Al-Neelain University Graduate College The Effect of Drying on Chemical Composition and Antioxidant activity ofCymbopogon citratus and Cymbopogon schoenanthus (L.) spring (proximus) Growing in Sudan A Thesis Submitted for Fulfillment of the Requirements of Master Degree in Chemistry By Balgies Abd Alrouf Abd Alrahman Sudan University of Science and Technology (B.sc) of Science Section of Chemistry Dec. / 2006 Supervisor: Prof. Dr.Saad Mohamed Hussein Ayoub Khartoum October . / 2017 1 ﭧ ﭨ ﭽ ﯱ ﯲ ﯳ ﯴ ﯵ ﯶ ﯷ ﯸ ﯹ ﯺ ﯻ ﯼ ﯽ ﯾ ﯿ ﰀ ﰁ ﰂ ﰃ ﭼ الكهف: ٩٠١ I DEDICATION I would like to dedicate my thesis to my Father, Mother, Brother and my friends II ACKNOWLEDGMENTS All praise be to Allah, who gave me the ability to complete this I would like to express my deepest gratitude to my work. Supervisor Prof. Saad Mohamed Hussein Ayoub My thanks are also to Dr. Itmad Awad Elhassan and special thanks to Dr.Mubark El-Siddig Elamin Special thanks to Dr.Zeinab Eldosh and my friends III Abstract The essential oils prepared by water distillation of fresh and dried leaves of Cymbopogonproximus and C.citratus family (Poaceae) were investigated by GC-MS analysis to determine the effect of drying on their chemical composition and antioxidant activity. The oil content based on weight of sample was different: 1% and 2.87% in fresh and dried leaves of C.proximus; 0.5% and 1.5% in fresh and dried leaves of C.citratus.Physico- chemical properties of the oils prepared from fresh and dried samples were not compatible with published data due to different geographical locations and seasonal variations. The antioxidant activity of the oils prepared from fresh and dried leaves of C.proximus was below 50% (08±03 and 28±01%), while that of C.citratus has increased significantly from 43±0.03% for fresh leaves to 66±0.0% for the dried leaves. This was confirmed by GC-MS analysis of the four oil samples from the two plants. In C.proximus oil the proportion of mono and sesquiterpenes was not effected by drying (2:1) with regard to pipertetone, 2-carene and D- limonene. The case was different in C.citratus oil where new compounds emerged and their content was high: (+)-3-(2-Hydroxy-2-methyl propyl)-2,2- Dimethylcycloprpanone-trans-1-Carboxylic acid (10.84%) and 2,2- Dimethyl-1-oxa-Spiro(2,4)heptanes (15.08%), with regard to β-Citral, α- Citral and β-myrcene. IV ملخص البحث فً هذا البحث حن اسخخﻻص الشيج هي ًباحً الوحزيب وحشيشت الليوىى عي طزيق الخقطيز البخاري وهي ثن اجزي الخحليل اﻻولً لوعزفت هكىًاث الشيج باسخخذام حقٌيت كزوهاحىغزافيا الغاس الولحق بالوطياف الكخلً لوعزفت حاثيز الخجفيف فً كﻻ الٌباحيي وححذيذ فعاليت الشيج هي ًاحيت هضاداث اﻻكسذة وًسبت الشيج فً الٌباحيي وحخزاوح قيوخها بيي )-1 %8.8( V Table of Contents I اﻻيت Dedication II Acknowledgments III English Abstract IV Arabic Abstract V Table of contents VI List of Tables X List of Figures XII Chapter one 1 Introduction and literature review 1 1.1 General introduction 1 1.2 Literature review 5 1.2.1 Essential oils 5 1.2.2 Methods of preparation of essential oils 5 1.2.2.1 Distillation 5 1.2.2.2 Hydro- distillation 5 1.2.2.3 Hydro- diffusion 5 1.2.2.4 Solvent extraction 6 1.2.2.5 Super critical carbon dioxide 6 1.2.3 Chemical constituents of essential oil 6 1.2.4 Uses of essential oils 8 VI 1.2.5 Cymbopogon proximus plant 8 1.2.6 Taxonomy 9 1.2.7 Botanical characteristics 10 1.2.8 Geographical distribution 10 1.3 Chemistry of the C.proximus 10 1.3.1 Chemical composition of C.proximus 10 1.4 Medicinal uses of C.proximus 14 1.4.1 Antimicrobial activity 14 1.4.2 Antioxidant activity 15 1.5 Cymbopogon citratus plant 15 1.6 Taxonomy 16 1.7 Botanical characteristics 16 1.8 Chemistry of C.citratus 17 1.8.1 Chemical composition of C.citratus 17 1.9 Pharmacological 23 1.9.1 Anti-amebic effect 23 1.9.2 Antibacterial activity 23 1.9.3 Ant diarrheal activity 23 1.9.4 Ant filarial activity 23 1.9.5 Antifungal activity 23 1.9.6 Anti- inflammatory activity 24 1.9.7 Anti malarial activity 24 1.9.8 Antimutagenicity 24 VII 1.9.9 Antimyco bacterial activity 24 1.9.10 Antinociceptive effect 24 1.9.11 Antiprotozoan activity 25 1.9.12 Ascaricidal activity 25 1.9.13 Antioxidant capacity 25 1.9.14 Hypocholesterolemic effect 25 1.9.15 Hypoglycemic and Hypolipidemic effects 25 1.9.16 Larvicidal Activity 25 1.9.17 Neurobehavioral effect 25 1.10 Traditional uses of Cymbopogon citratus 26 Chapter two 2 Materials and methods 27 2.1 Materials 27 2.1.1 Plant materials 27 2.1.2 Solvents and chemicals 27 2.1.3 Glassware 27 2.1.4 Apparatus and instruments 28 2.2 Methods 28 2.2.1 Determination of essential oils content 28 2.3 Determination of physicochemical properties of C.proximus 28 and C.citratus essential oils 2.3.1 Specific gravity 28 2.3.2 Refractive index 29 VIII 2.3.3 Optical rotation 29 2.3.4 Determination of acid value 29 2.3.5 Determination of saponification value 30 2.3.6 Determination of ester value 30 2.3.7 Chromatographic method 30 2.3.7.1 GC-MS Analysis of essential oils 30 2.3.8 Determination of antioxidant activity of the prepared 31 essential oils 2.3.8.1 DPPH radical scavenging assay 31 Chapter three 3 Results and Discussion 32 3.1 Pysico-chemical properties of the prepared essential oils of 32 C. proximus and C.citratus 3.2 GC-MS Analysis of essential oils 33 Chapter four 4 Conclusion and Recommendations 49 4.1 Conclusion 49 4.2 Recommendations 50 References 51 IX List of Tables 1 Table (1) Showes GC-MS-profile of components of 10 C.proximus essential oil obtained by hydro-distillation in Egypt 2 Table (2) Showes GC-MS Profile Major Constituents of 12 C.proximus oil obtained by hydro distillation. 3 Table (3) Chemical composition of the essential oil of 13 Cymbopogon schoenanthus (L.) spring of Burkina Faso 4 `Table (4) Optical activity of the essential oil of 13 Cymbopogon schoenanthus (L.)Spreng of Burkina Faso. 5 Table (5) Physical properties of Cymbopogon essential oil 14 6 Table (6) Chemical composition of Cymbopogon citratus 18 essential oil identified by GC-MS analysis 7 Table (7) Chemical composition (%) of essential oils from 19 fresh leaves of Cymbopogon citratus Staph natives from Brazil and Cuba 8 Table (8) Chemical compounds of essential oil of dried C. 20 citratus obtained by steam distillation 9 Table (9) Chemical composition of Egyptian and Madinah 21 lemongrass volatile oils 10 Table (10) Antioxidant activity of Egyptian and Madinah 22 lemongrass volatile oils 11 Table (11) Physico-chemical properties of the prepared 32 essential oils of C proximus and C.citratus 12 Table (12) Chemical composition of essential oil prepared 34 from C.proximus fresh leaves 13 Table (13) Chemical composition of essential oil prepared 37 from C.proximus dry X 14 Table (14) Chemical composition of essential oil prepared 41 from C.citratus fresh 15 Table (15) Chemical composition of essential oil 43 prepared from C.citratus dry XI List of Figures 1 Figure (1) Some chemical structures of essential oils 7 2 Figure (2) Cymbopogon proximus plant 9 3 Figure (3) Cymbopogon citratus plant 17 4 Figure (4) Chemical structure of the major constituents 17 of lemongrass 5 Figure (5) Gas chromatogram of essential oil prepared 33 from fresh leaves of C.proximus 6 Figure (6) Gas chromatogram of essential oil prepared 37 from dried leaves of C.proximus 7 Figure (7) Gas chromatogram of essential oil prepared 40 from fresh leaves of C.citratus 8 Figure (8) Gas chromatogram of essential oil prepared 42 from dried leaves of C.citratus XII CHAPTER ONE Introduction and Literature Review 1 1. Introduction and Literature review 1.1General introduction The term of medicinal plants include a various types of plants used in herbalism and some of these plants have a medicinal activities. These medicinal plants considered as a rich resources of ingredients which can be used in drug development and synthesis. Besides that these plants play a critical role in the development of human cultures around the whole world. Moreover, some plants considers as important source of nutrition and are recommended for their therapeutic values such as ginger, green tea, walnuts and some others plants. Other plants their derivatives consider as important source for active ingredients which are used in aspirin and toothpaste. (Rasool Hassan, 2012) Medicinal plants frequently used as raw materials for extraction of active ingredients which used in the synthesis of different drugs. Like in case of laxative, blood thinners, antibiotics and anti-malaria medications, contain ingredients of taxol, vincristine, and morphine isolated from foxglove, periwinkle, and yew, and opium poppy, respectively. (Rasool Hassan, 2012).Medicinal plants contain physiologically active principles that over the years have been exploited in traditional medicine for the treatment of various ailments. These medicinal herbs constitute indispensable components of the traditional medicine practiced worldwide due to the low cost, easy access and ancestral experience.(Abdalla et al, 2013) Cymbopogon is one of the most important essentialoil yielding genera of the Poacea.The genus comprises 140 species that are widely distributed in semi –temperate to tropical regions of Asia, Africa and America.