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A Dissertation Submitted To PHARMACOGNOSTICAL, PHYTOCHEMICAL AND MEMORY STRENGTHENING ACTIVITY ON THE SEEDS OF Abelmoschus moschatus Medic. A Dissertation submitted to THE TAMILNADU Dr. M.G.R. MEDICAL UNIVERSITY CHENNAI – 600032 In partial fulfillment of the requirements for the award of degree of MASTER OF PHARMACY IN PHARMACOGNOSY Submitted by Reg No.261220656 Under the guidance of Dr.R.VADIVU M.Pharm., Ph.D., Department of Pharmacognosy College of Pharmacy MADRAS MEDICAL COLLEGE Chennai – 600003 April - 2014 Dr .A. JERAD SURESH, M.Pharm., Ph.D., Principal, College of Pharmacy, Madras Medical College, Chennai - 600003. CERTIFICATE This is to certify that this dissertation work entitled “PHARMACOGNOSTICAL, PHYTOCHEMICAL AND MEMORY STRENGTHENING ACTIVITY ON THE SEEDS OF Abelmoschus moschatus Medic.,” submitted by the candidate bearing the Reg. No. 261220656, final year M.Pharm (Pharmacognosy) student, in partial fulfillment of the requirements for the award of degree in Master of Pharmacy in Pharmacognosy by The Tamil Nadu Dr.M.G.R.Medical University, Chennai-600032, is a bonafide record of work done by her in the Department of Pharmacognosy, Madras Medical College, Chennai, during the academic year 2013 – 2014 under the guidance of Dr. R. VADIVU, M.Pharm.,Ph.D., Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai – 600003. Dr. A. JERAD SURESH, M.Pharm, Ph.D., Place: Chennai – 03 Date: Dr .N. Jayshree, M.Pharm., Ph.D., Professor and Head, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai- 600003 CERTIFICATE This is to certify that this dissertation work entitled “PHARMACOGNOSTICAL, PHYTOCHEMICAL AND MEMORY STRENGTHENING ACTIVITY ON THE SEEDS OF Abelmoschus moschatus Medic.,” submitted by the candidate bearing the Reg. No. 261220656, final year M.Pharm (Pharmacognosy) student, in partial fulfillment of the requirements for the award of degree in Master of Pharmacy in Pharmacognosy by The Tamil Nadu Dr.M.G.R.Medical University, Chennai-600032, is a bonafide record of work done by her in the Department of Pharmacognosy, Madras Medical College, Chennai, during the academic year 2013 – 2014 under the guidance of Dr. R. VADIVU, M.Pharm.,Ph.D., Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai – 600003. Dr. N. JAYSHREE, M.Pharm, Ph.D., Place: Chennai – 03 Date: Dr. R.Vadivu , M.Pharm., Ph.D., Tutor, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai- 600003 CERTIFICATE This is to certify that this dissertation work entitled “PHARMACOGNOSTICAL, PHYTOCHEMICAL AND MEMORY STRENGTHENING ACTIVITY ON THE SEEDS OF Abelmoschus moschatus Medic.,” submitted by the candidate bearing the Reg. No. 261220656, final year M.Pharm (Pharmacognosy) student, in partial fulfillment of the requirements for the award of degree in Master of Pharmacy in Pharmacognosy by The Tamil Nadu Dr.M.G.R.Medical University, Chennai-600032, is a bonafide record of work done by her in the Department of Pharmacognosy, Madras Medical College, Chennai, during the academic year 2013 – 2014 under the guidance of Dr. R. VADIVU, M.Pharm.,Ph.D., Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai – 600003. Dr. R.VADIVU, M.Pharm., Ph.D Place: Chennai – 03 Date: ACKNOWLEDGEMENT The study was carried out at the Department Of Pharmacognosy, Madras Medical College, Chennai, during the year 2012-2013. With immense gratitude I acknowledge the following persons for their pivotal role in completion of the study. I whole heartedly express my high esteem and deep sense of gratitude to the respectable Dean Dr .V. Kanagasabai., M.D., Madras Medical College, Chennai, for providing me all facilities and support during the period of academic course work. It is with the great pleasure, I record my thanks and gratitude to our Principal Dr. A. Jerad Suresh., M.Pharm., Ph.D., College of Pharmacy, Madras Medical College, for providing the lab and administrative facilities to carry out the project work. I express my deep sense of gratitude to Dr. N. Jayshree, M.Pharm., Ph.D., Professor and Head, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, for her support in completion of the project work. I like to thank my guide Dr. R. Vadivu, M.Pharm, Ph.D., for her invaluable support and guidance throughout the project. I am greatly indebted to her for the excellent advice, patience, moral support and help. Her right suggestions at right time have rescued me from various red tape crisis. Without her support, the project would not have been materialized and completed. It’s a great pleasure for me to acknowledge my sincere thanks to all the staff members Dr. Radha, M.Pharm., Ph.D., Dr. P. Muthusamy M.Pharm., Ph.D., B.L., and Dr. R. Vijaya Bharathi, M.Pharm., Ph.D., of the Department of Pharmacognosy for their valuable support and excellent co-operation when needed. I acknowledge my sincere thanks to Dr. Joseph Dixon, B.V.Sc., special veterinary officer and Mr. Kandaswamy, Assistant of Animal Experimental House, Madras Medical College, Chennai-03 for providing the animals to carryout Pharmacological studies. I wish to show my gratitude to Mr.Chelladurai for his efforts in collection of plant material. A special word of thanks goes to the non-teaching staff members Mrs. T.S. Lakshmi and Mrs. M. Kumudhavalli, Department of Pharmacognosy, College of Pharmacy, MMC, Chennai-03. I have no words to express my joy in thanking the Almighty for giving me the fabulous group of friends in PG and all other who are behind me supporting my endeavours. I also extend my thanks to all those who have directly or indirectly helped me during this tenure. I wish to thank my father, who taught me perseverance in life and my mother who was the reason behind all my success. I would like to thank my sisters and brother for sharing all my happiness and challenges throughout the course period. The financial support provided by the Tamilnadu Government is gratefully acknowledged. CONTENTS S.No Title Page No 1 INTRODUCTION 1 2 REVIEW OF LITERATURE 11 3 AIM AND OBJECTIVE OF THE STUDY 13 4 RATIONALE FOR SELECTION 14 5 ETHNOBOTANICAL SURVEY 15 6 PLAN OF WORK 19 7 METHODOLODY 7.1 Pharmacognostical studies 20 7.2 Phytochemical studies 30 7.3 Selection of active extract 37 7.4 Pharmacological studies 39 8 RESULTS AND DISCUSSION 46 9 SUMMARY AND CONCLUSION 81 10 REFERENCES - LIST OF TABLES Table No Title Page No 1. Herbals used for treatment of memory disorders 10 2. Physico chemical constants 54 3. Qualitative estimation of inorganic elements 55 4. Quantitative estimation of inorganic elements 56 5. Quantitative estimation of Heavy metals 56 6. Percentage yield of successive extracts 59 7. Qualitative Phytochemical analysis 60 8. Behaviour of drug powder with different chemical reagents 61 9. Total flavanoid content of ethyl acetate and ethanolic extracts 62 10. Total phenolic content of ethyl acetate and ethanolic extracts 63 11. Quantitative estimation of phytoconstituents 64 12. Fluorescence analysis of powder 65 13. Fluorescence analysis of extracts 66 14. TLC profile of extracts 67 15. Solvent system for HPTLC 68 16. HPTLC profile of extracts 69 17. In-vitro reducing power assay 72 18. In-vitro hydrogen peroxide scavenging assay 73 19. Effect on Transfer Latency 75 20. Brain Acetyl cholinesterase level 76 21. Brain Malondialdehyde Level 78 22. Brain Reduced glutathione level 79 LIST OF FIGURES Figure No Title of figure Page No 1. Anatomy of brain 5 2. Three parts, Three pounds 6 3. The neuron forest and cell signaling 7 4. Alzheimer’s changes the whole brain 8 5. Whole plant of Abelmoschus moschatus 14 6. Seeds of Abelmoschus moschatus 14 7. OECD guidelines 41 8. Elevated plus maze model 43 9. Macroscopy of seeds 46 10. T.S of seed 47 11. Palisade like Scleride layer 48 12. Inner zone of seed 48 13. Inner zone of testa 49 14. Endosperm layer 49 15. Middle zone of seed 50 16. Embryo with folded cotyledons 50 17. Single layered endosperm cells 51 18. Outer zone contain polygonal epidermal cells 51 19. Sclerenchyma cells 52 20. Mucilage cells 52 21. Testa 53 22. Parenchyma and sclerenchyma cells 53 23. Total flavanoid content 62 24. Total phenolic content 63 25. TLC profile for ethyl acetate extract 67 26. TLC profile for ethanolic extract 67 27. TLC profile for ethanolic extract. 68 28. HPTLC profile of ethanolic extract 69 29. In vitro reducing power assay 72 30. In vitro hydrogen peroxide assay 73 31. Transfer latency on 7th and 8th day 75 32. AchE inhibitory activity 76 33. Brain Malondialdehyde level 78 34. Brain reduced glutathione level 79 Memory strengthening activity on seeds of Abelmoschus moschatus S.Nandhini, R.Vadivu*, N.Jayshree Department of Pharmacognosy College Of Pharmacy Madras Medical College Chennai, Tamilnadu, India. ABSTRACT Alzheimer’s disease is a progressive neurodegenerative disorder characterized by gradual decline in memory. The present study was undertaken to investigate the memory strengthening effect of Abelmoschus moschatus Medic. Ethanolic extract of seeds (100,200mg/kg. p.o) was administered for 7 successive days to young mice. Exteroceptive behavioral model such as elevated plus maze was employed to evaluate learning and memory. To delineate the mechanism by which AM exerts memory strengthening activity, the effect of AM on whole brain AchE, Brain Malondialdehyde content , Reduced Glutathione were also assessed. Piracetam (200mg/kg, i.p) was used as a standard drug. Pretreatment with AM (200,400mg, p.o) for seven successive days significantly improved learning and memory in mice and reversed the amnesia induced by diazepam (1mg/kg, i.p). AM also decreased whole brain AchE and Malondialdehyde content and increase the brain reduced glutathione. Hence Abelmoschus moschatus Medic. appears to be a promising candidate for improving memory, Anti cholinesterase activity and Anti oxidant property and it would be worthwhile to explore the potential of this plant in the management of dementia and Alzheimer’s disease.
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