1.Introduction================================================================== SCREENING OF ANTIMICROBIAL ACTIVITY OFBUTEA 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 ofAYURVEDA VACHASPATHI inDRAVYAGUNAunder 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. 0=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.)

2. Introduction==================================================================The Rajiv Gandhi University of Health Sciences, Karnataka, Bangalore. DECLARATION BY THE CANDIDATE I here by declare that this dissertation / thesis entitled SCREENING OFANTIMICROBIAL ACTIVITY OF BUTEA MONOSPERMA (LAM.). Is a bonafide andgenuine research work carried out by me under the guidance of Dr . S. K. HiremathM. D. Professor.Date : Signature of the CandidatePlace : Belgaum. Dr. Manjiri Kulkarni P. G. Scholar 1=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 3. Introduction==================================================================CERTIFICATE BY THE GUIDEThis is to certifythat this dissertation entitled SCREENING OFANTIMICROBIAL ACTIVITY OF BUTEA MONOSPERMA (LAM.) is abonafideresearch work done by Dr. Manjiri Kulkarni in partial fulfillment of the requirement for thedegree of AYURVEDA VACHASPATHI.Date :SignaturePlace : Belgaum.Dr. S. K. Hiremath, M. D.ProfessorP. G. DEPARTMENT OF DRAVYAGUNAK. L. E.s Shri. B. M. KankanawadiAyurved Mahavidyalaya,Post - Graduate Studies Cum Research Centre,Shahapur, BELGAUM. 2=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 4. Introduction==================================================================ENDORSEMENT BY THE HOD, PRINCIPAL OF THE INSTITUTION This is to certify that this dissertation entitled SCREENING OFANTIMICROBIAL ACTIVITY OF BUTEA MONOSPERMA (LAM.) is a bonafideresearch work done by Dr. Manjiri Kulkarni under the guidance of Dr. S. K. HiremathM. D. Professor.Dr. S. K. Hiremath, (M. D. Jamnagar) Dr. B. S. PrasadM. D. Ph. D.ProfessorPrincipal,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.3=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 5. Introduction==================================================================COPYRIGHT DECLARATION BY THE CANDIDATE I hereby declare that the Rajiv Gandhi University of Health Sciences, Karnatakashall have the right to preserve, use and disseminate this dissertation / thesis in print orelectronic format for academic / research purpose.Date :Signature of the CandidatePlace : Belgaum.Dr. Manjiri KulkarniP. G. Scholar 4=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 6. Introduction==================================================================CONTENTSTABLE OF CONTENTSSl. No. Contents Page No.1 Introduction 1-22 Objectives 33 Review of Literaturea) Drug review 4-27b) Microbiology review 28-41c) Extraction Procedure review 41-454 Methodology4.1) Collection464.2) Authenification 464.3) Collection of Test Drugs and storage464.4) Pharmacognostic Study 474.5) Physico Chemical Study48-534.6) Schematic Chart of extraction 544.7) Ethanol extraction55-574.8) Aqueous extraction57-584.9) Preliminary phytochemical screening 59-624.10) Qualitative confirmation (T.L.C., H.P.T.L.C.)63-724.11) Evaluation of Antimicrobial activity 72-785 Results79-866 Discussion 87-907 Conclusion 918 Summary929 Biblography references 93-9810Annexure I-V 5=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 7. Introduction================================================================== LIST OF TABLESl. No. Tables Page No.1 Showing synonyms according to different authors132 Rasa, Guna, Veerya, Vipak, Doshghanata 163 Prayojyange according to different authors 164 Karma of Palash175 Prayoga (uses) of Palash 186 Doshaghnata according to different authors 187 Prayojyanya according to different authors 268 Organoleptic characters469 Preliminary phyto-chemical screening81-8210Anti microgial results 8411E-coli 8512Staphylococcus Aureus8513Bacillus subtillis 8514Candida albicans 86 6=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 8. Introduction==================================================================LIST OF PHOTOS AND FIGURESSl. No. PlatesContentsPage No.1 Plates No. 1Palash plant with flowers, pods and seeds12 Plates No. 2Distillation apparatus 13 Plates No. 3Soxhlet apparatus14 Plates No. 4Muffle furnace 15 Plates No. 5Incubator16 Plates No. 6Ash of Palash seed 27 Plates No. 7Water soluble extractive 28 Plates No. 8Alcohol soluble extractive 29 Plates No. 9Cold water extract of Palash Beeja 210Plates No. 10 Culture medium 311Plates No. 11 Micro organisum312Plates No. 12 Micro Pippete313Plates No. 13 Micro scopic view of Palash seed 314Plates No. 14 Bacillus subtillus 415Plates No. 15 Candida albicans 416Plates No. 16 E-coli 417Plates No. 17 Staphylococcus 418Plates No. 18 Water sol. extractive519Plates No. 19 Alcohol soluble extractive 520Plates No. 20, 21 Water bath evaporation of Alc.and water sol. ext.521Plates No. 22 Evaporate extractive 522Plates No. 23 Water and alcohol extract of Palash seed 57=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 9. Introduction==================================================================ACKNOWLEDGEMENT A dissertation is an aim for every student. It is very tough and a field which is a veryailien for any student. Therefore during the compilation of this entire project. I have beenblessed with a lot of senior faculty members who have gone to extreme limits in helping mewhole heartedly to complete my research work project. I owe a lot of gratitude to them. Ihave strived to do my best of my knowledge. I hope I have achieved my success because ofeverybodys keen participation. It is my privilege for having worked under the guidance of Dr.ShivamurtiK.Hiremath M.D. (Jamanagar) (Professor) Head of Department, postgraduate department,DRAVYAGUNA for his valuable support and guidance. I express my deep sense ofgratitude for suggesting this study and for his constant encouragement with allmost patiencethroughout the course of my research work project. I express my sincere thanks to our Principal Dr B. S. Prasad M.D PhD, forprovidedmevaluable,timely given suggestionsand proper guidance andencouragement 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 mystudies. I am very much thankful to Dr. R.V. Savadi and Dr Kalpana Patil M Phram PhD, ofK.L.Es 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 outAntimicrobial studies. I also thank Dr Anand. S. Ammanagi M.D.DNB, MNAMS for their valuable guidance incarrying out antimicrobial studies. I thank Mr. Prakash Kokate NAFARI Pune. For carrying out TLC and HPTLCanalysis.8=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 10. Introduction================================================================== I express my sincere sense of gratitude to the management committee and PrincipalDr 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 juniorcolleagues Dr. Ramesh Konakeri, Dr. Deepak Mummigatti, Dr. Mahadev Gundakalle, Dr.Manisha, Dr. Poornima B, Dr. Poornima Undi, Dr Ajeet Herwade, Dr. Nayana.Patil whohelped whenever needed at the studies. I express my sincere heartfelt thanks to all the teaching staff members of otherdepartments for their valuable suggestion and support during my postgraduate studies. I Thank our college librarian Mrs. G.C. Gulla who provided me necessary books andjournals for my studies. I am very much thankful to my family friends Shailaja Katti and Mr. AnandDeshpande for their moral support and kind co-operation. I indebeted to my beloved parents and family members, family friends Shailaja Kattiand 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 CandidatePlace : Belgaum (Dr. Manjiri Kulkarni)9=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 11. Introduction==================================================================LIST OF ABBREVIATIONS1.A. H. - Ashtanga Hrudaya2.A. H. Ni-Ashtanga Hrudaya Nidana Sthana3.A. H. Su- Ashtanga Hrudaya Stura sthan4.A. K. -Amarakosha5.A. R. -Abhidana Ratnamala6.A. S. - Ashtanga Sangraha7.B. A. - Bruhat Dravyaguna Adarsha8.B. P. N / B. P. N - Bhavaprakash Nighantu9.B. R. - Bhaishajya Ratnavali10. Cha -Charaka11. Cha. Chi-Charaka Chikitsa12. Cha. Su - Charaka Chikitsa Sutra Sthana13. Cha. S. N.- Charaka Chikitsa Samhita Nidhana Sthana14. Cha. S. Vi-Charaka Chikitsa Vimana Sthana15. C. da/c. d. - Chakradata16. D. N. - Dhanvantri Nighantu17. D. G. H.-Dravya Guna Hastamalaka18. D. G. (V. M. G.)- Dravya Guna Hastamalaka Vijanana by V. M. Gogte19. H. S. - Haritha Samhita20. K. N. - Kaiyadeva Nighantu21. L. S. - Longitudinal Section22. M. N. - Madhanapala Nighantu23. Mau. N. - Mahaushadha Nighantu24. MIC.-Minimal inhibitory concentration25. N. A. - Nighantu Adarsha26. Sha - s - Sharangadhara Samhita27. S/k - Shloka28. Su- Sushruta29. Su. S. N. - Sushruta Samhita Nidan Sthana30. T. S. - Transverse Section31. T. L. C.-Thin layer chromatography32. H. P. T. L. C.-High performance Thin layer chromatography33. Y. R. - Yoga Ratnakara10=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 12. Introduction==================================================================ABSTRACTA. Background and Objectives : In modern medical acrence various diseases are treated with antimicrobials, whichplays 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 evaluateantimicrobial effect ofPalash 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 threeconcentrations.11=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 13. Introduction==================================================================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 extractshows also registrant for Candida albicans, but sensitive for remaining microbes.12=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 14. Introduction==================================================================13=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 15. Introduction================================================================== 1. INTRODUCTIONPotent substances are present in version plants and herbs. Modern medicines are notfree 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 treatvarious diseases. It is widely and abundantly available. Different parts of Palash are used in various diseases. According to the classicalreference having a prime property of "Krimighna"(1). It can be used to treat various skindiseases. An antimicrobial agent is one that inhibits or kills microbes but causes minimumharm to normal tissues of human being. As many micro organisms have become resistant tothe newest antimicrobial agent.So now we have taken furthers step towards inventing new antimicrobial agents fromthe field of Ayurveda without isolating active constituents. Thus we have conducted preliminary study of Palash seed by phytochemicalanalysis 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 littlehigher, 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 thepatient. We will be are giving these drugs without harming the patient and would be moreeffective without compromising with our basic principles.1=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 16. Introduction==================================================================A)Review of Liturature :- Drug review, Microbiology review, extraction procedurereview,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. 2=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 17. Objective==================================================================2=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 18. Objective==================================================================2. OBJECTIVES(2.1) Pharmacognostic and physicochemical study of Palash seed.To study regarding phyto constituents of the drugs so that it maybe helpful tointerpret 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 pharmacokinetic of the drug.(2.3) Anti microbial activity of Palash seed.To know the antimicrobial effect, so that we can interpret on the Krimighna effectwhich is stated in Ayurveda texts and then a step wise and complete antimicrobialstudy can be done for better therapeutic effects.3=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 19. Review of Literature==================================================================3=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 20. Review of Literature==================================================================(A) Drug Review(1) Vedic Review (2), (3) :We have got the references of use of Palash since vedic period. In this kala singledrug thearpy was present and since then single drug therapy is being usedThe use of Palash was common in vedic period not only to treat the ailments but alsoin routine life and in holy rituals. In vedic era leaves, stem and flowers were moreused but there are references regarding use of the seed. In vedic kala Palash tree wasknown as Shant Vruskha and Bramha Varchass. Samidha of this plant were used atthe time of different Homa and Yadgnyas.According to Koushika sutra, Palash isMedhajanan and lepa of Palash theseleaves was applied in Jalodar (Ascitis). Keshav told it was Sarvaroga Bheshaja andalso it was used in Krimi Roga ( Ke.P. 9. 4/25/20 ).In RigvedaKinshuk was the synonym given for Palash. Kinshuk means whoshines brightly. This synonym is given because of its bright attractive colour of theflower. In Rigveda kala, Palash leaves were used with Ashwath and we get thereferenes of uses of Palash leaves with Nyagrodha in Atharvaveda. In Upanayansamskar Dand (Stick) which is used by Brahmachari was also made from Palashwood. Palash was used frequently because it had a power to destroy the Rakshasasso also its stem was being used in Yadgnyas and patras were used to prepareAbhishek paatra. In Atharaveda Parnamani of Palash patra was used to gain Bala,Aayu, Samruddhi and fame. According to Shrout sutra patra valkala of Palash wasused in preparation of curds.The tree is considered sacred both by Hindus and Buddhist. Hindus consider it, asholy because of the trifoliate formation of leaves which represents the Holy trinity ofBrahma (the creator) on the left, Vishnu (the preserver) in the middle and Shiva (thedestroyer) on the right. The flower of this plant are offered especially to Goddess 4=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 21. Review of Literature==================================================================Kali. In Krishnasthami Vratam, wood of Palash is also being used. Because of curseof Goddess Parvati, Bramha was converted into the tree of Palash. In NavagrahaStotra 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 saidto be used in treating many diseases and in different ritual functions. We couldnot getany synonym for Palash in this samhita. Leaf, flower, skin, seed and kshar are theparts used in different yogas.Palash is used in kasa, Grahani, Arsh, Udararoga, kushtha and in skin diseases. Butseed 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 forPalash 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 forkids.2)Sangraha Kala :a)Ashtang Sangraha and Ashtang Hridaya : Vagbhata also includes Palash inRodharadi, Mustadi etc., (A.H.S 15/32, 15/38) Rogaghnata of Palash is given in 5=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 22. Review of Literature================================================================== 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)(4)c) Sharangdhar Samhita: 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.(5)d) Bhavaprakash : 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.(8)2) Shaligram Nighantu : 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.6=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 23. Review of Literature================================================================== (9)3)Dhanwantari Nighantu : In this Nighantu it is included in Amradi varg and 14synonyms are given for it. Krimihar property of seeds is mentioned.(10)4)Kaiyadeva Nighantu : In this Nighantu it is included in Aushadhi varga andtwelve synonyms are given for it.(11)5)Madanpal Nighantu: In this Nighantu it is included into the Vatadi Varga.Different Synonyms are given in this book. Types of flowers and its uses in differentRogas are explained.(12)6)Raj Nighantu : In this Nighantu it is included given in Karviryadi Varga 11synonyms are given for it. Nighantukar also mention that it is Krimihar and its seedsare used to treat various skin disorders. He has mentioned 4 types of Palash 1) Peeta2) Shweta 3) Rakta and 4) Neel pushpa.Though all 4 are having same quality, inthose 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 referencesof different authors regarding Palash are given in this Nighantu.9)Priya Nighantu (15): He mentioned it in Haritakyadi Varga. Its seeds are Krimiharin action and seven synonyms are given.ADHUNIK KALA : (16)1)Indian Mateia Medica : The author of this book has mentioned vernacularnames and chemical constituents of this drug.. Seeds are indicated in many skindiseases. Internally for worms, externally for ringworm, boils, pimples, bubbles,tumours and also in various other diseases. 7=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 24. Review of Literature==================================================================(17)2)Data base on medicinal plants used in Ayurveda Vol 1 : Detail explanationregarding plant Palash has been given such as - Family, Classical names (Synonyms)vernacular names, morphology, useful parts, uses and doses.Along with thatpharmacognosy, chemical constituents, pharmacological activities, toxicology andtherapeutic evaluation is explained.(18)3)Indian Herbal Pharmacopoeia: Detail Information regarding Palash seed isgiven i.e. macroscopic, microscopic examinations etc.,(19)4)Medicinal Plants Quality Standards of Indian : Description regarding plant isgiven and also all types of phytochemical tests are explained in detail.(20)5)Vanoushadi Nidarshika: Containts description of seed and Im edd ddddis given. (21)6)Indian Medicinal Plants : Detail description like names, vernacular namesdistribution etc., are given. Also properties and uses of seeds are given.7)Wild medicinal plants of India (22) : pharmacognasy and phytochemical descriptionand use of Palash seed in folk medicine8)Indian medicinal plant by Kirtikar and Basu. (23) : Properties of seed in detail havedescribed that seeds are not dry digestible, antihelmentic, aperient used in urinarydischarge, piles, cure, skin diseases, tumours, abdominal trouble and also used inscorpion stinge. (24)9)The Useful Plants of India: Herpes in dhobies itch seed are pounded withlemon 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 8=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 25. Review of Literature==================================================================Narahari Pandit the Author of Raja Nighantu has tried to arrange the synonyms in aproper way and elaborated them into 7 categories.The various synonyms of Palash as given in Nighantus as follows :DIFFERENT SYNONYMS :1) nbm (^m e m nbm ` & e: .)V{Z em `Leaves are more succulent and Fleshy.2) q$ H (^m "q$ Ho `_ ?> {V ^ O H ew w> nw dm & H $ .) H $ @B mV Z $ H g e n Vew :ew m p: $S V The flowers resembles beak of parrot3) H Z (gm H h , nbm rO ` H m `mV &$ : o {_.) $_rZ pV e~ oo om $ J J {_aThe seeds are used in krimirogahar4) j m o: (^m j m j o l : & al > .) a f w o d>Kshara prepared from this plant is superior.5) I a ({Z nf ` &nU .):nU _Leaves are rough.6) { n : (gm r{U n H` nU & .) o $ ` } mLeaflets are present 3 in No.7) nU (^m e m nU ` & : .) V{Z m`Plants having more leaves.8) nw (Y ny: n{d m w &V : .) V o _:This plant is used in Rituals.9) ~ o: (gm ~ Z _w &rO Zh .)o rO o h V` 9=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 26. Review of Literature==================================================================Seeds are oily and snehayukta.10) ~ d: (^m ~ Uo j : d{X $ $ o ` m dm & j .) m d H Ha w o` V g mf Wood of the plant utilized in yajnya kunda as fuel.11) `m $ (^m `ko Z & {kH: .)`w : `_mIt is used in yagnyas.12) a V nH (^m a V{Z nw ` & nw $ .) m nm : `Flowers in red in colour.13) dH nH (gm dH nw_ &$ : .) $ o n `nw $Flowers are curved.14) dm : (^m dm ` ha em $ dm m B A> {Z >"dm o: B gm o R Vha .) V : _H V o: {V m K Q V W {V ob nm:& aY J m nmT> > :It is used in vata vikara.15) dm W (n.) dZ m m o & Z :o W dgm `o@We get these plants more in forest.16) g{_: (^m `ko `w m m o > &a .) f w Z g{_Y R V ml :Wood is used in yadgnyakunda.17) dm o: dV nm`{V B & VW m onm W {V18) q$ H :- Its flowers resembele to nose ofH $ ew parrot in colour and shape.19) j m o :-a l > Its alkali is best among alkaline materials.20) nbm :-eIts leaves are fleshy and beautiful or it develops the muscle.10=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 27. Review of Literature==================================================================21)a V wH:- nn $ Its flowers are red in colour.22)da g{_ o Z B ( `V @ `m {V d.) It mesmerize every one with its qualities and it improves appetite. (25)DESCRIPTION OF PLANT ACCORDING TO ITS SYNONYMS: 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 (raktapuspaka) and curved (Vakrapuspaka) typical of papilionate resembling parrots beak(Kimshuka). Seeds are oily (bijasneha) and make a potent antihelmintic drug (Krimighna). Italso 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 :11=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 28. Review of Literature==================================================================a){e. {Z :- nw {e r, {V V rO $ H d ..db V n, ~~ H $ , V Ub)A{^. _:- d $$ _, a n n g{_, n w r, d .Hw , gy J wH a e H. WJ $ sc)H {Z :- q$ H H `m $ JU H dQU, o,$ Q. K w H $ $ , kH o $ w u rO Z>ew _m,, , >nh{ d, a V n, j m o, dm o, ~ d, g{_X am nw a V W ml > nm .dd)_X nm {Z :- nbm H H $ ,` m $ nm m ,a V n dm Z b K w e,q$ $ {_ {kH X a o nw,{ ,g{_V Q> ew,{H ,~n,j l > .e)Am {Z w nbm q$ H ~ nm , j m o, dm o, ~ d, g{_X a X K Q:-e > e, H $ X ew n a V W m ,l > nm d .TABLE NO. 112=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 29. Review of Literature==================================================================SHOWING SYNONYMS ACC. TO DIFFERENT AUTHORSSynonymSha ChaSu AS DN RN KN BP.N AS.H Sho.N + + ++ +++ ++ + + + ++ ++++++ + +++ +++ + + + + + +++ + + + + + +++ +++ +++ + + +++++ + + + + + +++ + + + + +++ + + +VERNACULAR NAMES:13=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 30. Review of Literature==================================================================Sanskrit- PalashHindi - Dhaka, Tesu, Palas, Chichra, Dhara, Faras Kankeri,Chini agondBeng- Palash, Ganch, KamarkasMar - Palasa.Tree Guj- Khasathi Khakra.Flower- Guj - Kemuda.Seeds Guj - Palash Paprha.Tam - Paras, Patsan, Camala, Paladula Modug Mooduga.Tel - Modhung, Midug chettu.Kann- Muttuga, Muttala, Muttagamara.Mal - Palashin Samatha, Camata, Pilacham, MuraklamarCV - nm aewd - I mQH> $rJm- n g y bm $ bA- Downy Branch buter.Flame of the forest, parrot tree, Judas tree (Myths.and traditions){b`adm- Beespaknm~Or- Tesh.Am [a ogm- Kinjuko Porasu.Cw X - Palash Papra.Bheda (Varieties) :According to Madanpal - Rakta, Peeta, Shweta and Neel( according to colour of the flowers)In Abhidhanmanjiri- 1) Palash and 2) Valli PalashShaligram - 1) Kinshulak 2) Hastikarnak.According to Raj Nighantu - Rakta, Peeta, Shweta and Neel.Guna and Doshagnata :14=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 31. Review of Literature==================================================================PN - Kashay, Shaligram -Katu, Tikta, Kashay,Katu VipakUshnavirya.Vatakaphahar.Taxanomical Classification :Kingdom- PlantaeDivision - Magnoliophyta,Class- MagnoliopsidaOrder- FabaleaFamily - FabaceaeGenus- ButeaSpecies- MonospermaLatin Name - Butea monospermaMeaning of Butea Monosperma (LAM) :Butea = Named after John Ear of Butea, patron of BotanyMono = One, sperma= seed.Before Butea monosperma, Butea frondosa was the name, frondosa means leafy.Palash is famous for its leaves.Flesh, like flesh or bloodOr - Demon.TABLE NO. 2Rasa, Guna, Verrya, Vipak, Doshaghnata 15=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 32. Review of Literature================================================================== Rasa B.P.N MN RNDN NAKNTikta + + +Katu+++ +Kashay+ + + +GunaLaghu + +Rukha + +Snigdha++VeeryaUshna + + +++ +VipakKatu+ +++ +Doshagnataa) Kaphaghna+ + +++ +b) Vataghna + + + + +c) Pittakar + + +TABLE NO. 3Prayojyanga according to different authors :Prayojyanga PN PV.S RN DG(VMG) DGH MN NA DN Sha.NBark+ + + + +Flowers + + + ++ ++ + +Leaves+ + + + + + +Seeds + + + ++ ++ + +Gum + + + ++ ++ + +TABLE NO. 4 : Karma of Palash16=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 33. Review of Literature==================================================================Karma Bpn DNRW MN KN PNDeepan + +++Vrushya+Krimihar + + ++++Sangrahi ++Sarak+ ++Vranahar + +++Gulmahar +++Grahani ++Arshya +Kusthahar+Rasayan +Veeryavardhak +USE OF "PALASH" SEED ACCORDING TO DIFFERENT AUTHORS :n eV H `m : H Xo {d m : & bm w$ o m ZeZ fmU $ f _rV rO n_m> X wXo ZeH & (em m {Z w X mH SV Xd m m & {bJ K Q~ $y f{ $V _ >)H ` a J n n gw` Z M~ nQ & ([`{Z w$ n m w a H rO _V n ^_ & fm {h ${_ > m KQ> )~ w$w pZY U H bm V &(Yd {Z > rO V H > g{O K QQ$ H_w $ {_~ . w )$ bKU _oe m $ m _&b w h : H V $ hm $ H n {_d{d m H w j w Jw o U & (^mnH $> H > X V & ..)$ Ho Q$$ _m waV rO H H>a `Z {h : &(gm .) X dg $ So gm o V~ $ {_{d { m mo.{Z$ rO MpZYo $> $ m O V & ({Z .) b~ U H w $ $ `o & .am QH {_H ZV rO H {h : H>a `Z &em . X dg V $ So gm o ob~ $ {_{d {mm T>TABLE NO. 517=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 34. Review of Literature==================================================================XII)Prayoga (Uses) :PrayogaBPN DN RN KNMNSNPrameha+ + ++Arsha+ ++Krimi+ + + + ++Kushtha+ + ++Gulma+ + ++Udar Roga+ +Twakvikar+ ++Kandu++Pleeha +Shool+Vatarakta+ ++Raktapitta+TABLE NO. 6XIII) Doshagnatha according to different authers : Doshaghnata ShaNKNPNBPN DNRNKaphavata Shamak ++ + + +Pittavardhak+ +(VII) INTRODUCTION OF PLANT (GENUS AND SPECIES) (27) : This plant was identified as Butea monosperma (Lam) belonging to family Fabaceae.It was found that this genus Butea consist of around 35 species. Out of which literatureregarding the phytochemical and pharmacological profiles was available on only fewspecies. Most of the reports available were on the Butea monosperma and very few reportsare available on Butea superba and Butea parviflora. But as per literature seeds are welldocumented for various activities,18=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 35. Review of Literature==================================================================BUTEA SPECIES : There are 35 species available in the Butea genus. Generally they are trees orscandent shrubs. The general characters of Butea species are as below :Leaves-Pinnately, 3-foliolate, stipellate.Flowers -Orange, purple, rose or white, densely fasciculate; Fascicles recemose or fasciculate paniculate, or ample paniculate.Calyx -Campanulate, the upper two teeth or lobes connate.Petals-Subequal or unequal; keen incurved and acute, or 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; stigma capitate or truncate.Pod -Pod is one seeded.Botanical description of the important species of Butea are as follows :1) Butea Superba : Butea superba is a gigantic climbing shrub and its characters are mentioned in the following paragraphs.Trunk -Trunk is climbing from left to right, attaining 0.6 0.9 m girth.Leaflets-The leaflets attain 30-45 cm and sometimes 50 cm in young plants.Flowers -The flowers are larger than other species, 4.5-6.3 cm long, orange or orange scarlet, borne in great profusion along the leaflets branches on racemes which are 30 cm long.19=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 36. Review of Literature==================================================================Pod- Pod is about 12.5 15 cm long.Distribution of Butea superba : Butea superba is distributed in the forest of Oudh and Bundelkhand, Chota Nagpur,Burma, Konkan, N. Kanara and Central. It is also found in South India.Distribution : Throughout India, in deciduous forests in areas up to 1,200 m elevation, also in openareas.The Plant : A medium-sized deciduous tree, with a somewhat crooked trunk,10-15 in height and5-6 in girth. The bark is bluish-grey or light-brown and yields a gum. Its bright orange-redflowers (1.5-2 long) bloom in great profusion at the beginning of the hot season before theappearance of new leaves. The pod contains a single seed (1x3/4) at its apex. Butea monosperma is common throughout India, Burma and Ceylon (up to 4,000),except in very arid parts. Generally, it grows gregariously on open grasslands and scatteredin mixed forests along with Shal (Shorea robusta). It is frost-hardy and drought-resistant andis a valuable species for reclaiming saline soils.Parts used : Bark, leaves, flowers, seeds, gums. Butea monosperma bark is white or yellowish-brown, but very liable to sap -stainand often turns greysh-brown or grey. It is a light wood and can be seasoned without muchdifficulty, but it contracts considerable during seasoning. It is not strong or durable inexposed situations, but is said to last much longer under water. It is easy to work either by 20=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 37. Review of Literature==================================================================hand or machine, and is used mainly for well-curbs, water-scoops and for fuel. It can also beemployed as a cheap board-wood . The leaves are much used throughout the country for making plates, cups, etc. Theyare eaten by buffaloes and elephants. The flowers yield a brilliant but very fugitive yellow colouring matter. This iscontained in the sap and may be obtained in the form of a decoction or infusion from driedflowers. The addition of alum, lime or an alkali deepens the colour to orange and also makesit less fugitive. The sap contains the chalcone, butein C15H12O5 (0.3%), orange yellowneedles, m.p., 2130c 2150c, and small quantities of butin, the colourless isomeric flavanone,and its glucoside, butrin.USES : Bark furnishes a very important exudation which hardens into a red brittle resinknown as butea-gum or Bengal kino or magugo, largely used as a substitute for the Kino inIndia and to a limited extent in Europe also. A red juice exudes from natural cracks and artificial incisions in the bark, and ithardens into a vitreous, ruby-red gum, known as Butea gum or Bengal kino. It isdistinguished from Malabar kino from Pterocarpus marsupium by its greater solubility inwater, and by the presence of corky particles. Butea gum contains a large proportion oftannin and mucilaginous material. On dry distillation, it is reported to yield pyrocatechin. Itis a powerful astringent and is given in many forms of chronic diarrhoea. (Dymock, Wardenand Hooper, I, 454) The seeds have long been valued as anthelmintic. Birdwood prescribes them forround worms and tapeworms. According to Chopra freshly powdered new seeds give fairlygood results against Ascaris, but old worm-eaten ones, like those frequently found in themarket, 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=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 38. Review of Literature==================================================================applied, the seeds act as a powerful rubifacient and they have been successfully used incuring a form of herpes, known as dhobies itch. (Dymock et al., loc. Cit.) The seed is alsouseful 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.,178; iod. Val., 67.2, Katti and Manjunath, J. Indian Chem. Soc., 1929, 6, 639). Fresh seedsare reported to contain proteolytic and lypolytic enzymes. The former is a mixture of plantproteinase and polypeptidase, and behaves like yeast trypsin (Chatterjee, Ghosh andChopra). The leaves are astringent, anti-inflammatory, anodyne and aphrodisiac, and are usefulin pimples, boils, flatulence, colic, worm infestations, inflammations, arthralgia andhaemorrhoids. The flowers are astringent, sweet, cooling, constipating, aphrodisiac, haemostatic,diuretic, febrifuge, depurative and tonic. They are useful in vitiated conditions of pitta andkapha, diarrhoea, haemorrhoids, menorrhagia, strangury, fever, leprosy, skin diseases,swellings, hyperdipsia, haemoptysis, arthritis, burning sensation, bone fractures, and are veryefficacious in birth control. Gum combined with other astringents and rock-salt is recommended byCHAKRADATTA, as an external application for Pterygium and opacities of the cornea. The seeds are purgative, ophthalmic, anthelmintic, rubefacient, depurative and tonic.They are useful in herpes, skin diseases, ringworm, ophthalmopathy, epilepsy, round worm,arthritis, flatulence, constipation and diabetes. As anthelmintic and aperient, BHAVAPRAKASH recommends new seeds to begiven in powder, 10 to 20 grains or as paste with honey added (because the seeds are veryunpleasant to take and often produce retching pain in the abdomen and occasionallyvomiting and giddiness) thrice daily for three successive days (especially for ascaris roundworms) and followed on the fourth day by a dose of castor oil. For this, the seeds are soaked22=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 39. Review of Literature==================================================================in water, shells removed and kernels powdered after being dried. Some medical menconsider that the seeds can be advantageously substituted for santonin against roundworms. Externally the powder is a remedy for ringworm; it may be applied better in theform 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 anthelminticactivity. Active principle of the nature of alkaloid, neutral principle or glucoside could not beisolated from the seeds.USE OF PALASH IN DIFFERENT YOGAS :- According to Charak Samhita. YogasIndicationReferences Palash beeja.Udar rog. Ch.chi (110/13.) Palash beeja.Arsha Ch.chi (92/14.) Choorna. Kasa. Ch.chi (77/18.) P. Beeja + Ghruta. Atisaara, Ch.chi (27/19.) P.Beeja. Visha Ch.chi (51/23.) Palash KsharaDwivrunaCh.chi (54/25.)ADHYAYA : According to Sushrut Samhita. YOGASIndicationReferences Palash ValkalVruna bandhan.Su.chi. (3/6) Palash Choorna.Vaata VyadhiSu.chi (4/32) PalashKshara.Ashmari Su.chi (7/22) Beeja lepa.Kushta. Su.chi (9/10) Beeja choorna. Prameha.Su.chi (11/8) Beeja choorna. Udar rog. Su.chi (14/13) Beeja choorna. Maha vyadhi.Su.chi (31/5)ACCORDING SHARANGDHAR SAMHITA :23=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 40. Review of Literature================================================================== Palash beeja choornaKrumihar,5/26. Palash beeja choornaYoni sankochanartha lepa. 11/110 (Uttarakhanda) Palash pushapa swaras.Netrapasadan 13/78. (Uttarakhanda)ACCORDING TO ASHTANG SANGRAHA. (A.S) : P. Vrunta Swaras. Kasa.A.S.Chi (3/65) P. Churna.Raj Yakshma. A.S.Chi (5/9) P. Churna ArshaA.S.Chi (8/63) P. ChoornaVaatshonita. A.S.Chi (22/45)ACCORDING TO ASHTANG HRUDAYA. (A.H) P. VruntaGhruta.Raktapitta.AH.Chi (2/44) P. Ghruta ArshaAH.Chi (3/101) P. Beeja. KrumiAH.Chi (20/26) P.Kshara. RasayanAH Kalpa Sthana(39/37)COLLECTION AND STORAGE OF DRUG : In the present study the matured seeds of Butea monosperma were collected in themonth of April and May near Belgaum forest area. The seeds were authenticated byTaxonomist. Soon after authentification all the seeds were dried at room temp until theywere free from the moisture and subjected to physical evaluation with different parameterslike nature, colour, taste, odour, size, shape width, length etc. Nearly 1.5kg seeds werecollected and kept in air tight jars to avoid fungal infections. Veerya period is 1 year.ADULTRATION AND SUBSTITUTION (28) : 24=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 41. Review of Literature================================================================== Butea monasperma seeds are adulterated with the seeds of B. Superba (Roxb). Thesecan be identified on the basis of the size of seeds i.e the seed of B. monosperma arecomparatively larger in size. But in this case the seeds were collected by own. So there is nochance of adulteration.(XI) CHEMICAL CONSTITUENTS REVIEW (29) : A Nitrogenous acidic compound (I) along with Palasonin isolated from seeds. Amyrin, Sitosterol, its glycoside and sucrase isolated from seeds Glycerides of Palmitic stearic, Lignoceric, Oleic, and, linoleic acids from seed oil. Seed contain 18% of fixed oil called moodooga oil small quantity of resin and largequantity of water soluble albuminoids. The compound (I) showed Antimicrobial activity against fungi which arepathogenius and gram +ve bacteria strepto coccus pyogences, staphylo coccus aureus,Bacillus substilis gram ve bacteria ,E. Coli, Pseudomonas, maximum inhibitary effect wasshown by gram +ve bacteria,ACTION AND USES IN SIDDHA : Uses and Action are mentioned in Siddha. Action and uses in Unani medicine. The fruits and seeds are bitter and oily and mare administered against piles, Eyediseases and Enlargement of spleen (Spleenomegaly).TABLE NO. 725=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 42. Review of Literature==================================================================Prayojyanga according to different authors:Parts usedNames of the authors Or S.H.N D.NK.N N.AP.N BP.N R.NPrayojyangaPatra (Leaf) + ++ ++ ++Stem +++ ++ Flower+ ++ +++ +Seed + ++ ++ ++ Kshara+ ++ +++ +NOTE : Studies carried out on Palash seed.Pharmacological activities and clinical trails as:- - Antihelminthic activity. - Antifungal and Antimicrobial activity. - Antidiabetic activity. - Antioestrogenic activity. - Ophthalmic disorder. - Anti implantation activity in female rats. - Anti hepato toxic activity. - Uterotropic and uterine perioxidised activities in female rats. - Antioxidant activity.Internet information (1)Kumari,N, Ehaum ,V, Mikosch, M, 2005 Seasonal photosynthetic performance and 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.Yadava R. N. Tiwali L.26=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 43. Review of Literature==================================================================(2) A potential antiwiral flavone glycoside from the seeds of Butea monosperma O.Kuhne J-Acian-Nal-Pro-Res-2005 Apr. : 7(2) : 185-8.Sexcena, Ajit Kumar, Gupta(3) A pharmacentical composition (extract of Butea monosperma flower) useful for thetreatment of the hepatocellular caricinoma world intellectual property organization2006?A. Khan, T. H. Prasad.(4) Butea monosperma and chemomodulation protective role against thioacetamidemedicated hepatic alternation in wistar rates. Physiotherapy & Phytopharmacology02/01/06.USES AND ACTIONS ARE NOT GIVEN IN SIDDHA :27=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 44. Review of Literature================================================================== (30)Actions and Uses in Unani : The fruits and seeds are bitter and oily and are administered against piles, eyediseases and enlargement of spleen. (Spleenomegaly).B) MICROBIOLOGY REVIEW :I. Microbes (Krimi) In Vedas (Ayurveda) (31) : From the vedic period itself acharyas have been aware of minute organisms of thenature / environment. They have also described krimis, their habitats types, synonyms andtreatment. The acharyas had knowledge about jantus etc. that had harmful and harmless tobeings. under the heading of krimis they have described about both > (drustha) A>H${_ (Adrushta krimis).HABITAT : `o n wZ w Y wewA & {H$_`: d o d o A r~ n f w gd mV f f f : `o _m Vd {d ew X {_ {H m & (A. 2/31/5) A H _m{d gd V $Y _ & $ _rU In atharvveda it is told that krimis are present in all factors of the nature. They arepresent on parvath, vana aushadhi, pashu, etc.Minuteness of Krimis :(1)gy_ o^d ` m&(C.S.VI 7/11) dm H : &M$ `(2)gm`m H X e: &(vaghbhata) V $ X Z &o {M m 28=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 45. Review of Literature==================================================================Here Acharyas says that Krimis are not visible to our naked eyes.(1)gm`m H X e: & V $ X Z &o {M mHere Drushta and Adrushta types are specified.Interpretation by Acharyas :1) H o {V B {H & $ `m {V & _o`$_r:One which survives on raw meat is krimi.2) H Vm$ omdOne which has desires.3) H om `m ga H : & $ d V U $_V _One which crawls and moves.CAUSES OF INFECTION BY KRIMS :1)C H ~ d mmnW _M$ f w & X o h : U: `m $ { bo Mgy_ `m ^ym V `{Z ^m & or{Z V{Z H JZ$a & Vn _Uo {n {Z m `o W $Y n ` : & m Am nV f m m H ` o Z V `o Z {d pV no {n Vo ZZ &Ao {d `m m w ~ m m f f O (` .d. 16/ 62) o2)gV Jm em {Z mV gh oZV & Jm g :dgm ^m m n VO gh `m m d _m Z o m &egZ {n w ZV &`m n bH > d MemMZ m X E M& $ w a o o{^` df Am H ao g_pV Z m & g{J m H aZ &n $ Jm $ _(gw . 5/ 33- 34).g.{Z3) e}Hha eX go m m o m& n $ m `m d V ``m : m r Z JXgV g[a Uo o H {d $ m eo : &} M Z m d Ha {d f VZ$ m (A.h.{Z 14/ 441). 29=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 46. Review of Literature================================================================== By dushita, vayu, jala annasevana, bhumi and by maithuna infected rogi gatrasparsha of infected pt. infection by nishwasa etc. use of contiminated clothes, bed etc. ofinfected patient / sahabhojana etc. krimis are said to be entering the body.ORIGIN OF KRIMIS :gbom {H XV ^dpV C m : &(M . 17/ 38) ${_`m` nhV_Z.g.gw Charaka says kleda in the body is one of main factors for the production of Krimis.ROUTE OF ENTRY : Am gw V e~ o n V `m {n M AeZ X &_o n o i {d o o emo o X` _m m V m m ` {n M {d `m V_JXo V & (E5/ 29)X O em _Z m `m m w V X`_ & Krimis enter the body through annapanadi and vruna mukhas.`m n gn m o m n gd & A`m [a o Zgo [a mV `m{Vd m o ` NV V {H O {g & (A. 5/ 23) `m J > ^`^m & V _{ $_rThere they also enter through eyes, nostrils mouth to produce diseases. In sharangdhar samhita, he mentioned main two types of krimis are given (1)external (2) Internal * Internal krimis are further divided into 20 types eg. Kaphaja, Raktaja,Pureeshaga etc.a V {h {` ao Z a V m dm d: &(A.h . 14/ 42) dm m m O OV o `m AU .gw Krimis presenting in blood are the minutest.VYAKT KRIMI LAKSHANAS :30=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 47. Review of Literature================================================================== em V m w bww>g_m g_w m m a V m o {U m o OZ V I H Z Z W d{h `m $: R W Z Y `: g m AU m V nnX gw m o_ Z do m ` d HWdm_m M$ ^d A : dU m h H S VopV `m JVZ Mf$ w mm > X g n m A{V YZ H {ga m _mV mW`m {_V & {Z dX m M d w m `w UpVU m$ Z g gU r &(M . 7/ 11).g.d gmm C o Ud w eo V b{gH Z n `_mm H _m {Ud {n $ g $m Ho H Xg o J: {H o {^_w>V$ $ X m bo d O X W $ _`m Np V ^ Jm &{H V JX V : &o `pV d XrZ $ w rMw_` da {ea: m M MV m X V & (M . 5/ 10)m Z`w mW U X o& .g.{Z p `m Raktaja krimis that are minute and because of the size mynuteners they aresometimes visible or invisible when vranagata they cause kandu, toda etc. when excessivelygrown they eat away twak, sira, snayu, mamsa tarunasthi. Thus krimis minutest in the twak, mamsa lasika, sharira in kleda and sweda. In vedas explanation regarding sun as a krimighna is stated (Ri/191/18), (A 2/32/1)(R1/28/1). Acharya Sushruta also clearly mentioned that 13 krimis are described as pureeshajaand shleshmaja are drushta krimis, while remaining 7 described as raktaja are invisible i.e.they are adrushta krimis, But acharya Charaka terms some krimis to be sukshma.INFERENCE : This brief explanation suggests that there was tremendous knowledge regardingmicrobes right from the vedic kala. Only the way of approach has been changed.31=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 48. Review of Literature==================================================================II)INTRODUCTION :Antimicrobials (32) :- The agents which have a capacity to kill the microbes but which causes minimumharm to the human body tissue. It is not sufficient to report the name of microbe to the physician after isolating andidentifying clinically in the laboratory. Although physicians may have knowledge of antimicrobial agents and a generalpattern of susceptibility of certain antimicrobial agents. Susceptibility of many bacteria, fungi, viruses to antimicrobial agents cannt bepredicted that many micro organisms including bacteria and fungi, become resistant to thenewest antimicrobial agent.SELECTION OF DRUG :-(1)Has the most activity against the pathogen.(2)Has the least toxicity towards the patient.(3)Has the least impact on the normal flora of the patient. Not only antimicrobial agents are made by natural products but also micro organismsthem selves. Antimicrobial activity can improve by antibiotics which are synthetics, semisyntheticand modification of natural compounds. 32=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 49. Review of Literature================================================================== Antibiotic which kills the organism is said to be cidal and which inhibit the growthof 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 broadspectrum antibiotic. High dose of antibiotics leads to disturb the natural defense mechanism in human.The normal flora can become disrupted in the intestine cauring diarrhoea. Vaginal flora canbe disrupted cauring fungal infections of genital tract. To over come infection antimicrobial agent must attain effective level in infectedsite. Some antibiotic enters easily in to C.S.F. Eye or prostate. Some organisms cannt be given through the mouth because of distruction by thestomach. Excretion of must of the druges are through kidney and some are broken down by theliver. Fixation of dose based on two criteria which kills micro organisms but doesnt harmto the patient. It is very difficult and tough to treat fungal diseases. Synergic action can be seen by giving drugs in combination than they actindependently.(3)Has the least impact on the normal flora of the patient. - Not completely destroying the intestinal flora. Not only antimicrobial agents aremade by natural products but also micro organisms them selves. 33=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 50. Review of Literature================================================================== One which kills the organism is said to be cidal and which inhibit the growth oforganism know as static. Antimicrobial agents have specific activity against bacteria, fungi and viruses. Theantibiotic which act on both gram +ve and -ve bacteria is known as broad spectrumantibiotic. High dose of antibiotics leads to disturb the natural defense mechanism inhuman. The normal flora can become disrupted in the intestine curing diarrhoea. Vaginalflora can be disrupted cauring fungal infections of genital tract. To over come infection antimicrobial agent must attain effective level in infectedsite, some antibiotic enters easily in to CSF. Eye or Pro state. Some organisms cannot begiven through the mouth because of distractions by the stomach. Exception of must of thedrugs are through kidney and some are broken down by the livers. Fixation of dose based ontwo criteria which kills micro organisms but doesnt harm to the patient. It is very difficultand tough to treat fungal diseases. Suberic action can be seen by giving drugs in combinationthen they act independently.MINIMAL INHIBITORY CONCENTRATION (MIC) (33) :- A test that determines the lowest concentration of the antimicrobial agent that inhibitthe growth of an organism.Sensitivity :- Sensitivity test, in which an organism is placed with antibiotics to determine whichantibiotic will effectively kill the organism with smallest dose, also refers to the ability ? Anantibiotic to inhibit growth of an organism.Resistance :- When an antibiotic dose not inhibit the growth of organism. 34=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 51. Review of Literature==================================================================Static :- Inhibiting growth of an organism.Synergy :- Two or more drugs in combinations having a great effect that the sum of the twoacting independently.III) BACTERIOLOGY (34) :-1) Scientific Classification :(i)Staphylococcus (Gram+ve) Domain - Bacteria Kingdom- Bacteria Phylum - Fermicutes Claso- Cocci Urder- Bacillales Family - Staphylococcaceae Species- S. Aures. S. Aureus is a gram + ve cocci which appears as grape like cluster when seen throughmicroscope and has lagered, round golden yellow colonies, often withy B-hemolysis whengrown on blood agar plates.35=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 52. Review of Literature================================================================== The bacterias name aureus means golden in Lactic. The most common cause ofstaph infections is a special bacterium, frequently living on the skin or in the nose of healthyperson, that can cause minor skin infection as pimples boils, c, Celluloids and abscesses. S. Aureus Has about 2600 genues. The species has been subdivided in to two subspecies S. Aureus and S. Aureus anaerobes. S. Aureus may occur as a commensal on humanskin 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. Skinor mucosa lining this leads to biols and carbuncle I infected it can cause severe diseasestaphylococcal scalded skin syndrome. Infection of this can be spread through contact with pus from an infected wound, skinto skin contact with an infected person and things. Which are used by this person.(ii) Bacillus subtilis Kingdom- Bacteria Phytum - Firmicutes Class- Bacilli Order- Bacillaceae Family - Bacillaceae Species- Subtilis It is a gram +ve, commonly found in soil. A member of genus bacillus. It is notconsidered as a human pathogen. It has approximately 4100 genus, only 192 were shown tobe indispensable.36=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 53. Review of Literature==================================================================(iii)Escherichia Coli (Gram -ve) : This genus is named after Escherich who was the first to be deseribe the colonbacillus under the name bacteritim coloc commune (1885) based on minor differences in biochemical characteristics, colon bacilli were described under various names, but in view ofthe mutability of the bio chemical properties in this group, they have all been included in theColi which is further subdivided in to biotypes and serotypes. A few other species are E.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 singly or in pairs.It is motile by peritricatate Hagelly, though some strains may be non motile capsules andfimbriae are found some strains, spores are not formed.VIRULENCE FACTORS : Theere are two types of virulence factors have been recognized in E-coli namely(1) Surface antigen (2) Toxins E-coli produces 2 types of toxins (a) Haemolysines and (b) Entero toxins. Heomolysines dont appear to be relevant in pathogenesis through they are producedmore commonly by virulent strains. Enterotoxins are very important in pathogenesis of diarrhoea. These distinct types ofE-coli, enterotoxins has been identified (i) Heat liable toxin (ii) Heat stable toxin (iii) Verotoxin. E-coli causes wound infection, abscesses and also septicemia.37=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 54. Review of Literature================================================================== Because of it UTI. diarrhoea pathogenic infection like meningitis, peritonitis mayoccur.(IV) CANDIDA ALBICANS :- Kingdom-Fungi Phylum -Ascomycota Subphylum-Saccharo mycotina Class-Saccharaomycetoes Order-Saccharo my cetales Family -Saccaromycetales Genus-Candida Species-C-albicans. Synonim-Candida stellatoidea It is diploid a sexual fungus (a form of yeast. It causes (skin), oral and vaginalinfection in humans. These are many species of candida. The clinical spectrum of candida albicans rangefrom superficial infection of the skin to the life thereatening infection. They can be isolatedfrom healthy mucosal surfaces of oral cavity, vagina, G.T. tract and malaria.CLINICAL SYNDROMES :- The infection caused by organisms in the genus candida includes localised disease ofthe skin and nails, diseases that affect the mucosal surfaces of oral cavity, vagina,oesophagus and brochealial trees.38=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 55. Review of Literature==================================================================Because of this fungal infection causes most areas or skin causing itchy lesions onthe perinium, including deeper rash in. Enterotoxins are very important in pathogenesis of diarrhoea. The distinct types of E-coil entero toxins has been identified .(a) Heatliable toxin (b) Heat stable toxin (c) Vero toxinE-coli causes wound infection , abscesses and also septicemia . Because of UTI,diarrhoea, pathogenic infection like Meningitis peritonitis may occur .(IV)CANDIDA ALBICANS :-Kingdom- FungiPhylum - AscomycotaSubphylum- Saccharo mycotinaClass- Sacch aromycetesOrder- Saccharo mycetalesFamily - Sacchro mycetaceaeGenus- CandidaSpecies- C - albicans.Scaling and ulceration in candida is also notorious for invading subcutaeneous tissuesand causing serious infections. It resides in most health care facilities.It will grow in most laboratory media, including blood agar and sabouroud agar. Itappears after 24 hours has tiny dots and by 48 hours the dots are soft, creamy colonies. Itsmells like bakers yeast.39=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 56. Review of Literature==================================================================(V) SKIN DISEASES (35) :-Severel infectional diseases are seen because of the various types of microorganisms. Palash seed is used in many skin diseases like Kustha, Pama, dadru and kandu.and also infection can be seen in.So in skin diseases ring worm infection is also present. This infections are commanlyfound in male, female and in children also. According to our references palash seed isusesful in skin manifestation. Fungal infections are difficult to cure fast. There is indirectrole of Bacillus subtilis in skin infetion.(VI)Microorganisms Frequently Encountered In Body Sites :1)Blood : Staphylococci, Coliform bacilli, Pseudomonas sp.,streptococci, Haemophilus influenzae.2)Genital Tract : Coliform bacilli, Enterococci, Bacteriodes sp., Lactobacillus,Gardnerell vaginallis, Trichomonas vaginalis, Candida albicans, B-Hemolyticstreptococci A,B and D. Neisseria gonorrohoeae, Treponema pallidum, Chlamydiatractomatis, Herpes simplex.3)Wounds :Staphylococcus aureus, Streptococcus pyogenes, Coliform bacilli,Bacteriodes sp., Pseudomonas sp., Clostridium sp., Anaerobic cocci.4)Respiratory Tract : Group A and B streptococci, Corynebacterium diptherise,streptococcus pneumonicae, Haemophilus influenzae, Mycobacterium tuberculosis,Klebsiella pneumoniae, Pseudomonas aeruginosa.5)Urinary Tract : Staphylococci, Diphtheroid Bacilli, Coliform bacilli, Enterococci,Proteus sp., Lactobacilli and B-hemolytic streptococci, Bacillus sp., E.Coli.40=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 57. Review of Literature==================================================================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.INFERENCE : Here we can see that E.coli and staph.aureus are commonly encountered organisms invarious sites of the body.(36,37)C) EXTRACTION PROCEDURE REVIEW:-There are there types of extraction procedures are present.(a)Solid-Solid in this type solid components extracted from solid substance. This extraction always carried out before any further separation / processing.(b)Solid liquid extraction :- Where the solid drug is extracted with a liquid medium.(c)Liquid - Liquid extraction - In this process any of the two liquids that arent miscible includes that substance to be extracted (solvent extraction). To get specific drug extract herbal drugs are extracting of certain particular size withsuitable extraction medium. For extraction fresh or dried parts of plants are used. The partswhich are used for extraction must be dried before the procedure taking place. To avoidchemical changes drying operation should be performed under controlled condition. Dry41=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 58. Review of Literature==================================================================these parts as quick as possible. Drying procedure must be going on in room temperature andin airy place High temperature must be avoid for drying. This part must be kept free frominfection. The extraction depends on the texture and water content of the plant material and onthe type of substance that being isolated. Alcohol is good solvent for preliminary extraction. After exhaustive extraction subsequently the material can be macerated in a blenderand filtered. After extraction of green tissues with alcohol chlorophyl is removed in to thesolvent at the extent. When the tissue on repeated extraction is completely free of greencolour, it can be assumed that all the low molecular wt. compounds have been extractedwhen 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 withpractice.SELECTIVITY OF SOLVENTS : Selection of the solvent is very important aspect, not only for the yield of one moreprinciple substances but also for the qualitative and quantitative composition of theacompanying substance.(i)Hot continuous extraction - soxhlation :- By the help of available soxhlet extractor one can prepare crude plant extract. Theprocedure is done by the help of pure solvents. The actual solvent proportions in the extracting chamber differ from that originallyused in the collector.42=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 59. Review of Literature================================================================== Advantages of this apparatus is that it is fully automatic continuous method thatdoesnt require further manipulation other than concentration of the extractive and saves.INSTRUMENTATION AND PROCEDURE : In this procedure the material to be extracted is placed in a thimble. This thimble ismade up off cellulose or cloth in a central compartment with a siphoning device and sidearm both connected to a lower compartment. Reflux condenser is attached above the centralsample compartment. Solvent container, sample compartment and reflux condenser are separate item ofglass which is assembled together with the appropriate contents, to make the completeapparatus. The solvent in the lower container which is usually round bottemed flask is heated toboiling, and the vapour passes through the side arm up into the reflux condenser. The vapourliquities and drips into the thimble containing the material to be extracted. The warm solventpercolates through the material and the wall of the thimble and the extract gradually collectsthe central compartment when the height of the extract reaches the top of the siphon, theentire liq. in the central compartment flows through this and back into the lower solventcontainer. The process gets repeated then. The extract is collected in the lower vessels, gradually be coming more and moreconcentrated. It is assumed that no volatile substance is present. The vapour rising from theheated extract is pure solvent vapour and so the liquid dripping into the material from thecondenser is essentially pure solvent, which get from the extract. Small volume of solvent isneeded, the effective volume of solvent used for the extraction is proportional to the time forwhich the process is allowed to continue.43=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 60. Review of Literature==================================================================The soxhlet processes is useful for exhaustive extraction of the plant material withparticular component is desired.LIMITATIONS :-(i) In this procedure solvent is recycled on and often the extract that collects in the lowercontainer is continuously being heated and it may suffer thermal degradationreaction.(ii)Total extract will exceed their solubility in that particular solvent so it mayprecipitate in the lower container and require much greater volume of solvent forlatter desolation.(iii) It is not possible when we are adopting the them procedure on a large scale it is notsuitable before use of solvents with relatively high boiling point such as methanol orwater, the whole apparatus below the condenser needs to be at this temperature foreffective movement of the solvent vapour.(iv)This procedure is carried out with single solvent and we are unable to extract withany mixture solvent.(38,39)(B) AQUEOUS EXTRACTION:- (Maceration Principle)Maceration is the process of extraction of drugs with a solvent with several dailyshakings at room temperature.44=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 61. Review of Literature==================================================================INSTRUMENTATION AND PROCEDURE :- A conical flask of required measurement is taken. Then required drug plant materialis choosen and poured stated quantity of water at room temperature to it and kept it for 7-8days. (Minimum 5 days). The vessel must be sealed highly. After 7 days residual liquid isdecanting and staining is expressed from the solid and extract kept to evaporate the watercontent. at certain temperature (300c-400c) on water bath. 45=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 62. Methodology==================================================================45=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 63. 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 andstored in moisture free container(ii)Then as required it is used.(4.4) PHARMACOGNOSTIC STUDY(i) Organoleptic Characters :Table No. 8Sl. No.ParametersI.P. / B.P. / As per LiteratureObservationI Physical TestsNatureFlat dull red nearly black Flat Dark BrownColourDull red nearly blackDark BrownOdour Characteristic CharacteristicTaste Bitter BitterIISeed Length 2.0 - 5.0 cm.2.5 - 4.0 cm.III Seed Width1.0 - 3.0 cm.1.5 - 2.5 cm. 46=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 64. 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 100seeds nearly 80-115gms. The hylum of the seed is conspicuous and situated near the middleof 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 (saltwater) 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 asingle layer of pigment cells. The endosperm region has parenchymatous cells containaleurone 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 arepresent.POWER - MICROSCOPIC STRUCTURE OF POWDER : Powder contain yellowish brown colour, acrid and bitter with oil flavor andtypical smell, small fragments of festa. Cotyledonary parmanchyma containing a few starchgrains, hour glass cells, abundant spiral protein bodies, mucilage and oil globules.Note :- It is tough to take seed section.47=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 65. 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 thecrude drug effects, total Ash value. Such variables are then by treated with acid andacid 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 airdried sample of the crude drug. ( Standard ash value of "Palash" seed is not morethan 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=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 66. 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 into 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 abeaker.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 residue7)Calculate acid insoluble ash of the crude drug with reference to the air dried sampleof crude drug.49=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 67. Methodology==================================================================ACID INSOLUBLE ASH VALUE :-Wt. of crucible= 40.775 gmsWt. of drug= 02.007 gms.Wt. of ash = 01.73 gmAcid 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 chloroformEquipments:- 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 MachinePROCEDURE :-1)Weigh about 5 gm of the powered drug in a beaker and transfer it to a dry 250 ml. ofiodine 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 thereminder of the solvent into the conical flask.50=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 68. 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 gmWt. of drug and beakar after drying= 52.175 gm0.484 gm L 25 ml gives =0.484 gm100 ml gives =0.484 x 41.936 gmL 5 : 100 : 1.936 gm 100 x 1.936= 38.720 %5 (standard value = not less than 25 % )51=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 69. 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 machinePROCEDURE :-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=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 70. 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 gm2)Wt. of beaker + 25 ml of substanceevaporated = 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 : 1.036100 x 1.036 = 20.72 %5 (Standard value not less than 19.70)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=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 71. Methodology==================================================================(4.6)Methodology (41)Extraction (Schematic Presentation)Extraction Aqueous Extract Alcohol Extract Hot Water ExtractCold Water extractHot continuous extractAbbreviations for extract sample :-Cold water - A Hot Water - BEthanol Extract - CExtractions :-INTRODUCTION :-As it is preliminary study, extractions selected were cold water extracted bymaceration and hot continuous extraction by soxhlation (Ethanol)To start with we have taken alcohol i.e. ethanol from distilling rectified spirit bydistillation procedure. Then the procured ethanol was used for the extraction of thefollowing drug.(i) Seed powder of Palash : 54=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 72. 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 fromthe inactive components by using selective solvent in standard extraction. The process at atemperature approximately that of the boiling point of the solvent apparatus permits theuniform percolation of the drug and the continuous flow of vapour of the solvent around thepercolator is best for this type of extraction.The process is generally applied to the removal of natural products from driedtissues originatingfrom plant fungi etc. The non stream volatile components may beremoved by solvent extraction using a batch or continuous process of extraction of a solid bya hot solvent it is better to use a soxhlet apparatus.55=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 73. 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 thesoxhlet app. The apparatus is then fitted to a round bottom flask of appropriate sizecontaining the boiling chips and then to a reflux condenser (preferably the double surfacetype) Before joining condenser or even after joining, pour the solvent in the thimble slowlyand 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 withappropriate quantity of solvent. The solvent is boiled gentely, the vapour passes up through the tube and is condensedby condenser and then condensed solvent falls in the thimble and slowly fills the body ofsoxhlet. When the solvent reaches the top of the tube, it siphons over into the flask and thisportion of substance, which it has extracted, gets collected in the round bottom flask. The process is repeated automatically, untill complete extraction is attained. Fifteento 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. 56=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 74. 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 gmProcedure - soxhlationSolvent - 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. (43)AQUEOUS EXTRACTION (WATER EXTRACTS) :Equipments :- Conical Flask - 2 lit capacity.Measuring cylinder - 25 ml capacitySilver foil and filter paperEmulsion cloth.57=============================================================================== SREENING OF ANTIMICROBIAL ACTIVITY OF PALASH SEED (BUTEA MONOSPERMA LIM.) 75. 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