Hepatoprotective rs018 gdg

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Preparation, physico- chemical analysis of shankha nabhi bhasma and evaluation of its hepato protective activity, -an experimental study, JAYASHREE S. , Department of rasashastra, Post graduate studies and research center, Shri D. G. Melmalagi Ayurvedic Medical College, Gadag

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Hepatoprotective rs018 gdg

  1. 1. “PREPARATION, PHYSICO- CHEMICAL ANALYSIS OFSHANKHA NABHI BHASMA AND EVALUATION OF ITS HEPATO PROTECTIVE ACTIVITY, -AN EXPERIMENTAL STUDY”. BY DR. JAYASHREE S. Dissertation Submitted to the Rajiv Gandhi University Of Health Sciences, Karnataka, Bangalore. In partial fulfillment of the requirements for the degree of AYURVEDA VACHASPATI (DOCTOR OF MEDICINE) IN RASASHASTRA Under the guidance of Dr. M.C. Patil M.D. (Ayu) Professor & H.O.D. Dept. of Rasashastra and Co-guidance of Dr. G.N. DANAPPAGOUDAR M.D.(Ayu) Asst. Professor Dept. of Rasashastra POST GRADUATE DEPARTMENT OF RASASHASTRA D.G M. AYURVEDIC MEDICAL COLLEGE AND RESEARCH CENTER, GADAG – 582103 2005 - 2008
  2. 2. Rajiv Gandhi University Of Health Sciences, Karnataka, Bangalore. DECLARATION BY THE CANDIDATE I here by declare that this dissertation / thesis entitled “Preparation, Physico-Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepatoprotective Activity - An Experimental Study” is a bonafide and genuine researchwork carried out by me under the guidance Dr. M.C. Patil M.D.(Ayu), Professor &H.O.D. Post graduate department of Rasashastra. and under the Co-guidance of Dr.G.N. Danappagoudar M.D.(Ayu), Asst. Professor Post graduate department ofRasashastra, D.G.M.A.M.C, PGS & RC, Gadag.Date:Place: Gadag. Dr. Jayashree S.
  3. 3. SHRI D. G. MELMALAGI AYURVEDIC MEDICAL COLLEGE, POST GRADUATE DEPARTMENT OF RASASHASTRA. CERTIFICATE BY THE GUIDE This is to certify that the dissertation entitled “Preparation, Physico-Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepatoprotective Activity - An Experimental Study” is a bonafide research work done byDr. Jayashree S. in partial fulfillment of the requirement for the degree of “AyurvedaVachaspathi M.D (Rasashastra)”, Under Rajiv Gandhi University of HealthSciences, Bangalore, Karnataka.Date: GuidePlace: Gadag. Dr. M.C. Patil M.D.(Ayu) Professor & H.O.D. Dept. of Rasashastra, Post Graduate Research Center D.G.A.M.C. Gadag
  4. 4. SHRI D. G. MELMALAGI AYURVEDIC MEDICAL COLLEGE, POST GRADUATE DEPARTMENT OF RASASHASTRA. CERTIFICATE BY THE Co - GUIDE This is to certify that the dissertation entitled “Preparation, Physico-Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepatoprotective Activity - An Experimental Study” is a bonafide research work done byDr. Jayashree S. in partial fulfillment of the requirement for the degree of “AyurvedaVachaspathi M.D (Rasashastra)”, Under Rajiv Gandhi University of HealthSciences, Bangalore, Karnataka.Date: Co GuidePlace: Gadag. Dr. G.N. Danappagoudar, M.D. (Ayu) Asst. Professor, Postgraduate department of Rasashastra. D.G.M.A.M.C. Gadag.
  5. 5. J.S.V.V. SAMSTHE’S ENDORSEMENT BY THE H.O.D AND PRINCIPAL OF THE INSTITUTION This is to certify that the dissertation entitled “Preparation, Physico-Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepatoprotective Activity - An Experimental Study” is a bonafide research work done byDr. Jayashree S. under the guidance of DR. M.C. Patil M.D. (Ayu), Professor & H.O.D,Postgraduate department of Rasashastra and co-guidance of DR. G.N.Danappagoudar M.D. (Ayu), Asst. Professor, Postgraduate department of Rasashastra,in partial fulfillment of the requirement for the post graduation degree of “AyurvedaVachaspati M.D. (Rasashastra)” Under Rajiv Gandhi University of HealthSciences, Bangalore, Karnataka.DR. M.C. Patil M.D. (Ayu) Dr. G. B. Patil.Professor & H.O.D. Principal.Post graduate department of Rasashastra. D.G.M.A.M.C, GADAG.D.G.M.A.M.C, GADAG.Date: Date:Place: Gadag Place: Gadag
  6. 6. COPYRIGHT Declaration by the candidate I hereby declare that the Rajiv Gandhi University of Health Sciences,Karnataka shall have the rights to preserve, use and disseminate this dissertation /thesis in print or electronic format for academic / research purpose.Date: Signature of ScholarPlace: Gadag Dr. Jayashree S. © Rajiv Gandhi University of Health Sciences, Karnataka.
  7. 7. ACKNOWLEDGEMENT  I salute to Lord Vighneshwara and His Holiness Shri Abhinava Shivanandswamiji to have bestowed their blessings throughout my carrier. I acknowledge my parents Shri Shantaveerappa Pattanshetti and Smt.Annapurana S. Pattanshetti being my inner strength to achieve every milestone in mylife. With pleasure I express my full respect and regards to my Grand Mother Smt.Gangamma Mahajanshetter and Doddamma Smt. Gouramma Yadavannavar whomade me proficient and gave constant support and encouragement. I am always at remembrance of my life partner Dr. Anilkumar V and hisfamily members who are spirit behind my enthusiasm and for their needful support. With deep sense of pleasure, I acknowledge my gratitude to my beloved guideDr. M. C. Patil MD (Ayu) Professor and HOD, PG Dept of Rasashastra, DGMAMC,Gadag, for his scholarly guidance, supervision, creative criticism, constantencouragement and high inspiration at every stage of this work. My gratitude is greatest towards my co-guide Dr. G. N. Danappagoudar MD(Ayu) Lecturer, PG Dept. of Rasashastra, DGMAMC, Gadag, who gave me timelyadvises and suggestions during the entire period of this effort. I express my sincere obligations to our beloved Principal Dr. G. B. Patil, PGS& RC, DGMAMC, Gadag, for his encouragement and provision of amenities requiredduring this toil. I offer my sincere thanks to Dr. R.K. Gacchinmath, Professor and H.O.D, UGDept of Rasashastra, DGMAMC, Gadag, for his constant support and valuabledirections. I wish to convey my regards to Dr. Jagadeesh G. Mitti, Lecturer, PG Dept ofRasashastra, DGMAMC, Gadag, for his enormous co-operation. I owe my heartfelt thanks to Dr. Dilipkumar B, Asst. Professor, PG Dept ofRasashastra, DGMAMC, Gadag, for his critical views and precious suggestions. I express my earnest gratitude to Dr. G.S. Hiremath, Dr. Varadacharyulu, Dr.Avvanni, Dr. Prushottamacharyulu, Dr. K.S.R. Prasad, Dr. R.V. Shettar, Dr. S.N.Belawadi, Dr. Rajshekar, Dr.Kuber Sankh, Dr Shashikant B. Nidagundi & Dr M.D.Samudri for their great co-operation. I
  8. 8. I am greatful to Dr. Shreevatsa MD (Ayu), Lecturer, TGMAMC, Bellary, Dr.Anjaneya murthy, Dr. Gopi Krishna for their inspiration and constant supportthrough out my carrier. With pleasure I extend my sincere gratitude to Dr S.D. Yarageri RMO, Dr. B.GSwamy, Smt. P.K. Belwadi, Smt. Sarangmath, Tippanagoudar, Kallangoudar,Biradar, Smt. Ekbote, Smt. A.C. Patil, Shri Shankar Belwadi, Shamshad, Mangala &Manju for their co-operation and help during the study. I extend my gratitude to Shri V.M. Mundinmani, Shyavi and Karur forproviding the required books during the study. I am deeply indebt to Dr. Sharnu Angadi, Dr. Pattanshetti, Dr. B.Y. Ganti, Dr.Pradeep, Dr. Sobagin, Dr. Shakuntala, Dr. Anita, Dr. Suvarna, Dr. Anand, Dr. Teggifor their friendly affection. I ackwoledge my sincere thanks to Shri Shivakumar Inamdar for his statisticalwork, Dr. Revati, Shri Sarfaraz, Shri Angadaki, Subhash for their kind co-operationand help in analytical study. I am blessed to have precious supervision, pinpoint suggestion and constanthelp of Shri. Shivakumar Inamdar Lecturer, Dept. of Pharmacology, J.T. College ofPharmacy, Gadag & Dr. Shashikant B. Nidagundi M.D. (Ayu), Lecturer, Dept. ofDravyaguna, DGMAMC Gadag who guided and accompanied me duringexperimental study. I am grateful to Shri Chaitrakumar (Sadguru Computers) for his kind co-operation and immense help to complete the dissertation work. With great pleasure I offer my recognition to my friends Dr. Ashwini V, Dr.Rudrakshi P.D, Dr. Suma J, Dr. M. Kattimani, Dr. Prasanna Mathad, Dr. SunitaSundaran, Dr. Shri Mukund for their friendly affection and amiable attitude duringmy study period without which it would never be complete. I offer my sincere thanks to my friends and colleagues Dr. Madhushri, Dr.Ashok, Dr. Siba Prasad, Dr. Shivaleela, Dr. Kamalakshi, Dr. Sulochana, Dr.Payappagoudar, Dr. Budi, Dr. Prasanna, Dr. Amnish for their immense help andaffection. I am also thankful to my junior friends Dr. Shivakumar, Dr. Ravindra, Dr.Kavita, Dr. Anupama, Dr. Sarvamangala, Dr. M. Hiremath, Dr. Deepa, Dr. Praveen,Dr. Jadhav, Dr. Ghorpade, Dr. C.C. Hiremath, Dr. Jaya, Dr. Kala, Dr. Savita, Dr.Mukta Y, Dr. Mukta H, Dr. Shailej, Dr. Natraj, Dr. Uday Ganesh, Dr. Adarsh, Dr. II
  9. 9. Joshi, Dr. Veena, Dr. Vijayalakshmi, Dr. Sanjay, Dr. Trupti for their support andaffection. I am happy to express my regards to my in-laws, Shri Veerappa. M & Smt.Chitravati. C for their moral and cordial support during the study. It would be my privilege to convey my regards to families of Mr. Mallikarjunand Mr. Timmaraj for their affection and care during my study. I express my affectionate love to my Sister Rajeshwari S. Martur & BrotherBasavaraj Pattenshetti for their moral and cordial support during my study. I am ever thankful to my family members Mr. Ashok Yadavannavar and Smt.Anita Yadavannavar, Mr. Ulavappa Kondikoppa & Smt. Drakshayani Kondikoppawho stood with me all the way at my turmoil. I thank my kiths and kins Rashmi, Smita, Prajval, Sangamesh for theiraffection. I am very much greatful to all the lecturers, house surgeons, Hospital staff andnon teaching staff for their timely assistance in completion of this work. I express my thanks to all those who have helped me directly and indirectlywith apologies for my inability to identify them individually. I dedicate this work done as partial fulfillment of postgraduate degree to myever remembering respectful parents and my husband Dr. Anilkumar V. Dr. Jayashree S.Date:Place: III
  10. 10. LIST OF ABBREVIATIONS USEDRT RasataranginiAP Ayurveda PrakashRRS Rasa Ratna SamucchayaRSS Rasendra Sara SangrahaBRRS Brihat Rasaraja SundaraBP Bhava PrakashaKN Kaideva NighantuSN Shaligram NighantuRN Raja NighantuSh.N Shodala Nighantu.MN Madanapala NighantuBR Bhaishajya RatnavaliYR Yoga RatnakaraCS Charaka SamhitaSS Sushruta samhitaSha. S Sharangdhara SamhitaAH Ashtanga HridayaMN Madhava NidanaCCl4 Carbon tetrachlorideSGOT Serum Glutamic – Oxaloacetic TransaminaseSGPT Serum Glutamic – Pyruvic TransminaseALP Alkaline PhosphataseT-Bil Total BilirubinS.Alb Serum AlbuminSNB Shankhanabhi BhasmaSNB – I Shankhanabhi Bhasma in 1 Karsha MatraSNB – II Shankhanabhi Bhasma in 2 Ratti Matra‘+’ Mentioned‘- ’ Not Mentioned IV
  11. 11. ABSTRACTBACKGROUND In the present Scenario change in life style of human beings like excessivealcohol consumption, smoking, stress, irregular food habits have increased theprevalence of Yakrit vikara. Liver plays a mojor role in detoxification and excretion of many endogenousand exogenous compounds, any injury (due to systemic drugs, food preservativesagrochemicals and addiction to alcohol) to it or impairment of its function may lead tomany complications on one’s health. There is no rational therapy available for treating liver disorder and is still achallenge to the modern medicine. Modern medicine have little to offer for alleviationof hepatic ailments where as most important representatives are of natural origin. Before evaluating efficacy of any drug, it is essential to carry out anexperimental study and to find out potent therapeutic form.Objectives: 1. Preparation of Shankhanabhi Bhasma. 2. Physico-chemical analysis of Shankhanabhi Bhasma. 3. Evaluation of Hepato-protective activity of Shankhanabhi Bhasma.Methods:Pharmaceutical study: 1) Shankhanabhi shodhana according to Rasatrangini 12th chapter sholka no.10. 2) Shankhanabhi marana according to Rasatarangini 12th chapter sholka no.17-19. V
  12. 12. Analytical study Shankhanabhi bhasma is subjected to physico-chemical analysis likeorganoleptic characters, determination of pH values, loss on drying at 1100C, loss onignition, determination of total Ash, Acid insoluble ash, water soluble extractive,alcohol soluble extractive, finess of particles, particle size, flow property, flow rate,Solubility text, Estimation of Ca, CaCO3, Fe, Mg, S and NPSTest.Experimental study: Shankhanabhi bhasma was evaluated for Hepato-protective activity againstcarbon tetrachloride (CCl4) induced Liver toxicity in albino rats. The trial drug wasadministered in two different doses and biochemical parameters and histopathologicalreports was observed, recorded and statistically analysed.Results: The Shankhanabhi bhasma has shown significant hepato protective effet (with“p” value <0.001) by reduction in elevated serum enzyme levels such as serumglutamic oxaloacetic transminase (SGOT), serum glutamic pyruvic transaminase(SGPT), alkaline phosphatase (ALP), Total bilirubin and Serum Albumin. Thesebiochemical observations were supplemented by histopathological examination ofLiver sections.Interpretation and Conclusion: 1. The dravyas which are mentioned in the classical procedure of Shankha shodhana and marana convert the Shankhanabhi into quick absorbable form and induces the disease curing property. 2. Ayurvedic bhasma pareeksha and modern physico-chemical analysis are confirmation tests for the complete formation of bhasma and its genuinitity. VI
  13. 13. 3. Shankhanabhi Bhasma is one of the ideal Hepato protective drug which has been proved experimentally by reducing the LFT values and histopathological report.Keywords: Shankhanabhi Bhasma, Analytical study, Yakritodara, Hepatoprotectiveactivity, Carbon tetrachloride, Significant results. VII
  14. 14. TABLE OF CONTENTSSl. Name of Topic & Sub Topics Page No.No. 1 INTRODUCTION 1-52. OBJECTIVES 63. REVIEW OF LITERATURE Drug Review 7-32 Disease Review 33-574. MATERIALS AND METHODS Pharmaceutical Study 58-68 Analytical Study 69-84 Experimental Study 85-905. RESULTS 91-1016. DISCUSSION 102-1157. CONCLUSION 1168. SUMMARY 117-1189. BIBLIOGRAPHY 120-12810. ANNEXURE – SHLOKAS VIII
  15. 15. LIST OF TABLESSl Tables Page N0No01 Showing full name of texts mentioned about Shankha 902 Showing full name of Nighantu’s mentioned about Shankha 903 Showing Synonyms of Shankha according to various texts 11-1204 Showing Shodhana procedure of Shankha according to 16 various Rasa classics05 Showing Rogaghnata of Shankha according to various texts 17-1806 Showing classical yogas of Shankhanabhi and Shankha 18-1907 Showing brief introduction of animal of Shankha belonging 20 to Phylum mollusca08 Showing correlation between Ranjakapitta and Bile 3709 Showing Nidana of Pleehodara and Yakritodara according to 38 various texts10 Showing Roopa of Pleehodara and Yakritodara according to 38-39 various texts11 Showing principle alterations of hepatic morphology 51-52 produced by some commonly used drugs and chemicals.12 Showing Laboratory Evaluation of Liver disease 5713 Showing quantity of liquid during preparation of kanji. 6014 Showing the changes in wt. of shankhnabhi before shodhana 62 and after shodhana.15 Shwoing physical features of raw and shodhita shankhanabhi 6316 Showing drugs used for shankhanabhi marana. 6417 Showing Time and Temperature of gajaputa 1 6618 Showing Time and Temperature of gajaputa 2 6719 Showing Physical features of Shankhanabhi bhasma after 68 each puta.20 Showing the loss of Shankhanabhi during its marana. 6821 Showing Analysis of Shankhanabhi bhasma by ancient 70 method.22 Showing Experimental Protocol. 8723 Showing Summary of Biochemical values of all groups 9124 Showing Intermediate calculation, ANOVA table SGOT 9425 Showing one way analysis of variation (ANOVA) 9426 Showing Intermediate calculation, ANOVA table SGPT 9527 Showing one way analysis of variation (ANOVA) 9528 Showing Intermediate calculation, ANOVA table ALP 9529 Showing one way analysis of variation (ANOVA) 9630 Showing Intermediate calculation, ANOVA table T- bil 9631 Showing one way analysis of variation (ANOVA) 9632 Showing Intermediate calculation, ANOVA table Albumin 9733 Showing one way analysis of variation (ANOVA) 9734 Showing the comparison of effect of toxic and natural group 100 with treated groups (By means of t values). IX
  16. 16. LIST OF GRAPHS: Sl. No Graphs Page No 1 Mean SGOT of all the groups 92 2 Mean SGPT of all the groups 92 3 Mean ALP of all the groups 93 4 Mean T-Bil of all the groups 93 5 Mean Serum Albumin of all the groups 94 6 Comparison between Biochemical parameters of 97 G2 and G3 7 Comparison between Biochemical parameters of 98 G2 and G4 8 Comparison between Biochemical parameters of 98 G2 and G5 9 Comparison between Biochemical parameters of G3 99 and G4 10 Comparison between Biochemical parameters of G3 99 and G5 11 Comparison between Biochemical parameters of G4 100 and G5LIST OF PHOTOGRAPHS: Fig.No Photographs 1 Shankha 2 Shankhanabhi 3 Pachana in Kanji 4 Pieces of Shankhanabhi after shodhana 5 Marana of Shankhanabhi 6 Shankhanabhi pieces after I Gajaputa 7 Mardana of Shankhanabhi bhasma with Kumari swarasa 8 Chakrikas of Shankhanabhi 9 Shankhanabhi bhasma after II Gajaputa 10 Suspensions of Shankhanabhi bhasma 11 Grouping of animals 12 Weighing of Rat for selection 13 Administration of CCl4 0.7 ml /kg i.p 14 Administration of trial drug orally. 15 Opening of abdomen for liver isolation. 16 Phatomicrograph of Normal Healthy liver (G1) 17 Phatomicrograph of Damaged liver with CCl4 (G2) 18 Phatomicrograph of Toxic control/ Natural recovery liver (G3) 19 Phatomicrograph of liver (G4) treated with SNB – I 20 Phatomicrograph of liver (G4) treated with SNB – II 21 Alcoholic extraction of Haridra 22 Haridra paper prepared 23 Haridra paper with standard spot of Shankhanabhi bhasma X
  17. 17. XI
  18. 18. Introduction INTRODUCTION Ayurveda is science of life and its aim is for prevention and cure of diseasesand also attainment of chaturvidha Purushartha i.e Dharma, Artha, Kama, Moksha.Though we can prevent diseases just by following sadvritta and regimen ofDinacharya, Ritucharya etc. yet for the cure of diseases, drugs are essential. In Ayurveda, acharyas have explained different charyas to maintain the health.The diseases are manifested mainly because of neglecting these routines.Change inlife style of human beings like excessive alcohol consumption, smoking, stress,irregular food habits have increased the prevalence of Yakrid vikara. From the time immemorial mankind has been in search for plant, animal andother materials that could be used to take care of the pains, deformities and diseasesthat affect some of the unfortunate members of the society.Among them one of themajor problem is the hepatic disorders. Liver is the largest organ in the body, whatever taken orally or parenterally,eventually it has to pass through the liver, either by portal or systemic circulation andgets altered, destroyed or detoxified by it. In this process of protecting the body fromharmful substances, liver cells themselves get destroyed and are usually regeneratedwith the same speed provided, the toxins are not in abundant quantities andcontinuously pass through it. Liver plays a pivotal role in regulation of physiological process likemetabolism and excretion. It is constantly endowed with the task of detoxification ofxenobiotics, environmental pollutants and chemotherapeutic agents. Liver diseases aremainly caused due to an exposure to toxic chemical substances like antibiotics,analgesics, antipyretics, chemotherapeutics, peroxidised oil, carbon tetrachloride,chlorinated hydrocarbons and also due to chronic alcoholism, viral infections and auto 1 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  19. 19. Introductionimmunity disorders. Most of the chemotoxic chemical effect on liver cells mainly byinducing lipid peroxidation and other oxidative damages takes place in liver. It has been estimated that about 90% of the acute hepatitis are resulting due toviruses. The major viral agents involved are hepatitis B,C,D,E & G of these, hepatitisB infection often results in chronic liver diseases and cirrhosis of liver. It has beenestimated that approximately 14-16 million people are infected with this kind of virus. In India alone over 2,00,000 people die due to liver disorders in every year.Consumption of spirits/ liquors is the prime cause for liver damage, drug toxicity isanother cause for liver disorders. On survey studies around 2-3% of Indian populationare carrying hepatitis B & C type of viruses, worldwide these figures are increasingwith an alarm. In Ayurveda also Acharyas have given utmost importance to the yakrit. It isthe seat of Ranjaka pitta, Bhutagni, Rakadhara kala and mula of Raktavaha srotas.Any damage to liver ultimately disturbs the digestive system, which is the maincausative factors of all the diseases according to Ayurveda.Among theAshtamahagadas the udara roga is one. Among the eight udara rogas pleehodara andyakritodara are described. In the derangement of yakrit and pleeha, raktakshaya andother symptoms are manifested.Thus liver and spleen plays vital role in udara andraktavikaras. In spite of the tremendous advances made in modern medicine, no effectiveand safe hepato-protective medicines are available. Hence an effective and safehepato guard formulations are need of the hour. About 80% of the world populationrelay on the use of Ayurvedic medicaments which has the treasure of uniqueRasoushadhis. 2 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  20. 20. Introduction In present scenario several hepato-protective drugs as well as compoundformulations explained in classics should be pharmacologically evaluated for theirefficacy, safety profile & world wide acceptance. Drugs used in Ayurveda can be broadly classified into three groups as herbal,animal and minerals.1) Herbal Drugs: In vedic literature and in Ayurvedic classics mostly vegetable drugs wereprescribed for the treatment of different categories of ailments. Very few animalproducts and still fewer metals and minerals are described in those texts. During the fifth century B.C and thereafter the important branch of Ayurvedanamely shalyatantra was viewed as a form of hinsa or violence. Ahinsa or non-violence was the cardinal rule of the religion prevalent in thosedays. The religion which was adopted by the rules and subjects alike, discouraged thepractice of surgery and it was almost legally banned. Therefore the physicians in thathad to find alternative for curing their obstinate surgical conditions. This provideddevelopment of Rasashastra. Metals and Minerals were processed and extensivelyused therapeutically in Bhasma form in that time.According to classics therapies are of three categories viz; a) Asuri - which includes surgical therapies. b) Manushi – which is performed by the use of decoctions etc of vegetable drugs. c) Daivi - which is performed by the administration of metallic and mineral preparations. The succeeding one are superior to the preceding categories of therapies.2) Mineral Drugs: a) These are usefull in obstinate and incurable conditions. 3 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  21. 21. Introduction b) Mineral remedies are therapeutically effective even when administered only in a small dose (unlike vegetable products which are generally required to be administered in a much larger dose). c) These mineral products are not unpalatable (unlike some of the vegetable remedies which are sometime very unpalatable because of bitter, astringent and pungent taste) d) Mineral products produce their therapeutic effects instantaneously (unlike vegetable products which take larger time because they have to pan through the process of digestion and metabolism) before they become therapeutically active.3) Drugs of Animal origin- The drugs of animal origin like Sudha varga dravyas do posess actions like minerals. Many drugs have been mentioned in sudha varga, among them Shankha is one. Shankha is a shell of an aquatic insect called Turbinella pyrum. These shells are available in various sizes and colours, probably according to the habitat. In India these shankha are available almost throughout its vast sea shore, but in different sizes and in various shades. Shankha is the base for therapeutically active drug called shankha bhasmawhich is effective medicine in various diseases such as Amlapitta, Shula, Grahani,Atisara, Yakrit vruddi, Pleeha vruddi etc. Calcium, apart from forming an essentialconstituent of bones, plays an important role in body homeostasis and disturbances incalcium metabolism are associated with derangement of various cellular functions. Shankhanabhi bhasma is one of the cost effective Rasoushadhi commonlymentioned both for pleeha and yakrit vikaras in Ayurvedic classics. With this 4 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  22. 22. Introductionperspective the study is taken to evaluate the Hepatoprotective activity ofShankhanabhi bhasma.Plan of suudy:The entire study was done in following chapters: 1. Introduction : Involves need of the present study, emphasis on its importance and plan of the whole study. 2. Review of Literature: The detailed classical Ayurvedic literature and relevant modern literature about the drugs Shankha, Nimbuka, Kumari, Kanji and about the disease Yakrit vikara were reviewed. 3. Methodology: a. Pharmaceutical study: In this, preparation of Kanji, method of shodhana of shankhanabhi, marana of shankhanabhi is incorporated. b. Analytical study: This includes physico-chemical analysis of shankhanabhi bhasma. c. Experimental study: This includes evaluation of hepato protective activity of Shankhanabhi bhasma with nimbu swarasa in albino rats. 4. Results: In this part the obtained results are systematically presented, which include data related to biochemical parameters, data related to histopathological examination and data related to response to treatment. 5. Discussion: In this chapter observation, findings and results of various studies have been found out with possible explanation for its effects. 6. Conclusion: The essence of the whole study is mentioned in this chapter. 7. Summary: It contains the information of the overall work in a nut shell. 5 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  23. 23. Objectives AIMS AND OBJECTIVES 1.Preparation of Shankhanabhi Bhasma. 2.Physico-Chemical analysis of Shankhanabhi Bhasma. 3.Evaluation of Hepato protective activity of Shankhanabhi Bhasma. 6“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  24. 24. Drug review DRUG REVIEW Rasaoushadhis by virtue of their unmatched properties like Alpamatra,Shigraphaladayi,Rasayanas, Bahu upayoga etc,have been leading the field of chikitsa sincetime immemorial. Shankhanabhi Bhasma along with Jambeera swarasa is a uniquecombination to treat Yakrit vruddi and Pleeha vruddi. In Yogaratnakara under Udara roga chikitsa, Shankhanabhi Bhasma withJambeera swarasa is mentioned for Yakrit vruddi1. The author of Bhaishajya ratnavali also explained the combination of .Shankhanabhi Bhasma with Jambeera swarasa under Pleehayakrit roga chikitsaprakarana to treat Yakrit vruddi2. This combination is also mentioned in texts like Vangasena inUdararogadhikara3, Rasa Chikitsa4 and Bharata Bhaishajyaratnakara5. Drug : Shankhanabhi Bhasma. Anupana : Jambeera swarasa.SHANKHA : As Shankhanabhi is the part of Shankha itself, review of the same is done.Aurvedic literature of Shankha :1) Utpatti : Relation with God Shankha is related with Lord Dhanwantari as it has been held by one of thehands of God.2)Historical aspect :a) Vedic kala- In Atharva veda we get references of Shankha many times.Accordingto it Shankha is mangalakaraka on dharana6 and krumighna on sunada7.b) Mahabharata : In Mahabharata and Bhagwata granthas also we get references ofShankha. 7 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  25. 25. Drug reviewc) Samhita kala : Brihatrayee’s1. Charaka samhita-Charaka explained shankha in 27th chapter of sutrasthana under Varishaya varga8.Also mentioned it in various places as a medicinal use.2. Shushruta samhita-Shushruta explained Shankhanabhi in 11th chapter Ksharavidhi adyaya in preparationof madyama kshara9. Further he has used Shankha churna for lomashatana10.Shankhanabhi Bhasma is extensively used in many yogas in uttaratantra to curevarious eye diseases in the from of Netra varti’s11.3.Ashtanga sangraha-In Ashtanga sangraha shankha is used as anjana paatra12 and in treatment raktapittaand various netra rogas.4.Ashtanga hridaya-In Ashtanga hridaya shankha is used in the treatment of shvitra13 and various netrarogas.Laghutrayee’s :1.Sharangdhara samhita-Sharangdhara explined the shankha shodhana by giving bhavana with jambeeraswarasa and drying in sunlight14. Shankha bhasma has been used in many yogas andlepas.Putapak yoga has also been explained15. Shankhanabhi Bhasma is used inpreparation of various netra varti’s.2.Bhavaprakasha samhita-The synonyms,guna and doshaghnata of shankha has been explained under dhatu-upadhatu varga16. 8 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  26. 26. Drug reviewTable-1 Showing Full name of texts mentioned about shankhaSl.No Full name of texts mentioned about shankha 1. Ayurveda prakasha. 2. Brihat rasaraja sundara. 3. Bhava prakasha. 4. Rasarnava tantra. 5. Rasamrutam. 6. Rasendra chintamani. 7. Rasendra chudamani. 8. Rasa hrudaya tantra. 9. Rasa jalanidhi. 10. Rasa kaumudi. 11. Rasa kamadhenu. 12. Rasa tarangini. 13. Rasa paddati. 14. Rasa ratna samucchaya. 15. Rasa prakasha sudhakara. 16. Rasendra sara sangrha. 17. Yogaratnakara.Table-2 Showing Full name of Nighantu’s mentioned about shankhaSl.No Full name of Nighantu’s mentioned about shankha 1. Bhava prakasha Nighantu. 2. Dhanwantari Nighantu 3. Kaideva Nighantu. 4. Madanapala Nighantu. 5. Raja Nighantu. 6. Shaligrama Nighantu. 9 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  27. 27. Drug reviewBhaishajya Ratanavali: Acharya Govinda Das Sen has explained the shodhana and marana ofShankha. Shaodhana by swedana in Kanji for one hour and by swedana in Jambeeraswarasa. He explained common process of Jambeera swarasa swedana for both shankhaand shankhanabhi (Khulwaka) in a single sutra17. He has mentioned the shankhagunas and indicated for udara roga18. Author has explained many yogas containingshankha and shankhanabhi.Yoga Ratanakara: In yoga ratnakara shankha has been mentioned under Koshasta varga ofAnupa mamsa19. Many yogas of shankha are explained. Shankhanabhi bhasma is acontent for many Netra varti’s.Rasa Grantha: In Rasa Shastra Shankha is included for the first time in Rasarnava in “Shukla-Varga”20 & then in all granthas under “Sudha-Varga”.Shukla Varga: vÉÑYsÉ uÉaÉïÈ xÉÑkÉÉMÔüqÉï vÉÇZÉ zÉÑÌ£ü uÉUÉÌOûMüÈ| (UxÉhÉïuÉ – 5/40) Based on Shukla varnata these drugs are classified in one group.Sudha Varga: As they contains lime (Sudha), they are included in sudha varga. Almost alldrugs in shukla varga contains lime and majority of them are of aquatic orgin &shankha is one of them.Properties: 1. These drugs contains high percentage of calcium.Many of them are formed from Kshariya jala of samudra & they are alkaline in nature. 10 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  28. 28. Drug review 2. Almost all drugs are hard in nature. Usually they are formed by the skeleton of marine animals.Other drugs in sudha varga: 1. Shukti 2. Kapardika 3. Pravala 4. Kukkutanda twak 5. Mruga Shrunga 6. Godanti 7. Badari Pashana 8. Khatika 9. Shambuka 10. Kurma Prushta 11. Ajasthi 12. MuktaTable No. 3 Showing Synonyms of Shankha21,22,23,24,25,26,27,28,29,30Synonyms R.T A.P RRS RSS BRRS BP KN SN RN M.NArnobhava - - - - - - - - + -Bahunada - - - - - - - - + -Deerghanada + - - - - - + - + -Dhaval - - - - - - + - + -Dirghanisvana - - - - - - + - - +Drushtidravi - - - - - - - - - -Haripriya - - - + - - - - + -Jaladhara - - - - - - - - - +Jalaja - - - - - - + - + -Kamboja + - - - - - - - - - 11 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  29. 29. Drug reviewKambu + + + + + + + + + +Kshudra - + + - + - - - -- -Kutilanta - - - - - - - - + -Mahanada - - - + - - - - + -Mangalaprada - - - - - - - - + -Pavana dhwani - - - + - + - + + -Putha - - - - - - - - + -Samudraja + + + - + + - + - -Shankha + + + - - + + + + +Shankhaka + - - - - - - - - -Shankhanaka + + - + - - - - - -Shrivibhushana - - - - - - - - - -Strivibhushana - - - - - - + - - -Sunada + - - - - + - + + -Suswara - - - - - - + - + -Trirekha + - - - - - - - -- -Varibhava - - - - - - - - - -Varichara - - - - - - + - - -Varija - - - - - - - - - + 12 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  30. 30. Drug review NIRUKTI31Based on Nada / SoundShanka - zÉÉqrÉÌiÉ AzÉÑpÉqÉxqÉÉÌSÌiÉ | Pecifies ashubhata xÉqÉÑSìÉå°uÉeÉliÉÑÑÌuÉzÉåwÉÈ | Special kind of Jantu found in samudra.Deerghanada - SÏbÉÉåï SÕUaÉÉå lÉÉSÉåSxrÉ | ( UÉ. ÌlÉ) Produces deep sound.Pawana dhwani - mÉÉuÉlÉÈ mÉÉÌuɧÉeÉlÉMüÉå kuÉÌlÉrÉïxmÉ | (UÉ. ÌlÉ) Produces devine sound.Mahanada - qÉWûÉgcÉxÉÉæ lÉÉSgcÉåÌiÉ | qÉWûÉzÉoSÈ CSqÉÎ°È xÉqÉÇ mÉëÉmiÉÉ rÉå MåüÍcÉSè kÉëÑuÉeÉ…ûqÉÉÈ | mÉëÌuÉzÉliÉÉå qÉWûÉlÉÉSÇ lÉSÎliÉ pÉrÉmÉÏÌQûiÉÉ || (UÉ. ÌlÉ) Produces strong sound.Bahunada - oÉWÒûqÉïWûÉlÉç lÉÉSÈ zÉoSÉå rÉxrÉ | (UÉ. ÌlÉ) Produces many sound.Based on Swaroopa:Dhavala - μÉåiÉaÉÑhÉrÉÑYiÉåå | (AqÉU) White in colourKutilanta - MÑüOûÌiÉ uÉ¢Çü uÉëeÉiÉÏÌiÉ | Curved insideBased on Origin:Kambu - MüqooÉgcÉ¢üzÉUcÉÉrÉaÉSÉÍxÉcÉqqÉï urÉaÉëæÌWïûUhrÉqÉmÉpÉÑeÉæËUuÉ MüÍhÉïMüÉUÈ || 13 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  31. 31. Drug review A shell, one marked with three lines.Kamboja - mÉgcÉlÉSÇ xÉqÉÉUprÉ qsÉåcNûɬͤÉhÉmÉÔuÉïiÉÈ | MüqoÉÉåeÉSåzÉÉå SåuÉåÍzÉ | Collected from the kamboja desha.Varibhava - uÉÉËUhÉå lÉå§ÉeÉsÉÉmÉ pÉuÉÌiÉ mÉëpÉuÉiÉÏÌiÉ | Found in water.Jaladhara - kÉUiÉÏÌiÉ kÉUÈ | eÉsÉxmÉ kÉUÈ | Found in water.Varija - uÉÉËUÍhÉ eÉÉrÉiÉå CÌiÉ | (CÌiÉ WåûqÉcÉlSìÈ) Originated from waterVarichara - uÉÉËUrÉÑ cÉUiÉÏÌiÉ | Moves in water.Vernacular Names32:Latin Name : Xanchus pyrum or turbinella RapaSanskrit : ShankhaEnglish : Conch shellHindi : ShankhKannada : ShankhaGujrati : ShankhaMarathi : ShankhaTelugu : SankhamBengali : SankhTamil : SankhaChemical Composition : Calcium Carbonate (CaCo3) 14 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  32. 32. Drug reviewSource33 Shankhas are of different varities according to its animal. These are widelydispersed in marine water. It can occur in all the sea shores. It is the outer covering of “Mollusca group” of aquatic animal which are seenin sea. It is collected from the sea and put in boiling water. The animal which ispresent inside dies and the outer portion, shankha is obtained. It is sold in market. Chanks are gregarious and exclusively marine animals occurring in largenumbers on muddy sand bottom thirteen metres in depth in Tamilnadu shores andAndaman waters.Grahya Lakshana34: In Rasatarangini, grahya lakshanas of shankha have been described as Vrutta : It should have round shape. Snigdha : It should have snigdha sparsha Sukshma mukha : Having small mouth (opening at apex) Nirmala : Having clean surface. Chandusundara : Being like moon looks beautiful. Dirghakaya : It must be large Guru : It must be heavy Sunada : It must produce good sound.It is of two types according to classics351) Dakshinavarta : It is rare and not preferred for preparation of Bhasma. It is mangalyakaraka & krimighna2) Vamavarta : It is found abundantly in all sea shores. It is preferred for preparation of Bhasma. 15 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  33. 33. Drug reviewShodhana of shankha: Shodhana of shankha is done by doing pachana in Dola yantra containingbelow mentioned liquids for particular time. Shankha pieces are first kept in pottali.Which is suspended in Dola yantra for pachana. After complete cooling shankhapieces are removed from pottali and washed with warm water.Table No. 4 Showing Shodhana of Shankha36,37,38-42,43,44,45-46S.L. Procedure & Liquids used RRS AP RT BRRS Sh.S BRno1. Swedana in Kanji + + + +2. Pachana in Kanji for 1 Yama +3. Swedana in Jambeera swarasa +4. Pachana in Jambeera swarasa + for 4 Yama5. Made fine powder, Bhavana is + done with Jambeera swarasa and kept one day in sunlight for drying.6 Pachana in Jayanti patra + swarasa for 1 Yama7 Pachana in Tanduliya patra + swarasa for 1 Yama8 Pachana in Nimbuka swarasa + for ½ Yama Almost all Granthas have mentioned shodhana of Shankha in Amala Dravyas.In the present study procedure of shodhana in kanji is selected.Shankha Marana47: Pieces of shodhita shankha are dried well, then they are placed in sharava,closed with other sharava & sandhibhandhana is made. After complete drying it is 16 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  34. 34. Drug reviewsubjected to Gajaputa. After swangasheeta the sharava is removed and the pieces arecollected, powdered and given bhavana with kumari swarasa & chakrikas are madeand they are once again subjected to Gajaputa such 2-3 putas will yield good whitebhasma of shankha. The pieces of shodhita shankha are put into the fire and subjected for samyaglaghu puta till they becomes bloomed.48,49 One pala of shodhita shankha along with half masha of tankana is heated inblind crucible to get shankha bhasma.50Pharmacological Properties51:Rasa : Katu Rasa (Kshara)Guna : Laghu, SheetaVeerya : SheetaKarma : Grahi, Balya, Vilekhana, Agni deepana, Vishagnhna, Varnya, Hridya.Dosha karma : Tridosha shamakaRogaghnata :Table No. 5 Showing Rogaghnata of Shankha bhasma52,53,54,55,56,57,58Rogaghnata R.T A.P R.S.S B.R.R.S R.N Y.R B.RAmlapitta + - + - + - -Grahani + + - + + + -Parinamashoola + - - - - - -Tarunyapidika + + - + - + -Netra pushaphara - + - - + + -Gulma - - + - - - -Swasa - - - - - - -Meha - - + - - - -Udara shoola - - + - - - - 17 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  35. 35. Drug reviewSarva ruja - - - - - - +Udaramaya - - - - - - +Matra59 : 2 Gunja (250 mg).Anupana60 : Nimbu – Swarasa, Trikatu churna or according to disease.Table No. 6 Showing Classical Yogas of Shankhanabhi and ShankhaS.L. Vishishta Yoga Indications ReferenceNo1 Muktadi choorna Hikka, Swasa, Kasa, Netra Cha.Chi. 17/125 rogas2 Usheeradi choorna Raktapitta, Tamaka swasa, Cha. Chi 4/73 Trushna etc Y.R. Raktapitta Chi / 45-463 Shankhapishti Prelepa Visarpa Cha. Chi 21/824 Shankha Pravaladi Varti Sarva Akshi Roga Cha. Chi. 26/2465 Lomashatana yoga Loma shatana Su. Chi. 1/1056 Anjana varti Kaphaja Netra Roga Su. Chi. 12/87 Ksharajana Balasagrathita Su. Chi. 12/338 Shankadyanjana Arma pidika Su. U. 15/259 Rasakriya Kaphaja Timira Su. U. 17/4310 Lekhana putapaka Kaphaja netraroga Su. U. 18/24-2511 Revati Pratisheda pradeha Revatighaha badha Su. U. 31/6 Y.R Baharoga Chi./412 Shankha dravaka (1) Pleehodara etc R.T. 12/35-3913 Shankhodara Rasa Grahani etc R.P.S. 8/26-2814 Hemagarbha Pottali Rasa Mandagni shwasa etc R.P.S. /80-8315 Lokanatha Rasa Cures 29 diseases with R.P.S. /89-93 different Anupana.16 Krutrima pravala nirmana R.P.S. 11/13317 Kaphaketu Rasa Peenasa, swasa, shiroroga B.R.5 /743-74518 Vishamajwarataka loha Jwara, Pleeha, Yakrit B.R.5. /1162 18 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  36. 36. Drug review19 Agnisuno Rasa Udara B.R.5. 8/51920 Shankha vati (1,2,3) Agnimandya B.R.5. /18721 Mahashankha vati (4) Pleeha, Udara B.R.5. /202,20422 Ratnagarbha pottali Rasa Rajayakshma, Udara B.R. 14/182 Y.R. Rajayakshma /1-623 Sarvangasundara Rasa Rajayakshma B.R . 14/19524 Shankha choorna Yakrit shula B.R. 30/7725 Vaishwanara louha Shula B.R. 30/13726 Shankhanabhi bhasma Pleeha, Yakrit B.R. 30 /112-11627 Chandrodaya varti (1&2) Netraroga B.R. 64/203 Y.R. Netraroga Chi, Sha. U. 13/75-7728 Chandraprabha varti Netraroga B.R.64/20329 Pradarantaka louha Netraroga B.R 64/72 (R.S.S)30 Sutikaharo Rasa Sutika roga B.R. 69/9931 Dantodbhedagadantaka Dantaroga B.R 71/123 Rasa32 Trilokyachintamani Pleeha, Jalodara, Rasayana B.R 73/136 Y.R Rajayakshma / 1-1233 Sutashekhara Rasa Amlapitta, Ajeerna B.R Anubhuta/223 Y.R Amlapitta Chi/1-534 Pratapalankeshwara Rasa Sutika roga B.R.Anubhuta /26635 Hemagarbha pottali Kasa Y.R. Kasa Chi /1-7 Sha. M. 12/10736 Muktadi mahanjana Netraroga Y.R. Netraroga chi/1-3, (B.P)37 Panchabana Rasa Pleehodara, Jalodara Sha.U. 13. Vajeekarana/1-4 Vajeekarana38 Tuttadi Rasakriya Netraroga Sha.U. 13/87-8839 Krishna sarpa vasanjana Netraroga Sha.U. 13/105 19 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  37. 37. Drug review MODERN DESCRIPTIONOrigin of Shankha (Shell) 61 According to Zoology, Shankha is exo-skeleton formed by secretion ofGastropoda class snails which belongs to phylum mollusca.Table No. 7 Showing Brief introduction of their animalsPhyllum MolluscaClass GastropodaSubclass Prosobranchiata (Strepto neura)Order Pectini branchiata (Mono tocardia)Sub order Tenio glossaTribe PlatypodaFamily FilaSpecies Globosca or Xancus Pyrum or Turbinella pyrum. There are thousands of species of Gastropoda class.Structure of shell: The typical Gastropoda shell is a conical spire composed of tubular whorls &containing visceral mass of the animal.Vertical section of shankha showing its part:1) Whorls : The term whorl is complete revolution of the “Spiral cone”2) Apex : The Apex is topmost or initial whorl which is the first to be formed.3) Aperture : It is the opening of the lowermost end of spire.4) Collumela : It is the central axis to which whorls of spiral closing applied together coiling round it.5) Sutures : It is the line that separates several whorls. 20 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  38. 38. Drug review Starting at apex which contains smallest & oldest successive larger whorls arecalled about central axis called collumella. The larger whorl eventually terminates atopening or Aperture from which head and foot of living animal protrude. The collumella itself is called Shankhanabhi.Identification of Right & left handed shells: A shell possesses a Right handed spiral when the aperture open to the right ofcolumella if shell is held with spire up and aperture facing observer and left handedopens to left. Most of Gastropodas are right handed. Few are left handed and some specieshave both Right handed and left handed individuals.Varieties of shells: Gastropoda shells display an infinite variety of colours, patterns, shapes. Inconsiderable number of gastropodas, the shell is conspisously spiralled only injuvenile stage. The coil nature disappears with growth and adults shell represents asingle large expanded whorl.Size of shells: The height of shell is the distance between the apex and the lower margin ofaperture. In some species of these little snails it does not exceed 1.5 mm. The largestshell which may reach a height of 2 ft. are found.Chemical composition of shell: According to modern science a typical Gastropoda shell is composed of threelayers, an outer peri-ostracum, a middle pre-mastic layer, a inner nacreous layer. Periosteoum is thin & composed of horney organic material called concolin.The two layers consists of calcium carbonate. 21 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  39. 39. Drug reviewCalcium Carbonate62: Calcium carbonate occurs in large quantities in nature as chalk, marble andlime stone. However enough CaCO3 is observed to cause systemic and renal effectsbut it has mainly considered to be the non-systemic antacid.Ref: B.S. Bahl & G.D. Sharma – Inorganic chemistry uses of CaCo3, P-470.Absorption And Excretion63:CaCO3 Ca+2 + CO3-2 H2O+ H2CO3 H2O + CO2 The Calcium cations formed in reaction and present as the water solublecalcium chloride salt can be either absorbed or precipitated as the insoluble calciumphosphate salt in the intestine or as insoluble calcium soaps from the hydrolyzedglycerides resulting from digested food. Calcium excretion varies directly with thecreatinine clearance.Preparation64 It is obtained in the laboratory by the action of soluble carbonate on a calciumsalt or by passing CO2 through lime water. CaCl2 + Na2CO3 CaCO3 + 2NaCl Ca (OH)2 + CO2 Ca CO3 + H2OUntowards Effects65 Constipation and chalky taste of calcium carbonate are clinical disadvantages.Nausea is an occasional complaint, mere seriously infrequent instances ofhypercalcimia with alkalosis, caleinosis and azotemia occur during chronic calciumcarbonate usage. 22 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  40. 40. Drug reviewContraindications Patients with renal disease, history of calculi, gastro intestinal haermorrhage,hypertension or dehydration and electrolyte imbalance due to excessive vomiting.Properties66 1) It is soluble in water containing carbon dioxide forming calcium bicarbonate. CaCO3 + H2O + CO2 Ca (HCO3)2 2) It is fine white, odorless, tasteless, microcrystalline powder which is stable in air. 3) It is insoluble in alcohol, water and dissolves with effervescence in diluted hydrochloric & diluted Nitric acids.Uses: 1) It is used as Diuretic, Emmenogogue, Astringent, Antacid, Local sedative and Antiseptic. 2) For the manufacture of lime. 3) As a flux in the smelling of ores. 4) In the preparation of tooth pastes and face powders. 23 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  41. 41. Drug reviewReview of Researches carried out on Shankha Bhashma67: 1) PL-31 Adarsh kumar /1984 Shankha Bhasma Nirman Evam Amlapitta par Ahdyayan. 2) Pune – Tilak – 901 Kulkarni (Ms) Manisha M /1995 A comparative study of Shankhanabhi Bhasma and Shanka Bhasma (Avarta) from the point of view of their chemical composition. 3) JM- 554 Tank ZG /2000. A Pharmaco – clinical study of Shankha Bhashma alone and Shankha Bhashma along with Amalaki churna in the management of Amlapitta. 4) JM – 563 Rameshchandra A/ 2002 Shankha Bhashma evan Yuvana – Pidika lepa ka Nirmanatamaka evam Yuvana – pidaka vyadhi par prabhavatmaka Adyayan. 5) Belgaum – 109 Benade shakhar V / 2003 Comparative analytical study and standardization of different samples of Shankha and shankha Bhashma. 24 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  42. 42. Drug review REVIEW OF LITERATURE OF KUMARIClassificationKula : Rasona KulaFamily : LiliaceaeLatin name : Aloe veraEnglish name : Indian aloeSynonyms68: 1) Kumari 2) Grita kumari 3) Gruhakanya 4) Deerghapatrika 5) Ajara 6) Veera 7) Taruni 8) Rama 9) Kapila 10) Sthaladala 11) Mata 12) Mandala 13) Akshayakaraka 14) AtipittalaBotanical Description69:Shrubs : 30-60 cms highLeaves : 1-2 ft in length 2-4 fingers in circumference. 25 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  43. 43. Drug review The margins of the leaves have spikes. Leaves contains a thick andmucilaginous juice. Once the shrub becomes old a straight stalk emerges from its centre whichbears red flowers in cluster. The plant flowers and fruits at the end of winter.Kala bol: The solid gum obtained after boiling the juice of kumari is called as aliabol,kala bol, kumarisara, musabbar etc. It is called aloes in English.Chemical composition70 1) Aloin ie anthraquinone glycoside i.e barboloin as crystalline glucosidee. 2) Aloe amodine 3) Resins : 16 to 63% 4) Water soluble substances 50%Properties71:Guna : Snigdha, picchilaRasa : Tikta, madhuraVeerya : SheetaVipaka : KatuPrabhava : BhedanKala bol : Laghu, Ruksha, Teekshna, UshnaDosha : Tridosha shamakaDhatu : Rasayani, Rasa, Rakta, Meda, Shukra vardhakaMala : Purisha bhedaka, Arthavajanaka.Uses: Gulma, Pleeha Yakrit vruddi, Kaphajwara, Visphota, Vishahara, Kushtagna,Swasa, Pittaja kasa. Aloe is used as a purgative, effect is mainly on colon.Upayukta Anga: Juice from leaves, Kala bol. 26 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  44. 44. Drug review REVIEW OF LITERATUER OF JAMBEERAClassificationKula : Jambeera kulaFamily : RutaceaeLatin Name : Citrus limonSynonyms72 1) Jambeera 2) Dantashata 3) Jamba 4) Jambeera 5) JambalaBotanical Description73Small tree : Strangling, bushy, 3-4m high with thorny branches.Leaves : Ovate, Petiole margined or wingedFlowers : Small, white or pinkish, sweet-scented.Fruit : Oblong or ovoid, bright yellow, rind thick, pulp acid, pale yellow.Macroscopic character74:Colour : Fresh outer surface, bright green yellow, internally white.Dried : Outer surface is yellow and inner surface pithy whiteOdour : Strong, fragnant, aromatic & characteristicTaste : BitterDistribution Cultivated /grown in U.P, Maharashtra, Tamil Nadu, and Karnataka. Foundwild in the north-west regions of India upto 1300m. 27 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  45. 45. Drug reviewMajor Chemical constituents75The Volatile oil of the drug contains mainly limonene (about 90%), citral (about 4%)and other aromatic compounds like geranyl acetate and terpineol.Pharmacological properties76Rasa : AmlaGuna : Guru, TeekshnaVeerya : UshnaVipaka : AmlaDoshaghnata : VatashleshmaharaUses/ Rogaghnata: Shoola, Kasa, Chardi, Trushna, Ama, Asyauairasya, hritpeeda, Vanhimandya, Krumiroga. 28 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  46. 46. Drug review REVIEW OF LITERATURE OF KANJINames in different languages:Sanskrit : KanjiEnglish : Sour gruelMethod of preparation77: The liquor prepared by the fermentation of manda obtained by boilingkulmasha i.e broken masha or dhanya i.e raktashali or yavachurna etc is called kanji.Pharmacological properties78:Guna : Laghu, Teekshna, Ushna, Sparshat sheetaVeerya : UshnaKarma : Dahanashaka, Klamahara, Deepana, Pachana, Bhedi, Rochana, Koshtashuddikara, Bastishodhaka, Rechaka, AmaharaDoshaghnata : VatakaphaharaRogaghnata : Trushna, Shula, Ajeerna, Jwara.Use : For swedana of parada For shodhana of Rasa dravya’s. 29 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  47. 47. Drug review PHARMACEUTICAL PROCEDURES AND YANTRAS USED IN PRESENT STUDYShodhana79: The process by which the malas or doshas of rasadravyas are eliminated iscalled as shodhana. Shodhana is accomplished by applying various methods likemardana, swedana etc.By shodhana rasadravyas becomes soft and brittle for further steps.Shodhana does notmean the mere elimination of impurities but also it is potentiation making the drugsbiologically acceptable.Main objectives of shodhana are : ■ Detoxification ■ Therapeutic potentiation ■ Making rasadravya suitable for next process. In present study shankhanabhi shodhana is done by pachana in dolayantra andmarana by subjecting to gajaputa.Pachana : The process of pachana (boiling) of any rasadravya along with kshara or anydravya swarasa is called as pachana.By doing pachana the mala or impurities loosetheir adhesiveness.Dolayantra80:In present study dolayantra is used for pachana of shankhanabhi. At the neck of earthen pot two holes are made and a iron stick is inserted.Thepot is half filled with required liquid and pottali made of three fold cloth containingrasadravya is suspended such that it is four angula above from the bottom of pot and 30 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  48. 48. Drug reviewimmersed in the liquid.Then the pot is subjected to mrudvagni.This apparatus isknown as dolayantra.Marana : qÉÉUrÉiÉå lÉzrÉiÉå pÉxqÉÏ¢üÏrÉiÉå CÌiÉ | Marana means “killing” and converting a metal into irreversible and final form i.e bhasma.Definition: The processes by which a metals, minerals or any hard substance is subjected to soaking, drying and ignition to convert into bhasma is known as Marana.This marana process converts metals into fine state of smaller molecules and makes them so light as to be highly absorbable and assimilable after oral administration. 1) Marana is process by which metal looses its original state (metallic) & still retains its originality (medicinal value) 2) By marana process drug is converted into a biologically acceptable form. Stages of marana: 1) Mardana: The shodhita dravya is put into khalva and mardana is done withswarasa of specified plants or kashayas of drugs mentioned for a particular mineral ormetal.Mardana is done for a specific period. 2)Chakrika: The small cakes are made which are called as chakrika.The sizeand thickness of cakes depends on the heaviness of the drugs.The heavier the drug thethinner are the cakes.These cakes are dried well under sunlight. 31 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  49. 49. Drug review 3)Sharava and Sandhibandhana : These chakrikas are placed in one singlelayer in shallow earthen plate called as sharava.It is closed with same anotherplate.The edge is sealed with clay smeared cloth in seven consecutive layers anddried.This process is called sandhibandhana. 4)Puta : The puta is the measure of heating arrangement meant for preparingvarious kinds of bhasmas of maharasa,uparasa,loha,etc.We should not give more orless quantity of heat.Gajaputa: Its an arrangement of heating in a pit 90cms (one Rajahasta) inlength,breadth and depth. 32 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  50. 50. Disease review DISEASE REVIEWReview of yakrit in Ayurveda In Ayurveda yakrit is considered as one of the Koshtanga and it is a matrujaavayava formed from samana vata, dehoshma and rakta. (Arunadatta)81The origin and development of the word Yakrit82rÉ qÉç A M×ü : rÉ xÉÇrÉqÉ MüUÉåÌiÉThe meaning is that the liver controls various physiological events. (Ayurved & Hepatic disorders)In veda yakrit is called as Takima or yakna. (Shabda stoma mahanidhi).The other synonyms are 83: Kalakhanda – Dalhana on Su.Sa.Sha.4/25 Jyotisthana / Agnisthana – Su.Sa.Sha.4/57-58 Raktadhara – Dalhana on Su.Sa.Sha.4/17 Raktashaya – Su.Sa.Sha.5/9, Yakritpinda - Parishadya shabdartha shareera.Origin of Yakrit: In garbhavastha yakrit & pleeha are formed from shonita. (Su.Sha.4/25)Position of Yakrit84: The anatomical site of the yakrit is mentioned as below and right to thehridaya,which is hard in texure.Site of Blood85: Sthana of Shonita is Yakrit, Pleeha. 33 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  51. 51. Disease reviewRaktavaha Srotas86: Raktavaha Srotas are two in number & their mula is yakrit,pleeha &raktavahini dhamanis.Yakrit87: An Abdominal organ Yakrit is one among 15 koshtanga’s.Site of Ranjaka pitta88: The pitta present in yakrit & pleeha gives ranjana.Raktadhara kala89: It is the second kala present inside the muscles,within which shonita is present,especially in siras localized in yakrit and pleeha. Just as milky sap flows out when trees with milky sap are cut ( or either barkis bruised), similarly when muscles are cut , blood flows out quickly in great quantity.Purishadhara kala90: This kala extends throughout the koshta and yakrit.Its function is to separatethe faeces from food at the level of unduka (the caecum). The functions of yakrit are mentioned as follows91 It is the moola of the rakta vaha srotas It is the seat for the ranjaka pitta Primary seat for the formation of the rakta Gives gati to the rakta Serves as a seat of raktadhara kala. The mula of raktavah srotas is considered as yakrit and pleeha. Also it ismentioned that when rasa reaches yakrit and pleeha it gets coloured and then it iscalled as rakta. From above description it follows that yakrti, rakta have an Samavayarelation. Therefore for every vitiation of Rakta there will also be derangement in 34 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  52. 52. Disease reviewfunctions of yakrit and vice versa92. The involvement of pitta in this pathology shouldalso be considered as rakta and pitta bear Asraya and Ashrayi bhava and also achapitta is derived from yakrit. In many disease conditions, where Rakta involvement oryakrit involvement is explained in ayurveda. From the description of the digestionprocess mentioned in ayurveda it is evident that yakrit also takes an important role indigestive process. Like this in conditions such as agnimandya, aruchi, hrallasa,uttklesha, ama etc, the involvement of yakrit should be considered.Agni vyapara and Yakrit: 13 types of Agni carry out the process of digestion according to Ayurveda.Jatharagni paka leads to the breakdown of different proximate components of the foodand renders them fit for absorption. Bhutagni paka processes and converts thenutrients absorbed from adhoamashaya as pre- homologous of substances, which aremeant finally to be utilised for the upachaya, or building up of the sthayidhatus. Shri Dwarakanathji (Digestion and metabolism in Ayurveda. Edition-First,1971) has concluded that the bhutagni paka takes place in the Liver from itsanatomical and physiological relationship to the koshta. This clearly indicates the roleof Yakrit in production of “Ama” in its impaired condition and hence produces aruchi,agnimandya, and ajeerna etc conditions. Further according to Dwaraknathji, eventhough the metabolism of asthayidhatu into sthayi dhatus takes places in dhatus itself, but yakrit has an important rolein action on dhatwagnis also.93Yakrit in Raktapitta: Both rakta and pitta are mutually vitiated in this disease. Dravatwa, ushnatwain rakta and pitta will be increased, and it is clearly mentioned by Charaka, that yakrit 35 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  53. 53. Disease reviewpleeha and raktavahi siras are affected in this disease94 (adhistana). While explainingraktapitta chikitsa, Sushruta advised consume Aja yakrit (raw) along with pitta.95Yakrit in Pandu roga: Pandu roga has been counted both in pittaja and raktaja diseases. Pandumanifests because of raktakshaya. There may be sanga in rasavaha, which inhibits thenourishment to rakta etc dhatus96. For the formation of rakta, proper functioning ofrakta dhatwagni and ranjaka pitta is necessary and it takes place at yakrit and pleeha.If ranjaka pitta and rakta dhatwagni are deficit there will be alparakta formationleading to raktalpata. Functional deficiency/vitiation of Yakrit is mentioned in thisdisease indirectly97.Yakrit in Kamala: When the person suffering from pandu, intakes more pittaja substances, theexcessively increased pitta burns rakta and mamsa leading to kamala98. Yakrit is rootof rakta vaha srotoses. In this disease the increased mala pitta vitiates rakta and henceproduces the kamala. It is also evident from investigations that there will be functionaland structural derangement of yakrit in this disease. The symptoms like haridranetra,twak,nakha, anana, raktapeetashakranmutra, bekhavarna, daha, avipaka, aruchi etcmentioned in koshta shakhashraya kamala are similar to the disease jaundice99. In second variety of this disease shakhashrita kamala, there will be sanga ofpitta by kapha, the symptom mentioned such as tilapista nibha varchas etc, are similarto obstructive jaundice. The investigations suggest that there will be sanga of pitta inyakrit, which moves to shakhas to produce the disease condition100. 36 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  54. 54. Disease reviewToxins and Yakrit: Garavisha, a special type of toxin described in Ayurveda affects liver &hepatomegaly may take place. The reason for hepatomegaly is that the blood has special affinity towardstoxins.The toxins immediately get spread in the body through blood.101Table No. 8 Showing Correlation Between Ranjakapitta And Bile. Ranjaka pitta BileSite Yakrit/Liver LiverDerived from Pitta Breakdown products of haemoglobinFunction Imparts colour to purisha, rasa Imparts colour to stool, helps in emulsification of fatsObstruction results in Tila pishta nibha varchas Clay coloured stoolYakrit in Udara roga: - In this condition the direct involvement of Yakrit is mentioned. There will beenlargement of Yakrit in this disease (sparsha gamya –kathinavastha). As theAgnimandya is the root cause of all the Udaras, the functional derangement of Yakritcan be inferred, because it takes part in digestion process102. All the nidana, linga andchikitsa etc. mentioned for the disease Plehoodara should be taken similar forYakritoodara. All the types of Udara rogas mentioned if neglected would turn toJalodara. Yakritodara can be taken, as one of the ajatodakavastha of Jalodara, which ifneglected becomes Jalodara along with morphological and functional changes inYakrit, and this condition resembles ‘Ascites’. All the measures described in Pleehodara are to be adopted inYakritodara103,104,105,106. The causative factors of Pleehamaya are just applicable to 37 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  55. 55. Disease reviewYakritamaya also. The description of Yakrudulyudara in all the text of Ayurveda ismixed up with that of pleehodara. Dalhana while commenting on Sushruta Nidana Sthana 7/16 categoricallystated that Pleehodara itself as Yakrudulyudara. But Bhavamishra describesYakrudulyudara as a variety of Pleehodara.Table No. 9 Showing Nidana of Pleehodara and Yakritodara:107,108,109,110,111 Nidana C.S S.S A.H M.N Y.RAshitasya atiyana sevana + +Atichesta + +Ativyavaya + +Bharavahana + +Adhwa + +Vamanavyadhi karshana + +Vidahi bhojana + +Abhishyandi bhojana + +Ajeerna + + +Malina anna sevana + + +Mala sanchaya + + + The etiological factors in modern science also given more importance to thediet such as exposure to toxic chemical substances, contaminated food, consumptionof spirit/liquors and through drug injury, is at large producing liver disorders.Table No. 10 Showing Roopa of Pleehodara and Yakritodara:112,113,114,115,116Roopa C.S S.S A.H M.N Y.RDourbalya + +Aruchi +Avipaka +Varchograha +Mutragraha + 38 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  56. 56. Disease reviewTama pravesha + +Angamarda +Chardi + +Murcha + +Angasada +Kasa + +Gourava +Admana +Daha +Shwasa + + + + +Mrudujwara + +Anaha +Agninasha +Karshya +Asyavairasya + +Parvabheda +Koshta vata shula + +Aruna varna udara + +Vivarna udara + +Neelarajimudara +Haritarajimudara +Haridrarajimudara + +Katinya +Dakshinaparshwa parivruddi + + +Seedati + + +Mandagni + + +Kaphapitta lingopadruta + + +Atipandu + + 39 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  57. 57. Disease review Doshasambanda Vivechana in Pleehodara and Yakritodara117: If the Yakritodara is associated with - Udavarta, Ruja, Anaha it is Vataja Moha, Trushna, Daha, Jwara it is Pittaja Gourava, Aruchi, Katinya it is Kaphaja 40“Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  58. 58. Disease review LIVER DISEASES - A MODERN APPROACH118 Liver plays a pivotal role in regulation of physiological processes. It isinvolved in several vital functions such as metabolism, secretion and storage. Furthermore detoxification of a variety of drugs and xenobiotics occur in liver. Liver diseasesare mainly caused due to an exposure to toxic chemical substances like antibiotics,chemotherapeutics, peroxidised oils aflatoxin, carbon tetra chloride, chlorinated hydrocarbons, varied infections, auto immuno disorders and also due to chronic alcoholism.Most of the hepato toxic chemicals damage liver cells mainly by inducing lipidperoxidation and other oxidative damages takes place in liver. It has been estimated that about 90% of the acute hepatitis is resulting due toviruses. The major viral agents involved are Hepatitis A, B, C, D, E & G. Of theseHepatitis B infection often results in chronic liver diseases and cirrhosis of liver. Consumption of spirits/liquors is the prime cause for liver damage in India, soalso been through drug injury is at large producing liver disorders. On survey, studiesaround 2-3 % of Indian population are carrying Hepatitis B or C type of viruses.World wide these figures are increasing with an alarm. (Ref: Indian Journal ofPharmacology 1999; 31:166-175)Physiology and its affections119 It is the largest organ in the body weighing from 1200 gm- 1500 gm. Itoccupies the whole of the right hypochondriac region and greater part of the epigastricregion; some times it may even extend to the left hypochondriac region. It is roughlypyramidal or wedge shaped with the base towards the right and apex towards the left. The liver is both exocrine and endocrine in nature. The exocrine part secretesbile, which is carried by bile duct and endocrine part secretes plasma proteins, glucoseand heparin. 41 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”
  59. 59. Disease review Its various function interrelates to each other. This becomes particularlyevident in clinical abnormality of the liver because many of its function aredistributed simultaneously but in different combinations, depending upon the natureof the disorder. The basic function of the liver can be divided into:1. Its vascular functions for storage and filtration of blood.2. Its metabolic functions concerned with the majority of the metabolic system of the body.3. Its secretary and excretory functions that are responsible for forming the bile that flows through bile ducts into the gastrointestinal tract.Physiologic Anatomy of the Liver120 The basic functional unit of the liver is the liver lobule, which is a cylindricalstructure several mm in length and 0.8-2mm in diameter. The human liver contains50,000-1,00,000 individual lobules. The liver lobule is constructed around a central vein that empties into thehepatic veins and hence into the inferior vena cava. The lobule itself is composedprincipally of many hepatic cellular plates that radiate centrifugally from the centralvein like spokes in a wheel. Each hepatic plate is 1-2 cells thick and between theadjacent cells lie small bile canaliculi that empty into bile ducts in the fibrous septaseparating the adjacent liver lobules. Also in the septa are small portal venules that receive their blood from theportal veins. From these venules blood flows into flat, branching hepatic sinusoidsthat lie between the hepatic plates, and then into the central vein. Thus the hepaticcells are exposed continuously to portal venous blood. 42 “Preparation, Physico- Chemical Analysis of Shankha Nabhi Bhasma & Evaluation of its Hepato protective Activity - An Experimental Study”

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