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Nausheen Chemistry 2019 Uom Malakand Prr.Pdf BIOASSAY GUIDED ISOLATION OF SECONDARY METABOLITES FOR ANTIDIABETIC AND ANTI-AMNESIC POTENTIAL OF ELAEAGNUS UMBELLATA THUNB. PhD Thesis By Nausheen DEPARTMENT OF CHEMISTRY UNIVERSITY OF MALAKAND 2019 BIOASSAY GUIDED ISOLATION OF SECONDARY METABOLITES FOR ANTIDIABETIC AND ANTI-AMNESIC POTENTIAL OF ELAEAGNUS UMBELLATA THUNB. By Nausheen A THESIS SUBMITTED TO THE UNIVERSITY OF MALAKAND IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY (PhD) IN BIOCHEMISTRY 2019 In the name of Allah, the most beneficent, the most merciful. CERTIFICATE OF APPROVAL This is to certify that the research work presented in this thesis entitled, “BIOASSAY GUIDED ISOLATION OF SECONDARY METABOLITES FOR ANTIDIABETIC AND ANTI-AMNESIC POTENTIAL OF ELAEAGNUS UMBELLATA THUNB.” conducted by Mrs. Nausheen under the supervision of Dr. Muhammad Zahoor, Assistant Professor. No part of this thesis has been submitted anywhere else for any other degree. This thesis is submitted to the Department of Chemistry, in partial fulfilment of the requirements for the degree of Doctor of Philosophy (PhD) in the field of Biochemistry, Department of Chemistry, University of Malakand is hereby approved for submission. Student Name: Mrs. Nausheen Signature---------------------- Examination Committee: External Examiner 1: Dr. Signature--------------------- External Examiner 2: Dr. Signature--------------------- Internal Examiner: Dr. Signature--------------------- Supervisor: Dr. Muhammad Zahoor Assistant Professor, Signature---------------------- Department of Chemistry, University of Malakand. Chairman: Dr. Manzoor Ahmad, Signature--------------------- Associate Professor, Department of Chemistry, University of Malakand. AUTHOR'S DECLARATION I Mrs. Nausheen hereby state that my PhD thesis titled “BIOASSAY GUIDED ISOLATION OF SECONDARY METABOLITES FOR ANTIDIABETIC AND ANTI- AMNESIC POTENTIAL OF ELAEAGNUS UMBELLATA THUNB.” is my own work and has not been submitted previously by me for taking any degree from University of Malakand or anywhere else in the country/world. At any time if my statement is found to be incorrect even after my graduation, the University has the right to withdraw my PhD degree. Nausheen Date: PLAGIARISM UNDERTAKING I solemnly declare that research work presented in the thesis titled “BIOASSAY GUIDED ISOLATION OF SECONDARY METABOLITES FOR ANTIDIABETIC AND ANTI-AMNESIC POTENTIAL OF ELAEAGNUS UMBELLATA THUNB.” is solely my research work with no significant from any person. Small contribution/help wherever taken has been dually acknowledged and that complete thesis has been written by me. I understand the zero tolerance policy of the HEC and University of Malakand towards plagiarism. Therefore i as an author of the above titled thesis declare that no portion of my thesis has been plagiarized and any material used as reference is properly referred/cited. I undertake that if i am found guilty of any formal plagiarism in the above titled thesis even after award of PhD degree, the University reserves the rights to withdraw/revoke my PhD degree and that HEC and the University has the right to publish my name on the HEC/University website on which name of students are placed who submitted plagiarized thesis. (A copy of the plagiarism report performed at the time of thesis submission, by quality enhancement cell (QEC) of the University is also attached). NAUSHEEN I am dedicated my thesis to my beloved parents and husband, whose prayers, moral and financial support enabled me to conduct and complete this achievement. Nausheen i ACKNOWLEDGMENTS Beginning with the greatest name of Al-Mighty Allah, the most gracious, sympathetic and ever merciful, who gave me mental and physical strength that enable me to complete this difficult task in time. It was great blessing of Allah that I got loving parents, inspiring and talented teachers and friends who provide me the intellectual guidance, moral support and all-round help for the fulfilment of my work. I offer my humblest thanks from the core of heart to Holly prophet Muhammad (PBUH) who is forever a torch of guidance and knowledge for humanity as a whole. I have no words to express my deepest sense of gratitude to the Holly prophet Muhammad (PBUH), who has deep sense of love, affection, sympathy, kindness and sincerity for whole humanity and all creatures leaving on the earth. I deem it utmost pleasure to avail this opportunity to express my heartiest gratitude to my respectable research supervisor, Dr. Muhammad Zahoor, Associate Professor, Department of Chemistry, University of Malakand, for his professional advice, valuable guidance, technical approach, consistent encouragement, sympathetic attitude, art of making useful suggestion and inspiring attitude made it very easy to undertake this work and to write these thesis. I wish to express from the core of my heart very special thanks to Dean of Sciences Prof. Dr. Rasheed Ahmad Department of Chemistry and Dean of Biological Sciences Prof. Dr. Mir Azam Khan Department of Pharmacy, University of Malakand for their tremendous knowledge of the subject and incentive guidance throughout my research work. I am greatly thankful to express heartiest gratitude to Dr. Manzoor Ahmad Chairman Department of Chemistry whose unwavering support, keep interest and encouragement due to which I was succeeded to complete this task and led to the completion of this scientific work. I would like to express my special appreciation and thanks to Dr. Mohammad Nisar Chairman Department of Botany you have been a tremendous mentor for me. Your impeccable research skills, clear guidance, administrative support and advice on both research as well as on my career have been invaluable. I would like to pay sincere thanks, to Dr. Waqar Ahmad Department of Pharmacy, Dr. Ezat Khan, Dr. Abdul Lateef, Dr. Mumtaz Ahmad, Dr. Sultan Alam, Dr. Naveed Umer, Dr. ii Najeeb and all faculty members of Department of Chemistry, University of Malakand, for their useful comments, support, kind guidance and encouragement and helping during my research work. I also offer special thanks to Dr. Nasiara Karim, Dr. Shoaib, Dr. Rukhsana, Dr. Wadud Ali Shah and Dr. Farhat Ullah, Department of Pharmacy University of Malakand and Dr. Imran Khan, Department of Pharmacy, University of Swabi, Sajjad Ahmad Department of Pharmacy, Sarhad University of Information Technology, Peshawar, Zia Uddin Department of pharmacy, COMSATS University Islamabad, Abbottabad Campous whose encouragement, impeccable research skills and support help me throughout my research work. I am thankful to all the faculty members, technical and administrative staff of the Chemistry, Botany and Pharmacy departments, University of Malakand, who contributed to the successful completion of my research work and providing me lab facilities. Finally, I pay my regards and duly acknowledge the co-operation, guidance and support of my loving Parents, Brothers and sister. Words are meaningless to what they have given me. It is, believe their prayers, sincere encouragement, and moral support which were a constant source of strength and inspiration to me that despite, many constraints, I was able to complete my Ph.D. work. And most of all for my loving, supportive, encouraging, and patient husband Muhammad Nazir whose faithful support during my Ph.D. is so appreciated. Thank you. NAUSHEEN iii LIST OF ABBREVIATIONS Abbreviations Full Name % SAP % Spontaneous Alternation-performance µg Micro gram µL Micro Litre 2D Two dimensional AAR Alternate Arm-returns ABI Acquired Brain Injury ABTS 2, 2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) ACh Acetylcholine AChE Acetylcholine esterase ACTH Adrenocorticotropic hormone AD Alzheimer's disease ADH Alcohol dehydrogenase AGEs Advance Glycation end-products ALDH Aldehyde dehydrogenase ALP Serum alkaline-phosphatase ANOVA Analysis Of Variance APP Amyloid precursor protein Aq. Ext Fraction Aqueous As Peak area of standard Ax Peak area of sample Aβ β-amyloid BBB Blood brain barrier BChE Butyryl cholinesterase iv But. Ext Fraction n-Butanol C13NMR Carbon-13 Nuclear Magnetic-resonance Chf-Ext Fraction of chloroform CNS Central nervous system COX-2 Cyclooxygenase-2 CRF Corticotrophin releasing factor CRP C-reactive protein Cs Concentration of standard CVA Cerebrovascular accident Cx Concentration of sample DI Discrimination index DM Diabetes-mellitus DMSO Di-methylsulphoxide DNA Deoxyribonucleic Acid DNSA 3, 5-dinitrosalicylic acid DPPH 2, 2-Diphenyl,1,picrylhydrazyl E. umbellata Elaeagnus umbellata EDTA Ethylene Diamine Tetra Acetic acid EI-MS Electron Ionization Mass Spectrometry EtAc-Ext Fraction of ethyl acetate FAB-MS Fast Atom Bombardment Mass Spectrum FDA Food and drug administration FTIR Fourier Transform Infrared-spectroscopy GC Gas-chromatography GC/MS Gas-chromatography/Mass-spectrometry v GIP Glucose insulinotropic polypeptide GLP-1 Glucagon like polypeptide-1 GLUT4 Glucose transporter 4 GSK3β Glycogen synthase kinase 3β HDL High Density-lipoprotein Hex. Ext Fraction n-hexane fraction HFD High fat diet HMBC Hetero-nuclear multiple-Bond Coherence HNMR H+-Nuclear Magnetic-resonance HPA Hypothalamic pituitary adrenal HPLC-UV High performance liquid chromatography- ultra violet HPTLC High Performance thin-liquid-chromatography HSAM Highly-superior Autobiographical Memory HSQC Hetero-nuclear single-quantum Coherence i.p. Intraperitoneal IAPP Islet amyloid polypeptide IC 50 Minimum inhibitory concentration IDDM Insulin dependent diabetes mellitus IDE Insulin-degrading enzyme IL-1β Interleukin-1β IR Insulin
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