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DEVELOPMENT and EVALUATION of HERBAL OINTMENT from Justicia Procumbens Linn., for INFLAMMATION on EYE

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DEVELOPMENT and EVALUATION of HERBAL OINTMENT from Justicia Procumbens Linn., for INFLAMMATION on EYE DEVELOPMENT AND EVALUATION OF HERBAL OINTMENT FROM Justicia procumbens Linn., FOR INFLAMMATION ON EYE A Dissertation submitted to THE TAMILNADU Dr. M.G.R MEDICAL UNIVERSITY CHENNAI-600 032 In partial fulfilment of the requirements for the award of degree of MASTER OF PHARMACY IN PHARMACOGNOSY Submitted by A.BABU REG. NO: 261520651 Under the guidance of DR. R.VADIVU, M.Pharm., Ph.D., Department of Pharmacognosy College of Pharmacy Madras Medical College Chennai-600 003. MAY 2017 DR. A. JERAD SURESH M.Pharm., Ph.D., M.B.A., Principal, College of Pharmacy, Madras Medical College, Chennai-600003. CERTIFICATE This is to certify that the dissertation entitled “ DEVELOPMENT AND EVALUATION OF HERBAL OINTMENT FROM Justicia procumbens Linn., FOR INFLAMMATION ON EYE " submitted by Reg. No: 261520651 in partial fulfilment of the requirements for the award of the degree of MASTER OF PHARMACY IN PHARMACOGNOSY by The Tamil Nadu Dr. M.G.R Medical University, Chennai, is a bonafide record of work done by him in the Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai-600003, during the academic year 2016-2017 under the guidance of DR.R.VADIVU, M.PHARM., Ph.D., Asst.Professor, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai-600003. DR .A. JERAD SURESH M.Pharm., Ph.D., MBA., Place: Chennai-03 Date: DR. R. RADHA M.Pharm., Ph.D., M.B.A., Professor and Head, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai-600003. CERTIFICATE This is to certify that the dissertation entitled “DEVELOPMENT AND EVALUATION OF HERBAL OINTMENT FROM Justicia procumbens Linn., FOR INFLAMMATION ON EYE " submitted by Reg. No: 261520651 in partial fulfilment of the requirements for the award of the degree of MASTER OF PHARMACY IN PHARMACOGNOSY by The Tamil Nadu Dr. M.G.R Medical University, Chennai, is a bonafide record of work done by him in the Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai-600003, during the academic year 2016-2017 under the guidance of DR.R.VADIVU, M.PHARM., Ph.D., Asst.Professor, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai-600003. Dr. R. RADHA M.Pharm., Ph.D., M.B.A., Place: Chennai-03 Date: DR. R.VADIVU M.Pharm., Ph D., Assistant Professor, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai-600003. CERTIFICATE This is to certify that the dissertation entitled “DEVELOPMENT AND EVALUATION OF HERBAL OINTMENT FROM Justicia procumbens Linn., FOR INFLAMMATION ON EYE " submitted by Reg. No: 261520651 in partial fulfilment of the requirements for the award of the degree of MASTER OF PHARMACY IN PHARMACOGNOSY by The Tamil Nadu Dr. M.G.R Medical University, Chennai, is a bonafide record of work done by him in the Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai-600003, during the academic year 2016-2017 under my guidance and supervision. DR.R.VADIVU, M.PHARM., Ph.D., Asst.professor, Place: Chennai-03 Date: ACKNOWLEDGEMENT I express my first and foremost respect and gratitude to GOD with whose blessings I was able to complete my project work. I wish to acknowledge my sincere thanks and express my heartfelt gratitude to the following persons for their help and encouragement.. I am grateful to express my sincere thanks to Dr.Narayana Babu, Dean, Madras Medical College, for giving an opportunity to carry out my project work. I acknowledge my sincere thanks to Prof. DR. A. JERAD SURESH M.Pharm., Ph.D., M.B.A., Principal, College of Pharmacy, Madras Medical College, Chennai, for his support in carrying out my dissertation work in this institution. I express my heartfelt gratitude to DR. R. RADHA M.Pharm., Ph.D., M.B.A., Professor and Head, Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai, for providing me with all the necessary facilities and valuable guidance for my project work. I am very much privileged to take this opportunity with pride and immense thanks expressing my deep sense of gratitude to my guide DR.R.VADIVU, M.Pharm., Ph.D., Assistant Professor, Department of Pharmacognosy, College of Pharmacy, Madras Medical College Chennai, for her innovative ideas, strong support, encouragement and valuable guidance throughout my project work. It’s a great pleasure for me to acknowledge my sincere thanks to the Teaching staff members DR. P. MUTHUSWAMY M.Pharm., Ph.D., DR. R.VIJAYA BHARATHI M.Pharm., Ph.D., and Mrs. B. KUMUDHAVENI, M.Pharm., of the Department of Pharmacognosy for their valuable guidance and excellent co-operation. I thank Mr. RADHA KRISHNA REDDY, Siddha College, Arumbakkam, for the detailed literature review of the plant. I thank Prof. P. JAYARAMAN .Ph.D., Director (PARC) and Mr. MANIKANDAN (PARC), West Tambaram for the Histological studies of this plant material. I thank Mr. KRISHNARAJ, Apex Laboratories, Guindy for doing in vitro studies. I acknowledge my sincere thanks to Dr. S. K. SEENIVELAN, B.V.Sc., Special Veterinary Officer, Animal Experimental House, Madras Medical College, Chennai for providing the animals to carry out the Pharmacological studies. I also thank Mr. KUMAR and Mr. KANDASAMY for their assistance Animal Experimental House, Madras Medical College, Chennai during the animal studies. A special word of thanks goes to the non-teaching staff members Mrs. T.S. LAKSHMI and Mrs. M. KUMUDHA, Lab Technicians of Department of Pharmacognosy, Madras Medical College and R. INDIRA, Chennai for their help during my research work. Also, I thank LAB INDIA CHEMICAL, MICRO FINE CHEMICALS for providing the necessary chemicals for the project work. My deepest heartfelt thanks to all my classmates ELAYASURIYAN, KARTHIK, KOPPERUNDEVI, NARAYANASAMY, PREMKUMAR, RAJKUMAR, SHALINI, SHANMUGAPRIYA AND SARAVANA RAJ for their help, co-operation, motivation, ideas and encouragement. Special thanks to KOPPERUNDEVI for guiding me to choose this plant and SHALINI for providing this plant. Also thanks to my lovable friends DINESH, VIVEK AND DEEPAN for always being there for me !!! I extend my sincere thanks to ALL OTHER BATCHMATES, SENIORS and JUNIORS for their help during the research work. My heartfelt thanks to Tamil Nadu Government for providing financial support for my post graduate studies. Last but not least, My special lovable thanks to my MOTHER , FATHER AND SISTER for their continuous overwhelming support. CONTENT PAGE S.NO TITLE NO. 1. INTRODUCTION 1 2. REVIEW OF LITERATURE 13 3. PLANT PROFILE 15 4. RATIONALE FOR SELECTION OF THE PLANT 19 5. AIM AND OBJECTIVES OF THE STUDY 20 6. PLAN OF WORK 21 PHARMACOGNOSTICAL STUDIES 7. 7.1 Materials and methods 22 7.2Results and discussion PHYTOCHEMICAL STUDIES 8. 8.1 Materials and methods 44 8.2 Results and discussion SELECTION OF ACTIVE EXTRACT 9. 9.1 Materials and methods 60 9.2 Results and discussion FORMULATION OF HERBAL EYE OINTMENT 10. 10.1 Material and Methods 66 10.2 Results and Discussion PHARMACOLOGICAL EVALUATION OF HERBAL EYE 11. OINTMENT 70 11.1 Material and Methods 11.2 Results and Discussion 12. SUMMARY AND CONCLUSION 79 13. REFERENCES 82 LIST OF TABLES S.NO TITLE PAGE NO. 1. Plant description 18 Histochemical colour reactions of Justicia procumbens Linn., 2. 39 3. Linear measurement of trichomes 40 4. Stomatal number and stomatal index 40 5. Physiochemical analysis of the leaves of Justicia procumbens 41 Linn., Qualitative estimation of inorganic elements of Justicia 6. procumbens Linn 42 Quantitative estimation of inorganic elements of Justcia procumbens 7. 42 Linn Quantitative estimation of Heavy metals 8. 43 9. Percentage yield of successive extracts of leaves of Justicia 51 procumbens Linn., Preliminary phytochemical analysis 10. 51 11. Determination of Total Flavanoid Content 52 Determination of Total Phenolic Content 12. 53 13. Quantitative estimation of phytoconstituents 53 Fluorescence characteristic of powdered 14. 54 leaves of justicia procumbens linn., 15. Fluorescence analysis of various 55 extracts of Justicia procumbens Linn., 16. Thin layer chromatographic studies of all three extracts 55 DPPH Radical Scavenging activity of various extract 17. 62 Hydrogen peroxide Radical Scavenging activity of various extract 18 64 In vitro anti-inflammatory activity of various extracts of Justicia 19. 65 procumbens Linn., S.NO TITLE PAGE NO. Physical Appearance 20. 67 pH of ointment 21. 68 22. Extrudability of ointment 68 23. Descriptive rating of intial pain reaction 71 24. Acute irritancy-parameters studied 72 Intial pain reaction observed 25. 72 Grouping of animals 26. 74 Changes in the chemosis 27. 76 Changes in the eye discharge 28. 77 Changes in the redness 29. 78 LIST OF FIGURES S.No. TITLE PAGE NO. 1. Cell mediated immunity 6 2. Whole plant of Justicia procumbens 16 3. Morphology of leaf 33 4. T.S of leaf through midrib 33 5. T.S of Midrib – enlarged 34 6. T.S of Midrib with vascular bundle 34 7. Nonglandular trichomes from the submarginal part of the 35 lamina 8. Epidermal peeling of the leaf showing stomata 36 9. Fibres 37 10. Cystolith isolated from the leaf 38 11. Total Flavonoid content 52 12. Total Phenolic Content 53 13. TLC of three extracts of leaves of justicia procumbens linn 56 14. HPTLC of ethyl acetate extract 57 15. Histogram of ethyl acetate extract of leaves of 57 justicia procumbens linn., 16. HPTLC fingerprint of ethylacetate extract of 58 Justicia procumbens Linn., 17. HPTLC of Ethanolic extract 58 18. Histogram of ethanolic extract 59 19. HPTLC finger print of ethanolic extract of justicia 59 procumbens linn., 20. DPPH Radical Scavenging activity of various extract 63 21. Hydrogen peroxide Radical Scavenging activity of various extract 64 22. Anti-inflammatory activity of various extracts of Justicia 65 procumbens Linn., Introduction 1.1 INTRODUCTION 1 Allergic conjunctivitis is an undesirable inflammatory reaction of the conjunctiva resulting from acquired immune response against an allergen (a foreign antigen to which the individual has been previously sensitized).
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