Phytochemical Screening for Leaves,Cortex and Pith of the Cactus Euphorbia trigona L. Ola Ali Abdelmageed Boshara B.Sc. (Hons.) in Pharmacy, Faculty of Pharmacy, University of Gezira , 2004 A Dissertation Submitted to University of Gezira in Partial Fulfillment of the Requirements for the Award of the Degree of Master of Science in Biosciences and Biotechnology (Biotechnology) Center of Biosciences and Biotechnology Faculty of Engineering and Technology University of Gezira May 2014 1 Phytochemical Screening for Leaves, Cortex and Pith of the Cactus Euphorbia trigona L. Ola Ali Abdelmageed Boshara Supervision Committee Name Position Signature Dr. Mutaman Ali Kehail Main Supervisor ………………………… Dr. Nizar Sirag Eltayeb Co-Supervisor ………………………… Date: May 2014 2 Phytochemical Screening for Leaves,Cortex and Pith of the Cactus Euphorbia trigona L. Ola Ali Abdelmageed Boshara Examination Committee Name Position Signature Dr. Mutaman Ali Kehail Chair Person ……………………..… Dr. Eltayeb Mohamed Teyrab External Examiner ……………………..… Dr. Elnour Elamin Abdelrahman Internal Examiner ……………………..… Date of Examination: 16 May 2014 3 Dedication To me.. “ Success is the journey, not just the destination.” ----------- 4 Acknowledgements I would like to aknowledge Dr. Mutaman Ali Kehail for his great assistance with his knowledge and experience. I also aknowledge Dr. Nizar Sirag for his great assistance in this work. Deepfull thanks for my future husband Ahmad Abdelgaffar for support,encouragement, and helpfull opinions. 5 Phytochemical screening for Leaves,Cortex and Pith of the Cactus Euphorbia trigona L. Ola Ali Abdelmageed Boshara Abstract The vast majority of people on this planet still rely on their traditional medicine for their everyday health care needs. The present study aimed to run a phytochemical screening for the leaves, stem cortex and pith of Euophorbia trigona L. Plant materials, were collected from within Wad Medani City. The targeted parts were washed while they were in their original plant. The collected parts were transferred directly to the Faculty of Engineering and Technology, University of Gezira, where all of the laboratory tests were done. The experimental tests involved: proximate analysis (percentage moisture, ash and protein contents), the percentage polar and apolar contents and the phytochemical screening for the main components in the three plant parts. The results of this work revealed that, the proximate analysis of Euphorbia trigona parts indicated that, stem pith had significantly, the highest percentages of moisture contents (95.76%), while the moisture in the stem cortex (89.91%) were also more than that of leaves (82.72%). Stem pith was found to possess significantly, the lowest percentages of ash contents (3.05%) than that of the stem cortex (7.70%) and leaves (7.71%). Stem pith showed the lowest percentages of protein contents (0.91%), whereas, the protein content in leaves (2.77%) were the highest. The percentage mean polar content of the leaves (10.47) was more than other parts collectively (6.77 + 2.88%). Meanwhile, the apolar content was relatively high in stem cortex (1.37%) than the other parts (0.81and 1.07%). The preliminary phytochemical screening of E. trigona leaves revealed the presence of saponins, alkaloids, flavonoids, flavonones and flavonols, glycosides and sterols and triterpenoids while tannins were not detected while the stem cortex and stem pith contained all phytoconstituents except tannins and flavonones. It can be concluded that, E. trigona could be a potential source of active principles which may help to discover new chemical classes of drugs that could serve as selective agents for the maintenance of health, and hence, this study recommends to run a further studies to identify the active constituents, as well as to verify the biological activities of this plants. 6 المسح الكيميائي النباتي لمختلف أجزاء نبات صبار اﻹيوفوربيا ترايقونا عﻻ علي عبد المجيد بشارة ملخص االدرسة يعتمد غالبية سكان هذا الكوكب علي الطب التقليدي في اﻻحتياجات اليومية للمحافظة علي الصحة. هدفت هذه الدراسة ﻹجراء مسح كيميائي نباتي ﻷوراق, قشرة ساق ولب ساق نبات صبار اﻻيوفوربيا اتريقونا. جمعت العينات النباتية من داخل مدينة ود مدني. غسلت ا ﻻجزاء المستهدفة أثناء وجودها في النبات اﻻصل. تم تحويل ااﻻجزء المجمعة الي كلية الهندسة والتكنولوجيا, جامعة الجزيرة حيث أجريت جميع اﻻخاتبارت المعملية. شملت اﻻخاتبارت المعملية: التحليل التقريبي )النسبة المئوية للرطوبة, الرماد والبروتين(, النسبة المئوية للم كونات القطبية والﻻقطبية, إضافة الي المسح الكيميائي النباتي للمكونات الرئيسية في ااجزء النبات الثﻻثة. أوضحت النتائج, أن التحليل التقريبي قد دلل علي أن لب الساق يحتوي بصورة معنوية علي أعلي نسبة رطوبة )95.76%(, بينما كانت الرطوبة في قشرة الساق )89.91%( ايضا اعلي من اﻻوراق )82.72%(. وجد أن لب الساق يحتوي معنوياً علي أقل نسبة رماد )3.05%( عن قشرة الساق )7.70%( واﻻوراق )7.71%(. يحتوي لب الساق علي أقل نسبة بروتين )0.91%(, في حين أن بروتين اﻻوراق )2.77%( هو اﻷعلي. متوسط نسبة المكونات القطبية في اﻻوراق )10.47( كانت اكثر من ما في بقية اﻻج ازء مجتمعةً )6.77 + 2.88%(. أما بالنسبة للمكونات الﻻقطبية, فقد كانت عالية معنوياً في قشرة الساق )1.37%( عن اﻻجزاء اﻻخري )0.81 و 1.07%(. أوضح المسح الكيميائي اﻻولي لﻻوارق وجود المواد الصابونية, القلويدية, الفﻻفونويدية, الفﻻفونولية والفﻻفونونية, الجﻻيكوسيدات واﻻستروﻻت والتريبينويدات, في حين أن التانينات لم يتم الكشف عنها, بينما احتوت قشرة ولب الساق علي كل المكونات الكيميائية النباتية عدا التانينات والفﻻفونونات. يمكن وضع الخﻻصة بأن نبات اﻻيوفوربيا اتريقونا يمكن ان يكون مصدر محتمل لمكونات فعالة يمكن ان تساعد في اكتشاف اصناف دوائية كيميائية جديدة يمكنها أن تشكل عامل انتقائي للمحافظة علي الصحة, وعليه, فقد اوصت هذه الدراسة بإجراء المزيد من الدراسات المستقبلية للتعرف علي المكونات الفعالة وكذلك تحديد الفاعلية الحيوية لهذا النبات. 7 List of Contents Subject Page Dedication iv Acknowledgements v Abstract vi Abstract (Arabic) vii Table of contents viii List of Tables x List of Figures and Plates xi Chapter One: Introduction 1 Chapter Two: Literature Review 3 2.1. Cactus 3 2.1.1. Adaptations 3 2.1.2. Morphology 3 2.1.3. Growth habit 4 2.1.4. Stems 5 2.1.5. Areoles 5 2.1.6. Leaves 6 2.1.7. Spines 6 2.1.8. Roots 7 2.1.9. Flowers 7 2.1.10. Adaptations for water conservation 8 2.1.11. Taxonomy 9 2.1.12. Uses of cactus 10 2.2. Euphorbia: 14 2.2.1. Description 15 2.2.2. Xerophytes and succulents plants 15 2.2.3. Irritance 17 2.2.4. Uses of Euophorbia 17 2.2.5. Systematics and taxonomy 17 2.2.6. Euphorbia trigona 18 2.2.6.1. Scientific Classification: 18 2.2.6.2. Medicinal benefits: 18 8 2.2.7 Researches on Euphorbia species 19 2.3. Phytochemicals 21 2.3.1. Clinical trials and health claim status 21 2.3.2. Food processing 21 2.3.3. Screening and Extraction: 23 2.3.4. Plant material 26 2.3.5. Choice of solvents 26 2.4. Proximate Analysis: 28 2.5. Chromatography: 28 2.5.1. Plate preparation in TLC 29 2.5.2. Procedure: 29 2.5.3. Separation Process and Principle 30 2.5.4. Analysis 31 2.5.5. Applications 31 Chapter Three: Materials and Methods 33 3.1 Samples of plant material: 33 3.2. Preparation of plant material 33 3.3. Approximate analysis 33 3.3.1. Moisture content 33 3.3.2. Ash content 34 3.3.3. Protein content 34 3.4. The polar and apolar contents 35 3.5. Phytochemical screening for the different parts of E. trigona: 36 3.5.1. Qualitative analysis Test for glycosides 36 3.5.2. Qualitative analysis Test for flavonoids 36 3.5.3. Qualitative analysis Test for saponins: 36 3.5.4. Qualitative analysis Test for tannins: 36 3.5.5. Qualitative analysis Test for sterols and or triterpenes: 37 3.5.6. Qualitative analysis Test for alkaloids and /or nitrogenous bases 37 3.7. Statistical analysis 37 Chapter Four: Results and Discussion 38 4.1. The proximate analysis of Euphorbia trigona parts 38 4.2. The polar and apolar contents of Euphorbia trigona parts 44 4.3. The phytochemical screening in Euphorbia trigona parts 47 Chapter Five: Conclusions and Recommendations 49 References 50 9 List of Tables Table No. Title Page 2.1 Proximate and phytochemical composition of Euphorbia heterophylla 16 2.2 Phytochemical analysis of E. trigona extract 20 2.3 Activities of various phytochemicals from plants 22 2.4 Solvents used for active component extraction 25 4.1 The moisture contents (%) in Euphorbia trigona parts 40 4.2 The ash contents (%) in Euphorbia trigona parts 41 4.3 The protein contents (%) in Euphorbia trigona parts 42 4.4 The polar contents (%) in Euphorbia trigona parts 45 4.5 The apolar contents (%) in Euphorbia trigona parts 46 4.6 The presence of some phytochemical in E. trigona parts 48 10 Chapter One Introduction Cacti are commonly grown as houseplants. They are pretty and easy to grow. Some cacti are grown in gardens, especially in dry areas. Cactus can be used as a living fence. The wood of dead cactus is sometimes used for building. People eat the fruit of some kinds of cactus, such as dragon fruit and prickly pear. Ciouvhiul insects also eat prickly pears. These insects produce a red coloring used in food and lipstick (Anderson, 1999). The Euphorbia plants are annual or perennial herbs, woody shrubs or trees with a caustic, poisonous milky sap (latex). The roots are fine or thick and fleshy or tuberous. Many species are more or less succulent, thorny or unarmed. The main stem and mostly also the side arms of the succulent species are thick and fleshy, 15–91 cm (6–36 inches) tall. The deciduous leaves are opposite, alternate or in whorls. In succulent species the leaves are mostly small and short-lived. The stipules are mostly small, partly transformed into spines or glands, or missing.