PHARMACOGNOSTICAL, PHYTOCHEMICAL AND PHARMACOLOGICAL EVALUATION OF THE LEAVES AND STEM OF Ipomoea sepiaria koen. ex Roxb. Dissertation submitted to The Tamilnadu Dr. M.G.R. Medical University, Chennai In partial fulfillment of the requirement for the Degree of MASTER OF PHARMACY IN PHARMACOGNOSY Submitted BY 26108670 MAY – 2012 DEPARTMENT OF PHARMACOGNOSY MADURAI MEDICAL COLLEGE MADURAI – 625020 PHARMACOGNOSTICAL, PHYTOCHEMICAL AND PHARMACOLOGICAL EVALUATION OF THE LEAVES AND STEM OF Ipomoea sepiaria koenig ex Roxb. Dissertation submitted to The Tamilnadu Dr. M.G.R. Medical University, Chennai In partial fulfillment of the requirement for the Degree of MASTER OF PHARMACY IN PHARMACOGNOSY Submitted BY VAITHIYANATHAN K. MARCH – 2012 DEPARTMENT OF PHARMACOGNOSY MADURAI MEDICAL COLLEGE MADURAI – 625020 CONTENTS CHAPTER TITLE PAGE.NO. I Introduction 1-11 II Literature survey 12-18 III Aim and scope 19-20 IV Pharmacognostic studies Section –A: General description of the plant, Ipomoea sepiaria 21-23 Section- B : Microscopical study of the leaf Ipomoea sepiaria 24-26 Section- C: Powder microscopy of leaves 27-28 Section-D: Quantitative microscopy 29-32 Section- E: Physical parameters 33-37 V Phytochemical studies Section-A: Organoleptic evaluation 38-42 Section-B: Preliminary phytochemical screening 43-44 Section-C: Estimation i) Estimation of total phenolic content 45-47 iii)Estimation of total flavonoid content 48-49 Section-D: Isolation and characterization of flavonoids 50-54 VI Pharmacological screening. Section-A: Invitro antioxidant activity of Ipomoea sepiaria i) Hydrogen peroxide scavenging activity 55-56 ii) DPPH scavenging activity 57-58 iii) DPPH scavenging activity for isolated compounds 59-60 Section-B: Pharmacological evaluation of Ipomoea sepiaria on 61-70 acrylamide induced neuropathic pain in rats. Section-C: Pharmacological evaluation of Ipomoea sepiaria on invitro 71-76 anticancer activity in HepG2 liver cancer cell lines VII Results and discussion 77-92 VIII Conclusion 93-94 IX References i-xiis Dr. Mrs. AJITHADAS ARUNA, M.Pharm., Ph. D., Principal& Head of the Department, Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai-625 020 CERTIFICATE This is to certify that the dissertation entitled “Pharmacognostical, Phytochemical and Pharmacological evaluation of the leaves of Ipomoea sepiaria koen. ex Roxb. was done by Mr. K.VAITHIYANATHAN, in the Department of Pharmacognosy, Madurai Medical College, Madurai-20, in partial fulfillment of the requirement for the Degree of Master of Pharmacy in Pharmacognosy. This dissertation is forwarded to the Controller of Examination, The Tamilnadu Dr. M.G.R. Medical University, Chennai. Station: (Mrs. AJITHADAS ARUNA) Date : Mr.T. VENKATARATHINAKUMAR, M.Pharm., (Ph. D)., Assistant Reader, Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai-625020 CERTIFICATE This is to certify that the dissertation entitled “Pharmacognostic, Phytochemical and Pharmacological evaluation of the leaves of Ipomoea sepiaria Koen . ex Roxb. was done by Mr. K.VAITHIYANATHAN, in partial fulfillment of the requirement of the award of DEGREE OF MASTER OF PHARMACY IN PHARMACOGNOSY at the DEPARTMENT OF PHARMACOGNOSY, MADURAI MEDICAL COLLEGE, MADURAI-20 affiliated to The Tamilnadu Dr. M.G.R. Medical University, Chennai is a bonafide work carried out by him, under my guidance and supervision during the academic year 2011-2012. Station: (Mr.T. VENKATARATHINAKUMAR) Date: ACKNOWLEDGEMENT It is my privilege and honour to express my sincere thanks to our respectful sir Dr. Edwin Joe M.D., Dean, Madurai Medical College, Madurai and Dr.T. Meena M.D.Vice Principal, Madurai Medical College, Madurai for providing me with all the necessary facilities to do my project work. I am very thankful to Dr. Mrs. Ajithadas Aruna, M.Pharm., Ph.D Principals, Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai for her excellent encouragement, guidance, boundless enthusiasm, motivation and valuable advice for the successful completion of my project. I wish to place on record here my indebtedness and heartfelt thanks to Mr.T.Venkatarathinakumar M.Pharm., (Ph.D) Assistant Reader, Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai for his enthusiastic co-operation as my project guide & for all the constant valuable suggestions and encouragement to improve and complete the project work. I extend my special and sincere thanks to Ms.R.Gowri, M.Pharm, Assistant Reader, Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai for her diligence and for all the consistent encouragement, suggestions, contribution and support extended during the project work. I express my sincere thanks to Dr. Mr. k. Periyanayagam M.Pharm., Ph.D Assistant Reader, Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai for his friendly and cheerful guidance during the course. I am thankful to Dr.Mr. Jonat M.V.Sc. Assistant Surgeon, Central Animal House, Madurai Medical College, Madurai, for their constant support for permitting me to utilize animals for pharmacological study of this dissertation. I owe my special thanks to Mr.Arunachalam Muthuraman M.Pharm (Ph.D) Department of Pharmaceutical Science and Drug Research, Punjab University, whose valuable suggestion regarding the pharmacological studies and timely help made this work as a successful one. I am thankful to Mrs.A.Sedhuramani M.Pharm (Ph.D) and Mrs.A.Krishnaveni. M.Pharm (Ph.D) Tutors in Pharmacognosy, Madurai Medical College, Madurai for their friendly and encouragement to improve and complete this work. I extend my special thanks to Miss.G. Shanthini nachiar M.pharm,department of pharmacognosy,college of pharmacy, Madurai medical college Madurai. I extend my special thanks to Prof. Dr.P. Jayaraman, Ph.D, Director (PARC) Chennai for his help regarding the Microscopy and authentification of the plant. I extend my special and sincere thanks to Dr.Mr.Sethuraman M.phill, Phd, ASSISSTANT MANAGER IN BAL pharma pvt ltd, his diligence and for all the consistent encouragement, suggestions, contribution and support extended during the project work. I express my gratitude to Mr.M.BARANI ,Msc Assisstant manager in MEDRICH pharma pvt ltd Bangalore for his support and help in carry out the analytical calculations of this work. I place on record my sincere thanks to Mr.Sureshkumar B.pharm, MBA,R&D in ORCHID ltd Irunnkaatukotai chennai for hiscontinuous support on thesystem applications at appropriate timings I express my sincere thanks to Mr. N. Cheransenguttuvan for his generous help in accomplishing this scientific work. I thank Mr.P.Sivakumar M.sc Lab supervisor and Mrs S.Kalpana sivakumar M.Sc. Jayanithi uro labs I am thankful to Mr. Johns Universal Scientific Suppliers for his timely supply of chemicals to carry out this work. I am thankful to my juniors for their help for this project work. I also thank the faculty and non faculty of the pharmacognosy department for their help during the course of my dissertation work. Most of all I would like to thank my lovable , mother,sisters and their family members all the support and love they have given to me, always they are the reason behind my every successful moments. CHAPTER-I INTRODUCTION Medicinal and aromatic plants have been used since ancient times in the traditional practices for health care. it is well known that plants are an important source for a variety of bioactive compounds for a variety of purposes including pharmacy ,medicine and industry, According to WHO report nearly 70-80% of the world population relies on traditional medicine largely which are produces plant based for primary health care needs. the annual international market of herbal product is estimated to be US$20 million. with increasing popular demand for medicinal plants both in india and internationally. Human being had depended only on natural sources to a larger extent particularly on plants of his vicinity for getting rid of these ailments. Besides for his other needs Several potent medicinal plants have seen the light of the day due to his continous usage of the effective drugs, Some of these plants have attracted the herbalist who have put these plants in to use for centuries together and included the more potent one in the materia medica of their region. Scientific study of folk medicine is very essential because the efficacy of a drug depends upon various factors such as genuinity, preparation administration and dosage of drugs. Pharmacognosy is the study of crude drugs obtained from plants, animals and mineral kingdom and their constituents. Even though science of Pharmacognosy is practiced since a very early period. 1 The term Pharmacognosy was first used by C.A. SEYDLER, a German Scientist, in 1865 in his book Analecta Pharmacognostica. It is derived from two Latin words Pharmakon, “a drug” and gignosco, “to acquire knowledge of”. It means knowledge or science of drugs.1,2,3,4,5 NEUROPATHY A classical term for any disorders affecting any segment of the nervous system. NEUROPATHIC PAIN Neuropathy is a collection of disorders that occurs when nerves of the peripheral nervous system (the part of the nervous system outside of the brain and spinal cord) are damaged. The condition is generally referred to as peripheral neuropathy, and it is most commonly due to damage of nerve axons. Neuropathy usually causes pain and numbness in the hands and feet. It can result from traumatic injuries, infections,