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CHEMICAL STUDY OF EFFECT OF SHODHANA ON

CHEMICAL STUDY OF EFFECT OF SHODHANA ON
TOXIC PRINCIPLE OF GUNJA BEEJ - by Nilima Narayanrao wadnerwar 2006-2009, Dept. of Agadtantra and Vyavhar Ayrved, Shri Ayurved Mahavidyalaya, Nagpur

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    Gunja chem study-agada Gunja chem study-agada Document Transcript

    • MAHARASHTRA UNIVERSITY OF HEALTH SCIENCES, NASHIK Name of candidate - vd. Nilima Narayanrao wadnerwar Name of College - Shri Ayurved Mahavidyalayq Nagpur. Name of Guide Name of Co-Guide - Vd- R C. Wegheye, - Vd. U. D. Duragkar Dr. S. G. Jyotishi Course - Speciality/Subject - Admittedin/ M.I). (Ayurvede Vechrrpti) Agadtantra - 2006-2007 Academic Year - Topic of Dissertation - "CHEN{ICAL STTJDY OF EFFECT OF SHOI}HANA ON TOXIC PRINCIPLE OF GT]NJA BEE '
    • MAIIARASIITRA TJNIVERSITY OF IIEALTH SCIENCES, NASHIK . Course . M. D. (AYURVED) ' Speciality / Subject . AGADTANTRA . Admitted in / Academic Year o 2006-2007 o Topic of Dissertation r .,CHEMICAL STIIDY OF EWECT OF SEODEANA ON TOXIC PRINCIPLE OF GI]NJA BEEI'
    • . r.f,'tllft -lasrrrof rcDI. lBlPg!ir.".r.o*," ru*ao REcocrsED !y c.c.r.r. r ra^na$fi^'a;;ili*iir- This is to cerriS that, Name of Candidate - Vd. Nilima Narayanrao Wadnerwar Name of College Shri Ayurved Mahavid yalaya,Nagpur. Name of Guide Vd. U. D. Duragkar N4me of Co-Guide - Vd. R- C. Waghaye, Dr. S. G. Jyotishi Course M.D. (Ayurveda Vachaspati) Speciality / Subject Agadtantra Adrnitted in I Academic year. 2006-2007 Topic of Dissertation C'CHEMICAL STUDY OT ETSECT OF SIIODHANA ON TOXIC PRINCIPTE OF'GUNJA BEEJ,, I have checked the data and observation of the work from time to time. I recommended this dissertation to be submitted to the adjudicator for the partial fulfillment of the degree of M.D. Agadtantra through the proper channel. Date: l8ll zlzso9 Place : Nagpur d. U. D. Duragkar H.O.D. Dep1. of Agadtantra and Vyavahar Ayurved Shri Ayurved Mahavid yalaya,Nagpur. PROF"E*qsoR & H.O.D. &p*rtrrcnl of ngdr*n+lyavheAn{rrrruda _ AndVtdsyaidy*Si$tra ranvantarillarg,*anulnanflqgrr,ilagpur.44gggg Phone: (cdbgBl :zrl?sl20rocpftar) :272l1i6rtn. *"*r, oo! rr**i reregarn."i*t--' g4 -Xao
    • d" il|. [1. Jumb tHClpAt rh.D.{rilnocD ^m""tsIffiilt CERTIF'ICATE This is to certifu that, Name of Candidate - Vd. NilimaNarayanrao Wadnerwar Name of College - Shri Ayurved Mahavidyalaya, Nagpur. Name of Guide - Vd. U. D. Duragkar Name of Co-Guide - Vd. R. C. Waghaye, Dr. S. G. Jyotishi Course - M.D. (Ayurveda Vachaspati) Speciality / Subject Agadtantra Admitted in / Academic year - 2006-2007 Topic of Dissertation "CHEMICAL STT]DY Or.Erf,.ECT OI'SHODHANA ON TOXIC PRINCIPLE OF GUNJA BEE , I recommended this dissertation to be zubmitted to the adjudicator for the partial fulfillment of the degree of M.D. Agadtantra through the proper channel. Date : te I tz- l*"g Place : Nagpur Vd. R C. Waghaye Reader ""i#ffiffifrtril;il#:,ffiT:. *.:rys""rlt}r:i,lii.f ',",,"'i[ETf hanvanffi Phone ll*g, Ilanumen l{agpr, llqF,r - 4a0 m9 Fi*qFt, T;ffi rrq, ;fFntt-rrco ooi :{CegF}:2?,t2591(}tcpitd}:ttlf!1i6(p.G.hd.} :ttffigg Tetegrranr:Rragdrye
    • M. M, Jumls :lpAl, rh.D.(Ayundf CERTIFICATE i This is to certifr that, Name of Candidate - Vd. NilimaNarayanrao Wadnerwar Name of College - Shi Ayurved Mahavidyalaya, Nagpur. Name of Guide - Vd. U. D. Duragkar Name of Co-Guide - Vd. R. C. Waghaye, Dr. S. G. Jyotishi Course - M.D. (Ayurveda Vachaspati) Speciality / Subject Agadtantra Admitted in / Academic Year- 2006-2007 Topic of Dissertation 66CHEMICAL STUDY OF'EFF'ECT OT' SHODHANA ON TOXIC PRINCIPLE OX'GT]NJA BEEJ' I recommended this dissertation to be submitted to the adjudicator for the partial fulfrllment of the degree of M.D. Agadtantra through the proper channel. Date : lS I la-l LA Place : Nagpur o g o-Guide Jyotishi M.A, M.Sc.PhD. S. G. Head, Centrai Reseirch l-abmaqr, Shri Ayurved MahavidyatEra l.Iagrr- lt4nr, ilag;xrr -4t0 oB {td0qd, €lffq phorc: {Golhgs} :?,l1lFfil2(Hc&l :?it?i116 (P.G- H.l :?IttF nfrr: hanvantari ffug,lhnrmran q-.ttro ftrgneq6 oog
    • M. M. Jurnle :lpAl PtLD'aYurwq EE D EII.NEN !ilTgIY, HAIIAnAIHIXA tnTVEI$rV Of IfrATIH n n Gtrretc.cll. r uAil nAlittlA coYlftmfr This is to cLrtify that, Name of Candidate - Vd. Nilima |rfaalrysBWadnerrrar Name of College - Shri Ayurvod Name of Guide - Vd- U. D. Name of Co-Guide - Vd- Course - ilt-D R C. tkhavidFlrya, Nagnrr. Drr,agh trrqhnF,Ih- S- G. Jyorishi Va&rymi) Speciality / Subject aea&ma Admitted in / Academic year- am6.il0l Topic of Dissertation *cEEltlrcar, srrlDy or nDrBcr oF SHODHANA oN TOXIC PRINCIII^G (lF GITI{JA BEET' I recommended this disstui.n the partial fulfillment of the b be o'bmittd to the adjudicator for dryE of trtf-D- Agdtantra through the proper channel. Date: lgllz-fa-o"1 Place : Nagpur Vd. U. D. Duragkar of Agadtantra and Vyairahar,Ayurved Shri Ayurved Mahavidyalaya, Nagpur. DS. PROFESSOR & H.O.D. Drprftncntof - Arrd Hilg; Hanurnan ltrgar, gFrlr -al|0 Phone : {Co@al : l?tAFEl gfo.F.rr} : rnvantml t *S * 'ff,i/Z,tlSFS. hsr} : El4g66g -lu:o oo3 SCFIICI
    • Yd. M. M. Jumlg papc;pAt HLD{Alnn'to CERTIFICATE This is to certifi that, Name of Candidate - Vd. Nilima Narayanrao Wadnerwar Name of College - Shri Ayurved Mahavidyalaya, Nagpur. Name of Guide - Vd. U. D. Duragkar Name of Co-Guide - Vd. R. C. Waghaye, Dr. S. G. Jyotishi Course - M.D. (Ayurveda Vachaspati) Speciality / Subject Agadtantra Admitted in / Academic year _ 2006-2007 Topic of Dissertation "CIMN4ICAL STUDY OF EF'f,'ECT OF'SHOI}HANA ON TOXIC PRINCIPLE OF'GUNJA BEEJ, I recommended this dissertation to be submitted to the adjudicator for the partial fulfilrment of the degree of M.D. Agadtantra through the proper channel. Date : lS ltz-12-a as Place : Nagpur I*#ilffiq Shri Ayurved Mahavid5nhyr, hl W *lalgl* Fkr. Aur,"'ed ter llnile;rtryl Dhanventari r*rrg, Hanurnan fagaa phorc: ilagtrr.4{o o0g mnl rpqr r;tFr rd,l:rnl3 rrr:H* {ffigf,t : zro2g2-6ffit :E!EllAt".c. o.t
    • M. ltl. Jumks llpAl ?h.D.{Ailrnlc4 AFFrnFErs*.sErnaryv,@ c.c, -. r rrrrssmi'ciffid;i ,yfrgp EqEF r This is to certiS that, - Name of Candidate Name of College Name of Guide Nameof Co-Guide Course Vd. Shri Ayurvorllferdrtyalala,Nagnrr. Wadnercrar Vd-U.D.Drr4h Vd- R II,LD. Speciality/ Subject Admittd in / Academic NilimaNrayarrc C- W+h.lln'Ih- $. c,Jldisei (At'EtcftVtCfqillJ egfdhrrrl yer- arFanm Topic ofDissertation ..CHEDIICAL SflIDT TOXIC FRTNCIII^C . The prresd study hrs Institutional Ethical Colrrmife ffi f,DIrcr [F ffiDTDEANA ON ffi GIINXA TNNF . ba d fu E @hly ttuougfu the drGtuy ftough respective E ethics.. Institutional Eftical Cmfuc rmds dissertation wort for adjudicdim Date: md forwards this o fu uiwndty Authority. tSlttlzao3 Place: Naglur ph.D. (Ayurved) Cnairpenon, Committee -Etr9ul Shi Ayuwed Mahavidyalaya, Nagpur nlartr Phom: kg' ltarnrman lsar, hFr-aaeos FinfrqFi, ulIrIFI:TrR; qFr[t-rro {G*gr}: naiEftz(rbeftd} :triZ'|rii(p.G. hll}: 2?{Osn ngsan: Rugnarap oog
    • ACKNOWLEDGEMENT The impulse behind everything I do is the blessing of God and constant moral support of myparents and mybrothers. I have ventured to complete fulfillment of my post graduation my research project in Agadtantra rmder for the partial able guidance of Dr. U. D. Duragkar, H.O.D. Dept. of Agadtantra and Vyavhar Ayrved, Shri Ayurved Mahavidyalaya, Nagpur. I express my sincere and profound gratitude towards him for providing me his excellent guidance, ge,nerous help and persistent encouragement for completing my project. I am deeply indebted for his kind support and valuable zuggestions drring the entire oourse of present work. I wish to express my heartful thanks to Dr. Ramesh ilaghaye, Reader, Dept. of Agadtanfa and Vyavhar Ayurved, Shri Ayurved Mahavidyalaya, Nagpur for his timely counsel and encouragement at every moment during the course of this study. With deep sense of gratitude in all humanity, I wish to express my heartful thanks to Dr. s. G. Jyotishi, co-guide, Lecturer and Head of central Research Laboratory, Shri Ayurved Mahavidyalaya, Nagpur in conducting Laboratory works. His experience and guidance hefued me a lot to complete this dissertation. I feel lack of words while expressing my profound and regards to sense of gratitude Dr. M. M. Jumle, the Principal, Shri Ayurved Mahavidyalaya, Nagpur for granting me permission to carry out this project. I express my profound sense of gratitude and regards to respect Late Dr. Shivkaran Sharma chhangani for their inspiration at every step helped me in completion of my prese,lrt work- At this juncture, it is great pleasure for me to eryress thanks to my f,ather Late Mr. Narayanrao lYadnemar who is my inspiration. I feel lack
    • of words to express thanks to my mother Mrs. Ratnamala Wadnerwar and my brothers Mr. Prashant wadneryvar and Mr. Nitesh wadnerwar who always tried to exclude every difficult situation. They are e,nergetic persons behind whole education of my life and making me liable. I express my immense gratitude to my best friends Dr. Vanmalq Dr. Vikram, Dr. Viren, Dr. Nilam, Dr. Swati, Dr. Shweta, Dr. Manisha, Dr.Nitm" Dr. Pradny4 Dr. Sunil, Dr. Babit4 Dr. Shital, Engineer Sonu Lohi, Monika Gajbhiye, Amrapali Gaikwad whose inspiration love and moral support strengthened me in every I difficult circumstances of my life. express my thanks to my colleagues in the Departrnent Dr. Biradar, Dr. Dipak, Dr. Parmeshwar for their timely help. I wish to place thanks to my colleagues in l.aboratory Dr. Shweta, Dr. Sayali, Dr. Kanchan, Dr. Deepali, Dr. Giridhar, Dr. Ramakant, Dr. Dnyaneshwar, Dr. Amol for their help and Co-operation. I express my thanks to Lakaswar and Shankhdarwar family for their moral support. I am very much thankful to Ramakant Sharma, Attendent of Agadtantra Departrnent for day to day work. Special credit must go to Mr. Nilesh Mankar, proprietor, Sunita Cyber club, Reshimbag Road, Nagpur for his entiring hours at computdto bring out this dissertation with distinguished and novel ornamentation. I extend my thanks to all those who were associated directly and indirectly in accomplishing this work. Ilr. Nilimt N. Wadnerwar M.D. (Scholar)
    • INDEX 1. 2. 3. INTRODUCTION AIM AND OBJECTTVES REVIEW OF LITERATURE a) Agadtantra b) Agadtantra in Veda and Ayurveda c) Toxicology Modern Review d) Visha - Ayurvedic and Modern Review e) Shodhana 0 Drug Review i) Ayurvedic literature of Gunja ii) Abrus precatorius modern review iii) Abrin g) Law Relating to Poisons 4. MATERIAL AND METHODS A) Pharmaceutical Study . Shodhana of Gunja Beej B) Analvtical Study a) Physico-chemical Analysis b) Protein Assay c) Amino Acid Test d) Thin layer chromatography C) Toxicity in Vitro Studv 5. OBSERVATION AIYD REST'LTS 6. DTSCUSSTON 7. SUMMARY 8. CONCLUSTON 9. BIBLIOGRAPHY 10. AINEXURE a) Abbreviations b) Blood Investigation Report 14 5 G57 58-81 TL93 9+9t 99-l0l to?-l04 rOSlO9 110-111
    • INTRODUCTION
    • TNTRODUCTTON Surrounding and Environment are always crucial in upbringing of any system, influencing it in many ways. The systern of medicine closest to nature is Ayurveda. Ayurveda studies human physiology and pathology in relation to the nature. In real sense it cares and cures the living beings in the most natural ways. Ayurveda is the encyclopedia of the Indian Medicinal system which itself is a reflection of various laws of nature because they are inherent to life of all the sensorial. The first ever documented knowledge of human race is the vedas and these Vedas are accompanied by a large corpus of ancillary literature. One of them is Atharvavcda which imposes the foundation of medicinal science. As the time passed, our great sages using their empirical knorvledge. developed our own tradation of medicine'oAyurveda which is said to be the derivative of Atharvaveda". There are more than 250 different systems of medicine in the world today. Oniy a few of them me considered important and recognised by rvorld health organisation. Ayurvda being one of the,rr. Research and development are essential to progress in any field. To improve our hclrizon of knowledge, the conc.ept need to be focused deeper in order to make it rnore elaborative, to avoid enors and to open the horizon to greater limits for exploration. This process involves thinking, exploring, evolving hypothesis, expedmenting and evaluation of the results with peers of
    • successful conclusion of the process. This is an essential yardstick in the modern world for the growth of knowledge in any field and particularly in the field of science. This helps in development of a particular branch of science reaching the frontiers of knowledge. Agadtantra is one of the important branches of Ashtang Ayurveda. It has its own importance in lndian system of Medicine. It is practiced extensively especially in rural and tribal areas. As defined by Acharya Sushruta, it is the branch of Ayurve.da which deals with Agada (Medicine) that combat toxic effects of visha (Poison). It includes the study of prevention, method of detection and estimation harmful effects, fatal doses of poisoning and its management. sTrrEtq'qrq@r M-wqrPfffirr Su.Su. l/14 The ancient Indian healers were the first to use poison as a medicine. They considered poison and ambrosia to be equivalent and it is the application which makes the differe,nce. rnFmrr*qrffitw,Et'*or r ffift1?qrcq+qrflr(d*qfrf+R.rl (Shodashang Ilridaya 14/ l, Hyavat Sharma) ,rd rl.r' srw +qqrft.rn:, ilqsf TqqIKrq{ 1 (Su.Su. l/4 Commentory)
    • eFrE: r1_(frFrsr q r1( q Fft qr r "rq5@qT Visha (Poison) is the dravya, which when enters the body spreads tirstly and vitiate Doshas causing acute damage to Dhatus like Rasa, Raktq etc" and there by destroys life. Though these poisons are harmful and dangerous to life, Ayurveda has mentioned the uses shodhan procedure. of poisonous drugs therapeutically as medicine after It is stated that strong poisons can be best medicine when used properly in correct therapeutics dose, formulation and a good medicine can also effect adversely ifnot used for proper person in proper dose etc. ffiCqfrepisrq cH et-id slqd qrfr S{rr" *epi fuT frqq r rl q.q q/q?q But to remove the harmful part of the drug and to retain the therapeutic properties of the toxic substances, Ayurveda describes various pharmaceutical processes ; one of thern is a process of purification i.e. the "shodhan Samskar". This particular process affects the morphological part of the crude drug. Hence it becomes necessary to study these chmges q,stematicaily and scientifically with the help of modern methods, so as to give a scie,lrtific base to this process, being used since avery long time. Scientific study becomes more essential if the drug is toxic. By means of purification, toxicity of a crude drug should decrease. Taking all these
    • points into consideration it was decided to select the seeds of Abrus precatorius to study purification process systematically and scientifically. The foundation of a good research is based on appropriate and adequate literature search of relevant revierrs and studies. The initiate step towards this study was a through compilation of Ayurvedic as well as modem literature regarding the concerned topics. Sushruta clearly mentions, "For explanations of truths and principles quoted from their branches of science, the student should refer to expositions made by the masters of those sciences since branches of science at it is impossible to deal with all once. He further adds, "By the study of a Shastra, a man can never catch the true import single of this science of medicine (Ayurveda). Therefore a physician should study as many allied branches of science as possible". s;q{nffiffir qF.arenffiq@rnreffiry q trsTiTr r TIrs frqrftqrc frG;tw*^: n Su Su 416,7 Hence the laboratory work in this study is done mder the guidance of the experts of related specialities. This dissertation work is an eamest and honest "attempt" to put forth some valuable information and interpretations few quotations from the Ayurvedic literature. of
    • AIM AID OBJECTIVE The aim of this Project is 1) - The study of changes taking place in the effects of toxic the basis pincipFon of chemical analysis before and after ffhm qrr**r of Vanaspatic Visha, Gunja Beej (Abrus precatorius). 2) To develop a laboratory test to differentiate shrddha Gunja Beej. al rcmdfha
    • RBWEW OF LITERATURE
    • REVIEW OF LITERATT]RE AI AGADTANTRA: The word "Gada" literally means a disease, pain or a poison Therefore "Agada" stands for something which is meant for the defend of disease or to combat with toxins. qlFGFnfrvdw6Slrrqr: g*: *or r ffifr1M$qftfin:n (Shodashang Hridaya l4l1, Priyavd Sharmd ,rdfrr, er.r+ffi6n:, Tqf Taqrq (Su-Srrl/4 I mcdily) The word oAgada' is ofteir used to denote 'antidote', a medicine th* inactivates the poison. Tantra i.e. the science which mainly deals with the toxins, toxicding condition and the methods of intoxication (i.e. treatuent with antidotes) is known as 'Agadtantra'. Sushruta defines describes the symptoms and treatment it is a branch of Ayunreda wftich of- (a) Bites of snakes, insects, spiders, scorpion, etc. .. .. and (b) Various artificial poisons which are man made by compoundingtwo more substances. $rqTfqqrq@r M-wqrrfffirr Su.Su. 1/6 m
    • a?ren Tr€'?niTrtr s..iqRfrR+r, Tilfrar, ?fitqFrrftr{ | 3r{Kdq{rTrarqqrffifr r Su.Su 1/7 sFtE: T(fr$rsr q rrq q Eft q|' r Sabdakalpadrum Ayurvedic toxicology is one of the eight classical disciplines of It was called as ooDamshtra Chikista" by Vagtrhd4 "Vishagaravairodhika prashaman" by Charaka, "Jangaloe (hftig" by Ayurveda. Kautilya and Agadtantra by Sushruta. frIqrg@ ffi; ; a€Ier anqfrGffi; rnctm qrerrE-{*i TilAar, EtqnlIgd-, r*rrr{, srffifr r CbaSu- 3OEt 6-REld.r6r*& aiftis-r sTuq-ETft Ter3"tsRftqr vflrqqFt tg rft*ar I tr (AJr$- ll5l iRrTKs qr$frR+ frqqr€rsflr: €: u (Kmnrfm*aqleqa) It is the branch which deals with Agad (MoAirlrc) rrrr efilects of Visha (Poison) of animate and odd toxic inenim4cmigin- &dameans disease and thus Agada means medicine, antidotes.
    • In western medicine, Agadtantra is named as Toxicolory. Toxicology is the science which deals with poisons with reference to their sources, properties, mode of action, symptoms which they produce, lethal dose, nature of fatal results, treatrnent, method of their detection and estimation and autopsy finding. It is concerned with law regarding their sale and description. Forensic toxicology deals with medical and legal aspects of harmful effects of poisonous substances on the human body. Bl Agadtantra in Veda & Ayurveda : The Ayurveda stands for the medical tradition of India which began with the beginning of the Aryan race, continued through the hymns and charms of Vedas and the literature of different epochs. This ancient Indian Medical Tradition has passed through successive stages. Charaka and Sushruta summarized, perhaps the high ideal achievement of this tradition during their respective ages. ' The magtc formula of Atharva veda should, therefore, be regarded as an indisputable link in the ancient Indian Medical Tradition medicine proper. Ayurveda is also calld as Upaveda and even as of Athanraveda The classification of the subjects of the Ayurveda is done into eight parts. One of its parts is Agadtmtra Ag&ntra deals wift meftods of diagnosis and treatrnent of poisonous bites of srakes, imects also herbal or other poisons.
    • The Atharvaveda contains charms against poisons like scorpions, insectsplants andarrows (IV- 6,7;YI - 100; V- snakes, 13; VI -121' r4-56;VI-56;VII-88) The Rigveda also has the hymn I VII - 50 against snake poison and - l9l against scorpions and venomous vermin. The Atharvan charms are to be accompanied by the practices of Kautilya Sutra (28.I-4; I'141' 29.28, 29; 50.17 -22, 3l .26; 32.5 -7 ; 32.20-25, etc.) According to Atharvan conception, there is position in fire, in the sun, in the earth and in the plants (V.B.I.) There is also Kandavisha (X'4.22) poisonous plants are found on mountains (V-6.8) snakes like Kasarnila, Suitra" Asita, Ratharvi, Pradaka (adder X-4.5), Aghashva, Svaja, Adyavant4 Triaschiraji, Darvi, Karikvata, etc. are poisonous. Some of these live in grass (He,nce the proverbial sake in the grass). The poison of the snakes is either in their top, middle or bottom. Garutrnan Suparna appears in connection with all sorts of poison and also medicinal plants (l-24.1, ll-27.2, lV-203, V-14) Kairata, Prushna, Upastrnya, Babhra, Taimata, Aligi, Vligi, Urgula seem to be varieties of poisonous snakes (V-13, VII-56), Kankaparvan, Sarkota, Vrishchika and Babhruare terms for scorpion in the A.V. Its poison is in the tail (VU-56.8), Mashakas or bitting insects (VII-56.2) are also poisonous. The Atharvaveda thinks of ants Upajika (termites or white ants) and particulruly wate produced by them (VII-100) as an efficient antidote against poison So the horse of pedu (X-4) considered very useful against poison. Amory plants, many are praised without being identified by name, But meutim fu mrl
    • l0 be made of Taudi, Ghritachi (X-4.24). Kandvisha seems to be some poisonous root (X-4.22,rv-6) makes a clear reference to the poisoned flrow. In the later samhitas of Ayurveda attention is paid to 'Kalpana' and antidotes. Poison is divided by them in to Sthavara (Plant of mineral) and Jangam (Animate). Sushruta deals with this topic more particularly. The description of Agadtantra is described Ayurveda which are as follows Cl in different soction in : Toxicolory- Modern Review . Toxicology is that branch of medical science which deals with poisons with reference to their sources, characters and properties, the symptoms which they produce the lethal dose, tho nature of the fatal results, the remedial measures which should be employed to combat their actions or effects, the method of their detection and estimation and cldlqpgy findings. It also concem law regarding their sale and prescription. C.K. Parikh - T.B. of M.J. & Toxicology Toxicology deals with diagnosis, symptoms and treatment of poisons and the method of detecting them. Modi J.P. - T.B. of M.J. & Toxiology Toxicology is the science dealing with properties, actions, toxicity, fatal dose detection and estimation and treatnent ofpoisons. Dr. K.S. Narayan Reddy and Toxicology. - T.B. of the essentials of Forensic Medicine
    • ll Dl vrsHA (PorSoN) : Visha is the substance which spreads readily and possesses a property of quick and extreme assimilation, vitiates dhatus and desuoys life. ffiftfrqrffifiqrrrq$yqft6R: r (Shodshang[ridalml E€dRqRftRq{r€ilsrRq'q {t ) I rtqr@ftq{1 (R.r.2ry1) a) Visha Nirukti : q{qfuaprffiffi*r (chi.ffii_zrro frvgqtfiF{iTET(til: r (ChiOti 23fr-flitupnt) ftqrq{ffiqffiffiFlqtr (SN!-Kd 3ll) Visha causes Vishad (sorow) to liviry bciry hry 1 3 Visha. Vishad is severe, depression of spoech, bodymdmfud- b) Definition of Visha : ffi qlFf r{Eqft{ffi'E{rgryf6ffi, dgseiqr1; frq{ dr4rft ftef n (Cha.Chi.23t24) terrrred as
    • t2 W Tftd TS TdePi q6tr r(I{ Rtq t Wll T€rxfrfufrffirffiqrrTuf (Cha.Chi.24l3t) Visha is defined as the Dravya which when enters the body spreads rapidly and vitiates Dhatus like Rasa" Rakta, etc and there by destroys life' The gunas of visha are exactly opposite to the gunas of oja. Symptoms in poisoning and action of poison in the body and its treatment are described in details in the Samhitas. t€qlrdqGRq@l srs+,t' E{rrJUr q{ffirrffi tt (Cha.Cln.2316) Charak Samhita has aptly described poison as originated from water, two types, similar to fire, produces eigtrt stages of toxicit5 has ten properties and is heated c) with Chaturvinshati Upakramas. Synonyms of Visha : Kshweda Garal Visha Mugar Kalkuta Kalkul qisq rRFi N I Gara Gada KalkalPa (Amarkosha frf q+-.qvwio,rcqatrqtrq ll9l9 I (R.T.2412
    • l3 Gunas (properties) of Visha : @frepiffi{eilq1 gsi qRfu Eflgsrgo- frq Ffi, rr (Cha.Chi.23lt4) €qrgwr frer trqrr qeil sTrT qqrfr i{ r ffifuiqdEqqrfui{TilTTftr{ u (Su. Kal. 2/19) ffepfrsroefAaq qgomqc*r qn (A.H.U. 40n2) r{tqq frr+rffirm r qffiqffiqrr rrftrrqq€tqqmqfriF{iftq sTrTdrqr{r a<R q r r{.fr rr*a rr q{r+q@r ffifiddEBfuqHF|?lkr il gdrfuffifrffr (Su KaL Al&zt)
    • 't4 Properties of Visha and their corresponding Action. Characteristic 1) Mechanism of Action Spreads without digestion making (Lish| 2) Laghu diffi cult for treatment. Rooksha a Vitiates Vata situation (Dry) 3) Aashukaree - Act very rapidly ; cause acute toddty d takes away life fastly. 4) ofpdm b fu minute orifices and thus aids ryread Vyavayee Makes it indigestiblg e,nter (Spreading without without proper bio-transfumdirn digestion) Teekshna tifrAffects functions ofthebmincnq -' (Sharp) 6) Facilitates unopposed entry (Clear) 5) Vishada of varying degree; vital ofpdm d odlulrhrd d ffiy affects the basic constitrfion ofthe rll* poiffi offtbody (marma) also get shattered- 7) Vikasi Deranges humors, tissues md (Muscle Relaxant) (Dosha, Dhatu and Mata) *e* crpcild mdddfie*tbi function. poim b fu Sookshma Permits free entry of (Minute) e) Ushna (Hot) Vitiates pitta and blood- l0) Apaki Makes its elimination r@ portions of the body 8) diffiolt ft is not
    • l5 (Indigestible) 1l) metabolized properly due to ftis YirfiE Avyakta Ras Capable of vitiating Kaphq als albfrG ryGad (of undifferentiated of poison through the esse,nce of food taste) (Annarasa). t2) Anirdeshya Ras (Undetectable taste) - e) Unidentifiedbyis' *" Classification of Visha : qf,T errfirqi ifrfr aqr @ RIrT{qf,{+{RfreifrqgErt I (cha.chi. z3ttsl t (Su. Kal.2/3) €rrd{qf,{tftftS*orft Tfr{ rrtri g ftqt frfreftqe}: I tt (A.H.U. 35/4) Acharya Charaka and Acharya Sushrut classified Visha in tw,o Sthavara (immovable) and Jangama (movable). Acharya detailed classification as follows Vagbhdabdm : Kritrimvisha Akritrimavisha (Artificial) (Natural) Sthavar Visha Jangam Visha (Immovable) (Movable) gqr
    • t6 Further classification in Ras Shastra texts is available according their potencies i.e. Mahavisha (Visha) and Upavisha with vrying to numbers in different texts. Kritrim visha (artinUal) Akritrima visha (Natural) Sthavar Visha Mahavisha Jamgam Visha DuSi Visha Gara Visha Upavisha ffixqlg{r Tffrq rruS'*{rq I il (Cha. Chi.23lt4) Tfr{ Afrei dm'rcgftt-fi. r (Sha.Sam-Pn lUt/zOU Action of Visha : About action of Visha, Acharya VagbhatahavedcscfrGdfuhice ftq tr f,6 qqq urrgwftail@ ilFffisn{sqdt'qryl r*qwqr€Imtft#<r"qw* n (AS-,lmfl r
    • 17 visha after entering in body, first of all vitiates blood (Rakta Dhatu). Then doshas like Vata, Pitt4 Kapha and then after all Ashayas gets vitiated at the time. Then enters in heart and stays there and becomes a reason for destroying life. Hr+q rrcid iagqsta q qrRft: BE ftqR'q I Mqrun+tq ftrls{ft 11 (A.S. 40t2s) At last Visha spreads in whole body restricts all connection of Shira, Dhamani and Strotasa and by final ends life. g) Use of Visha in Medicinal Preparation qrrt{riqem q : I frftReft qq qrnrnrrf *qd: rr (A. Hri. S.9) There is no drug in this world which do not have medicinal value. qrdq*Td qqft r drdrgfrqf qrnqfr*m r (Cba Su- 2d12) No substance in this world which cmot be usod as modicinc condition that they are used rationally and with a de,fineobjdive. frt'qru6{qqffiWl (cha Cllni-z4tffi) m the
    • l8 daqgfut'UqruErfrr{q I (A.Pra.6/46) If used properly, life threatening Visha can also be used as life saving medicine. fu sTfr frsftpivnt' sH er*tr r (Cha. Su. 1/126) After knowing its properties and mixing properly in medicinal preparation, fatal Visha becomes good medicine. q!{frqgs-qr{qETR6gui@r €-cTrfm geflgdt genQ6 gul aen rr (Ra.Ra. Sa.......) Any drug with properties of Visha is more efficient than other drugs Arnrut mixed cleverly and finely with Halalal Visha is more sffici€nf then Amrut alone. It means if we use Visha dravya in medicinal pepr*im th* preparation become more active and more efficient Drring use of dravyas in medicinal preparation one should consider its dosq q;rqr qrfti tfr r ftqrtrErw{f, rTrdrft*,, r q{rit €ffif B frS sra( rr (R.R. S.2et2t) yisha
    • 19 If as used properly in proper dose, Visha can also be as good lift rilLt Amrut. After considering above things, a conclusion can be done u w d' vishadravya in Medicinal preparations can be done only by knowing ile { properties and proper dose. But before using in medicine, these Vishadrav5po should have to be purified. El vrsHA (POISON) rN MODERN VrEW : There is not legal definition of poison. Anything which when usod internally or on the body surface in a dose or in repeated doses, if acts chemically and physiologically causing disturbances of body functions and leads to disease or death is a poison - Principles of Forensic Medicine Its is a substance - Apurba Nandy which acts on the body chemically and physiologically causing in toxic doses, disturbances of functions which may result in illness or death. Any substance when introduced into living being produces morbid, noxious, deadly effect. Any substance which through its chemical action usually kills, injures or impairs a living being. Any substances gaining access into a living being produces i[ness' disease or death. Dr. A.C. Mohanty's Legal Medicine-
    • 20 Broadly speaking, a poison may be defined as a substance of the nature of a drug which is administered in a way and in amount in which it is likely to be administered will produce deleterious effects of a serious nature. - Modi J"P. T.B. of MJ. & Poison is a substance (Solid, liquid or gaseous), which the living body or brought into contact with any part there of if Toxiolqgr introduoed in will poduce in health or death by it's constitutional or local effects or both. - Essentials of Forensic Medicine ILS-IiL RGddy A poison is defined as a substance which when arlministere4 ifrdod or swallowed is capable of acting deleteriously on the body. - Parikh C.K.T.B. of MJ. & Toxtonlpgr Fl SIIODHAN (Purification) : Before therapeutic use, there is need of pure drug besmsc arnount of unwanted material in natural form of drug. The drug useful only after removing these impurities. Othemrise ca d nrro h it can bc Hm rather than beneficial. Some drugs are highly concenfiated. So their omffiiin ffi have to be minimized and Shodhana is the best method that toxic drugs should also get purified before Rasshashtra gives method theiruseinnqf;I-' of purification- h Ras$ilfift4 therc is gven a method of purification of all dravyas like Rc, Sadharanras, Visha, Upvisha, Ratna" etc. forfta Bs-'e of Ilprc, Uanrasas,
    • 2t - There are three types of man impurities 1) Physical, 2) Chemical, 3) Natual Natural impurities is also called Visha Dosha. Definition of Shodhana rilert' qrf : frid qarclqfrqnq{ | (Prifiaum Shodhan is a process of removal of impurities sGftqP): €F{ frq} tqunFd,,r TdTRfuatugTiltr{ Shmr) ofdrry; r ffi rr G-T- 52) With the help of any dnrg uthich is metallic substance to remove its Kshalan, Nirvapan are done. It is Shodhan Prayojan fu fu ffi pocess, for imfitics sre foffics like Mrdan, calld c fufu- : ,r ggun frqs@ i q-6Tqr GdAd- | rsGsq+iS"ilF*dqfrfrm'r (Akr- 6t4T Those impurities are present Shudha in Ashdh drsvJra re nil fdmd in dravya. So use of Visha drav-ras in mdicine shmld be after Shodhana process.
    • 22 eTR{Etf ft$ ............ iwrir qft{kriq ' (R. T. 24n8) If Visha is taken or given without shodhan, it creates Dah4 Mundh4 Hrudgati Avarodha, etc. symptoms in the body. Even after using in amount, it exmive can be life threatening. That's why for avoiding ftesc s51rytons Visha should be used after Shodhan. Acharya has said rhd proper dose after Shodhana in medicinal preparations if Vifr it cm d is uscd in as gmd r Amrut. AYURVEDIC LITERATT'RE GT]NJA : Classification : Mulvisha - Su.Ka.2l5 Upavisha - Bha.Ni, Dh.Ni., AP-, RT- Guduchyadi Varya - BhavprakashNigfim, not mentioned used Aaragwadhadi Varga - fffif it in ffirr it in Vajikran bythc A. H. Su. Cffihd ttiftrrF h re *tl.*tr--. lstll.Itismeatiarodbyfunme "Kaktikta". hild
    • ZJ Gunja is also described in various granthas like shodhal Nighantu, Raj Nighantu, Kaiyadev Nighantu, Nighantu Aadarsh, Sharangdhar samhit4 Harit Samhita, Bhaishajya Rafiravali, Yogaratnakar, Rasaratna Samucchaya, Rasendrasara Sangraha, Vangasen, Chakradatta- R. il{ ffitr+' srfirqn 5qr grrrr.rrft*. ftgfuqrffi{dffir ?. m gqffi a. ry t(Su. K.2/s) ffifrdrT{qRgrflT ....... I (A.S.U. 40/7) qfrefr{ gfrrGi'il tnltn-a Erfl.dr q:rftr*': r srqt; sqmscTrilq: tt (Bh.Ni.Dhatwadi Varga, Upvisha Y. et.({1q qgrfgr{ R?n Tqr r+ ffiqr - 206) I AvgfuH!fi- q rrur$qfrqr(qq rr (Dh.Ni.Misrakadi V-ga, Upvisha-l I 56) q" sddgus erq1 Fn{.fr wt*rq'r: t lqr$i,tffitr: sffiror*q: il (A.p. Upvisha: Gl08) q. frqftg-+'tr-qiqe*-{srtqa,q eftW-ffi I ftqqr{qr qffirfrq,{+q: r 3r#qfttq-e$rffi*,r*r+q sqRilr$quils4'gpffi* r n (RT.2+163, 164)
    • 24 Description : A perennial, high climbing, twining or trailing woody vine with altematively compound leaves indigenous to India. Leaves: Leaves are alternate, 5-13 c,m. long, even pinnately compound 5-15 pairs of leaflets, oval in shape l.8cm long with margins entire. These are exactly similar to those of Tamarindus Indica (Amlika) Flowers: Flowers are shaped like pea flowers and are small, pale violet to pink are affanged in clusters. It flowers in August- Septernber. Fruits: Fruits are shorto oblong pod, splitting before falling to reveal 3-8 shiny hard seeds,6-7 mm long, scarlet with black bases. The seeds are much valued in native jwellery for their bright colouration. one third of the bean with hilum is black while the rest is bright red suggesting a ladybug. The seods were traditionally used to weigh jewellery in lndia. The measure Ratti is equal to the weight of the seed. Fruits ripe during winter. The seeds are white in colour also. frETrffrlRIFffirUrffi Fsr{gnffi {qr str r frRdr qdr il (RT. 24l43e) ier+drgr(It-ffigFrfuortr qunour T}6 ......... (Shivdarta)
    • 25 Habitat: qgt €-d-r^fiS I (Shivdatta) .It is found all over India. It is wild plant hence known as Vanya. It is found in Himalayas at the height of about 3000 feet. Synonyms: Gunja Chudamani Krishnachudika Rakta Shilftandi Sheetpaki Raktika Kakanati Bhillabhushanika Tamrika Ucchata Shyamakhuda Aruna Kanchi Kakashimbi Vanya Raktala Raktaphallika Krishnala Kambhoji Vaktra-Shalya Durmogha Vayasadani Dhwankshanakha Chutaki Tulabija Ksrishnaraktika Kakpilu Tamrika Kakanantika Kakavallari Kak Kaktikta Sughata Tamra Kakachinchi Kakini Ratti Anganranfri Kaksha Kanichi Sushimft Shangustha Chinchapatri Kakjsngha Kakadanti Manchooda Saumya Kakadani Dhwankshadani Kaktudi
    • 26 t. Tqr ffir rRn-*.r q arfr*r Swqf*,,r sqarMqftwTqfufrToun t tt qgrwmqlqlMF'srTraqT q,rcrurfr v q,,rrffi d?a qfiffiar I rr (R.T .24 I 437, 438, P age 7271 ? rfrffir TRaT 5qr a,,r*,,sfqr E.srfir q,,rRiuft mqn dR 1Mft t q,r*qfu*T tt (Sha.Ni.Guduchyadi Y atga, {-qtwr R. fffr Tgrmr ffi qar P age 34, 341) t "ndT rkTr €r qrmffi a,w{ft a.r+,ft-q: Til rTfrr sranq'urfr q rfu*'r qr*qeqft ll rr (Bha.Ni. 125,126; Page 354) Y. Tsrqfr: ?ffilMTq?Trqflr s€{er frrtrfir gcr vrfr I q,FrTTreqqr ll (Dha. Ni' Karveeradi Varga 126,P t". gqT rfsrqfrRrql rGf,-fir a'mqfu61 ffis+r?ffigraF'srdrrffi - 187) , rr (Kai. Ni., Aushadi Varga, 793-794, P-148)
    • 27 u'r*r{ft arqilftg: a,r*.,Rrd q *ftFn ens.qTdi *Frfl-etr gtr6r qrqsrqft 6T{Etrs qrq.ail-{d qra$ edrssf-f.ftft Gffinarq I sg *eftor r qrr qrfren rr eT (Ra.Ni.Guduchyadi Varga gqr Asrq0r' 9. *qr fiTqus EEIF{rs€qr ilfror Eftdqrft Rlilqcr qpwrqRm TfrTr r q friqr *eflr{qr I r q rGn-*l *q q,,rnt* ftfufEun ?rangT qrf,{sT r n (Ra.Ni.Guduchyadi Varga Characters - 109, I l0) - ll2, ll3) : Characters Guna Root Seed Laghu (light), Rooksha Ruksh4 sheet (dry), Tikshana Gharp) Veerya (Potency) Tikta @itter), Kashap Madhr(Sweet), (Ashingent) Ras (taste) Tilta Ushna (Hot i.e. Shcct thennogenic) Vipaka (Metabolic Taste) Kattu (Pungent) Itdadhra Doshaghnata Kaphavata Shamaka Tridoehaghma
    • 28 q. Tqr FqTr ten ft-ffr ddqT q trfrftftfi I (Dh.Ni.Karveeradi Varga ?" 1sqr rtwn r+ ftdar 6qrqr a,,m.frr6r - 27, P-187) 7 95, 796) r (Kai.Ni.Aushadhi Varga - (Sha.Ni. Guduchyadi Varga, P -341, 342) a. {trgqrtu (Dh.Ni. Karveeradi Varga) Y. q,,rfrr(ftq..^Nnqftqffir1Mr qftrdqf{r vqr qffirft qr rr (Ra.Ni.Guduchyadi Varga - 111) . IFcTr{dqgffitrqrffiFrmrr "tf,q frs-fil qt' Td+ r+F{ arst rr (Ra. Ni. Guduchyadi Varga Prayojyanga (Officinal Part) : Leaves, root and seds are useful formedicinal 1rtxpose- rrt'rd{STd{trqrqrgw} r (R.T.z4tffi,P -znl - I 15)
    • 29 Properties (Guna) of Gunja : TqrfuqfrFf s€'GrElr6r a,r+{frwrTtr{q dq r€KiqdT q{q lt Ttrrrf, Tilrrdt ErFanrronfr;q relq q eqrqrf +Eqrc{{I"tq r u TwtlmrTrffi r(fr: rr?=fitferT gtli-qqr{nr' ailerd tfit{r: Tqr{ngqtt6,,frft:qr{fr.. ffiiqqurynhqfrqruom I rr grrhqlr+raqrqrRfr{Fuq ?fi,r+rq r u TfrqTTvq'ffiqETffi qerqunqti *a Gffififif6q TmT{dfrniurvgu$guilwm 3mWf{rrcTflr+sgq} I l q c€rdffiq I r 11 r r (kT.24tffi-45?-? -675)
    • 30 Medicinal Uses : Both red and white types of Gunja are beneficial for hairs, qlre diseases due to vitiation of Vata and Pitta, fever, dryness of difficulty in breathing, thirst, excitement, and mod, gieincss, of eyes, impovc diseases w-vigou bodily shength and is beneficial or useful in pruritus, ulca, destructirn of wofins and similar parasites, alopecia and other skin diseases. U€r{trgffiqqrdfrtrq{q6{ gq{iq qrrfi[Fr iIUn rr€{rtlrrr |71qqEtEq'qq'frugTunqq E-frqqcrg;srfrrffi+g I tt I qqffiq (Bha.Ni. Pur. 126 - 11 128, Pags- 35,[,:155] its urrc b ffit Kaiyadev Nighantu additionally . mentions rakshograhavisha. qqcqr{fidr a'flr Fralr 5qtiffi|-(t Fdqwc{rft{r€AqrrfrE Efr{qtrffif I t qgsffi+u (Kai. NI. AuSadhi Varyt ?9S'?!16, P- l{Sl ffic, ffi sciatica, erysepalous and few other dermmoses, tilinnncs$, fiqm of b4 Various other texts explain its use in dental caries, etc. mryhid cr
    • 31 Chakradatta - qffituehref{_ ffisTrtqG+$tvgmrr Tdfrt' irflryffi q'ofrndftqeffir oufffiqRiasrqffi rr r Bhavaprakash' qrsurb - TsGrrm-S: {d id Tfl.{rd{+{ q rt Vangasen - rrrsqlqlqq tt @u{r rrrsqk{i qErsqTt Tsrr{+{ "rqt( Tersm - Afren+S rfrsr frrtrrwrifun r Td{ilrfrdffi frqr q' sa-si{qfr *Eqrq u Harita - ffi - ffiqrTswqftTdqqq Shodhal - gS - e{trqii Esgiqquf tffi{ qruRdgqaTfrTB+ufilgq r r
    • 32 qR g ctqR fu sft€il-qqr+ tl @Ud,rlEn rftgq frftt - 1_oqT{trq*n$qfrsrTr*t's{dfr"t s{-qqrrki €dq ffi: elqt+ n Frfrtt - Fdfu{ Rrt}+ Tqrr1r{rr n-ffi{ E.Rfir*'- @: vgqeycqerrr ffi *-Tqr qfrsmrqrffi qrr"'gufdri Uqr r rl I rr Fd{qrar r Kai. Ni. Aushadi Varga 795,796,P-148 Sha. Ni. Grrduchyadi Varga wrn6qdqrwrftar!T: !T: 1.dqrs-df$t @ I +11r$ rrrlEftr: p Bhai. Rat. 60171, P-604 Chakradatta : Kshusdraroga Chikitsa, 55197, P -431 3{-drn6G'ffiarg-{: g.r: I Twrudf$t@ffi:gr1'iliT: Vangasen n Kshdruogtdffin T6fl lsrg-drq-dwfr 139, P-1ZL {rft rarrersfi Vangasen : Kshdraogadhikil 1n,P-522 uqrwf6'tr q]f 4:-firqf: rsFrf,if: I rr
    • 33 t{ diq 6nrg qrqrfti gg:F{ u R.R.S.24188,P-504 1-wttuffid'qtunqtud@r ftaqfueerqrounrhQrdqil{ Fqgwqef-q$raEgeu@ n r R.T.24/454,455,P -730 Mention of Toxicity: If the powdered seeds are consumed in their original form and in excess quantity, vomitings and severe form of diarrhoea starts. The symptoms resemble exactly to those of cholera and hence to exclude or to avoid these harzardous effects, purification of its extemal use has been seeds before advised in ayurvedic texts. TqrftqqrfuffifitrdenI rftffiggwrftwmil@rr ffi-dq{qqRtq-tfrRfrqfrq efidqfiTnPf Tiqffirilt& r rr R.T. 24/441,42
    • 34 Treatment of Toxicity : 'Meghanada Rasa' if taken along with water and sugar can relive toxic symptoms produced by Gunja. ffiTr€q: I "r&r{ftraFftr: crtrqrfrfiRs:@ll Ni.Aad.P-457 Forrnulation: 1) Guniadva Tailam : It is used as a local application in scarf of the scalp, prurigo and other skin diseases like leprosy. It also reduces the pain. 2) Guniaieevan Ras : Guniabhadra Ras : lt is grven in doses of about eight grains with It increases the strength of the body and sex- vigour. 3) rock salt and assafoetida in paraplagia and Urusthambha. 4) The seeds of Gunja reduced to a paste are recommended to be applied locally in sciatica, stiffiress of the shoulder joint, paralysis and other nervous diseases. 5) In white leprosy, paste composd of Grmja seed md plumbago root is applied as a stimulant dressing. In alopeia" a paste of Crrmja seed is recommended to be rubbed on the bare scalp. TqrRiq filrg rT€rm,fr: Titr Tf,rs{+{q qrurKss4qilqfrfirq{ I r sFnF6't-{r: I Bhavaprakash
    • 35 Tqtclflq: | frTfi'T{ ............ TTrq€.[ui g:ugormf TrEr€oFr I Rasendrasarasangraha ffiq€<arqRs1€Ttrd:m r iq6q1€qrftsrffirprSaur qffiqmqftsrytrffi*rm: r r Sharangdhar gqrE-dlFrqofq dftd' T+dSsEq I Sharangdhar TqrhqBr@ +flTfr strqtr. ffiT(qrg@rs+q.rflqq Fffi{qr++lgrqrffigqrfr Chakradatta qrGilFt-fi ffierdvgffi qqlrqm *r n ftdRurr r chakradana rfu'ff*&sprf.*@r qqftftrt T{q'qnTqtfi qfr-d{ | q,r*sq quRqr$; CRr{ ffitn"lq q I
    • 36 l) Gunjadya Tailam 2) Gunjajeevan Ras 3) Gunjabhadra Ras 4) Gunjagarbha Ras 5) Nili Bhringadi Taila 6) Mritsanjeevani Gutika. lt is also used in various formulations Dose of Gunja Yz as one of the constituent drug. : - l% Ratti for internal lqreCa: and external use in purified form. Trqrusqwtffifral {.dGilfugRrcirrg-frfrerqfril r rr R-T.241453,P -728 Mention in Brihattrayee 1) Charak Samhita i) : Vvhile describing ideal characteristics of blood in tw€aty fourft chapter of Sutrasthana. q{qrFffiffifuq I TqftqgquhftT€{l@rr Cha. Su. 24122
    • 37 ii) Description of Menstrual blood in Chikitsa Sthana" Chryter 30. T-qrsdsquhll-{qrerffifi Trfu Tqt FEriq'fi-firflqriq' {t<qrmtr rr Cha. Chi. 301226 iii) Formulation of Kanak-Kshiri oil in chikitsasthana, chapter 7/t12. 2) Sushrut Samhita : i) While describing scientific procedure of making pratisaraniya Kshara (caustic for local application) Su. Su. ii) Medication for external application Ilft2 on haemorrhoids Sa.Chi.6ll2 . iii) Treatment of Dooshyodar Su.Chi iv) Local application of Kaphaja Visarpa Su.Chi. v) Treatment of Tumours (Granthi) with one medicated oil. .l4lll lTlls Su.Chi.18/19. 3) Ashtang Hridayam: i) Insertion of Varti p€r anum forhaemorrtoids A. H. Chr.8/2A ii) Lepa on haemorrhoids (external rylicdim) A. H. Chi.8/22 iii) Kakadani / Gunja in extreme stage of Samipdaj Udtra A. H. Ch| tst78
    • 38 iv) Ingridient of one medicated oil "Bhallatakadi Taila" for Leucoderma. A. H. Chi. 20/16 v) Treatment of Kaphaja Galaganda A. H. Chi.22/70 Methods of purification of Gunja seeds : l) Gunja seeds should be crushed and tied in a piece of cloth in the form of pottali (Round bolus). It should be cooked in Dolayantra by adding cow's milk for six hours. Then these seeds should be washed with warm water and dried" 2 same procedure, if carried out in Kanji for three hours also detoxity the seeds of Abrus precatorius. R. ftnfr Tqrffi qstrFR +ffiffifuTqgqr: e-zlFftt: I I FeigffiEfurqFs{aqtq rr R:t-2U43 - 441 R. T-"qrfu qWiei +dFr* sotfrq;q qrq-fr ffiq qfur-qrtr{dqrq r 11 R.T.2414p.5
    • 39 i. 55qT*lR*TRdT {RTRTft errftr: I TqrtqtrqrueTreH:@ 6tsn-qqT Fffiq fffi;rEunqdr Yogratnakar el({sqqRqqqffir*qqtr ildelrqtM $qfserqr*t - tt rr Uttarardha Vishadhikar r rr R. S. S. l/388 Dhattur beej and other (Kalihari kand, Root of Kaner and Gunja) should be treated with cow milk in Dolayantra for detoxification. Literature About Godugddha : In Ayrveda, various dravyas like Godugddh{Cow -ilk), Gomutra(Cow urine), Kanji etc. are used in the Shodhma Prrooo&rre bme of @ their useful properties and detoxifuing nature. These dravyas of toxic substances. purification Placement of Inclusions Utility : Gunja Shodhan : Charaka : Gorasa Varga Sushruta : KshiraYarga in
    • 40 Properties : pH:6.5 -6.7 Titrable acid : 0.12 - 0.15% Specific Gravity: 1.030 - 1.035 at 15.5" C Refractive Index of Godugddha - 1.3440 - I .3485 at 20o C The properties are derived from these values depend on the presence Casin, CO2, proteins, fats, Phospholipids and free fatty acids in it. Composition: Solid-12.63% Solid (not fat) 0.341% Fat-03.69% -04.82% Ash-00.71% Protein Lactose - KzO - 23.54% NazO - CaO - 22.57% MgO - 02.84% -00.31% PzOs - Fe2O3 cl- ts.ot% 11.44% 27.68% 8.94% of
    • 4t Abras precatorius- Modern Review Vernaculars: Sanskrit Hindi - Gunja Ratti, Ghunghchi Bengali - Kunch Marathi - Gunj Gujarathi - Chanothi Tamil Kuntumani - Telagu Farsi Guriginj - Chashme Kharosh Gu'alior - Ghumachi Punjabi - Mulati, Ratak Kashmiri - Shangir Urdu & Arab - Ain-ed-dik (seeds) Asam Liluwani, Kuntumani English - Wild liquorice,Indian liquorice, Jequirity, Beed seed tree Binomial / Botanical Name Abrus precatorius :
    • 42 Scientific classification : Kingdom Plantae Division Magnoliophyta Class Magnolipsida Order Fabales Family Fabaceae Sub family / Leguminosae Faboideae Tribe Abreae Genus Abrus Species Abrus precatorius Derivation of Name and Historical Aspects : Abrus precatorius is derived from the Crreek word Ab,rus uftich means delicate and refers to the leaflets, precatorius refers to pctitiffiiqg chosen because of the use Linnaeus in md was in rosaries. Abrus precatorirn was described by 1753 as Glycine abrus (the genus of Soya bems). ln lxi7, Adanson desoribed the genus Abrus. Much later, in 1970 ve,nlioorrt usod two 'zubspecies for the African and plants as they differed in their genus of 13-18 fruit. Abrus is a species in the family Fabaceae. Cultivation of Abrus precatorius : Abrus precatorius is propagated by seed which germinates faster if soaked in water. Cuttings of strong shoots in sand and keep darnp with plastic @nes.
    • 43 Microscopic View of Seed : Transverse section of seed shows testa about 75p thick, greater parts being formed by epidermis, composed of radially, much elongated arranged irregularly and measures 45-50 p consists of collapsed cells forming endodermis composed cells, in lengtb, inner region sf thin testa a hyaline layers bout 25 p thick, of thick walled cellulosic parenchyma, isodiametric of hemicellulose and swell cells, larger towards inside walls mainly considerably in water, outer one or two layers of cells of endodermis (pseudo epidermis) formed of rather smaller cells, walls of which swell to less extent in water. Introduction : Abrus precatorius is commonly known as Rosary pea, John crow Bead, Precatory bean, Saga tree or Giddee. The seeds are used as beads and in percussion instuments. The plant is native to Indonesia and grows in tropical and sub tropical areas of the rvorld where it has been introduced. It has a tendeircy to become weedy and invasive where it has been inhoduced- It has been a symbol of love in china and its name in chinese is xiang si dou.
    • 44 Uses: Abortifacient Anodyne Aphrodisiae Antimicrobial Diuretic Emetic Expectorant Febrifuse Hemostat Iaxative Purgative Refrigerant Sedative vermifuge paste of seed is applied locally in sciatica, stiffness of shoulder joints and paralysis. Seeds are highly poisonous. Roots are used for gonorrhoe4 jaundice, haemoglobinuric bile. Powdered seeds are said to disturb the uterine functions and prevent conception in women. The oil extracted from seeds is said to promote growth of human hair. Medicinal Properties : The roots, leaves and seeds are used in medicine. l) The roots and leaves contain glycerrhizin, the principle cmstitrmt liquorice and are used as substitute for liquorice of in cou$ md catu6al affection. (Hence the plant is known as Indian Liquorice). 2) The roots posses diuretic, tonic and emetic properties and ae usd in preparations prescribed for gonorrhoea, jaundice and haemoglobinrric bile. 3) Petroleum ether and alcoholic extracts of the roots given orally to rats 100 mg/kg per day for 1 to 5 days post coitus have been shown to prevent
    • 45 nidation by fiA %. The alcoholic extract also showed anti oestrogenic activity. 4) A decoction of the leaves is widely used for cough, cold and colic. The leafjuice is employed as a cure for horseness. 5) A paste of the seeds and of the roots of Plumbago zeylanica made with water is a stimulant dressing when applied over leucoderma patches. 6) A paste of the seeds is used as a rubaeficient in Sciatica, stiff shoulders, paralysis and other nervous diseases. 7) For the cure of alopecia, paste of seeds is rubbed on the exposed skin of scalp. 8) Ethanolic extract of the seeds inhibited the growth of micrococcus p)'ogenes, enteric and dysenteric group of microorganisms, se'€,ral other bacteria and some pathoge'nic fungi. 9) Abrin the chief constituent of the seeds, has been shrdied intensively for its antitumour activity. 10) Powered seeds are said to disturb the uterine finrc{ion md prevem conception in women. Petroleum ether extract f€rtility activity in rats. The of the seeds shwed mti- aqueous extract adversely influenced presnancy and development of the foetus in mice. The oily steroidal fraction sepratea from the seeds, when fed orally for t'*'enty consecutive days before marin& showed anti-fertility activity on albino rats and swiss mice. Injection of a single dose of this fraction on the post coital period produced 80 % sterility in rats.
    • 46 11) Extemally (paste of seeds) is fungistatic against cryptococcus neoformans. 12) Seeds are also used as abortifacient. 13) Half boiled seeds taken as tonic. crushed root to cure white eye of cattle. 14) seeds poor antihelmintic, extract cNS depressant, analygesic, uterine stimulant. 15) Plant extract one of the constituents of long acting oral contraceptives preventing implantation of fertilized ovum. Chemical Composition : The chief poisonous constituents of the seeds is 'abrin' a toxalbumin similar to ricin of castor seed. albuminosae. A Haemagglutin It has been resolved into globulin and an and a glucoside abralin are also reported. In addition the seeds contain the alkaloids bases, abrine [crzHr+ozNz mp295(decom)l hlpaphorine, Choling trigonelline, precatorine (Cr+tInNOo mp218-20) and methyl ester of N, N{im€th},ltrypo'phm mefhcation lry 272 (decomp)1. Abrine is a major alkaloid which is not conftsed wift Abri4 a toxic albuminoid product isolated from the seeds of Abrus precatorius. The leaves, stems and root also furnish these bases. 5 p-Cholanic acid is present in seeds, stigmasterol, p-sitosterol and trvo other steroidal fractions, one crystalline [CzrH:oOz mp 124] and the other oily: have been separated from the seeds.
    • 47 The seeds yield a light reddish oil (2.5 %) with the following characteristics D2a, 0.9108 ; rD 1.4702; acid value 4.8 ; sap. value 187.5 ; iod value 90.64; acetyl value 7.2 and rmsapon matter 1.3 o/o. The fatty acid composition of the oil is as follows : Palmitic 1.2; Stearic4.g;arachidic 5.4; behenic 4.6 ; lignoceric2.6; oleic 48.5 ; linoleic 13.3 and linolenic l9.S %. The amino acids presents in the seeds are (9/169 N) : aspartic 10.60 threonine 3.87 ; glycine 1.28; valine 5.95 ; methionine ; l.l1 ; leucine 7.20; tyrosine 5.15 ; arginine 15.77 ; phenylalanine 6.80 ; lysine 3.13 and histidine 2.77. Shell of seeds contain a red colouring matter of the seed coat contains a monoglycoside anthocyanin, abranin other anthocyanins identified are celphinidin - 3, 5 diglucoside and cyanidin - 3 - glucoside. The presence of gallic acid is also reported. The powdered root contains precol lCezHazOs mp 305-061, glycemhizn (lsV) [C37H76Oa, mp 78-80], abrol and two alkaloids viz. abrasine [C sH2 N3 O 3 mp 218 -20] and precasine (as picrate mp 23 4-326). 1 1 The leaves contain glyce'rrizin (9.6yA a saturatod alcobol" a crystalline compound [CrsHzoOs mp 280 (decomp.)J and pinitol.
    • 48 ABRIN chief poisonous constituent of abrus precatorius is abrin which is a toxalbumin similar to recin of caster seeds. It is a powerfirl irritant produces oedema ecchymoses at the site of inoculation. Abrin resolved into a globin and an albuminose, both of which are poisonous and are inactivated by heat. seeds contains toxic protein Abrin and non toxic proteins abrus agglutinin. Both the toxin and the agglutinin are referred to as lectins. More than one variety of Abrin with haemaagglutinating activity and binding affinity for D-galactose with or without such properties have been obtain. Abrin is highly toxic protein (LD 50-0.029 mgkg body weight of mice) present to the extent of 0.15 o/o in the seed. It consists of Abrus agglutinin and toxic lectins. Abrin [a] to [d] are the fine toxic glycoproteins found in the seeds (Budavari 1989). Five glycoproteins have been purified from the seeds. They are abrus agglutinin (a haemagglutinin) and toxic principles abrin [a] to [d]. Abrus agglutinin is tetramer with a molecular weight of 134900. It is non toric to admals cell and a pote,nt haemagglutinator. Abdns ta] fuough tdl (mol.wt. 63000 - 67000) are composed of two disulphide linked polypeptide chains. The larger sub unit *'hich is a neutral-B chain has a mol. weight of approximately 35000. The other sub unit an acidic A chain has molecular weight of approximately 30000. a
    • 49 Pure abrin is a yellowish white amorphous powder. The toxic portion is heat stable to incubation at 600 for 30 minutes. At 80o most of the toxicity is lost in 30 minutes. (Budavari 1989) Abrin is soluble in sodium Chloride solution usually with turbidity. It is also soluble in glycerin when taken by mouth, gastric juice has some inactivating action on it. Abrin has been studied inte,nsively for its antitumour activity. In experiments conducted on mice, abrin suppressed Enrlich ascitis fumour growth. Intraperitoneal injection of 20 ugkg for 3 days after tumour inoculation destroyed tumour growth as evidenced by decrease in the mouse weight and absence of tumour cells in the peritoneal cavity after 5Odays. The protein extract of the seeds has also been shown to exhibit antitumour activity on Yoshita Sarcoma (solid and ascitis form) in rats and on fibrosarcoma in mice. The extract had a direct cytotoxic effects on the tumour cells. The tumour cells incubated with the extract showed cellular pathology, decreased viable cells counts and prolongation of survival periods transplanted animals. Two toxic and possibly neoplasm abrin A and abrin C have been isolated from the drid Iodinated abrin is seen to be less toxic in rats. of the tumour - inhibitory proteins seeds.
    • 50 Signs and Syinptoms of Toxicity: when taken orally, there are signs and symptoms of severe gastrointestinal irritation with nausea, vomitings and diarrhoea which often bears evidence of intemal haemorrhage. 'lhere rnay be flushing of face, rapid pulse, full of blood pressure, spsf,, convulsion, muscular weakness, dilated pupils, trenors, tetanic coma and death due to circulatory failure. In some cases there rnay be hallucinations. In a case of atternpted suicide where the powdered seeds precatorius were taken with groundnut of abrus oil, the symptoms were vomiting, feeble pulse, cold clammy skin and sunken eyes with normal pupils. No deep sleep, no tingling of the skin or throat, no convulsions, twitching or delirium was noticed. In a few hours after an extract of the seeds injected under of the skin of an animal, inflammation, edema and possibly necrosis surrounding the site of the injection occur. The animal is disinclined to take food and 3 to 4 days after it drops down and is unable to move. It then gets tetanic convulsions and dies in 24 to 48 hours. The symptoms are very much similar to viperine snake poisoning. Hence the peasants think that the animal died from the effects of a snake bite. In hurnan poisoning a painful swelling with ecchyrrosis occurs near the site of injection. The srvelling rapidly increases and inflammation and necrosis supervene. 'Ihe patient suffers from faintness, vertigo, vomiting, dyspnoea and general prostration with cold clammy skin, small frequent
    • 5l irregular pulse, hemolysis, oligouria and uraemia. convulsions may precede death which occurs from cardiac paralysis within 3 to 5 days. Abrin exerts its toxic action by attaching itself to the cell mernbrane. Abrin's toxic effect is due to its direct action on the parenchymal cells (eg. liver and kidney cells) and red blood cells. Necrotising action of toxin causes liver damage. At the cellular level abrin inhibits protein synthesis, there by causing cell death. Many of the features observed in abrin poisoning can be explained by abrin induced endothelial cell damage which causes an increase in capillary permeability with consequent fluid and protein leakage and tissue oedema. (So called mascular leak syndrome). when the seeds are swallowed raw or after cooking, they are not poisonous. Fatal Dose : One or two crushed seeds for an av€rage adult. of about 0.001 mg to 0.002 mg of abrin per kg body weight Doses injected subcutaneously are said to be poisonous. Thirty mg of the porvdered seeds rubbed up with l0 minims of distilled water and injecte.d subcutaneously into cats killed the,m in Fatal Period lg rtto4o hogrs. : The average fatal period is 3 to 5 days. The shortest is 24 hours.
    • 52 Treatment: If anti abrin is available that should be used (anti abrin can be produced by repeated, small and gradually increasing doses which can be used curatively in Abrus poisoning). However, in any case, treatment is synptomatic stomach perfonned and demulcents are grven, if wash is poisoned by mouth. Spikes are taken out from the site of parental injection. calcium gluconate is gven to combat tetany. circulation is maintained and other symptomatic treatment glven. Post Morteum Appearance : when taken by rnouth, there may not be any remarkable external sign, when used in the form of spikg then the site of injection will be swollen, inflamed and necrosed w'ith presence of spike in the tiszue and the site of ksion. Intemally when taken by moutlr, the mucus membrane of the stomach md intestine is highly congested with numercus hae,morrtagic patches on its srface as well as in the interior of the organs such as lrrngs, liver and spleen.
    • 53 Medico-Legal Points : Most of the poisoning cases are accidental in children who being athacted by the colour may chew seeds. Homicidal poisoning occurs mostly through parenteral route. For this, the seeds are crushed to powder. This alone of mixed with datura, opium and onion is rnade to paste. Spikes of lmgth of about 1.5 cm, weighing about 100 mg each, commonly known as *Sui" are made out of the paste and dried under the sun. These spikes are held in between fingers and the victim is forcefully slapped to insert spikes in the flash. The spikes are also used to kill cattles. For this the spikes are fixed on rvooden frames and the animal is forcefully hit on the buttock to inject the spikes in its body. crushed seeds are often taken to commit suicide. Abnrs seed paste is used as alrow abortion. Seeds dust poison. This has also beeir usod locally to cause is used by malingeres to produce conjrmctivitis. This may cause permanent damages of the eyes. OTHER STUDY REFERENCE In the study by M Pugalenthi, et.al.(2007) It is shown that fte maftrre seeds were found to contain 21.99% of protein, 8.9% of lipid 6.a% of fibs, of ash and 57.3% of carbohydrates. Although, various antinrtritional compounds were present in the raw seeds, the autoclaving tneah€nt 5.3o/o effectively reduced their maximum levels without affecting the nutritional value of Abrus precatorius seeds. When considering the biological value, the rats fed on autoclaved seeds exhibit better growth pe,rformance zuch as higher feed intake (FI) (2399) and body weight gain (BWG) (67.a9. Moreover, the
    • 54 protein quality parameters such as protein efficiency ratio @ER), true digestibility (TD), biological value (BV), net protein utilization (Npu) and utilizable proteins (UP) of Abrus precatorius seeds were also significantly improved by autoclaving treatment when compared to other processing methods such as soaking, cooking and roasting. LAW RELATING TO POISONS A poison is commonly defined as a substance which, when administered, inhaled or swallowed, is capable of acting deleteriously on the body. Thus, almost anything is a poison. There is really no boundary betwen a medicine and a poison for a medicine in atoxic dose is a poison and a poison in a small dose may be a medicine. In law, the real difference between a medicine and a poison is the intent with which it is given. If the substance is given with the intention to save life, it is a medicine but if it is given wift trc intention to cause bodily harm, it is a poison. Section zB4 rpc lap down lhe punishment for careless handling of poisonous substances, while section 299, 302, 304A, 306, 307, 309, 326, 328 and 498A deal with offe,nces relating to administration of such substances.
    • 55 FOSSESSTON, SALE AND CONTROL OF POTSONS (Revised by Dr. B.N. Mattoo, Director, Matrarashtra State, Bombay, and Dr. Forensic science l,aboratories, c.K. Parikh, Medicolegal conzultan! Bombay). These are governed by the Poisons Act 1919, the Drugs Act 1940 the Drugs and Magic Remedies (objectionable Substances Act 1985, and the PharmacyAct 1948) of 1919) is designed to regulate the importation, possession and sale of poisons. It empowers State Governments to make rules The Poisons Act (12 regrrlating these matters in their own States. In Maharashtra these rules have been framed in the year 1972 (Maharashtra Poisons Rules, 1972), and these include a Schedule grving a list of poisons, class A and class B, covered by fte Poisons Act. Class A poisons generally are those which have medicinal use while Class B poisons do not have anymedicinal use. The Drugs and cosmetics Act 1940, cosmetic means and article intended to be rubbed, poured, sprinkled or sprayed on, or introduced into, or otherwise applied to, the human body or any part there of for cleansing beautifying, pomoting attractiveness or altering the appearance and includes any article intended for use as a compone,nt of cosmetic but does not include soap. The Act has been further ame,nded by Drugs (Amendment) Act" 1964 (13 of 1964) to include Ayurvedic and unani drugs. The Act provides stringent punishment in respect of offences concerned with dnrg adulteration.
    • 56 schedule *SCHEDULE H and L drugs are required to be labelled with the words H DRUG" and *SCHEDULE L DRUG", Warning - To be sold by retail on the prescription of a Registered Medical Practitioner only. The prescription shall mention the name and eddress heatment of the person for whose it is given. Any such prescription must be compounded in the authorised premises (chemist's shop) only by or under the supervision of a qualified compounder and must not be dispensed more than once unless specifically asked for. At the time of dispensing, a note should be made of the name and addressed of the patient, the date on which the prescription is dispensed, the serial number under which it is entered in the register (in case of poisons) the batch number, and the date of expiry of potency. In addition, the poisons are subject to the following regulations : (1) A person must possess a licence to stock, sell or distribute poisons listed zupply in the rules. (2) The of listed poisons must be entered in a register which must be maintained for this purpose. (3) These poisons must be kept in authorised premises only in a cupboard or drawer reserved solely for this purpose and to which customers are not permitted to have access. (a) They must be kept in leak proof containers labelled with the word *poison" Certain poisons are in red le{ters. not allowed to be k€pt in more tbm fte concentration while others have to be coloured. (5) given
    • II/TATERTALAND METHODS
    • 57 MATERIAL AND METHODS It involves : Al Pharmaceutical Study Bl Analyical Srudy Cl Toxicity in Vitro Study PHARMACEUTICAL STUDY It includes the following procedures. 1) 2) 1) ' Selection of the drug (Gunja) ShodhanaofGunjaseeds Selection of the drug (Gunja) : Fully developed seeds of Rakta Gunja with black spot and shwet Gunja were selected for the complete course of the study. r r ' Both types of 500 gm Gunja seeds were purchased from the markef. Foreign matters were removed and @arse powder was prepaod in grind€r. Four groups each weighing 125 gm of Shwet md RatG ftnja beej powder were made. 2) Shodhana ofGunja seeds: r Among eight groups, one group each of Shwet and Rakta Grmja soed was kept untreated as Ashuddha sample. ' Remaining six groups of sample were treated separately with Godugddha (cow's milk), Kanji and Distilled water for Shodhana.
    • 58 Shodhana with Godugddha r : Two litres fresh, unboiled and untreated milk of Indian breed variety of cow was taken. ' 125 gm coarse powder of Ral:ta Gunja beej was taken separately in a piece of cotton cloth and tied with cotton thread tiStly to form Pottali (Round Bolus). I one litre cow milk was taken in an earthen pot md the pottali was hanged with thread to glass rod kept horizontally on tbe pd in pottali immersed in milk was not in contact ' I wiftbffi This unit named Dolayantra was then kryt on low cow milk was added further srfi a way that the ofthe euthen pol flme for sir hours. as and wb€n requir€d fu svedaa of Rakta Gunja Beej. ' After six hours, Pottali wa-s takelr out of flolaymtra and ftnja Bej were under shade. I After drying sufficiently, weight was taken and powder Same procedure was repeated Shodhana with Kanji henaration of Kanii r wc pepued- with Ashuddha Shwet Gunjapowder. : : various methods of preparation of Kanji are mentioned in Ayurrredic .literature. One of them was used for Shodhana of Gunja seeds. . 250 gm of Shashtishali Rice was taken and cooked to make r It was smashed and kept in an earthen pot. it soft.
    • 59 r water was added to it in quantity three times to the total quantiV of cooked rice. ' It was well mixed and the mouth of pot was closed with piece of cotton cloth,tightly and kept undisturbed in . a dark place for ten days. After ten days, on passing all the Ayurvedic tests of preparation, Kanji was used for Shodhana. IFFqrqqF{wusrR €frt' a.rfufi ftg' r Sharangdhar Samhita Madhyam Khanda l0/lz Shodhana: ' 125 gm coarse powder of Rakta Gunja beej was taken and pottali was prepared. r Swedan hours. r ' r of Gunja seeds was done Dolayantra using Kanji for three ' After three hours, seeds were dried under shade. Powder was pre,pared after measuring the weigfot of dried Gmja seeds. Same procedure was repeated Shodhana with Distiled water r in with Shwet Gunja Beej. : For comparative study, swedana of Gunja seeds was done in Dolayanha for three hours with the help of Distilled water. . For this, distillation of water was done in the laboratory. r After three hours, Gunja after weighing the seeds. seeds were dried and powdered was prepared
    • 60 ANALYTICAL STT]DY It includes : l) Physio-chemical Analysis 2) Protein Assay 3) Qualitative Test for Amino acids 4) Thin Layer Chromatography f) Physio-chemical Analysis Analytical tests like total ash value, water soluble ash value, acid insoluble ash value, Aqueous extract value, Alcohol extract value, pH value md conductivity of all the Shodhit and Ashuddha samples of Shwet and Rakta Gunja seeds were done in the laboratory. r) I Clean and well dried empty crucible was weighed. . 2.5 gm coarse powder of Gunja seed was taken into the cnrcible. r The crucible was kept on low flame burner Total Ash Value : till the whole powder of Gunja seed tumed into the ash. I Then cool and weigh it. ' The same procedure was repeated ' observations were noted and calculation was done samples. till the constant wei$t was obtained. for all the eight
    • 6T Formula: Total Ash value ino/o: - weight of empty srucible crucible Constant weight of ash with W"tght : (W? "f r"-ptdt" x 100 gut) - W1l x 100 % Calculation: Total Ash value inYo: (.16.5282 - 16.4340) x 100 2.4308 : 0.0942 x 100 2.4308 : 3.88%o Result: Total ash value of sample I, II, 0.64 yo, 1.27 yo,2.04 yo,2.26 oh,2.4 oA, ilI, fV, V, VI, VII and VIII is 3.88 %, l.l4 yo, l.l8 % respectively. b) Water soluble ash value : . Weight of clean and dry empty cintered glass crucible was taken. r The total ash prepared in transferred into a 100 ml beaker. Then add 25 ml distilled water and then is boiled for 15 minutes. ' Filteration was done with suction pump. Cint€red glass crucible was then dried into oven at 450oC for two hours. r It was then cooled into desicator for half an hour and then weigfot was taken. I Theprocess was repeatedtill constant obtained. I Observation were noted and calculation was done.
    • 62 Formula: Total Ash Water soluble Ash value - Water soluble Ash Wt. of sample (%): Value : (Wr=_Sld_ x value x 100 N, V, VL VII and 100 w, Water Insoluble Ash : ash weight of crucible with residual W: : Wz-Wr Wr : 30.3288 weight of empty crucible Calculation: Water soluble Ash value (%) : -30.2074:0.1214 : 0.1686 - 0.1214 x 100 2"43A8 0.472 x 100 2.4308 = 1.94%o Result: Water soluble ash value of sample I, II, UI, VIII is 2.95 oh,0.080 ,0.62Vo,0.49 yo,2.O5 oA, 1.95 oh,0.24 yo,0.68 % respectively. c) Acid Insoluble Ash value : Procedure: ' weighed accurately cintered glass crucible with water soluble ash. ' Added 50 ml of dilute HCI and boiled it on burner for 15-20 minutes. ' cooled and washed pump. it with distilled water and sucked by using suction
    • 63 r It rn'as dried in oven and then cooled in desicator. weight was takeir. I Repeated the procedure till constant weight was obtained. Formula: Weight of cintered - weight of cintered % of Acid Insoluble: glass crucible glass crucible after Ash water insoluble ash filteration of with value W"tght : (Wr-Wd "f x m-ptdi" HCI x 100 gr") 100 w. Calculation: '/oof Acid Insoluble : 30.2110 -30.2074 x for Ash value Sample I 100 2.4308 : 0.0036 x 100 2.4308 : 0.15 Yo Result: Acid Insoluble ash value of sample I, II, UI, fV, V, VI, VII and VIII is O.l5 aA, 0.5 yo, 0.04 yo, 0.02 yo, O.08 yo, 0.12 yo, 0.04 % and 0.12 respectively. O Aqueous Extract value : . 5 gm Gunja beej powder was taken into dry and clean conical flask. ' r chloroform water was added and mouth of the flask was closed tightly. It was shaked for six hours and then kept undisturbed for eighteen hours. %o
    • 64 r Then filteration of the supernatant was done and collected into the beaker. I Weight of empty and dry china dish was taken. ' 25 ml extract was taken into the weighed china dish and kept over hot water bath r till all the exhact evaporated. The China dish was kept into oven for two hours. It was then cooked for half an hour into the desicator and weight was taken. r This process was repeated . Observation were noted and calculation was done. ' pH and conductivity of the exhact was examined and observations were till constant weight was obtained. noted. Formula: extract : (%) Aqueous value of Constant weight - Weight of empty china dish with extract china x Weight of sample (in gn) dish 4 x 100 : (Wz-Wr)x4 xl00 w, tCrlculation : Aqueous extract value Sample l(in%) of : : (0.1902) x 5 gm 4 x 100 15.22Yo Rcsult: The Aqueous extract value for sample I, IL III, IV, V, VL Vtr and i! VIII 15.22yo, 8.08yo, 8.32yo, 30.52yo, l4.6gyo, 6.99%,,, B.3Syo, 2l.g% rcspectively.
    • 65 c) Alcohol Extract value : kocedure: I 5 gm Gunja beej powder was taken into conical flask and alcohol was added. I It was then shaked for six hours and kept undisturbed for eighteen hours. r Filteration was done and 25 ml extract was collected into the weighed China dish. r It was evaporated on hot water bath and was kept into oven for two hours. I Then it was cooled into desicator for half an hour and weight was taken. ' Same procedure of heating, cooling and weighing was repeated till constant weight obtained. r Observations were noted and calculation was done. Formula: Alcoholextract value (%) : Constant weight of - Weight of empty chinadish x4 chinadishwithextract Weight of sample (in gn) : (Wz-Wr)x4 xl00 % Celculation: Alcohol extract value Sample I of : (in% (.70.2720 -70.2358) 5 : 0.0372x4 x 100 5 : 2.98 o/o x4 x 100 x 100
    • 66 frult: The Alcohol extract value for sample I, II, III, IV, b2.98%o,1.70h,6.64yo,l.50 ,4.3gyo,0.ggoh,0.gg%, and vIII and S.S}yorespectively. plf value : o a v, vI, vII pH of aqueous extract of all the eight samples was taken by using pH meter. fccult: The pH value for sample I, II, ilI, IV, V, VI, VII 5-83 yo,5.12yo,4.86 yo, 5.61 oh,5.5 yo,5.g7 d r % and 4.72 0/o and VIII is 4.g3 %o, respectively. Conductivity: conductivity of aqueous extract of all the eight samples was taken by using conductivity meter. Result: Conductivity of sample I, II, III, N, V, VI, VII and VIII is l0830pmoles, 3705pmoles, 4503pmoles, r3965pmoles, l0260pmoles, 2850pmoles, 3 I 3 5pmoles and I 3395pmoles respectively.
    • GUNJA (Abrus precatorias) RAKTA GT]NJA GT]NJA SI{WET GT]NJA GT]NJA SIIODHANA SAMPLESAFTER SHODIIANA -i.,2 3 tl $s $t {111 AQUEOUS EXTRACTS Rakta Gunja Shwet Gunja ALCOHOLIC EXTRACTS Rakta Gunja Shwet Grmja
    • 67 PROTEIN ASSAY : Gunja Beej (Abrus precatorius) contains Abrin which is a protein and toxic principle. Hence protein assay was done to estimate the percentage of protein contained in Ashuddha and Shodhit samples of Gmja Beej. Requirements: 1) Tris HCI Buffer a) : Solution A : 0.5 M solution ofTris (0-1513 gE) + 25 ml Distilled water. b) SolutionB: 0.5NHCI 'Iris HCI Buffer:25ml soldimA+m.?mlffiB Dihfrd to 100 nl Distillod 2) Reagent A -.* wc{fl : ?.4) : 2 % Sodium Carbonate in 0.lN NaOH. 3) Reagent B : 0.5 % Copper Sulphate 4) inl Yo Potassium SodiumTuffi. Reagent C : Along with biureate reaction i.e. protein reacts wift alHire cupric titrate. Alkaline CuSO+ solution:> 50 ml Reagent A + I rnl ReagctrB 5) Reagent D : Folin Ciocalteau's reagent (diluted with distilled wil€r itr fte ratioof 1: l)
    • 68 6) Stalk Standard (Protein Solution) : 50 mg Bovine Serum Albumin in Distilld water made upto 50m1. 7) Working Standard : Dilute 5 ml of Stalk Solution with 25 ml of Distilled water. Procedure: . 100 mg of Ashuddha Rakta and Shwet Gunja and 500 mg of Shodhit Gunja B-eej powder was taken and triturated with 10 ml Tris HCI buffer solution. i Centrifusion was done and supernatant was separated. . 5 ml supernatant was taken and I rnl Trichloroacetic acid (TCA) was added, then centrifused and collected the precipitate. t ) ml NaOH was added into the precipitate, then centrifused and supernatant was collected. . Pipetted out 0.2 ml of extracted sample I and V and 0.5 ml each for other samples. . 0.1, 0.2,0.4,0.6 ml of working standmd was pipefted out. r The volume of all the samples was made to I ml by adding . Blank solution was prepared by adding I ml distilled wder in a test tube. ' 5 ml Reagent C was added in all the extracted samples, worrting standard and Blank sample. distilld wat€f,.
    • 69 After 10 minutes, 0.5 ml Reagent D was added in all the sarrrples and placed in a dark place for half an hour. Spechophotometer was set at 660 nm with Blank sample and optical density of all the samples was measured. Observations were noted and calculation was done. Formula: : Total protein 7o (For Ashuddha Ralcta and Shwet Gunja samples) OD x 462.5712x4x2 Where, OD: Optical density measured with spechophotometer 462.5712: Value calculated from standard graph. O.2 : Quantity of extracted sample in ml. Others are constants obtained during procedure. Calculations: o/o : Total protein (For Ashuddha Rakta Gunja sample) 4.76o/o o/o Total protein = (For Ashuddha Shwet Gunja samples) 0.274x462.5712x4x2 0.2x0-lxld 5.07 o/o
    • 70 Formula: protein : Shodhit Total for All OD x 462.5712 x 2 0.5 x 0.5 x 104 samples (%) : OD x 462.5712 x2 1250 Calculations: protein : (%) : Total for D/IV SRG protein : (%) : Total for Kanji SRG for : (%) : Total protein Godugddha SRG protein : (%) : Total for DAM SSG protein : (%) : Total for Kanji SSG for : (%) : Total protein Godugddha SSG 0.131 x 462.5712 x2 l2s0 0.097 % 0.045 x 462.5712 1250 x2 0.033 % 0.161x462.5712x2 1250 0.119 % 0.164 x 462.5712 x 2 1250 0.121% 0.015 x462.5712x2 1250 0.011 % 0.234x462.5712x2 1250 0.173 %
    • 7t 0F against concentration of Frotein 0.27 824 F 0.21 E 018 t o1b E 012 v ot o'ffi 0.06 003 B 4U 60 FrutHn ffg!
    • 72 aua[tative Test for Presence of Amino Acids: This test was done to detect the prese,nce of nnino acids in the extracted samples of Ashuddha and Shodhit Rakta and Shwet Gmja Beej. Procedure: . 0.1 ml. extract of the sample prepared for protein assaywas tak€o- . I ml Ninhydrin Reagent (Ninhydrin 15 mg n-Brffiol 5 ml ild uic acid 0.15 ml) was mixed. I After heating for 2 seconds, Blue colour develo@ in varying intensity except in extact of Distillod ydcr dl fu smpl€s wi6 Sbft n*h md Shwet Gunja Beej. The colours observed in all the samples are as Sample follom: Colourobserned I Dark Blue il Colourless ilI Very Faint Blue ry Faint Blue V Dark Blue VI Colourless VII Very Faint Blue VIII Faint Blue This observations show that amino acids are present in less quantity in Shodhit samples as compared to Ashuddha samples of Gunja Beej.
    • 73 Thin Layer Chromatography (TLC) Preparation of Samples for TLC Sample I Sample : II 500 mg powder of Ashuddha Rakta Gunja Beej was taken. : 500 mg powder of Distilled water Shodhit Rakta Gunja Beej was taken. Sample III : 500 mg powder of Kanji Shodhit Rakta Gunja Beej was Sample IV : 500 mg powder of Godugddha Shodhit Rakta Gmja Bcej taka- was taken. Sample V : Sample VI : 500 mg powder of Ashuddha Shwet Gunja Beej was mkeo500 mg powder of Distilled water Shodhit Shwet Cnmja Beej was taken. : Sample VII Sample VIII : 500 mg powder of Kanji Shodhit Shwet G4ia Beej sa tdrco- 500 mg powder of Godugddha Shodbit Shwcr @i!Bod was taken. Extraction Procedure : Powder of Gunja Beej was triturated with get homogenized. Then l0 ml of 80 % Elhmol till it it was cenfrifused and the supernatmt was seprmed- The extract was spotted on Silica gel TLC plate and widi nm using following solvent system. l) Butanol 2) Methanol 3) Acetic acid 4) Distilled water
    • 74 Proportion : 3"2 : 0"8 : 0.2 :0.8 (B : M : Aa : W) The plate was dried after running and obsernations were noted after spraying iodine for 2 minutes. Table No. 1 : Showing RF values with Iodine vqxxrrs R.F. + Value 0.08 0.21 0.34 0.s6 0.67 o.75 0.9 I VF * Brown DB Brown + Brown il VF * VF VF VF * Brown m VF * VF VF VF * VF ry VF * VF VF VF * VF V VF VF VF VDB Brown Brown Brown VI VF !'F VF VF VF F VF VII VF VF VF VF VF F VF VIII VF VF VF VF VF F VF Samples t I Note: VD - Very Dark VDB - Very Dark Brown F - Faint VF Very Faint * Absent . The intensity of the spots is in comparison with Sample I and Sample V which are of Ashuddha Rakta and Shwet Gunja Beej respectively.
    • 75 Obsen'ations of Table No. 1 : (RF Value rvith Iodine Vapours) l) Five spots rvere found with extract of Ashuddha Rakta Gunja Beej (Sample I). Same spots were found in sample [, m and IV but they were very faint in intensity (nearly negligible). The Brown spot at Rf value 0.9 appeared in sample I and II but was very faint in sample Itr and IV. 2) Seven spots rvere found with extract of Ashuddha Shwet Gunja Beej (Sample V) But the spot at Rf value 0.56,0.67,0.75 and 0.9 were very faint in samples VI, VII and VIII. 3) The spots appeared at Rf value 0.21 and 0.75 in sample V, VI, VII and VIII were absent in samples I, II, III excess quantity and IV. This may be due to the of extract applied. (10 pL in Rakta Gunja samples where as 15 pL in Shwet Gunja samples) Results: Spots found with Ashuddha Rakta and Shwet Gunja samples were very faint (nearly negligible) in sample treated with Distiled water. Kmji Godugddha Shodhit Rakta and Shwet ed Gunja This chaNUB oocurred drr to Shodhana Samskar. The toxic principle has bee,n rcducd due to Shodhma Samskar.
    • 76 Table No.2 : Showing RF values after dipping in Ninhydrin Reagent R.F. Value 0.03 0.15 0.28 Orange VF Purple Purple 0.41 0.53 Samnlesl I WF Purple Intense Dark Orange II * * + * VF Orange ru VF * VF + VF Orange + VF Orange VVF Puple Intense Dark Pqple Orange ry :r * VF Purple V Orange VF Purple Purpte Orange VI :f * VF * VF VU :r * F a VF VIII * * F * VF Note: F - VF + - Faint Very Faint Absent WF Very Very Faint . The intensity of the spots is in comparison with Sample I and Sample V which are of Ashuddha Rakta and Shwet Gunja Beej respectively.
    • 77 ' spot with Rf value 0.53 in sample I appears very dark in the center with orange border because the plate was initially kept in iodine vapours. The plate without iodine treatrnent produces intense dark orange colour as shown in the plate treated with Ninhydrin reagent. ' l0 pL extract of Rakta Gunja samples was applied where as in the plate of Shwet Gunja, 15 pL of extract was applied. ' Spots appearing above Rf value 0.53 are not reactive to Ninhydrin as seen in plate treated with iodine. But they appeared here due to earlier treatrnent with iodine. I Spots appearing initially due to iodirre above Rf value 0.53 disappeared on standing for some time as the intensity was initially was very poor. Observations of Table No.2 : (RF Value after dipping in Ninhydrin Reagent) l) Five spots were found with extract of Ashuddha Rakta Gunja Beej (Sample I and V respectively). 2) The spots at Rf value 0.03, 0.15, 0.28 and 0.41 in Sample I were abseirt in Sample II where as the spot at Rf value 0.53 which was intense dark orange appeared very faint orange in colour because of Shodhana with Distilled water. 3) The spots appeared negligible) due respectively. to in Sample III and Shodhana Samskar fV u'ere very faint (nearly with Kanji and Godugddha
    • 78 {) The spot found at Rf value 0.03, 0.15 and 0.41 absent t in Sample V were in Sample VI, VII and VIII. The spots found at Rf value 0.28 and 0.53 in Sample V were very faint in Sample VI, VII and VIII. 6) The purple spots at Rf value 0.15, 0.28 and 0.41 in Sample I and V of Ashuddha Rakta and Shwet Gunja Beej respectively were of amino acids which were absent or very negligible VII and VIII. in Smple tr, ilL IV, VI, The absence of amino acid spots was due to Shodhana Samskar with Distilled water, Kanji and Godugddha Results : t) The Spots found with Ashuddha Rakta and Shwet Gunja samples were absent or very faint (nearly negligible) due to Shodhana Samskar with Distilled water, Kanji and Godugddha. 2) These three media principle. of Shodhana have destroyed the proteinous toxic
    • THIN LAYER CHROMATOGRAPITY After spraying with Afterdipping in Ninhydrin Reagent Iodine vapours After dipping in Running of Ninhydrin Reagent TLC plate Mff I tr mry vvlvllvm Spectrophotometer II Itr trI TV VVIVIIVII gample I Samples Extracted for Protein Assay IIMryVVIVIr V VI VII Qualitative Test for Presence of Amino Acid IImIVvvr V VI VII VItr VIII
    • 79 TOXICITY IN VITRO STI}DY AGGLUTINATION REACTION Definition of Agglutination : ftGtdlthdffi Agglutination is the collection of separate particles into clumps or masses. -'A' Agglutinat ion < -0"' Agglutinogen + alpha agglutinin Agglutinogen * beta agglutinin Agglutination occurs when an agglutinogen is mirGil fl' ill corresponding agglutinin (Isoagglutin)' Blood grouping is done on the basis of agglutination ofRBCr" Selection of the Candidates : Before performing this experiment, we selected the ffi S differentbloodgroq)(A,B,ABandORhpositive)'Th-!nr! laffiil' h Rh +ve) and cc* candidates of each blood goup (A, B, AB and O investigations like TLC, DLC was done in pathology eachofbloodgroupARhnegativeandBRhnegativeClbtdffiL limib number) whose bloo<l investigations were within normal the study. wUilnrh
    • 80 Procedure : ' Fresh extracts of all the eight samples were prepared separately by taking 500 mg powder of the drug and 3 ml of distilled water. . Every day fresh extract was used. . After trituration of the drug in distilled water, it was oennifused for 15-20 minutes. . The supernatant was separated and used for lbe Ody- Sample: I - Extract of AshuddhaRaktacunnrpomda(ARG) U - Exhact of Distilled water.Shodhit Ralda GrtsPmler (D^M sRG) m - Extract of Kanji Shodhit Rakta Gunja powder ry - Extract of Godugddha Shodhit Rakta Gunjapowder (Kmji SRG) (Godugddha SRG) V - Extract of Ashuddha Shwet Gunja powder (ASG) VI - Extract of Distilled water Shodhit Shwet Gunjapowda (D/w ssc) VII - Extract of Kanji Shodhit Shwet Gunja powder (Kmti SSG) VIII - Extract of Godugddha Shodhit Shwet Gunjapowder (Godugddha SSG)
    • 81 . 0.5 ml intravenous blood sample of candidate of blood goup A Rh +ve was taken" r Four blood spots were taken on two slides into which one drop of distilled water was added by dropper. . Then one drop extract of sample I was added to blood spot No. I and mixed with blunt end capillary immediately md time of qilication was noted. r Same procedure was repeated with blood spot No. Sample . L m md tV with II, III and IV respectively. mmy Observations and changes in blood sample spots were notod for minute. Photographs w€re taken at the end of ten minutes and twenty minutes. r Again four blood spots numbering V, VL VII another two slides and extracts of sample V, VI, VII and VIII were and VIII were taken on rylied to them respectively. . Time and observations were noted and photographs were taken- . Similar procedure was carried out with all the samples of blood gfoup A, B, AB, O Rh +ve and A and B Rh Negative. I In this way, three candidates of each blood goup positive) and one candidate each of group A (A g, AB and O p& P& Negative and Negative (total fourteen candidates) were included in the study. B F.h
    • oBsEI?mr:r.0tr &
    • 82 OBSERVATIONS AND RESTJLTS The colours of the samples observed after Shofu of Rakta and Shwet Gunja beej with various methods are as follows : Table No. I : Showing Colour of thc Sample $r+ltr k I Ashuddha Rakta Gunja powder(ARG) Y&dtrd II DAM Shodhit Rakta Gunja powder(SRG) Hdirtgny UI Kanji Shodhit Rakta Grmja powder Illditrgny IV Godugdhha Shodhit Rakta Gunja powder BrwnifiHrt V Ashuddha Shwet Gunja powder (ASG) Faid yE$or VI DAM Shodhit Shwet Gunjapowder whitishrrlh VII Kanji Shodhit Shwet Gunjapowder Whiti$tcilou VIII Godugdhha Shodhit Shwet Gunja powder Cremibhfdbr
    • 83 TableNo.2 Showing % Wcight Loss in samplcs after shodhan pmccss Sample % Weight Loss after Shodhaoa I ARG u D/W SRG l6 m Kanji SRG 16 rV Godugddha SRG 00 V ASG VI DAil VII Kaqii SSc l5 VIII Godugddha SSG 00 SSG 15 Gnaph Showing ?6 UtlblghtLctEioh 18 16 14 s o12 t jro is rfi s 6 g8 SRG ssc *6 s 4 2 0 D/WSRG KaniiSRG @rqddha D/WSSG KanjiSSG Godugdfie SRG Sempb ssc
    • E{ Table No.3 showing prr of the sampres before and after shodhan process I 4.83 II 5.38 m Kanji SRG rV 5.12 Godugddha SRG V 4.86 ASG 5.61 VI DAil ssc 5.50 VII 5.87 VIT 4.72 Graph showing pH of the sampree before and aftershodhan 7 6 5 T o, 4 3 2 1 0 KanjiSRG God€ddha ASG SRG Sampb! IARG IASG IDM/ SRG ID/VVSSG DWSSG Kdssc Gortqddlra ssc
    • 85 TableNo.4 Showing Conductivity of sempler before and after shodhan prccess Conductivity (in pmoles) Sample I ARG 10830 II DAM SRG 3745 m Kanji SRG 4503 rv Godugddha SRG 13955 V ASG 10260 VI DAM SSG 2850 VII Kanji SSG 3735 VIII Godugddha SSG 13395 Grapfi Showlng Conductivity of samplec beforc and afur shodhan prccosc 16000 ^ o 14(X)0 * rzmo E I romo i3 g 8@0 E 6000 E 4ooo o (' 2ooo 0 ARG D/WSRG l(giiSRG CoCldCta TIWSSG lG4iSSG SRG OoOngddra ssc SrAb rARG ID/WSRG BKanjiSRG trGodngddu$ IAS IIIWSSG fKaSSSG lGodr.tgddhaSSG
    • 86 Table No.5 Showing Total Ash values of the samples beforc & rftcr rhodhan pnocess Total Ash values (%) Sample I ARG 3.t8 u DAil SRG 0.4+ n Kanji SRG t.tt Iv Godugddha SRG LU v ASG z.fr VI DAH SSG LN VII Kaqii SsG l-14 VM GodugddhaSSG r.tt Gmph Showlng Totrl Ash valuec of $e eanpbc bfrtt tf 5h ptoc€.s 4.5 4 E s.s E 2.5 t3 a2 i r.s a ,3 1 0.5 0 D/WSRG KanjiSRG God4ddhe ASG SRG IyWS r(fi$ eoatgddre ssc $mpbs IARG TDrwSRG OKaniiSRG trGodugddhaSRG IASG ID/WSSG rKtf,SSG lGodtttddhaSSG
    • 87 Table No. 6 : Showing Weter Soluble Ach end vdn6 of the semples before rftcrrhodlen pnlocas Sample Water soluble AS I ARG 2.95 il DAV SRG 0.07 ilI Kanji SRG 0.62 ry Godugddha SRG 0.49 V ASG 2.05 VI DAil SSG 1.95 VII Ktrrji SSG a.24 VIII Godugddha SSG value (o/d 0.58 Gnph Showing Waler Soluble Adr value* of the samples boioro and after drodhan prccoso * 3.5 o E 2.5 E az t 1.5 5 I 61 o E .l o.s }o DWSRG Kan$Sffi Godqgddhe ASG IIWSSG l(utiSSG Godtlgrldha ssc SRG Sarpbr tARc IDltll SRG n l€ni SK g Godngddha SRG IASG I ttW S$ I l(gtt SSG r Godugddha SSG
    • 88 Table No. 7 : Showing Acid Insoluble Ash values of tte rrnphc bc{orc and after shodhan prosess gamplo Acid Insoluble Ash value O6) I ARG 0.15 u DAM SRG 0.50 n Katdi SRG 0.04 ry Godugddha SRG 0.(E V ASG 0.08 VI DAM SSG o.t2 VII Kanji SSG 0.04 VM Godugddha SSG 0.12 Graph $hoMng Acld lnsoluble Ash value of the tamples beforc and afbrshodhan prccess € 9 E o.o o.s f, o.n E E o.z Io. o.r 5o t s" auf **f g" €" "d o*d *sf 'f d"tr rARG ID/W SRG trKd1iiSRG IA,SG rDnrssc rKilii SSG trGodugddha ssG
    • 89 Table No. I: Showing Wrter Effmct valucc of thc ranples before and after shodhan proccsr WderExtrctwhs Sample I ARG t522 II DAM SRG ffi-m m Kanji SRG mjrl IV Godugddha SRG 3(t52 v ASG t{19 VI DAM SSG Itr9 vu Kanji SSG Inat VIII Godugddha SSG (%) 2[-m Graph Showing Water Efrrct vahrc of 0nr*5-- $odhen prccGr. 35 E30 lru E,o €ru H,o g5 0 D/WSRG KanjiSRG Godugddte ASG SRG IHS |(I;SSG GulryHha ssc SrrTtr IARG lD/W SRG trKanjiSRG trGodugddha SRG IASG rtlWs$ rrriSSG rGodqrldha SSG
    • 90 Table No. 9 : Showing Ncohol Extrac't values of the ramples before and rfter shodhan process Alcohol E)fiact values (7o) Sample I ARG 2.98 n DAM SRG t.70 m Kanji SRG 6.4 ry Godugddha SRG 150 V ASG 43t VI DAM SSG ott VII Kanji SSG 0.tt VUI Godugddha SSG 5.s0 Gnph Showing Alcohol Ertnctvalues of ffre samplee beforc and afur shodhan prccless 7 to o 65C E 3r I a, is |rl 62 o .Jlt 0 O/W SRG KaniiSRG eoOqtste ASG IIWSSG lGriSSG Godr8ddha ssG SRG! Srnff ID/|l/SRG BKanjiSRG trGodt{dfiaSRG rASG rt)w$ rKsiiSSG lGodtgddhatSG
    • 91 Table No. 10 : Result of Physico-chemical Analysis of Rakta Gunja Beej before and after shodhan prosess Rakta Gunjr DAM Kmji Crodugddha Shodhit Ashuddha Shodhit Shodhit 16 l6 00 4.83 5.38 s-t2 4.86 10830 3705 4503 13965 Total Ashvalue (%) 3.88 0.& 1.27 2.M Water soluble Ash value(%) 2.95 0.08 0.62 o.49 Acid Insoluble Ash value(%) 0.15 0.5 0.04 0.02 Aqueous extract value(%) t5.22 8.08 8.32 30.52 Alcohol extract value(%) 2.98 1.7 6.64 1.5 Wt. loss after Shodhana(%) PH Conductivity(pmoles)
    • 92 Table No. 1l : Result of Physico-chemical Analysis of Shwet Gunja Beej before and after shodhan process Shwet*i" DAil Kanji Godugddha Shodhit Ashuddha Shodhit Shodhit l5 15 00 5.61 5.50 5.87 4.72 10260 2850 135 t3395 Tertal Ash value (%) 2.66 2.4 r.t4 1.18 Water soluble Ash value(%) 2.05 1.95 0.24 0.68 Acid Insoluble Ash value(%) 0.08 a.t2 0.04 0.12 Aqueous extract value(%) 14.69 6.99 8.35 2t.9 Alcohol extract value(%) 4.384 0.88 0.88 5.50 Wt. loss after Shodh ana(%) PH Conductivity(pmoles) 3
    • 93 Table No. 12 : Showing Total Protein content bdorc and efter Shodhana Weight Opticat Total (7o) Loss (ms) Density Protein inhotein Conten(7o) Sample Conte,nt I ARG 100 0.257 4.760 II DAM SRG 500 0.131 0.097 97.96 m Kaqii SRG 500 0.045 0.033 99.31 rV Godugddha SRG 500 0.161 0.119 97.s0 V ASG 100 0.274 5.070 VI DAil SSG 500 0.164 0.121 97.6t VU Kanji SSG 500 0.015 0.01I 99.78 vm Godugddha SSG 500 0.234 0.173 96.s9 Graph thowing (%) Loor ln Prctaln Content 101 E too Eee c 8sB g o 897 e E90 ge5 94 D/WSRG KaniiSRG CoOugddte IyWSStt l(a{aSSG GoduCrklha ssG SRG SmCc r D,W SRG I Kanii SRG tr Godt{ddha SRG E I}rW SSG rKtnii SSG I Godugddha SSG
    • Observation And Result Of Toxicity In Vitro Study The observation and result of the toxicity in vitro study was very significant. Agglutination of RBCs found in all the blood spots treated with extract of Ashuddha Rakta and Shwet Gunja beej. This agglutination occurred in all individuals of blood group A, B, AB and O Rh +ve and A Rh -ve and B Rh - ve. Agglutination of RBCs was not found in all the sample of blood treated with extracts Gunja of distilled water, kanji and Godugddha Shodhit Rakta and Shwet Beej. The results was same in all the individuals of different blood group. The time required for agglutination of RBC's in samples treated with extract Ashuddha Rakta Gunja beej was two to four minutes while it of was less than thirty seconds in samples treated with extract of Ashuddha Shwet Gunja Beej.
    • AGGLUTINATION REACTION SAMPLE OF'A RhPOSITTYE GROT]P AT TIIE EID OT' TEN MIIYUTES AT THE EID OF' TWENTYMIIIUTES Iv SAMPLE OF B Rh POSITIVE GROUP tr ATTHE END OF' TEN MINUTES ATTM, END OF TWENTYMINUTES
    • AGGLUTINATION REACTION SA]VIPLE OT'AB Rh POSITIVE GROT]P nV AT THE EID OF' TEN MIIIUTES ATTEN.END OF TWENTYMIIIUTES IV SAMPLE OF O Rh POSITIVE GROT]P tr ATTHE EID OF TEN MIIYUTES AT flTn EIID OI' TWENTYMINUTESI
    • AGGLUTINATION REACTION SAMPLE OFARh IYEGATIVE GROT]P ATTHE EIID OF TEN MNruTES AT THE EIID OF TWENTYMIITUTES :;:^! Itr utr I VI AT TITN END OF TEN MIIIUTES AT THE END OF TWENTYMINUITS :.'naB
    • 94 DISCUSSION The excellence of Ayurvedic pharmacology is glorified with the use of toxic drugs for medicament. One such drug is studied thorougflytrrough this treatise with signifi cant observations. An analytical experimentation to determine the toxicity pofile after shodhana of Gunja seeds is supplemented with its toxicity in yito stndy on individuals of different blood groups. In the previous sections, we hane dealt with the actual experimentation along with observations and resulb obtainert The essence of this study is presented sequentially through this disrcion- Gunja, as a drug, when studied for its properties, tbrough both modem and Ayurvedic view, was found to contain some toxic princifle* undoubtedly accepted amongst "Upavisha". In It is various Ayrrvodic tcrtq Rasatarangini has specifically described the toxic slmptoms po&rced by consumption of crude drug. By the influence of Tikta Ka$aya rasa, Ufua Veerya and rooksha, tiksha guna, Gunja beej acts as a Visha- ftvitides Kryha and Pitta dosha, thereby causing vomiting, diarrhoea md otrcr iritding symptoms as described to be produced after consumption of seeds. Though Gunja was accepted as a single drug therapy for various diseased mditions and was included in different "Kalpa" eg. Gunjabhadra Ras4 it is dvised to be used after proper "shodhana Prakriya". Gunja, an ancient medicinal remedy is biologically accepted as Abrus precatorius, a species of leguminosae family. By the various complex
    • 95 analytical procedures carried out worldwide, of it is reported to contain number active ingredients with predominant toxicity of seeds by virtue of an albuminoid substance "Abrin". While observing the toxic effects produced by Gunja seeds, Abrin is considered as a potent active principle. The animal studies have explored the antitumour activity of abrin. Abrin is a highly toxic protein. It exertS its toxic effect by attaching itself to the cell membrane. Its toxic action is due to its direct effect on parenchymal cells and red blood cells. ln terms of classification, Abrus is included amongst the vegetable, organic initant poison, nevertheless its toxicity can be significantly observed with subcutaneous infilteration through the symptoms like ecchymosis, swelling, rapidly increasing inflammation followed by necrosis. The ingested poison is reported to cause the toxic symptoms as described in Ayurvedic literature. A detailed review of literary aspect of Gunja as well as Ab'rus precatorius through the whole ancie,nt and modern literafire has p'rovided ild initiative to study the ancie,nt literature with modem amlytical m dfuical tools. Toxic drug is admired as *Amril Sahachd in Ayunodic after the process of purification i.e. *Shodhma conhibuted as a unique feafure Rasatarangini, a book of Smff- fmlmiom This phrmaoother4afics gorc has in Ayunreda of twentieth century has described Grmja as a single drug regime in number of diseases after proper purification process. A basic
    • 96 therapy pertaining substances i.e. a the hlpothetical principle of purification decrease in percentage of toxic of toxic content after poper purification. The crude drug was subjected for the analytical waluation of toxic content through proper laboratory procedure. Under the expert guidance, various prescribed procedures were carried out to standardize the chemical behaviour ofpowdered Abrus seeds before and after the treatment. Taking into consideration the proteinous nature of toxic ingredient, the set up for thin layer chromatography, protein assay, qualitative test for amino acids and toxicity in Vitro study was assembled. This analytical and toxicity in vitro study before and after the detoxification of abrus seeds revealed some interesting facts. ' r After the process, nature of powdered drug remained to be coarse. Reduction of values was observed in weight of samples treated with Kanji and distilled water. The weight of samples treated with Godugddha increased to some extent because of the moisture and fat content due to boiling with cow milk. ' The determination of pH value after detoxification process revealed sonre increase though the nature of the solution remained acidic. ' Conductivity of the solutions of sample treated with kanji found to be reduced while it increased in distilld wdrr d sample tneded with Godugddha. ' Total ash, water soluble and acid insoluble ash value showed a decrease after Shodhana.
    • 97 The result of protein assay showed that the total protein content of rte Shwet and Rakta Gunja Beej has decreased significantly after shodhana Samskar. ' The Loss in total protein content of the sample ranges from 96.s9 vo to 99.78 %owhich is maximum in Kanji Shodhit Rakta and shwet Gunja Beej as compared to Distilled water and Godugddha Shodhit Shwet and Rakta Gunja Beej samples. ' This clarifies the reduction in Toxic principle after Shodhan Samskar. ' The qualitative test for amino acids showed the presence of amino acids in all the samples except sample treated with Distilled water Shodhit Rakta and Shwet Gunja Beej. In thin layer chromatographS samples extracts of .Ashuddha and shodhit of Rakta and Shwet Gunja Beej were prepared. Running of sample (dummy) plates was done to find out solvent system. After running about 15-20 plates, we got satisfactory solvent sy*em. Then by using that solvent system running of plate with all the eigftt samples was done. After satisfied running of all plates obsernations wcre akea after sprayrng with Iodine vapours and dipping in Ninhy&in rcagEdRf valoe of each spot was measured. ' Shodhana Samskar has showed lhe trmendous dnqgF wirh 6e heip of TLC. The spots found in Ashrddha sqles of Rakta md shw€r Gunja
    • 98 Beej were absent or very negligible in sample treated with Distilled water, Kanji and Godugddha. The concentration of amino acids has reduced in Shodhit samples as compared with Ashuddha samples. The observation and result of the toxicity in vitro study was very significant. Agglutination of RBCs ford in dl the blood spots treated with extract of Ashuddha Rakta and shwd Grnjabej. This agglutination occurred in all individuals of blood gloup A" B, AB md O Rh *ve and A Rh -ve and B Rh -ve. ' Agglutination of RBCs was not found in all the sryle of blood trcatod rvith extracts of distilld water, kanji and Godugddha shodhit Rakta and Shwet Gunja Beej. The results was sarl€ in all the individuals of diffe,relrt blood group. ' The time required for agglutination of RBC's in samples treated with extract of Ashuddha Rakta Gunja beej was two to four minutes while it was less than thirty seconds in samples treated with extract of Ashuddha Shwet Gunja Beej. The process of Shodhana may help to decrease the poisonous nature of the substance. Further analysis with advanced sophisticated methods will precisely conclude the contribution of this purification process to detoxi&/ Gunja seeds. Further study should also focus on procuring pure abrin sample and standardize these findings.
    • 99 SUMMARY The present thesis entitled, "chemical study of e,frect of shodhana on Toxic Principles of Gunja Beej" has been discussed in d€tdl sffiing frrom Introduction of the study, Aim and Objectives, Review of and methods used in the study. Analytical study with l^iffire,Irt*erial observrirns md rc$lts, Discussion and final section dealing with Summary md 116 ih* mdusim and Bibliography. Introduction: Toxicity of the seeds of Abrus precatorius is scientificall5rdommd in Ayurvedic texts and well elaborated by modern literature. Medicinal utility of poisonous substance is a chaactedsni: Ayurvedic pharmacopoeia and ftftc of it is accepted worldwide oday- Bd m ancestors described "shodhana Samskar" (a proc€ss of pificeilim/ detoxification) before their internal administration. However rirrrc i*rmrrae the scientific evaluation of such procedures on modern analyticd 8'wft. Extensive literary study of Ayurvedic texts gives a sfrong eyid6a63o,ftrilityo'f this drug in paralysis, Urustambha, Indralupta, Sciatic4 joint vrims q*iq c1c md diseases. Aim and Objectives : The purpose of this study was to putforth all related truths. veriff the claims of Shodhma ed to
    • lo0 Review of Literature : Literature review has been divided into i) Historical review of Agadtanha and Vish4 ii) Literary review of the drug and Visha. Historical review of Agadtantra describes about the term 'Agadtaffia' in Veda and Ayurveda. Literary revlew of Visha contains detailed description about definition and tlpes of visha. Drug review contains detailed description about Gunja (Abrus precatorius) as per Ayurveda and Modern science along with the description of Abrin which is the toxic principle of Abrus precatorius. It also contains various Shodhana procedures of Gunja. Material and Methods Coarse powder : of red and white Shodhana with the help seeded variety was subjected to of Godugddha, Kanji and distilled water. Physico- chemical analysis, Protein AssaS Qualitative Test for presence of Amino acid and Thin layer chromatography were done in the laboratory. Toxicity in Vitro study was done with extracts of Ashuddha and Shodhit Gunja Beej powder. Experimental Study : Shodhana of coarse of powder of red and white Gunja was carried qtr in Dolayantra with Godugddha distilled water for three hours. for six hours, Kanji for three hurs md
    • 101 Results of various physico-chemcial analysis like total ash value, water soluble and acid insoluble ash valug aqueorui and alcohol extract values, pH and conductivity of all the freated as well as rmtreated sarnples were obtained. To detect protein value Present in heated as well as untreated samples, protein assay was done. To identis toxic principlg drin layer chromaography was done. Toxicity in viho study was done with blood group individuals. Aggtutination of sryles of different RBCS formd in smples blood heated with extract of Ashuddha Rakta and Shwet Gunja boej where as no agglrtination of RBCs was found in samples treated with extract of Godugddha Kanji md distilled water shodhit Rakta and Shwet Gunja. Discussion : Probable mechanism of action and the results obtained are elaborated in a chapter of discussion.
    • coNcLusroJv
    • t02 CONCLUSION Conclusion drawn on the basis of analytical assessment and results toxicity in vitro study appear as follows ' of : Gunja seeds are accepted as vegetable toxin in ancient and modem literature as well. ' After Shodhana process, nature of the powdered drug remained the same i.e. coarse. ' colour of all the eight samples after shodhana has changed. Ashuddha Rakta Gunja beej powd€r was yellowish red in colour whereas after shodhana with distilled water and kanji it tumed to blakish gray. On Shodhana with Godugddha, colour of the powder was brownish black Ashuddha shwet Gunja beej powder was faint yellow in colour while it was whitish yellow after shodhana with distilled water and shodhana kanji. After with Godugddha Shwet Gunja beej powder turned to creemish yellow colour. ' Reduction of values was observed in weight of samples treated with and distilled water. The weight of Kmji sample treared with Codugddha increased to some extent because of the moishrre md ft oded due to boiling with cow milk. ' The determination of pH value after detoxificaim increase through the nature pro6 revealed some ofthe sohnion rmained to be acidic.
    • r03 ' Conductivity of the solutions of sample heated with distilled water and kanji found to be reduced while it incneased in sample treated with Godugddha. ' Total Ash value, water soluble and acid insohrble ash value showed a decrease after shodhana. ' The quantitative data of protein assay conchrdes fhrt the total protein content of the shwet and Rakta crrmja Beej has decreased significantly after Shodhana process. ' The loss in total protein content of the s*mple mnges fiom 96.59 o/o to 99.78% which is maximrmr in Kanji shodhit Rakta and Shwet Gunja Beej. ' Qualitative test for Amino acid showed the decrease in Amino acids after Shodhana as compared to Ashuddha samples. ' TLC showed the effect of Shodhana Samskar on Toxic principle of Gunja beej. The spots present Beej were absent in Ashuddha samples of Rakta and Shwet Gunja in Distilled water, Kanji and Godugddha shodhit Samples. ' The obsenration and result of the toxicity in vitro study was very significant. Agglutination of RBCs found in all the blood spots heated with extract of Ashuddha Rakta and shwet Gunja beej. This agglutination occurred in all individuals of blood grow A, B, AB, Negative and B Rh Negative. o Rh +ve and A Rh
    • IM ' Agglutination of RBCs was not found in all the blood samples heatod wift extract of distilled water, Kanji and Godugddha shodhit Rakta and shwet Gunja beej. The result was same in all the individuals of diffe,rent blood group taken in the study. r The time required for agglutination of RBCs in samples treated with extract of Ashuddha Rakta Gunja beej was two to four minutes while it was less than 30 seconds in samples treated with extract of Ashuddha Shwet Gunja beej. The study has provided arnple evidence of efficacy of Ayurvedic "shodhana Samskar" in the acfual detoxification of Gunja seeds. The honest efforts through this treatise build the concrete foundation for the eternal truths of Ayurvedic concepts.
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    • il0 ABBREVIATION A.H.Chi Ashtang Hridayam Chikitsasthana A.II.S. Ashtang Hridayam Sutrasthana A.H.U. Ashtang Hridayam Uttarsthana A.P. Ayurved Prakash A.S.U. Ashtang Sangraha Uttarsthana Bhai.Rat. Bhaishajya Ratnavali Bhar,.Ni. Bhavprakash Nighantu Cha.Chi. Charak Chikitsasthana Cha.Su. Charak Sutrasthana Dh.Ni. Dhanvantari Nighantu Kai.Ni. KaiydevNighantu Ra.Ni. Raj Nighantu R.R.S. Rasaratna Samucchaya R.S.S. Rasendra Sara Sangraha R.T. Rasa Tarangini Sha.Su.Pu. Sharangdhar Samhita purvardha Sho.Ni. Shodhal Nighantu Su.Kal. Sushruta Kalpasthana Su.Su. Sushruta Sutrasthana
    • llt REPORT ON BLOOD EXAMINATION Total Polyporphs Llmphocytes Eosinophills Monocytes Basryffib kucocyte o/ /o o/ /o % o/ /D ./t 40-75 2045 0l-06 02-r0 00{l Count Normal 4000- Range 11000/cmm Patient A 8300 63 3l 04 02 0 B 8200 69 28 03 02 0 c 7400 66 29 03 02 0 D 6400 58 35 03 04 0 E 6800 63 30 03 04 0 F' 5800 64 29 03 04 0 G 7800 62 3l 04 03 0 H 7200 65 31 02 02 0 I 7200 68 28 03 0l 0 J 7600 64 27 04 05 0 K 6500 59 34 04 03 0 L 7800 6l 32 03 o4 0 M 7200 65 28 o4 03 0 Note : A to M are individuals for toxicity in vitro study.