


42


Advantages of this apparatus is that it is fully automatic continuous method that

oesn't

In this procedure the material to be extracted is placed in a 'thimble'. This thimble is

ade u

ral

mple compartment.

nd reflux condenser are separate item of

lass which is assembled together with the appropriate contents, to make the complete

passes through the side arm up into the reflux condenser. The vapour

quities and drips into the thimble containing the material to be extracted. The warm solvent

n the central compartment flows through this and back into the lower solvent

ontainer. The process gets repeated then.

d require further manipulation other than concentration of the extractive and saves.

INSTRUMENTATION AND PROCEDURE :

m p off cellulose or cloth in a central compartment with a siphoning device and side

arm both connected to a lower compartment. Reflux condenser is attached above the cent

sa

Solvent container, sample compartment a

g

apparatus.

The solvent in the lower container which is usually round bottemed flask is heated to

boiling, and the vapour

li

percolates through the material and the wall of the thimble and the extract gradually collects

the central compartment when the height of the extract reaches the top of the siphon, the

entire liq. i

c

The extract is collected in the lower vessels, gradually be coming more and more

concentrated. It is assumed that no volatile substance is present. The vapour rising from the

heated extract is pure solvent vapour and so the liquid dripping into the material from the

condenser is essentially pure solvent, which get from the extract. Small volume of solvent is

needed, the effective volume of solvent used for the extraction is proportional to the time for

which the process is allowed to continue.

===============================================================================


43


The soxhlet processes is useful for exhaustive extraction of the plant material with

particular component is desired.

LIMITATIONS :-

(i) In this procedure solvent is recycled on and often the extract that collects in the lower

i) Total extract will exceed their solubility in that particular solvent so it may

the lower container and require much greater volume of solvent for

latter desolation.

(iii)

fore use of solvents with relatively high boiling point such as methanol or

water, the whole apparatus below the condenser needs to be at this temperature for

(iv) carried out with single solvent and we are unable to extract with

any mixture solvent.

) AQUEOUS EXTRACTION (38,39) :- (Maceration Principle)

cess of extraction of drugs with a solvent with several daily

akings at room temperature.

container is continuously being heated and it may suffer thermal degradation

reaction.

(i

precipitate in

It is not possible when we are adopting the them procedure on a large scale it is not

suitable be

effective movement of the solvent vapour.

This procedure is

(B

Maceration is the pro

sh

===============================================================================


44


===============================================================================


45

antity of water at room temperature to it and kept it for 7-8

ays. (Minimum 5 days). The vessel must be sealed highly. After 7 days residual liquid is

ecanting and staining is expressed from the solid and extract kept to evaporate the water

bath.

INSTRUMENTATION AND PROCEDURE :-

A conical flask of required measurement is taken. Then required drug plant material

is choosen and poured stated qu

d

d

content. at certain temperature (300c-400c) on water

Methodology ==================================================================

===============================================================================


45

Methodology ==================================================================

4. METHODOLOGY

(4.1) COLLECTION :-

Plant seeds were collected from around Belgaum area.

(4.2) AUTHENTIFICATION FROM :-

I C M R - Belgaum.

(4.3) COLLECTION TEST DRUG AND STORAGE.

(i) 1.5kgs of 'Palash' seeds were collected and dried in shade and

stored in moisture free container

(ii) Then as required it is used.

(4.4) PHARMACOGNOSTIC STUDY

(i) Organoleptic Characters :

Table No. 8

Sl. No. Parameters I.P. / B.P. / As per Literature Observation

I Physical Tests

Nature Flat dull red nearly black Flat Dark Brown

Colour Dull red nearly black Dark Brown

Odour Characteristic Characteristic

Taste Bitter Bitter

II Seed Length 2.0 - 5.0 cm. 2.5 - 4.0 cm.

III Seed Width 1.0 - 3.0 cm. 1.5 - 2.5 cm.

===============================================================================


46

Methodology ==================================================================

(ii) Macroscopic and Microscopic Characters.

Macroscopic characters of 'Palash' seed :

The seed is reddish brown thin, flat, reniform, longer axis, 3-4cms and shorter 2-

2.5cms. seed coat reddish brown waxy, faint odour, taste slightly acrid and bitter. Wt. of 100

seeds nearly 80-115gms. The hylum of the seed is conspicuous and situated near the middle

of the concave age of the seed.

Microscopic Characters of 'Palash' seed :-

The seed taken for the study were very hard. So they were soaked in brine (salt

water) till they set softened and enable for easy sectioning. The transverse section (T.S.) or

*Longi – Section as required are taken and viewed under microscope.

OBSERVATION :-

The seed has an outer festa and inner endosperm region. The outer festa contains a

single layer of pigment cells. The endosperm region has parenchymatous cells contain

aleurone grains and starch grains. The cells of the endosperm are filled with oil globules.

The starch grains are rod shaped or ovoid. Near starch cell spiral form of proteins are

present.

POWER - MICROSCOPIC STRUCTURE OF POWDER :

Powder contain yellowish brown colour, acrid and bitter with oil flavor and

typical smell, small fragments of festa. Cotyledonary parmanchyma containing a few starch

grains, hour glass cells, abundant spiral protein bodies, mucilage and oil globules.

Note :- It is tough to take seed section.

===============================================================================


47

Methodology ==================================================================

(4.5) PHYSIO-CHEMICAL STUDY (40) :-

(a) Ash value :

(i) Use to determine quality and purity of drug

(ii) Ash contains in organic radical like phosphates, carbonates and silicates of Sodium,

Pottasium Magnesium and Calcium etc.

(iii) Some times inorganic variables like calcium oxalate, silica, carbonate content of the

crude drug effects, total Ash value. Such variables are then by treated with acid and

acid insoluble Ash value is determined.

DETERMINATION OF TOTAL ASH VALUE :-

(i) Weigh the silica crucible

(ii) Powdered drug is weighed and put in to the crucible.

(iii) Then it is placed in the Muffle furnace at 4500c for about 1/2 - 1 hour.

(i.e. : untill all carbon particles get burnt off.)

(iv) Cool it in dessicator

(v) Then weigh the ash and calcutale the percentage of total ash with reference to the air

dried sample of the crude drug. ( Standard ash value of "Palash" seed is not more

than 8%).

ASH VALUE :- (OBSERVATIONS)

Crude powder :-

(a) Weight of empty crucible = 18.227 gms. - (A)

(b) Weight of drug taken (2 gms) = 2.000 gms. - (B)

(c) Weight of crucible + ash = 18.642 gms. - (C)

(d) Weight of ash = C - (A+B) = 1.485 gms

∴ For 100 gms of drug ash value = C - (A+B) * 100/2

= 7.425% %

===============================================================================


48

Methodology ==================================================================

FINE POWDER :-

(a) Wt. of empty crucible = 16.272 gms. - (A)

(b) Wt. of drug taken (2 gms) = 2.000 gms. - (B)

(c) Wt. of empty crucible + drug taken = A + B = (C) = 18.272 gms.

(d) Wt. of crucible + ash = 16.735 gms. = (D)

(e) Wt. of ash = C - D = 1.537 gms. 98

∴ 100 gms of ash value is 1.537 x 50 = 7.685 %

(B) ACID INSOLUBLE ASH VALUE :-

(i) Using 25 ml. dil HCL (0.5 N) wash the ash from the dish used for total ash in

to a beaker.

ACID IN SOLUBLE ASH VALUE :-

1) Using 25 ml of dil HCL (0.5N) wash the ash from the dish used for total ash into a

beaker.

2) Place wire gauze over a bunshen flame and boil for 5 minutes.

3) Filter through a Whatman filter paper - wash the residue twice with hot water.

4) Place it in a crucible Ignite the crucible (placing it in a muffle furnace)

5) Heat it until all carbon is removed (at 2500c for half an hour to 45 minutes)

6) Cool it and weigh the residue

7) Calculate acid insoluble ash of the crude drug with reference to the air dried sample

of crude drug.

===============================================================================


49

Methodology ==================================================================

ACID INSOLUBLE ASH VALUE :-

Wt. of crucible = 40.775 gms

Wt. of drug = 02.007 gms.

Wt. of ash = 01.73 gm

Acid insoluble ash = 0.030 gm.

(Standard acid insoluble ash value - > not more than 0.5 %)

C) DETERMINATION OF WATER SOLUBLE EXTRACTIVE :-

Ingredients :- a) Powdered drug - 5 gm.

b) Water + Chloroform - 100 ml.

c) 500 ml water + 1.25 ml chloroform

Equipments :- Iodine flask ( conical flask C stopper ) (250 ml.)

:- Beakers - 50 ml.

:- Funnel - 1 in no. graduated cylinders (100 and 25 ml)

:- Filter papers

:- Water bath

:- Incubators (oven)

:- Weighing Machine

PROCEDURE :-

1) Weigh about 5 gm of the powered drug in a beaker and transfer it to a dry 250 ml. of

iodine flask.

2) Fill a 100 ml graduated cylinder to the required mark with the solvent (water +

chloroform) wash out the weighing bottle and pour the washing together C the

reminder of the solvent into the conical flask.

===============================================================================


50

Methodology ==================================================================

3) Cork (stopper) the flask and set aside for 24 hrs. shaking frequently (maceration).

4) Filter it into a 50 ml cylinder - When sufficient filtrate has been collected transfer 25

ml. of the filtrate to a weighed 25 ml. beaker as used for the ash values

determinations.

5) Evaporated to dryness on water bath and complete the drying in an oven at 1000c for

about 10-15 min.

6) Cool in desiccator and weigh.

7) Calculated the percentage w/w of extractive with reference to the air dried drug.

OBSERVATION :-

Wt. of drug and beakar before drying = 52.659 gm

Wt. of drug and beakar after drying = 52.175 gm

0.484 gm

L 25 ml gives = 0.484 gm

100 ml gives = 0.484 x 4

1.936 gm

L 5 : 100 : 1.936 gm

100 x 1.936 = 38.720 %

5

(standard value = not less than 25 % )

===============================================================================


51

Methodology ==================================================================

Determination of alcohol soluble extractive :

Ingredients :- a) powdered drug - 5 gm

b) alcohol (90 % ) - 100 ml.

Equipments :- Iodine flask - 250 ml - 1 in no.

:- Beakers - 50 ml., 25 ml.

:- Measuring cylinder - 100 ml and 25 ml (graduated)

:- Funnel - 1 in no.

:- Filter papers

:- Water bath

:- Incubator (hot air oven)

:- Weighing machine

PROCEDURE :-

1) Weigh about 5 gm of the powdered drug in a beaker and transfer it to a dry 250 ml.

Iodine flask.

2) Fill of 100 ml graduate cylinder to the required. Mark the solvent (90% alcohol)

washout bottle and pour the washing, together with the remainder of the solvent into

the conical flask.

3) Cork (stopper) the flask and set aside for 25 hrs. Shaking frequently (maceration)

4) Filter into a 50 ml cylinder when sufficient filtrate has been collected, transfer 25 ml.

of the filtrate to the weighed 25 ml .

5) Evaporated to dryness on water bath and complete the drying in an oven at 1000c for

about 10 -15 mins.

===============================================================================


52

Methodology ==================================================================

6) Cool in desiccator and weight.

7) Calculate the percentage w/w of extractive with reference to the air dried drug.

Calculation :-

1) Wt. of empty beaker = 49.241 gm

2) Wt. of beaker + 25 ml of substance

evaporated = 49.500 gm.

Diff. = 0.259 gm.

L 25 ml gives = 0.259 gm.

100 ml gives = 0.259 x 4 = 1.036 gm.

L 5 : 10 : 1.036

100 x 1.036 = 20.72 % (Standard value not less than 19.70) 5

E) Determination of Extractive values :

* Useful for the evaluation of crude drug.

* Give idea about the nature of chemical constituent present in a crude drug.

* Useful for the estimation of specific constituent soluble in that particular solvent

used for extraction.

===============================================================================


53

Methodology ==================================================================

(4.6) Methodology (41)

Extraction (Schematic Presentation)

Extraction

Aqueous Extract Alcohol Extract

Hot Water Extract Cold Water extract Hot continuous extract

Abbreviations for extract sample :-

Cold water - A Hot Water - B Ethanol Extract - C

Extractions :-

INTRODUCTION :-

As it is preliminary study, extractions selected were cold water extracted by

maceration and hot continuous extraction by soxhlation (Ethanol)

To start with we have taken alcohol i.e. ethanol from distilling rectified spirit by

distillation procedure. Then the procured ethanol was used for the extraction of the

following drug.

(i) Seed powder of Palash :

===============================================================================


54

Methodology ==================================================================

4.7) ETHANOL EXTRACTION (42) :-

(i) Seeds Alcoholic Extraction :

Instrumentation : (i) Soxhlet Apparatus.

(a) Thimble - 400 ml.

(b) Reflux condenser

(c) 2 rubber pipes (inlet and outlet)

(d) 1 lit round bottom flask.

(ii) Heating Mantle :- 1 lit capacity.

Other materials : (a) Filter paper (b) 500 ml beaker (c) Glass funnel.

Ingredients : 1) Palash seed powder - 500 gms.

2) Alcohol (Ethenol) - 1000 ml.

PRINCIPLE :

Extraction involves the separation of bioactive portion of the plant tissues from

the inactive components by using selective solvent in standard extraction. The process at a

temperature approximately that of the boiling point of the solvent apparatus permits the

uniform percolation of the drug and the continuous flow of vapour of the solvent around the

percolator is best for this type of extraction.

The process is generally applied to the removal of natural products from dried

tissues originating from plant fungi etc. The non stream volatile components may be

removed by solvent extraction using a batch or continuous process of extraction of a solid by

a hot solvent it is better to use a soxhlet apparatus.

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55

Methodology ==================================================================

PROCEDURE :

The solid substance (the seed powder of palash) is placed in a porous thimble (i.e.

made of tough filter paper or cotton) and the latter is placed in the inner tube (thimble) of the

soxhlet app. The apparatus is then fitted to a round bottom flask of appropriate size

containing the boiling chips and then to a reflux condenser (preferably the double surface

type) Before joining condenser or even after joining, pour the solvent in the thimble slowly

and fill it. Then allow the thimble to soak along with the solvent over night.

Then next day pour solvent (alcohol/ethanol) and fill the round bottom flask with

appropriate quantity of solvent.

The solvent is boiled gentely, the vapour passes up through the tube and is condensed

by condenser and then condensed solvent falls in the thimble and slowly fills the body of

soxhlet. When the solvent reaches the top of the tube, it siphons over into the flask and this

portion of substance, which it has extracted, gets collected in the round bottom flask.

The process is repeated automatically, untill complete extraction is attained. Fifteen

to eighteen hours is needed for complete extraction of constituents.

2nd Part :

1) After extraction the solvent collected in round bottom flask is collected in a beaker

and then evaporated.

2) The extract was then concentrated on hot water bath and finally reduced to dryness

(or till the alcohol smell in lost completely).

3) After drying the respective extract was weghed weight and yield recorded.

4) The physical characters were noted. yield of extract : 20.72 gms.

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56

Methodology ==================================================================

OBSERVATIONS :

Seed powder taken - 500 gm. solvent ethanol - 1000 ml.

yield of extract - 20.72 gm.

Time taken - 22 hrs.

Cycles - 25 cycles.

Colour - Changes from - dark yellow to whitish yellow.

Inference - At first cycle extract is thick then thickness is

decreases andalso colour changes.

2) Hot Water Extraction :

Palash seed powder - 250 gm

Procedure - soxhlation

Solvent - Water Distilled 750 ml.

Yield of extract - 20.09 gm.

Time taken - 20 hr.

OBSERVATION :

Colour changes as ethanol extract but yellowish tinge is more than that.

AQUEOUS EXTRACTION (WATER EXTRACTS) (43) :

Equipments :- Conical Flask - 2 lit capacity.

Measuring cylinder - 25 ml capacity

Silver foil and filter paper

Emulsion cloth.

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57

Methodology ==================================================================

Solvent :- Water, chloroform ( Preservatory )

Ingredients :- Seed powder - 200 gm

Water C. 1000 ml.

Chloroform - 20 ml.

PROCEDURE :-

1) Required amount of the drug powder is taken in a conical flask.

2) Pour some water (required amount of water) in a certain ratio.

3) Add some chloroform as preservatory.

4) Close the mouth of the conical flask with a filter paper and silver foil.

5) Shake it vigorously with frequently intervals.

6) Then place the conical flask in a warm place for 7 days with frequent vigorous

shakings.

7) After 7 days filter the content and the filtrate is dried and the liquid part is kept on a

water bath and evaporated.

8) It is evaporated till the required concentrate is achieved.

9) And stored in a dry containers.

Seed powder - 200 gm.

Water - 1000 ml.

Chloroform - 20 ml.

7 days it is keep as it is kept.

After 7 days they were filtered respectively and later.

Were kept on a water bath and evaporated to their required concentration.

Observation - before evaporation - ml.

After evaporation - 25.32 mg.

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Methodology ==================================================================

(4.9) PRELIMINARY PHYTOCHEMICAL SCREENING (44) :

PROCEDURE :-

I. TESTS FOR PHYTOSTEROLS :

(a) Salkowaski Test : To the test extract solution added few drops of conc. H2SO4

shaken and allowed to stand, lower layer turn red indicating the presence of sterols.

(b) Liebermann - Burchard test : The test solution treated with few drops of acetic

anhydride and mixed, when conc. H2SO4 is added from the sides of the test tube, it

show a brown ring at the junction of the two layers and the upper layer turns green.

(c) Sulphur test : Sulphur when added in to the test solution, it sinks in it.

II. TEST FOR TRITERPENOIDS :

(a) Salkowski test : Few drops of concentrated sulphuric acid was added to the test

solution of the extract, shaken and on standing lower layer turns golden yellow.

(b) Liebermann Burchard Test : To the test solution of the extract, few drops of acetic

anhydride was added and mixed well. Then 1 ml of concentrated sulphuric acid was

added from the sides of the test tube, a red colour is produced in the lower layer

indicate Triterpenes.

(c) Tschugagiu test : Excess of acetyl chloride and pinch of zinc clroride added to the

test solution of extract, kept aside for reaction to subside and warmed on water bath,

eosin red colour produced.

(d) Briekorn and Brinar test : To the test solution of extract, few drops of

chlorosuphonic acid in glacial acetic acid (7.3) red colour is produced.

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59

Methodology ==================================================================

III. TESTS FOR GLYCOSIDES :

(a) Baljet's test : The test solution treated with sodium picrate gives yellow to orange

colour.

(b) Bromine water test : Test solution dissolved in brominoine water gives yellow

precipitate.

(c) Raymond's test : Test solution treated with dinitrobenzene in hot methanolic alkali

gives violet colour.

(d) Legal's test : Test solution when treated with Pyridine (made alkaline by adding

sodium nitroprusside solution) gives pink to red colour.

IV. TEST FOR ANTHRAQUINONES :

(a) Borntrager's test : Boiled test extract solution with 5 ml of 10% sulphuric acid for 5

mins. Filtered while hot, cooled the filterate shaked gently with equal volume of

benzene. Separated the benzene layer and when treated with half of its volume

solution of ammonia (10%). Allowed to separate ammoniacal layer aquires rose pink

colour due to the presence of anthraquinones.

V. TESTS FOR SAPONINS :

(a) Foam test : Test solution and shaken shows the formation of foam, which is stable

an least for 15 mins.

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Methodology ==================================================================

VI) TESTS FOR CARBOHYDRATES :

(a) Molisch's test : Test solution with few drops of Molisch's reagent and 2ml of

concentration H2SO4 added showly from the sides of the test tube shows a purple

ring at the junction of 2 liquids.

(b) Barfoed's test : Test solution treated with Barfoed's reagent, on boiling on a water

bath shows brick red colour precipitate.

(c) Benedict's test : The test solution treated with Benedict's reagent and boiling on

water bath shows reddish brown precipitate.

(d) Fehling's test : The test solution when heated with equal volume Fehling's A and B

solutions, gives orange red precipitate presence of reducing sugars.

(e) Selivinoff's test : A crystal of resorcinol is added to the test solution and warmed on

a water bath equal volume of conc. HCL a rose colour is produced that showed

ketone is present.

(f) Tests for pentoses : Heat the test solution with a equal volume of HCL containing

little phloroglucinol. Formation of red colour lidicates the present pentoses.

VIII. TESTS FOR ALKALOIDS :

(a) Mayer's test : Test solution with Mayer's reagent (Potassium Mercuric iodide) gives

cream coloured precipitate.

(b) Hager's test : The acidic solution with Hager's reagent (Saturated picric acid

solution) gives yellow precipitate.

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Methodology ==================================================================

(c) Dragendorff''s test : The acidic solution with Drogendroff's reagent (Potassium

bismuth iodide) shows reddish brown precipitate.

IX. TESTS FOR FLAVONOIDS :

(a) Shinoda test : Test solution with few fragments of magnesium ribbon and conc.

HCL shows pink to magenta red colour.

(b) Zn/HCL reducing test : Test solution with Zinc dust and few drops HCL shows

magenta red colour.

(b) Alkaline reagent test : Test solution when treated with sodium hydroxide solution

shows increase in the intensity of yellow colour, which becomes colourless on

addition of few drops of dilute acid.

IX) TEST OF PHENOLICS / TAINS :

(a) FeCl3 test : Test solution treated with few drops of FeCl3 solution gives dark colour.

(b) Gelatin test : Test solution treated with gelatin gives white precipitate.

X) TESTS FOR PROTEINS :

(a) Millon's test : Test solution when treated with Millon's reagent and heated on a

water bath, Protein is stained red on warming.

(b) Xanthoproteic test : Test solution treated with conc. HNO3 and boiled gives yellow

precipitate.

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Methodology ==================================================================

(c) Biuret test : Test solution treated with 40% NaOH and dilute CuSO4 solution gives

blue colour.

XI) TEST FOR AMINO ACIDS :

(a) Ninhydrin test : Test solution treated with Ninhydirn reagent gives blue colour.

(A) THIN LAYER CHROMATOGRAPHY (TLC.) (45,46) :

Introduction :-

With the help of Thin Layer Chromatography (TLC.) One can determine particular

value? Small amount of impurities. We get principle of TLC. and application of the

technique in pharmaceutical analysis in volume I of the internal pharamocopoeia (5) This is

the earliest method to perform. And equipments which are required is introversive, so the

technique is used frequently to evaluate medical plant material and their preparations.

Parameters should be determined on the basic of published pharmacopeia

monographs established experimentally for the analysis of each individual plant material.

* Type of absorbate and method of activation of no information on the letter can be

obtained, heat at 1100c for 30 minutes.

* Method of preparation and concentration of the test and reference solution.

* Volume of the solution to be applied on the plate.

* Mobile phase and the distance of migration.

* Drying condition (including temperature) and modern of detection.

For the spots obtained.

* Number of approximate position, or the Rf values if necessary.

* Fluorescence and colour.

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63

Methodology ==================================================================

Types of thin layer chromatography methods are described as follow :

(i) Classical method and (ii) Micro method.

Which uses different size of plates and hence different quantities of solvent.

Classical Method :

Recommended Procedures :

The method outlined below assumes that chromatographic plates prepared in the

laboratory are used but precoated plates, activated if necessary, may be used provided that

they have proved suitable for application concerned.

A powdered specimen of pharmacopoeial quality may be used as the reference

material, if a test for the presence of known active principle of a medicinal plant material is

to be carried out, a chemical reference substance identical to that principle should be used.

The test and reference solution should be prepared simultaneously in exactly the same way.

The reference solution should be of known concentration. If the relative concentrations of

the chemical substances in the reference solution are selected in accordance with the

composition of typical material, comparison of the spot size offers valuable additional

information. The solvent system should be specified in the test procedure of the individual

material being examined. A three colour mixture (e.g. 0.01% solution in toluene of

indophenol blue, sudan red G and dimethyl yellow) run together, permits a rapid check on

the prevailing chromatographical conditions.

If it is suspected that the materials being examined are unstable, the chamber in

which chromatography takes place should be protected from light. In any case, the

chromatographic chamber should always be kept out of direct sunlight. Otherwise, the rays

of the may be refracted to different degrees owing to imperfections on the glass walls of the

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64

Methodology ==================================================================

chamber, giving rise to area of elevated temperature on the chromatographic plate and erratic

flow of the mobile phase.

Preparation of samples :

Prior to testing, prepare an extract of the plant material to be examined, using a rapid

extract process, as follows, To 0.0-1.0 g of the powdered plant matetial add 1-10 ml of

solvent and extract either by stirring, shaking the mixture of 3-30 minutes, or heating to

boiling and allowing to cool. Remove the insoluble matter by centrifugation, or filter through

a small funnel with filter paper or a cotton plug. If necessary, evaporate the filtrate on a

water-bath for just as long as is required to remove the solvent, then re-dissolve the residue

in a small volume of solvent (e.g. 0.1 to 0.2 ml). If necessary, purify the test solution by

repeating the extraction with solvent at a different pH, or by sublimation, distillation, or

other appropriate method.

APPARTUS :

The equipment consists of :

Glass plates of uniform thickness throughout entire area, 15-20 cm long, and wide

enough to accommodate the required number of test and reference solutions.

A device for spreading uniform layer of coating material of desired thickness onto

the glass plates;

A rack to hold the prepared plates (normally 10 plates with set spacing) during the

drying period or for transportation and storage; the rack should be small enough to fit in a

drying oven and desiccators;

A chromatographic chamber of transparent material, usually glass, with a tightly

fitting lid, of suitable size to accommodate the test plates;

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Methodology ==================================================================

A suitable spraying implement with a fine spray nozzle, made of material resistant to

the reagents to be used; An ultraviolet light source emitting short (254mm) and ling

(365mm) wavelengths.

Before use, clean the plates scrupulously by immersing in a suitable cleaning liquid

and rinsing thoroughly until the water runs off the plates without leaving any visible

watermarks or oily spots, and then dry. The plates must be completely free of lint or dust

when the coating material is applied.

METHOD :

Preparation of the adsorbent :

Unless otherwise specified in the procedure for the plant material concerned prepare

a slurry of the coating material and water on an aqueous solution (see section 21,

"specifications for adsordents") and, using the spreading device, coat the cleaned plates with

a layer about 0.25 mm thick. Dry the coated plates in air, heat to activate 1100C for minutes,

and then allow to cool. Inspect the uniformity of the coating in transmitted light and the

texture in reflected light. If the plates are not to be used immediately, store them in a

desiccator containing silica gel, desiccant, R. To form and edge remove a narrow strip (2-5

mm) of the coating material from the sides of the plate.

If acid, alkaline or buffered layers are required, use diluted acid, base or salt mixtures

instead of water for the preparation of the slurry, As specified in the test procedure. An

aqueous solution of 5-7 g of sodium carboxymethylcellouse R could replace the water, if the

absorbent dose not already contain a binder.

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66

Methodology ==================================================================

Saturation of the chromatographic chamber :

Unless otherwise specified in the test procedure, the chromatography is carried out in

a saturated chamber. To achieve saturation, line at least half of the total area of the inside

walls of the chamber with filter-paper, into the chamber a sufficient quantity of the mobile

phase to saturate the filter-paper and form a layer about 5 mm dep. Close the chamber and

allow to stand for at least 1 hour at room temperature.

All operations during which the plate is exposed to the air should preferably be

carried out at a relative humidity of 50 - 60%, and the plates should be handled with care.

Application of the test and reference solution :

Using a micropipette or a syringe graduated in ml, place spots of the test and

reference solution onto the starting line, which should be parallel to and about 15 mm above

the lower edge. The spots should be at least 15 mm from the sides of the plate, and at least

15 mm. apart. They should be as possible, preferably not more than 4 mm in diameter, if

necessary, apply the solution in portions, drying between applications. Mark the distance the

mobile phase in intended to ascend as specified in the rest procedure, usually 10-15 cm from

the starting line.

The results of speration can often by improved by applying the solutions to form a

horizontal band 10-15 mm long and not more than 5 mm wide.

Development of chromatograms :

Allow the spots to dry, place the plate - as nearly vertical as possible - into the

chamber, ensuring that the points of application are above the surface of the mobile phase.

Close the chamber. Develop the chromatogram at room temperature, unless otherwise

specified in the test procedure, allowing the solvent to ascend the specified distance. Remove

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67

Methodology ==================================================================

the plate, mark the position of the solvent front and allow the solvent to evaporate at room

temperature or as specified.

Observation and interpretation of the chromatograms :

Observe the spots produced in daylight, then under short-wave and long-wave

ultraviolet. Mark the centre of each spot with a needle. Measure and record the distance from

the centre of each spot to the point of application, and indicate for each spot the wavalength

under which it was observed. If indicated in the test procedure, spray the spots with the

specified reagent, and observe and compare the spots with those of a reference material.

If required calculate the ratio of the distance traveled on the adsorbent by a given

compound to that traveled by the leading edge of the solvent (the Rf value) or the ratio of the

distance moved by a compound and a stated reference substance (the Rf value) as follows :

Rf = Rf = a b

a c

where

a = the distance between the point of application and the centre of the spot of the

material being examined ;

b = the distance between the point of application and the solvent front;

c = the distance between the point of application and the centre of the spot of

reference material.

Rf value may vary with each experiment depending on the saturation conditions in

the chromatographic chamber, the activity of the adsorbent layer, and the composition of the

mobile phase.

Note : The above mentioned procedures are as per W.H.O. criteria.

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Methodology ==================================================================

1. Thin layer Chromatographic - Analysis (TLC) of Drugs :

Of the many chromatographic methods presently available, thin layer

chromatography has become widely adopted for the rapid and positive analysis of drugs and

drug preparations.

There are several reasons for the popularity of this method :

- The time required for the demonstration of most of the characterstic constituent of a

drug by TLC is very short.

- In addition to qualitative detection, TLC also provides semi-quantitative information

on the chief active constituents of a drug or drug preparation, thus enabling as

assessment of drug quality.

- TLC provides a chromatographic drug fingerprint. It is therefore suitable for

monitoring the identity and purity of drugs, and for detection adulteration and

substitutions.

- With the aid of appropriate separation procedures, TLC can be used to analyses drug

combinations and phyto-chemical preparations.

- Thin layer chromatograms can be documented.

II. Documentation of Thin Later chromatograms :

Various methods of documentation are possible :

- Description of the Rf value and colour of the characteristic main zones, with

reference to a standard substance or test mixture. This method has been adopted in

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69

Methodology ==================================================================

the 8th edition of the German pharmacopoeia, the European pharmacopoeia, USP

XX and other.

- Construction of a scale diagram of the thin layer separation, showing migration

distance and intensities of the characteristic zones. The zones observed in visible

light (vis.) in UV-254 nm and UV-365 nm are described.

- Colour photography in daylight or UV-light, under conditions that give the most

authentic reproduction of the colours and intensities of the separated zones.

- Densitometry or fluromety of the chromatogram at certain wavelenghts. Under

favourable conditions, this procedure also yields a drug fingerprint, and enables the

quantification of certain chief constituents. If suffers from the disadvantage that a

calibration graph constructed at one wavelength is applicable to only some of the

constituents.

B) High Power Thin layer chromatography (H.P.T.L.C.) (47) :

The high performance thin layer chromatography commonly known as H.P.T.L.C. is

sophisticated from of thin layer chromatography. It involves the same theoretical principles

of T.L.C. where in substances are separated on the basis of their differential migration in

system of 2 phases of special types of plates.

The important steps involved are :

1. Sample preparation 2. Selection of chromatographic layers.

3. Plates. 4. Pre washing

5. Conditioning 6. Sample application

7. Pre conditioning 8. Mobile phase

9. Chromatographic development. 10. Detection of spots.

11. Scanning and documentation.

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Methodology ==================================================================

1. Sample Preparation : For normal phase chromatography using silica gel pre-coated

plates, solvents should be non polar of volatile type. For reversed phase

chromatography usually polar solvents are used for dissolving the sample.

2. Selection of chromatographic layers : The solution depends on the nature of

material to be separated commonly used material are silica gel 60F 253, alumina

cellulose etc.

3. Plates : Plates of 20 x 20 cms or 5 x 7.5 cms having 100-200 mm adsordent

thickness is used for qualitative analysis. Silica el 60F254 having a pore size 6 mm

with flurescent indicator is a coat material. The solvent layer is fixed with the help of

glue. The basic difference between T.L.C. and H.P.T.L.C. plates is partial size of

coated material, which is 5-20 mm of T.L.C. and 4-8 mm for H.P.T.L.C.

4. Pre-washing : Plates need to be washed to remove water vapours and volatile

impunities. The plates are cloned by methanol.

5. Conditioning : The pre-washed plates are placed in oven at 1200C for 15 to 20

minutes. This process is known as conditioning.

6. Sample application : Application of 1.5-5 ml is most satisfactory for H.P.T.L.C.

application of the sample.

7. Pre conditioning (Chambers saturation) :- For low polarity mobile phase there is

no need of saturation, however saturation is needed for highly polar mobile phases.

8. Mobile Phase :- The solution of appropriate mobile phase is by trial and error in

which chemical of analytic absorbent layer etc. are considered.

9. Detection of spots : Immediately after the development is completed. The plates are

removed from the chamber and dried to remove trees of mobile phase, Generally

detection can be known by iodine vapour in Jodive chamber.

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71

Methodology ==================================================================

10. Scanning and Documentation : The H.P.T.L.C. equipments are supplied with

computer, data recording and storing divice. The developement of H.P.T.L.C. plates

are scanned at selected U.V. regions wavelength by the instrument and the detected

spots are seen on computer in the form of peaks. The scanner converts bond into

peaks and peak heights or area is related to the concentration of the substance on the

spot. The peak heights and area under the spot (curves) are measured by the

instrument and are recorded as the printer.

INFERENCE :

These are the general procedures explained according to W.H.O. standards. In this

study we are concentrating only on the qualitative estimations of the sample extracts. i.e. By

their finger printing we are observing for their Rf values, which indicate the presence of

certain phyto-constituents in each sample extract. These can be further clarified by phyto-

chemical studies of their fractions which can be procured by fractionation.

4.11 EVALUATION OF ANTIMICROBIAL ACTIVITY (48,49) :

Principle :

The agar dilution technique is used to measure qualitatively the in vitro activity of an

Antimicrobial agent against the test bacterial.

In this method, the petridishes were filled with inoculated liquefied agar medium to

uniform thickness. Then graded amount of test samples (Ex. antibiotics) are incorporated in

agar plates and inoculated in spots with the organisms under study. If the organism under

study is susceptible to the incorporated test samples.

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Methodology ==================================================================

No bacterial growth is expected in agar plates with higher amount of the drugs.

Bacterial growth is observed as the test sample concentration in the agar plate diminishes.

Inhibition of growth at the minimum or lowest concentration of test sample.

Antimicrobial activity :

The antimicrobial activity of a drug is generally expressed as its inhibiting effect

towards the growth of bacterium in nutrient broth or on nutrient agar.

For this study, following conditions are observed for :

1) The substance or extract must be in contact with the test organism.

2) Conditions must be favourable for the growth of microorganisms in the absence of

Antimicrobial substances.

3) There must be means of estimating the amount of growth and thereby percentage of

growth of inhibition.

4) The activity of extract should be observed and determined by the growth response of

microorganisms.

Different methods employed in the study are agar streak. serial agar diffusion, paper

disc, cup-plate, strip diffusion, cylinder plate and turbidimetric methods.

Methods :- Tests was done by cup diffusion method.

In present study "Palash" seed powder were extracted with water and ethanol as

solvents and all extracts were subjected for antimicrobial activity.

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Methodology ==================================================================

Experimental Procedure :- (50)

In the present study, the antimicrobial screening was done by cup plate method or

cup diffusion method. This is one of the authentic procedure in I.P. where the antimicrobial

extract is differed from the cup through an agar layer in petriplate (dish). Petridish to an

extent such that the growth of added microorganism are restricted entirely in circular area or

zone arround cavity containings the solutions of the test sample. The antimicrobial activity

expressed as zone diameter in mm which is measured with a divider.

All the extract of the seed were screened for antimicrobial acitivity against broad

spectrum of microorganisms and the activity compared with appropriate standards.

Standards used in the study :- For bacterias (Gream +ve, Gram -ve)

(i) Amikacin (ii) Olfloxacin (iii) Penicillin

For fungi

(iv) Griseofulvin.

Test organism used for the study :-

(1) Bacterias :-

(a) Staphylococcus aureus (gram +ve)

(b) Bacillus subtillis (Gram +ve)

(c) Escherichia Coli (Gram -ve)

(2) Fungi :-

(a) Candida albicans

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Methodology ==================================================================

Preparation of Antimicrobial agent stock solution :

1) Remove the Antimicrobial agent from the freezer and warm to room temperature

before opening to avoid condensation of water.

2) Weigh appropriate amount of the powdered semi liquid Antimicrobial agent.

3) Dissolve the Antimicrobial agent powdered / semi liquid in solvent to make 2 mg /

ml and mg / ml concentrations.

Preparation of standard solutions :

1) Required amount of standard Ofloxacin - 2 micg / disc and norfloxacin - 10 micg /

disc solutions are prepared.

2) The concentration of standard equivalent to 100 micg/disc of griseofulvin was

prepared as standard for antifungal activity.

Preparation of Inoculum :

Stock cultures were maintained at 40c on slopes of nutrient agar. Active cultures for

experiences were prepared by transferring a loopful of cell from the stock cultures to test

tubes of nutrient broth for bacteria and that was incubated without agitation for 24 hours x at

370c an 250c respectively.

Culture Medium :

Different types of media have been used according to the types of organism In the

present investigations, Antibiotic medium No. 5 (Nutrient Agar-Himedia)

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75

Methodology ==================================================================

Nutrient agar composition :

Agar : 15. gm

Beef extract : 1.5 g

Yeast extract : 3.0 g

Peptone : 6.0 g

Distilled water to make : 1000 ml.

About 27 g of above readymade medium was dissolved in freshly prepared distilled

water (1000 ml) by gentle heating.

Sterlization :

Sterlization of the medium, tubes for slants, borer etc. was done by autoclaving at 15

lbs/spqure inch. for 20 min. The glasswares like syringes, petridishes, pipettes, empty test

tubes, were sterlized by dry heat in an over at a temprature of 1600C for 1 hr.

Preparation of agar plates :

The sterlised medium was cooled at 400C and 0.5 ml of inocula per 100 ml. of

medium were added to the conical flask. This was shaken gently to avoid the formation of

air bubbles and then transferred into petridishes so as to obtain 6 mm thickness of medium.

The medium in the plate was allowed to solidify at room temperature.

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76

Methodology ==================================================================

Experimental Procedure :

The sterile borer was used to prepare nine cups of 8 mm diameter in the medium of

each petri-dish. An accurately. measured 0.1 ml solution of each concentration of solutions

of extracts and standard samples were added to the cups with the help of micropipette. All

the plates were kept at room temperature for effecting diffusion of drug extract and standard.

Later, they were incubated at 37 + 10C for 24 hrs. The presence of definite zones around the

cup of any size indicated antibacterial activity. The control were run simultaneously to

assess the activity of petroleum ether, chloroform, acetone - water and water which were

used as a vehicle for extract. The diameter of the zone of inhibition was measured and

recorded.

The zone of inhibition for the antibacterial and anti-fungal activities of extracts were

calculated by measuring the inhibitory effect towards the growth of bacteria and fungus

around nutrient agar cup. The results for antibacterial activity were shown in table no. and

anti-fungal was shown no.

Conclusion :

From the results, it is concluded, all extracts are resistant to bacterials as well as

fungi, when compared to the standards.

M.I.C. Method :

First the stock solution of the test sample i.e. ethanol and water extracts of "Palash"

seed were prepared separatively i.e. 10 mg. ml. concentrated stock 501n is prepared as much

required.

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77

Methodology ==================================================================

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78

Produce :

(1) Sterile tubes were selected and serially labeled from 1 to 7.

(2) 0.5 mg. of Muller Hinton broth solution is added from tube no. 2 to no. 7.

(3) Then in tube 1 and 2, 0.5 ml. of stock 501n (10 mg/ml.) added.

(4) From tube 2, transfer 0.5 ml to tube 3 mix 8 transfer to tube 4 and transfer similarly

till tube 7 Discard 0.5 ml. from tube 7.

(5) Adjust the bacterial count 105 organisms/ml by comparing with Mac Farland's

Standards.

(6) Add 0.5 ml of culture broth to all 7 tubes

(7) Incubate the tubes at 370c cover night.

(8) Than observe for turbidity and record the values observation note - Turbidity in first

tube is the M.I.C. value for that microbes.

Result : M.I.C. value of the extract has been mentioned in Table No. 11 to 14.

Conclusion :

Sample share better results on e-coli and S. aureus and basilius subtillis but totally

resistant again candida albicans That means ethanol extract shows best activity against

bacteriaes but not effectively acting on fungis.

Results ==================================================================

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78

Results ==================================================================

RESULTS

(5.1) MACROSCOPIC AND MICROSCOPIC STUDY :

As per the figures and photos in Annexure :

(5.2) PHYSIO CHEMICAL STUDY :

(i) Total Ash value :

Sl. No. Parameters Observation

1. Seed powder of crude drug 7.425 %

2. Fine seed powder 7.685 %

(ii) Acid insoluble ash

Sl. No. Parameters Observation

1. Seed powder 0.03 gms

(iii) Water soluble extractive values 38.72 %

(iv) Alcohol soluble extraxctive value of "Palash seed 20.72 %

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Results ==================================================================

(5.3)

(a) Yeild of Extract :

Sl. No. Extract Wt. Yield W/W.

(1) Ethanol 20.72 gms. 20 %

(2) Hot Water 20.09 gms. 20 %

(3) Cold Water 25.32 gms. 25 %

(b) Organoleptic Characters of Extracts :

Organoleptic Alcoholic Hot water Cold Water

Extract Extract Extract

Colour Yellowish Brownish Brownish Dark Brown

Consistancy Oily Oily Oily

Odour Characteristics Characteristics Characteristics

Taste Bitter Bitter Bitter

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80

Results ==================================================================

TABLE NO. 9

5.4 PRELIMINARY PHYTO-CHEMICAL SCREENING :

Chemical Tests

Extraxts

Alcohol Aqueous

1) Test pf Sterols

a) Salkowaski test + −

b) Liebermann - Burchard test + −

2) Test for Steroidal glycosides + +

3) Test for Triterpenoids

a) Salkowaski test + −

b) Liebermann - Burchard test + −

c) Tschugajew test + −

d) Briekorn and Brinar test + −

4) Test for Glycosides

i) Cardiac glycosides − −

a) Baljet's test − −

b) Raymond's test − −

c) Legal's test − −

ii) Anthraquinone Glycosides − −

a) Borntrager's test − −

b) Modified Borntrager's test − −

5. Test for Saponins

a) Foam test −

6. Test for Flavonoids / Flavonoid Glycosides

a) Shinoda test + +

b) Zinc Hydrochloric acid reduction test + +

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81

Results ==================================================================

c) Alkaline reagent test + +

7. Test for carbohydrates

a) Molisch's test + +

b) Fehling test + +

c) Barfoed's test + +

d) Benedict's test + +

e) Selvinoff's test + +

f) Test for pentoses + +

8. Test for Alkoloids +

a) Mayer's test − +

b) Wagner's test − +

c) Hager's test − +

d) Dragendorff's test − +

9. Test for Phenolics and Tannins +

a) Lead acetate test + +

b) Ferric Chloride test + +

c) Gelatin test + −

10. Test for Lipid +

11. Test for Proteins − +

a) Millon's test − +

b) Xanthoproteic test − +

c) Biuret test − +

12. Test for free amino acid

a) Ninhydrin test − +

Note : " + " Present, " - " Absent.

Glycosides + in both extract. Carbohydrat + in both extract.

Sterols + in alcohol Terpenoids + in both extract

Alkaloids + present in aq. Tannins + present in both

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82

Results ==================================================================

HPTLC ANALYSIS :-

Qualitative analysis of aquesous extract and ethenolic extract :

Sl. Sampled Detail Amount of Rf values

No. extract on plate @ 500 m.

1. Ethanol extract of "palash seed" 200 mcg. 0.16, 0.30, 0.47

2. Aqueous extract of "Palash seed" 200 mcg. 0.08, 0.14, 0.24

Result :-

Rf values of ethanol extract are approximately double that of aqeous extract.

Remark :-

There were no bands observed @ 254 mm. and @ 366 mm, but, ethanol extract

showed bands @ 500 mm but we cann't observed any bands of aqueous extract @ 500 mm.

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83

Results ==================================================================

(5.5) ANTIMICROBIAL RESULTS : Cup and Diffusion Method (Mueller and Hinton Agar). TABLE NO. 10

Drug / Compound

Tested

Zone of Inhibition of

E-coli

Zone of Inhibition of

Staphylococcus Aureus.

Zone of Inhibition of of Candida Albicans.

Zone of Inhibition of of Bacilus Subtillus.

Ofloxacin (2micg/disc) 20 mm 15 mm - - Amikacin (10micg/disc) 14 mm 14 mm - - Penicillin (10micg/disc) 12 mm 24 mm - - Greseofulvin (100micg/disc) - - 10 mm - Cold water extract for 4 mg / ml - A1

0 0 0 0

Hot water extract for 4 mg/ml - B1

0 0 0 0

Ethanol extract for 4 mg/ml - C1

0 11.2 mm 0 10 mm

Cold water extract for 6 mg/ml - A2

2 mm 0 0 0

Hot water extract for 6 mg/ml - B2

0 0 0 0

Ethanol extract for 6 mg/ml. - C2

0 12.3 mm 0 10.5 mm

Cold water extract for 8 mg/ml. A3

6 mm 2 mm 0 0

Hot water extract for 8 mg/ml B3

3 mm 0 0 4 mm

Ethanol extract for 8 mg/ml - C3

12 mm 13 mm 2 mm 12 mm

Note : ‘Aqueous extract’ showed resistant activity to bacteria and fungi. ‘Ethanol extract’

showed resitant activity to fungi and sensitive to bacteries.

Indicates :

* A-Cold water extract * B-Hot water extract * C-Ethanol extract

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84

Results ==================================================================

ANTIBACTERIAL RESULTS :

TABLE NO. 11

a) Esherichia Coli :

Sl. No. 10 mg/ml 5 mg/ml. 2.5 mg/ml. 1.25 mg/ml. 0.612 0.3 0.15 MIC Value

A R R R R R R R > 10

B R R R R R R R > 10

C S S S R R R R 2.5

TABLE NO. 12

b) Staphylococcus Aureus :




C S S S R R R R 2.5

TABLE NO. 13

c) Bacillus Subtillis :




C S S S R R R R 2.5

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85

Results ==================================================================

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86

ANTIFUNGAL RESULTS :

TABLE NO. 14

D) CANDIDA ALBICANS :




C R R R R R R R > 10

NOTE : "S" - Sensitive "R" – Resistant

Discussion ==================================================================

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86

Discussion ==================================================================

6. DISCUSSION

"Palash" plant is wellknown in vedic period (are having lot of convining references.

"Kinshuk" is more popular synonym is refered to "Palash". In vedic kala "Palash" is used for

not only to treat the disease but day today life also.

In "Bruhatrayes" is also known as "Palash" and "Kinhuk". Controversy regarding this

plant is not seen. Different parts like stem, seed, leaves, flowers kshara are used to treat

various diseases.

In nighantus nomenclature of the plant is given according to their physical characters

and combating common ailement. "Palash was used widely in ritual functions like “yagaya’’

etc. That is why it is also given a name as 'yajnika' and it is supposed to have more holy

properties that is why it is termed as "Putadru" and "Sumidwara", "Krimighna" is another.

Synonym which is used to treat in krimi use of "Palash" patra is more common and

according to its physical character, it is said to be Kharaparna and Tripatra. "Palash beeja is

sneha yukta that is why it is termed as 'Beeja sneha'.

According to chassical texts seeds are used to treat various skin diseases, so in this

study more attention and concentration is given regarding this property of "Palash" seed.

Charakacharya not classified in four Ganas . But according to Sushruta Charya

and Vagbhatacharya "Palash" is included in same ganas like "Ambhasthadi, Nyagrodhadi

gana etc.

The use of "Palash" seed is given seperately and different yogas in different samhitas.

In nighantus "Palash" was explained under "Phala varga" Haritakyadi Varga, Vatadi Varga,

Aamradi Varga, Etc.

In "Raj Nighantu" references regarding "Palash" classification is done on the basis of

different colours of the flowers as Rakta, Peeta, Shweta and Neela.

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87

Discussion ==================================================================

"Palash" seed posses katu - tikta and kinchit kashaya rasa. Veerya - Ushna, Vipak -

Katu, Guna "Kapha – Vata Shamaka pittakar". Its main karmas are Krimighna,

Vaatashamak Udar rog, Arsha and It is also used in various skin disorders like Dadru,

Paama, Kandu and Kushta etc.

Botanically first it is known as "Butea frondasa" but now it was also termed to be as

"Butea monosperma Limm? Controversy regarding this plant is not present. Because "Butea

monosperma is a tree and Butea superba is another (type of) variety of this plant but that is

climber one ! Adulteration may taking place with Butea monosperma seed with Butea

superba seed, but seed of Butea monosperma is larger than that of Butea superba.

"Palash" seed has been considered as "Krimighna" by different acharyaas. The work

is intended to study on inference that correlates microbes explained in modern terms. In

antimicrobial study is needed for the time. As the bacteria, fungi etc. Microrganisms have

become resistant to the newest antimicrobial agents, antimicrobial agents have traditionally

been made of natural products in cluding microganisms themselves. Before starting actual

study the drug was authenticated by expert Botanist, and then pharmacognostic study of the

drug i.e. macro and microscopic study was carried out to study its morphology, internal

structures and their contents in detail.

After that physico - chemical analysis was done i.e. ash value, acid insoluble ash,

alcohol and water soluble extractives values were determined. The ash value of sample was

within normal range as given in Ayurvedic herbal pharmacoepoeia, this represents inorganic

salts present in drug like carbonates - bicarbonates etc. In this procedure the carbon is

removed by heating at low temperature.

Acid insoluble ash value was also near by standard by this process we ruled out

presence of excess foreign particles, that is sand or silica etc. which may not be absorbed by

acid media in body and may give rise to further complications.

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88

Discussion ==================================================================

Alcohol soluble and water soluble extractive values are done at pilot study. This is

helpful regarding the yield of extract, which can be acquired by doing extraction on extract,

which can be acquired by doing extraction on larger scales.

Then the extraction procedure were carried out by ethanol and water as solvent. The

procedure adopted was soxhlet extraction and cold maceration procedure. Ethanol was

selected because it is standard during the new drug extraction. The ethanol which was used

prepared in laboratory by distilling rectified spirit. Then the procedure ethanols percentage

95% was clarified by feasible laboratory procedure.

The preliminary phytochemical screeing was done that reveals the presence of

alkaloids, tannines, reducing sugars, proteins, flavanoids, glucosides in all alcohol and water

extracts.

Qualitative analysis of plant extracts were done by T.L.C. and H.P.T.L.C. In this test

presence of various compounds that were detected by their Rf. values. These Rf. values can

be directly and indirectly help for checking adultration in raw or compound drug.

In T.L.C. 0.2 gm. of extract was diluted to 10 ml. of ethanol and used for T.L.C. no

bands were observed with aqueous extract. But with ethanol extract bands were not observes

at 254 and 366. But band were seen at @ 500 nm. Rf values of ethanol extract are (0.16,

0.30, 0.47) It shows that ethanol extract of seed is more effective than aqueous extract.

Antimicrobial Activity :-

This study was carried out on Gram +ve bacterias (S. aureus and B. Septilis) Gram -

ve bacteria E-coli and fungi (Candia albicans).

It was carried out by cup diffusion method agar plates.

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89

Discussion ==================================================================

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90

The zone of inhibition were recorded. In our study aqueous extract was resistant

when compared to the modern standard drugs but ethanol extract is sensitive to bacterias but

it is also resistant to fungi.

MIC values against E-coli and S. aureus were 2.5 mg/ml. but against candida

albicans it was 10 mg or more than that.

Conclusion ==================================================================

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90

Conclusion ==================================================================

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91

7. CONCLUSION

1) The physico chemical results i.e. ash value, acid insoluble ash, alcohol and water

soluble extractive values are within parameters of Ayurvedic Herbal pharmacopoeia.

2) Ethanol extract yield of Palash seed is less than the aqueous extract.

3) The preliminary phytochemicals screening of both ethanolic and water extract show

presence of tannins, carbohydrates, Terpenoids, Glycosides and Alkoloid present in

aq. extract and Streroil present in alcohol extract.

4) Rf. values of ethanol extract are double than that of aqueous extract also height of

peak of ethanol extract in more than that of aqueous extract.

5) In antimicrobial activity aqueous extract showed resistance to all bacterias and fungi

but ethanol extract is sensitive to bacterias and resistant to fungi.

Thus ethanolic extract of palash seed can be used in bacterial infections

The aim of this study is to evaluate the antimicrobial activity of "Palash" seed and

not to check the incidence or prevalence of disease. The statistical analysis of date is not

considered.

Summary ==================================================================

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91

Summary ==================================================================

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92

7. SUMMARY

Seed of "Palash" is used as "Krimighna" but it is not included in krimighna gana or any gana by Charkacharya. It is used in to treat skin diseases and its antihelmentic property is proved. In the present study preliminary study was done in context to study its antimicrobial activity. The plant drug (seeds) were subjected to : (i) Pharmacognostic review. (ii) Water ethenol soxlet extraction. (iii) Preliminary phytochemical screening (iv) T.L.C. and H.P.T.L.C. (Finger printing) (v) Antimicrobial activities (Bacterias and Fungi) The phy to chemicals observed were Tannins glycosides, etc. Antimicrobial study by cup-diffusion method showed positive result of ethenol extract of "Palash" seed on bacterias but it is resistant to fungi and aqueous extract showed resistant to both bacteria and fungi. Scope of further study :-

(a) To study more for their single drug efficacy clinically. (b) Comparative study with patra, flowers can be done. (c) Extraction by different solvents to find extract base for their antimicrobial acitivity. (d) In vitro study for their antimplantation, aphro disiac and antioxidant property. (e) It will be better to repeat the same procedure for 3 to 4 times and then arrive to

conclusion.

NOTE : In a way to standardization we can check for the microbes in the crude / raw drug

and then proceed with antimicrobial study. Then we can compare the presence of amount of

microbes present in the drug with that to its antimicrobial activity.

Biblography ==================================================================

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92

Biblography ==================================================================

BIBLOGRAPHY

1. Triphathi Indradev; Raja Nighantu; Varanasi , Krishnadas Acadami

3rd edition 2003; Page No.304

2. Sharma P.V., Namarupa Vijnananam, Varanasi, Satyaprakashan,

1st edition, 2000, Page No.123

3. Gupta Shakti M , Plant Myths & Traditions In India ,

Chaukhamba Publications , edition 2001 , Page No. 11

4. Shrivastav Shailaja , Sharandhar Samhita , Varanasi ,

Chawkhamba Orientilia , edition 2003 , Page no. 172.

5. Chunekar Krishnachandr , Bhavaprakash Nighantu , Varanasi ,

Choukhambha Bharati Academy , edition 2004 , Page No. 335.

6. Shastri Shrilaxmipati , Yogaratnakar , Varanasi .

Choukhamba Sanskrant Santhan , 8th edition 2004 , Page No. 334,335.

7. Chunekar Krishnachandr , Bhavaprakash Nighantu , Varanasi ,

Choukhambha Bharati Academy , edition 2004 , Page No. 335.

8. Vaishya Lala Shaligramaji , Shaligram Nighantu , Bombay,

Khemaraj Shrikrishnadas Prakashan , edition 2002 , page 515.

9. Sharma P.V., Dhanavantari Nighantu , Varanasi ,

Chaukhamba Orientilia , 2nd edition 1998 , Page No. 177.

10. Sharma P.V., Kayadeva Nighantu , Varanasi ,

Chaukhamba Orientilia , 1st edition 1979 , Page No. 177.

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93

Biblography ==================================================================

11. Khemaraj Shrikrishnadas , Madanpal Nighantu, Bombay ,

Shri Venkateshwar., year 1966.

12. Triphathi Indradeva , Raja Nighantu , Varanasi ,

Krishnadas Academy, 3rd edition 2003 , Page No.304.

13. Sharma P.V. Sodhal Nighantu , Baroda , Oriental Institute

1st edition 1978 , Page No. 304.

14. Vaidya Bapalal , Nighantu, Baroda ,Varansi, Chaukhamba Bharati Academy,

third edition 2002, page355.

15. Sharma P.V. , Priya Nighantu , Varanasi ,

Chaukhamba Surabharati Prakashan , edition 2004 , Page No. 35.

16. Nadakarni A. K., Indian materia medica vol. 1 , Popular prakashan, page 222.

17. Sharma P. C.,Data base on Medicinal plants used in Ayurveda vol 1, Central council

for Research in Ayurveda and Siddha, page336.

18. Indian Herbal Pharmacopia, Indian drug Manufactures association , Mumbai ,

Edition 2002, Page 98.

19. Gupta A.K.; Indian Council of medical Research, Mew delhi, Quality standards of

Indian Medicinal plant, 2003.

20. Singh Ramshushil , Vanoushadi Nidarshika Ayurvediyapharmacopia Lahor, Uttar

Pradash Hindi Santhan 2nd edition 1983, Page –

21. Vaidyagrantham P.S. Arya Vaidya sala, Indian Medicinal Plants, Chenni, Orient

Longman private limited, Page – 314.

===============================================================================


94

Biblography ==================================================================

22. Dhiman Anil Kumar, Wild medicinal plant of India Deharadun, Bisher, sighnaendra

Palsigh.

23. Kirtikar K.R & Basu B.D., Indian medicinal Plant vol- I

2nd edition, Dehradun, International Book Distribution, 1999 Page- 785.

24. Ambasta S. P, The useful plants of India, New Delhi publications & information

directorate council of Scientific & Industrial Research, Page – 91.

25. Sharma P.V., Namarupa Vijnananam, Varanasi, Satyaprakashan,

1st edition, 2000, Page No.123

26. Pandey Gyanedra, Dravyaguna Vijnana(Vol III), Varanasi.

Krishna Academy, 1st edition 2001 Page-

27. Reviews on Indian Medicinal Plant. Vol –IV Page – 490.

28. Gupta A.K. , Quality Standards of Indian Medicinal Plants.

Sharma P.V., Kayadeva Nighantu , Varanasi ,Chaukhamba Orientilia ,

1st edition 1979 , Page No. 177.

29. Sharma P.C., Yelne M.B, Dennis T.J. , Data Base On Medicinal Plants Used In

Ayurveda Vol- I, New Delhi, Central Council for Research in Ayurveda and siddha,

Reprinted 2002, Page 338.

30. Shastri J.L.N. Ayurvedokta Aushadhi niruktamala, Varanasi Choukhamba oriental,

1st edition 2001, Page – 66.

31. Baghel M.S. Research in ayurveda past & present, article Ayurveda and

Microorganism, Page- No. 216, 226.

===============================================================================


95

Biblography ==================================================================

32. Marshall Jacquelyn, Microbilogy, The clinical laboratory, manual series, Tomson

Delmar Learning, 1st Reprint 2003, Sigapore.

33. Marshall Jacquelyn, Microbilogy, The clinical laboratory, manual series, Tomson

Delmar Learning, 1st Reprint 2003, Sigapore.

34. Gupta Satish, The Short Text book of Medical Microbiology, 8th edition, New

Delhi, Jaypee brothers-2002, Reprint 2004.

35. Gupta Satish, The Short Text book of Medical Microbiology, 8th edition, New Delhi,

Jaypee brothers-2002, Reprint 2004.

36. Makharjee Pulok K. Quality control of Herbal drugs New Delhi, And Business

Horizons- Pharmaceuticals publishers 1st edition 2002, Page – 381, 425

37. Choudhari R.D., Herbal drug Industry, 1st edtion 1996, New Delhi.

Eastern publishers India Tt58, 34,147,160,173,192,24,350,370,464,477,598.

38. Makharjee Pulok K. Quality control of Herbal drugs New Delhi, And Business

Horizons- Pharmaceuticals publishers 1st edition 2002, Page – 381, 425

39. Choudhari R.D., Herbal drug Industry, 1st edtion 1996, New Delhi.

Eastern publishers India Tt58, 34,147,160,173,192,24,350,370,464,477,598.

40. Controller of publication , Indian pharmacopeia, 1996.

41. Mukharjee Pulok K.Quality control of Herbal Press, new Delhi, Business Horizons –

Pharmaceutical Publishers.



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96

Biblography ==================================================================



44. R. Khandelwal K. Practical Pharmacogonosy, Techniques & Expriments, Pune ,

Nirali Prakashan, 13th edition, April 2005, Page- 10-18, 149-156.

45. Qulity Control Methods of medical Plant Materials, wt 0, Geneva, ATIBS publishers

& Distributors (Regd.) E-Mail- [email protected].

46. Wagener H. Bladt. S.E.M. Zganiski , PLANT DRUG Analysis , A Thin layer

Chromotography Atlas.

47. Kasture A.V. Mehadik R. et.al. Pharmaceutical Analysis, VOL 5 , Pune Nirali

Prakashan 8th edition May 2002, Page No. 18-13.

48. Gupta Satish, The Short Text Book of Medical Microbiology, 8th Edition, New Delhi,

jaypee Brothers – 2002, Reprint- 2004.

49. Harry w.et.al. , Aniseptic & disinfectant action , Microbes in Action. & lab Manual

of Microbiology 1982 Page No.75 – 76.

50. R. Khandelwal K., Practical Pharmacogonsy, Techniques and experiments, Pune,

Nirali Prakashan, 13th Edtion, April 1 -2005, Page No. 10-18, 149-156.

Other Books and References :

1) Pandey Gyanendra, Dravya Guna Vignana, Vol-I, 2nd Edition, Varanasi Krishnadas

Academy, 2002,

2) Jayengar M. A. Study of Crude Drugs, Manipal, 12th Edition – 2004.

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97

mailto:[email protected]

Biblography ==================================================================

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98

3) Rajpal V., Standardization of Botanicals, Vol – I, 2002, Reprint – 2004, New Delhi,

Estern Publishers.

4) Pammel L. H., A Manual of Poisonous Plants, Torch Press Cedar rapids, lowa, 1911,

Reprint 1991.

5) Shastry J. L. N. Ayurvedokta Aushadhi Nirukta Mala, Edition – 2001, Varanasi,

Chowkhamba Orientalia, India.

6) Singh Ram Shusheel Vanaushadhi Nidarshika Nirukta Mala, Edition – 2001,

Varanasi, Chowkhamba Orientalia, India.

7) Uniyal Mayaram, Medicinal Plants and Minerals of Uttarakhanda, Himalaya, 1st

Edition, May 1997, Bihar, Baidyanath Ayu. Shoodh Sanstan.

8) Mukhypadhyaya, Girindranath, History of India Medicines, Vol-I and II, Munshiram

Manoharlal Publishers, Reprint 2003.

9) Sharma P. V., Dalhana and his comments on drugs, Ist Edition, 1982, Munshiram

Manoharlal Publication Pvt. Ltd.

10) Raja Radha Kanta Deva, Shabdha Kalpa Druma Vol-II, Varanasi, Chowkhamba

Sanskrit Series, 3rd Edition, 1967,

11) Sharma P. V. Dravyaguna Vijnana Vol-II Varanasi, Chokhamba Bharati Academy,

Reprint – 2005.

12) Chopra R. N., Chopra I. C. et.al., Indigenous Drugs of India; (Tt) Culcutta, Academic

Publishers, 1st Edition, 1958, 2nd Reprint – 1994.

13) Singh Thakur Balwant, Chunenar K. C. et.al; Glossary of Vegetable drugs in

Brahatrayees, 2nd Edition, 1999, Varanasi, Chokambha Amarabharati Prakashan

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