بسم هللا الرحمن الرحيم AL Neelain University Collage of Graduate Studies Taxonomic Revision of the Family Euphorbiaceae A. Juss. Sensu lato in the Red Sea State-Sudan And Chemical Analysis of Three Selected Species from the Family A thesis Submitted to Al- Neelain University in Fulfillment of the Requirements for the Award of the M. Sc. Degree in Biology (Plant Taxonomy) By Zeinab Abd Ellatif Saleh Abd Ellatif B. Sc. Honour Supervisor: Dr. Alawia Abdalla Elawad Co-supervisor: Dr. Ragaa Satti Mohammed Abadi September, 2017 I بسم هللا الرحمن الرحيم جامعة النيلين كلية الدراسات العليا دراسة تصنيفية للفصيلة اللبنية بوﻻية البحر اﻷحمر-السودان والتحليل الكيميائي لثﻻثة انواع مختارة من الفصيلة اطروحة مقدمة لجامعة النيلين لنيل درجة الماجستير في اﻷحياء )تصنيف نبات( اسم الطالبة : زينب عبداللطيف صالح عبد اللطيف ماجستير شرف المشرف: د. علوية عبدهللا العوض مشرف متعاون: د. رجاء ساتي محمد عبادي سبتمبر 7102 II Dedication To my father, to my mother To my brothers, to my sisters To my teacher Dr. Alawia El Awad I Acknowledgments Iam grateful to thank Allh, who granted me life, the power, peace and courage to finish this study. Thanks and deepest appreciation to my supervisors Dr. Alawia Abdalla Elawad for her guidance throughout this research and Dr. Ragaa Satti for her encouragements and continuous help. Thanks for Dr. Somya Khidir Ali, Faculty of Marine Sciences and Fisheries Red Sea University for her continuous help and useful discussions. I would like to thank the staff of the research lab of EL Neelain University. I also extend my profound thanks to my colleagues in the Faculty of Education –Red Sea University and the Technical Chemistry lab and Biology lab for offering the facilities available for me. II Abstract Euphorbiaceae is a family of dicotyledonous flowering plant which includes a number of species distributed all over the Sudan. It contains species used as a source of food and others are medicinal that contain some bioactive compounds and are used in local medicine. The present study constitutes two parts. The first part comprises a systematic survey of the representatives of the family Euphorbiaceae s.l. Juss. in the Red Sea State based mainly on material deposited in the local herbaria in the Sudan and Kew herbarium- England and the author collections from Sinkat Locality. In addition to the species reported in the Sudanese floras and relevant publications. For each species standard up- dated scientific binomial, synonyms, a brief description, locations of species in the study area, habitat and economic uses are included when available. In the second part phytochemical analysis of (secondary) constituents in three species from the family namely: Euphorbia abyssinica Gmel. E. polycantha Boiss. And Jatropha glauca Vahl (stem, seed, latex or sap) in alcohol and water solvents were carried out using standard methods. The representatives of the family Euphorbiaceae s.l. in the Red Sea State contain 29 species included within 8 genera. According to the (APG IV) system of classification the studied taxa are presented in two families: Euphorbiaceae s.s. Marc.-Berti and Phyllanthaceae s.s. Martinov. Euphorbiaceae s.s. contains 3 subfamilies: Acalyphoideae with 3 genera including 6 species, Crotonoideae 2 genera with 4 species and Euphorbioideae one genus with 15 species. Phyllanthaceae contains 3 genera with 4 species. The life forms of the studied members show that most of the species are Phanerophyta (59%) including 21% succulents. Herbaceous species constitute 41% most of them are Chamaephyta; Therophyta are rare. Information on collected economic uses shows that most of the species in the study area are of medicinal importance. III The preliminary phytochemical screening resulted in the presence of different levels of the tested phyto-constituents: carbohydartes, alkaloids, saturated and unsaturated sterols, saponin, coumarins, tannins, cardenolides, flavones, flavonols, deoxy sugars and traces of cyanidin and leucoanthoyanins. Quantitative analysis of saponin in aqueous extracts of Jatropha glauca and Euphorbia polyacantha yields 20% in the seeds of the studied plants. The stem of E. polyacantha contains 20% saponin and the sap of Jatropha glauca scores the highest level (26.3%). Aqueous extracts of the latex of Euphorbia polyacantha revealed the presence of 60% alkaloids. In view of the above the family Euphorbiaceae in Sudan needs more studies using different taxonomic approaches and detailed chemical analysis. IV المستخلص انؼبئهخ انهجٍُخ يٍ انُجبربد انزْشٌخ راد انفهمزٍٍ، ًْٔ رضى ػذد يٍ اﻻَٕاع انزً رُزشش فً كم اَحبء انضٕداٌ. ْٔزِ اﻻَٕاع رضزخذو كًصبدس طجٍخ ٔانجؼض اﻻخش كغزاء ٔرحزٕي ػهً اَشطخ ثٍٕنٕخٍخ يشكجخ فً انذٔاء انًحهً. ْٔذح انذساصخ رزضًٍ خزئٍٍٍ. اندزء اﻻٔل ٌزؼهك ثبنززبثغ انزصٍُفً انمذٌى انذي يشد ثّ انؼبئهخ انهجٍُخ فً ٔﻻٌخ انجحش اﻻحًش. ٔلذ رى حصش اﻻَٕاع انًحفٕظخ فً انؼًشجبد انًخزهفخ فً انضٕداٌ ٔانًحفٕظخ فً يؼشجخ كٍٕ )Kew) ثبَدهزشا. ٔاﻻَٕاع انزً خؼًذ يٍ يحهٍخ صُكبد. ثبﻹضبفخ انً اﻻَٕاع انزً ركشد فً يشاخغ انفهٕسا انضٕداٍَخ ٔانًشاخغ راد انصهخ ثًٕضٕع انذساصخ. نكم اﻻَٕاع انزً شًهزٓب انذساصخ رى رحذٌث اﻻصًبء انﻻرٍٍُخ ٔانًشادفخ، ٔصف يٕسفٕنٕخً يخزصش ايبكٍ رٕاخذ اﻻَٕاع فً يُطمخ انذساصخ، ٔانجٍئبد انزً رًُٕ فٍٓب ْزِ اﻻَٕاع ٔ اﻻًٍْخ اﻻلزصبدٌخ نٓب اٌ ٔخذد .ٔاندزء انثبًَ ٌزؼهك ثبنزحهٍم انكًٍٍبئً انػًُٕ نهضبق ، انجزٔس، انًبدح انهجٍُخ أ انؼصبسح نثﻻثخ إَاع رُزًً نهؼبئهخ انهجٍُخ. ٔاﻻَٕاع ًْ: E. ٔ Euphorbia abyssinica Jatropha glauca ٔ Polyacatha ثبصزخذاو يزٌجٍٍ ًْب انًبء ٔانكحٕل حضت انطشق انمٍبصٍخ. رضى انؼبئهخ انهجٍُخ انمذًٌخ فً ٔﻻٌخ انجحش اﻻحًش 92 َٕع َجبرً رُضٕي رحذ 8 اخُبس. اػزًبدا ػهً انُظبو انزمضٍى (APG IV) فبٌ اﻻَٕاع انزصٍُفٍخ رحذ انذساصخ رمغ رحذ ػبئهزٍٍ: انهجٍُخ انحذٌثخ Euphorbiaceae s.s ٔانؼبئهخ فبٌﻻٍَضٍخ انحذٌثخ Phyllanthaceae s.s. رضى انؼبئهخ انهجٍُخ انحذٌثخ ثﻻثخ رحذ ػٕائم Acalyphoideae ٔثٓب 3 اخُبس ٔ 6 إَاع ، Crotonoideae ٔثٓب 9 خُش ٔ 4 إَاع ، Euphorbiodeae ٔثٓب خُش ٔاحذ ٔ 55 َٕع . ٔانؼبئهخ فبٌﻻٍَضٍخ انحذٌثخ رضى 3 اخُبس 4ٔ إَاع. ٔثٍُذ دساصخ اشكبل انحٍبح إٌ يؼظى اﻻَٕاع رحذ انذساصخ يٍ اﻻشدبس ٔانشدٍشاد ) %52( يُٓب )95%( يخززَخ ٔاﻻَٕاع انؼشجٍخ %45 يؼظًٓب اػشبة يؼًشح ٔانمهٍم يُٓب حٕنً. اظٓشد انؼًهٕيبد انزً رى خؼًٓب ػٍ اﻻصزخذايبد اﻻلزصبدٌخ اٌ يؼظى اﻻَٕاع فً يُطمخ انذساصخ راد اًٍْخ طجٍخ. أصفشد َزبئح انزحهٍم انكًٍٍبئً اﻻثزذائً ػٍ ٔخٕد يضزٌٕبد يخزهفخ يٍ انًكَٕبد انكًٍٍبئٍخ انزً رى اخزجبسْب ًْٔ: انكبسثٍْٕذسٌزبد carbohydartes، انمهٌٕذاد alkaloids ، اﻻصزشٔﻻد V انًشجؼخ ٔغٍش انًشجؼخ saturated and unsaturated sterol ٔ انصبثٍَٕبد saponin ٔ انكٕيبسٌُبد ,coumarins ٔ انزبٍٍَُبد tannins ٔ انضكشٌبد انمهجٍخ cardenolides ٔ انفﻻفٍَٕذاد flavones ٔ صكشٌبد دٌٕكضٍخ deoxy sugars ثًٍُب ظٓشد اثبس ثضٍطخ يٍ انضٍبٍَذاد cyaniding ٔنٍكٕاَثٌٕبٍٍَُبد leucoanthoyanins. أظٓشانزحهٍم انكًً نُضجخ انصبثٍٍٍَٕ نهًضزخهص انًبئً نجزٔس.Jatropha glauca Euphorbia polyacanthaٔ َضجخ )92%(. ايب انضبق نُجبد Euphorbia polyacantha فمذ اػطً َضجخ )92%(. ايب َضجخ انصبثٍٍٍَٕ فً ػصبسح َجبد Jatropha glauca كبَذ اﻷػهى )96.3%(. انًضزخهص انًبئً نهًبدح انهجٍُخ فً َجبد Euphorbia polyacantha اظٓش َضجخ 62% يٍ انمهٌٕذاد. يًب ركش أػﻻِ فبٌ انؼبئهخ انهجٍُخ فً انضٕداٌ رحزبج انً انؼذٌذ يٍ انذساصبد انزصٍُفٍخ ثبصزخذاو انذﻻئم انزصٍُفٍخ انًخزهفخ ٔانذساصبد انكًٍٍبئٍخ انًفصهخ. VI List of Content Page No. Content DEDICATION I ACKNOWLEDGMENTS II ABSTRCT ENGLISH III ABSTRCT ARABIC V List of Content VII List Tables XI 1-CHAPTER ONE:INTRODUCTION AND LITERATURE REVIW 1.1 General Introduction 1 1.1.1 Preamble 1 1.1.2 Objectives 2 1.2 LITERATURE REVIEW 3 1.2.1 Taxonomical Treatment of the Family Euphorbiaceae 1.2.2 Morphological Description of the Family Euphorbiaceae 5 1.2.3 Geographic Distribution of the Family Euphorbiaceae 5 1.2.4 Taxonomic Studies of Euphorbiaceae in Sudan 5 1.2.5 Euphorbiaceae in the Red Sea State 6 1.2.6 Economic Uses of the Family Euphorbiaceae 6 1.2.7 Phytochemical Screening of the Family Euphorbiaceae 8 1.2.8 Euphorbia abyssinica 8 1.2.9 Euphorbia polyacantha 9 2 CHAPTER TWO: MATERIALS AND METHODS 2.1 Study Area 01 VII 2.2 Taxonomical Study 01 2.2.1 Herbarium Collections 12 2.2.2 Specimens Collection 12 2.3 Chemical Analysis 01 2.3.1 Plant Materials 01 2.3.2 Chemicals and Reagents 01 2.3.2.1Chemicals 01 2.3.2.2 Reagents 01 9.3.3 Methods 02 9.3.3.5 Methods of Extraction 02 9.3.4 Phytochemical Screening Tests 02 2.3.4.1Screening of Sterols 13 2.3.4.1.1 Screening of Saturated Sterols (the Liebermann Burchard Test) 13 2.3.4.1.2 Unsaturated Sterols (the Salkowaski Test) 14 2.3.4.2 Screening for Flavonoids 14 2.3.4.2.1 Cyanidin Test 14 2.3.4.2.2 Test for Flavonols 14 2.3.4.2.3 Test for Flavones 14 2.3.4.2.4 Screening Leucoanthocyanins 14 2.3.4.3 Screening for Tannins 15 2.3.4.4 Screening for Saponin (Forth Test) 15 2.3.4.5 Screening for Alkaloids 15 2.3.4.6Screening for Coumarins 16 2.3.4.7 Screening for Carbohydrates 16 2.3.4.8 Screening for 2-Deoxy Sugars 16 2.3.4.9 Screening for Cardenolides (Baljects Test) 16 VIII 2.3.5 Quantitative Deterination of the Chemical Constituents 17 2.3.5.1 Determination of Saponin 17 2.3.5.2 Determination of Alkaloids 17 CHAPTER THREE RESULT AND DISCUSSION 3.1Taxonomy Study 18 3.1.1 Herbarium Collections 18 3.1.1.1 Check-list 19 3.1.1.2 Life forms 42 3.1.1.3 Economic uses 42 3.2 Phytochemical analysis of the plant extract 44 3.2.1 Qualitative Phytochemical screening Tests 44 3.2.2 Quantitative analysis 49 Conclusions 50 Recommendations 50 REFERENCES 51-59 APPENDIX (A) APPENDIX (B) IX List of Tables Table (3.1) Phytochemical screening of Euphobia abyssinica (E. a.), E. 45 polyacantha (E. p.) and Jatropha glauca (J. g.) stem in different solvent. Table (3.2) Phytochemical screening of Euphorbia abyssinica (E. a.), E. 46 polyacantha (E. p.) and Jatropha glauca (J. g.) seeds in different solvent. Table (3.3) Phytochemical screening of E.