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PHYTOCHEMICAL COMPOSITION and ANTIOXIDANT and ANTIMICROBIAL ACTIVITIES of Solanum Retroflexum LEAF EXTRACTS By

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PHYTOCHEMICAL COMPOSITION and ANTIOXIDANT and ANTIMICROBIAL ACTIVITIES of Solanum Retroflexum LEAF EXTRACTS By COPYRIGHT AND CITATION CONSIDERATIONS FOR THIS THESIS/ DISSERTATION o Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. o NonCommercial — You may not use the material for commercial purposes. o ShareAlike — If you remix, transform, or build upon the material, you must distribute your contributions under the same license as the original. How to cite this thesis Surname, Initial(s). (2012) Title of the thesis or dissertation. PhD. (Chemistry)/ M.Sc. (Physics)/ M.A. (Philosophy)/M.Com. (Finance) etc. [Unpublished]: University of Johannesburg. Retrieved from: https://ujcontent.uj.ac.za/vital/access/manager/Index?site_name=Research%20Output (Accessed: Date). PHYTOCHEMICAL COMPOSITION AND ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF Solanum retroflexum LEAF EXTRACTS By DAJI GRACE ABOSEDE STUDENT NUMBER: 201505331 A Dissertation submitted to the Faculty of Science, University of Johannesburg, South Africa In partial fulfilment of the requirements for the award of a Master’s Degree in Technology: Food Technology Supervisor: Dr. B. C. Dlamini Co-Supervisor: Dr. N. E. Madala September, 2017. EXECUTIVE SUMMARY Solanum retroflexum is a herbal plant that is consumed in some parts of the Venda region in Limpopo Province, South Africa. In this study, the phytochemical composition, antioxidant and antimicrobial activity of leaf extracts from the plant were investigated. Extraction of the metabolites was carried out with pressurized hot water extraction (PHWE) and different concentrations of aqueous methanol (40 %, 60 %, 80 %, methanol/water, v/v). Ultra-performance liquid chromatography coupled to quadruple- time-of-flight mass spectrometer (UPLC-qTOF-MS) technique was used to achieve an untargeted metabolite fingerprinting of the S. retroflexum leaf extracts. In total, 30 phytochemicals including alkaloids, flavonoids and cinnamic acids derivatives were identified and characterised from the aqueous methanolic leaf extracts. The identified compounds were from different classes namely, phenolics (22), flavonoids (6) and alkaloids (2). The different concentrations of aqueous methanol resulted in very similar phytochemical composition. As expected, chlorogenic acids were identified in S. retroflexum leaf extracts. Interestingly, both trans- and cis- isomers of chlorogenic acid were identified in this study. Among the flavonoids, quercetin and kaemferol were present, while two alkaloids, Solasonine and Solamargine were identified in the aqueous methanol leaf extracts. With PHWE, optimum extraction occurred at the highest temperature (250 °C). A total of 6 health-promoting metabolites were extracted and these included Quercetin-3-rutinoside, Kaempferol-3-0-rutinoside, Kaempferol-3-0-glucoside, 3- Caffeoylquinic acid, 5-Caffeoylquinic acid and 3, 4-di-Caffeoylquinic acid. Quercetin rutinoside was extracted at all extraction temperatures, while alkaloids were absent in the PHWE extracts. Solanum retroflexum aqueous methanol leaf extracts showed strong scavenging activities against DPPH and ABTS radicals. Similarly, the FRAP assay showed some antioxidant activity of the leaf extracts. The concentration of methanol did not substantially affect the antioxidant activity with all the assays, while the highest concentration of total phenol content did not exhibit high antioxidant activity in this study. The antimicrobial activity of S. retroflexum aqueous methanol leaf extracts was investigated with the disc diffusion and minimal inhibitory concentration assays. About 14 microorganisms constituting of 6 Gram-negative and 8 Gram-positive bacteria were used i in the study. In general, the S. retroflexum leaf extracts showed moderate inhibition against 6 (B. cereus, M. smegmatis, S. aureus, S. epidermis, K. pneumonia and P. vulgaris) of the investigated microorganisms. As expected, Gram-positive bacteria were more susceptible to the extracts than Gram-negative bacteria. Among the Gram-positive microorganisms, B. cereus was the most susceptible to the extracts followed by S. aureus, K. pneumonia and M. smegmatis. Better inhibition occurred with samples extracted with the highest concentration (80 %) of methanol. The minimum inhibitory concentration (MIC) as determined by micro dilution method was found to be 4-16 mg/mL. This study shows that S. retroflexum leaves contain phytochemical compounds that significantly contribute to the antioxidant and antimicrobial activity that are similar to those found in other Solanum plants. The extraction solvent affects the extract yield with aqueous methanol yielding more compounds than PHWE. The concentration of methanol does not affect the type of metabolites extracted. Understanding the type of metabolites in S. retroflexum will be useful in encouraging the use of the plant as food and its application in other foods to control foodborne bacteria. ii DECLARATION I, Daji Grace Abosede hereby declare the composition of this dissertation and the work herein described was carried out entirely by me unless otherwise cited or acknowledged. It has not been submitted for degree purpose at any other University or institution. I further declare that all cited sources are acknowledged by list of referencing. iii DEDICATION I dedicate this dissertation to God Almighty for his grace and mercy that He has bestowed upon me, my husband, our daughter and my parents who together have helped me to sustain my sense of wonder through a rather long scientific career. v ACKNOWLEDGEMENTS First, my utmost gratitude and honour goes to God Almighty for His grace and mercy upon my life during the period of my study at the esteemed University of Johannesburg. I would like to express my deepest gratitude to my supervisors Dr B.C. Dlamini and Dr N.E. Madala for their inspiring guidance, support, expertise and valuable advice. Words are not enough to express how grateful I am and I could not wish for nothing better. Thanks for being part of my success story. God bless all. Besides my supervisors, I would like to sincerely appreciate Dr V. Mavumengwana, Prof P. A. Steenkamp, Dr E. Kayitesi, Dr P.B. Njobeh, and Associate Prof O.C. Nwinyi for their encouragement, mentorship and insightful comments. Importantly, I wish to render my profound appreciation to my lovely husband Mr. A.O. Idris and our daughter Miss Esther Ebunoluwa Lerato Idris for emotional understanding and always cheering me up during the quest of this journey. I am indeed a million times better than I was before I started this journey. May God bless you abundantly. Special thanks to my parents MWO. (Rtd) and Deac. Mrs. Folorunsho and Glory Daji for their love, prayers and support and for bringing me to this earth of great discovery. I also wish to thank my in-laws Mr. and Mrs. Adisa and Abimbola Idris for your words of encouragement and best wishes. My Siblings Mrs. M. Idris, Mr. V.K. Daji, Mr. A. Idris, Mr. O.N. Daji, Mr. M.H. Idris, Mr. E.O. Daji and Mr. J.O. Daji for always supporting me in love and prayers. Furthermore, I appreciate my friends and colleagues for your companionship, assistance and Guidance: Miss. S. Moyo, Mrs. N.C. Uche-Okereafor, Miss. T.E. Sebola, Mr. O.A. Adebo, Miss. J.A. Adebiyi, Mrs. I. Olotu, Mr P. Akanji and Mr. S. Gbashi. And my colleagues at Biochemistry Department: Mrs. R.M. Ramabulana, Miss. K. Masike, Miss. P.S. Mudau, Miss. M. Makola and Mr. E. Ncube, thank you for making my stay worthwhile. God bless you beyond measures. Last but not the least, I wish to thank the University of Johannesburg and the National Research Foundation (NRF) for providing financial support for this study. vi PRESENTATIONS AND PUBLICATIONS Part of the work in this dissertation has been presented in International conference. CONFERENCE ORAL PRESENTATION: Daji, G.A., Steenkamp, P.A., Dlamini, B.C. and Madala, N.E. (2016). Phytochemical composition of Solanum retroflexum with the aid of Ultra-performance liquid chromatography hyphenated to quadruple time-of-flight mass spectrometry (UPLC-qTOF- MS). Oral presentation, 25th South African Society of Biochemistry and Molecular Biology. University of Fort Hare, 10th-14th July 2016. ARTICLES PREPARED FOR PUBLICATION Daji, G.A., Steenkamp, P.A., Dlamini, B.C. and Madala, N.E. (2017). Phytochemical composition of Solanum retroflexum with the aid of Ultra-performance liquid chromatography hyphenated to quadruple time-of-flight mass spectrometry (UPLC-qTOF- MS). Manuscript to be submitted for publication in Journal of Food Quality Hindawi. Daji, G.A., Madala, N.E., and Dlamini, B.C. (2017). Total phenolic, flavonoid, tannin content, antioxidant and antimicrobial activity of Solanum retroflexum leaves extracts. vii TABLE OF CONTENTS EXECUTIVE SUMMARY .................................................................................................. i DECLARATION ................................................................................................................ iii AFFIDAVIT ........................................................................................................................ iv DEDICATION ..................................................................................................................... v ACKNOWLEDGEMENT ................................................................................................. vi PRESENTATIONS AND PUBLICATIONS ................................................................. vii TABLE OF CONTENTS
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