Nutritional, Phytochemical and Toxicity Characteristics of Local Sana (Cassia senna L.) Compared to Samples from Erkweit, Red Sea State, Sudan Ayda Ali Khalifa Mohammed B.Sc. (Hon.) in Agronomy, Faculty of Agriculture, University of Zalingei, 1997 A Dissertation Submitted to the 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 March 2015 1 Nutritional, Phytochemical and Toxicity Characteristics of Local Sana (Cassia senna L.) Compared to Samples from Erkweit, Red Sea State, Sudan Ayda Ali Khalifa Mohammed Supervision Committee: Name Position Signature Dr. Mutaman Ali Kehail Supervisor …………….. Prof. Elnour Elamin Abdelrahman Co-Supervisor …………….. Date: March, 2015 2 Nutritional, Phytochemical and Toxicity Characteristics of Local Sana (Cassia senna L.) Compared to Samples from Erkweit, Red Sea State, Sudan Ayda Ali Khalifa Mohammed Examination Committee: Name Position Signature Dr. Mutaman Ali Kehail Chair Person …………….. Prof. Elnaeim Abd Allah Ali External Examiner …………….. Prof. Ali Osman Ali Internal Examiner …………….. Date: 25, March, 2015 3 Dedication To my Family 4 Nutritional, Phytochemical and Toxicity Characteristics of Local Sana (Cassia senna L.) Compared to Samples from Erkweit, Red Sea State, Sudan Ayda Ali Khalifa Mohammed Abstract Cassia senna plant is well known as sanamakka. It grown in many areas in the Gezira State and in other States. Many nutritional and pharmaceutical uses of this plant were reported worldwide. The comparative nutritional composition, qualitative phytochemical screening, thin layer chromatography and toxicity, for Wad Medani (W) and Erkweit (E) samples of C. senna seeds (S) and leaves (L) were the aims of this study. The study was conducted during September 2014. The collected samples were prepared for the study experiments in the University of Gezira, Faculty of Engineering and Technology. Larvae of Anopheles and Culex mosquitoes were also collected for the toxicity test. The conclusions of this study revealed that, the highest nutritional contents in all samples was the carbohydrates (61.75%) followed by protein (17.93%), then ash (7.19%), fat (0.31%) and lastly fibers (0.09%), but Anova proved no significant differences between the four samples in their nutritional contents. The phytochemical screening, showed that, the leaves of Erkweit sample, contained glycosides, flavonoids, flavonone and alkaloids, as same as Erkweit seeds, which contained also sterols, while the leaves sample of Wad Medani resemble completely Erkweit seeds, but Wad Medani seeds sample, lack sterols and glycosides. The thin layer chromatography detected a total of three spots from the leaves and four spots from the seeds of Wad Medani samples, but the case was not similar for Erkweit samples, leaves part separated four spots (two from the polar and two from the apolar extracts), while that, five spots were separated from the seed part (two spots from the apolar extract and three from polar extract). The study recommended to get benefit of the nutritional contents from C. senna, irrespective of their source, also the pharmaceutical importance of such plant should be studied. 5 الخصائص التغذوية, الكيميائية النباتية والسامة لنبات السنمكة المحلي مقارنة بعينات من أركويت, وﻻية البحر اﻷحمر, السودان عايدة علي خليفة محمد ملخص االدرسة يعرف نبات "كاشيا سنا" بالسنمكة. ينمو في عدة مناطق في وﻻية الجزيرة وفي الوﻻيات اﻷخري. غني بالمجتمعات الحيوانية المائية كنتاج للشبكة الكثيفة من قنوات مشروع الجزيرة. العديد من اﻹستخدامات التغذوية والصيدﻻنية تم نشرها علي مستوي العالم. مقارنة المحتوي الغذائي, المسح الكيميائي النباتي الكيفي, كروماتوغرافيا الطبقة الرقيقة والسمية لعينات بذور وأوراق السنمكة من ود مدني )وﻻية الجزيرة( وأركويت )وﻻية البحر اﻷحمر( هي أهداف هذا البحث. تمت هذه الدراسة في الفترة سبتمبر 2014. تم إعداد العينات المجموعة ﻹخاتبارت الدراسة في جامعة الجزيرة, كلية الهندسة والتكنولوجيا. تم أيضاً جمع يرقات بعوض اﻻنوفلس والكيولكس ﻹختبار السمية. خلص البحث إلي أن أعلي مكون غذائي لكل العينات هو الكربوهيادرت )61.75%( يتبعه البروتين )17.93%(, الرماد )7.19%(, الدهن )0.31%( وأخي اًر اﻷلياف )%0.09(, ولكن إختبار تحليل التباين أوضح أن ﻻ فرق معنوي بين العينات اﻷربع في محتواها الغذائي. المسح الكيميائي النباتي أوضح أن أوراق عينة أركويت إحتوت علي الجﻻيوكوسيدات, الفﻻفونويدات, الفﻻفونونات والقلويدات, مثل بذور عينات أركويت, والتي تحتوي أيضاً علي اﻹستروﻻت, بينما أو ارق عينة ود مدني تشبه كلياً بذور أركويت, ولكن بذور عينة ود مدني تفتقر لﻹستروﻻت والجﻻيكوسيدات. الدراسة حدد إختبار كروماتوغرافيا الطبقة الرقيقة مجمل ثﻻثة بقع من اﻷوراق وأربعة بقع من بذور عينات ود مدني, ولكن الحالة ليست متشابهة لعينات أركويت, حيث فصلت جزء اﻷوراق أربعة بقع )إثنان من المستخلص القطبي واثنان من المستخلص الﻻقطبي(, بينما تم فصل خمسة بقع من البذور )إثنان من المستخلص الﻻقطبي وثﻻثة من المستخلص القطبي(. أوصي البحث باﻹستفادة من المكونات الغذائية لنبات السنمكة بغض النظر عن مصدرها, وكذلك بادرسة اﻷهمية الصيدﻻنية لهذا النبات. 6 List of Contents Subject Page Dedication iii Acknowledgements iv Abstract v Arabic Abstract vi List of Contents vii List of Tables ix CHAPTER ONE: INTRODUCTION 1 CHAPTER TWO: LITERATURE REVIEW 3 2.1. Cassia senna 3 2.1.1 Scientific classification 3 2.1.2.Description 3 2.1.3 Uses 4 2.1.4 Chemical composition 5 2.2. Phytochemical analysis 7 2.2.1 Procedure 7 2.3. Thin layer chromatography 9 CHAPTER THREE: MATERIALS AND METHODS 3.1 Materials 10 3.2. Proximate analysis tests 10 3.2.1. Ash content 10 3.2.2. Oil content 10 3.2.3. Protein content 11 3.2.4. Crude fiber content 11 3.2.5. Carbohydrates content 12 3.3. Phytochemical screening tests 9 7 3.3.1 Test for the presence of alkaloids 12 3.3.2 Test for the presence of flavonoids and flavonones 12 3.3.3 Test for the presence of glycosides 12 3.3.4 Test for the presence of saponins 12 3.3.5 Test for the presence of steroids 12 3.3.6 Test for the presence of tannins 13 3.4. Thin layer chromatography test 13 3.5. Toxicity test of C. senna leaves and seeds powder 13 3.4. Statistical analysis 13 CHAPTER FOUR: RESULTS AND DISCUSSION 4.1 Proximate analysis of C. senna of Wad Medani and Erkweit areas 14 4.2. Qualitative phytochemical screening of C. senna 16 4.3. Thin layer chromatography test for C. senna 18 4.4. Toxicity of C. senna leaves and seeds powder against mosquito larvae 20 CHAPTER FIVE: CONCLUSIONS AND RECOMMENDATIONS 5.1 Conclusions 23 5.2. Recommendations 23 References 24 8 List of Tables Table Title Page No Profiles of the phytochemical screening, minerals and nutritional 2.1 6 contents of Cassia senna leaves and seeds 2.2 Qualitative phytochemical analysis by using different solvents 8 the proximate analysis of C. senna brought from Wad Medani and from 4.1 15 Erkweit areas 4.2 Qualitative phytochemical screening of C. senna leaves and seeds 17 Thin layer chromatography test for C. senna leaves and seeds brought 4.3 19 from Wad Medani area and Erkweit area Toxicity of 0.6 g (per 250 ml) powder of C. senna leaves and seeds 4.4 (from Wad Medani and Erkweit areas) toward Anopheles and Culex 21 larvae 9 Chapter One Introduction The name Senna is from Arabic sana. Cassia is an angiosperme widely distributed in Africa, in Latin America, and in Australia and Fiji. Firstly classified in Caesalpiniacae family, then in those of Leguminoseae, Cassia is now classified among the Fabaceae. This plant is a shrub which has a medium-size, 10-12 m tall, occasionally reaching 20 m. The bark is grey and smooth, and later with longitudinal fissures. The leaves are alternate, 15-30 cm long, compound, with 6-14 leaflets each ending in a tiny bristle. The flowers are bright yellow, in large, up to 60 cm long, upright, with pyramid-shaped panicles. The fruits are flat with indehiscent pod, 5-30 cm long, and constricted between the seeds. There are about 20 seeds per pod. The seeds are bean-shaped, greenish-brown, and 8-15 mm long (Gutteridge, 1997). The leaves, stems, roots, flowers and seeds of Cassia regardless the subspecies have been used for the treatment of several illnesses including mostly malaria, a tropical endemic disease with high morbimortality (Oyedunmade and Olabiyi, 2006). The preparation process of remedies was not clearly described and the dosages prescribed were approximate. Moreover, the treatments are supposed to be continued until recovery. According to the ethnic differences of populations from localities, the plant is used alone or in combination with other plants or with natural substances for the preparation, especially in decoction. For the treatment, people mostly used the preparations by oral administration route (Ejobi et al., 2007). The leaves are the most used parts’ the plant especially by African and Asian population in preparation of the herbal remedies. In Burkina Faso, fresh and dried leaves decoction is drunk with lemon juice or for body bath throughout the day to treat malaria and liver disorders (Nadembega et al., 2011). In Côte d’Ivoire, the decoction of leaves is administered orally (0.5 L, twice daily) for treating cough, stomach pains and malaria. Also, in Sierra Leone and Togo, the leaves decoction is drunk against malaria (Mbatchi et al., 2006) and used as antimicrobial (Souza et al., 1995). In Nigeria, the dried leaves are mixed with lemon’s leaves (Cymbopogon citratus), pawpaw’s leaves (Carica papaya), and the lime’s leaves (Citrus lemonum) and are boiled within an hour. The "tea" of the mixture is drunk against malaria (Ogunkunle and Ladejobi, 2006). 10 In Uganda, the leaves are picked, cleaned and chewed, and liquid swallowed to treat abdominal pains (Kamatenesi et al, 2011). In India, the leaves are cleaned thoroughly and boiled. The decoction is filted in which is added honey.