Tài liệu Brown algae as a source of bioactive compounds for pancreatic cancer treatment

Brown algae as a source of bioactive compounds for pancreatic cancer treatment Thanh Trung Dang B.Eng (Nha Trang University, Khanh Hoa, Vietnam) MSc (Nha Trang University, Khanh Hoa, Vietnam) A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy in Food Science School of Environmental and Life Sciences, Faculty of Science University of Newcastle Australia May 2018 STATEMENT OF ORIGINALITY I hereby certify that to the best of my knowledge and belief this thesis is my own work and contains no material previously published or written by another person except where due references and acknowledgements are made. It contains no material which has been previously submitted by me for the award of any other degree or diploma in any university or other tertiary institution. Thanh Trung Dang Date: 6/5/2018 i DECLARATION OF AUTHORSHIP I hereby certify that this thesis is in the form of a series of 8 papers. I have included as part of the thesis a written statement from each co-author, endorsed in writing by the Faculty Assistant Dean (Research Training), attesting to my contribution to any jointly authored papers. Thanh Trung Dang Date: 6/5/2018 ii ACKNOWLEDGEMENTS Firstly, I would like to give a great appreciation to my supervisors: Principal supervisor: A/Prof. Christopher J. Scarlett; Co-supervisors: A/Prof. Michael C. Bowyer and Dr. Ian A. Van Altena for their supervision and support during my PhD course. The suggestions and encouragement from the supervisor panel played an important role in my research achievements. I acknowledge the financial support from University of Newcastle; the Vietnamese Government through the Ministry of Education and Training; the Ministry of Agriculture and Rural Development for awarding a VIED-TUIT scholarship, which enabled me to study for a PhD at the University of Newcastle, with full cover for academic expenses, as well as living and travellingallowances. I highly appreciated the contribution of brown algae as the material for my PhD project from Dr. Maria Schreider (School of Environmental and Life Sciences, Faculty of Science, University of Newcastle), in particular for identifying algal species and allowing her students assist in the collection of the samples. I would like to say thank you to Dr. Quan V. Vuong, Dr. Danielle Bond and other PhD students in Food Science, technical staff and administrative staff for helping me to overcome the difficulties in the laboratory and administrative works. Finally, I would like to say thank my colleagues and friends from Nha Trang University, Vietnam for their encouragement. A very special thank you is given to my family (parents, young brother), who have always been behind me, encouraging and inspiring me during my PhD project. iii LIST OF PUBLICATIONS INCLUDED AS PART OF THE THESIS I warrant that I have obtained, where necessary, permission from the copyright owners to use any third party copyright material reproduced in the thesis, or to use any of my own published work in which the copyright is held by another party. 1. Paper I: Dang TT, Vuong QV, Schreider MJ, Bowyer MC, Van Altena IA & Scarlett CJ. (2017). The Effects of Drying on Physico‐Chemical Properties and Antioxidant Capacity of the Brown Alga (Hormosira banksii (Turner) Decaisne). J. Food Process Pres 41(4): e13025. doi.org/10.1111/jfpp.13025. 2. Paper II: Dang TT, Bowyer MC, Van Altena IA & Scarlett CJ. (2018). Comparison of chemical profile and antioxidant properties of the brown algae, Inter. Food Sci. Technol 51(1): 174-181. doi: 10.1111/ijfs.13571. 3. Paper III: Dang TT, Vuong QV, Schreider MJ, Bowyer MC, Van Altena IA & Scarlett CJ (2017). Optimisation of ultrasound-assisted extraction conditions for phenolic content and antioxidant activities of the alga Hormosira banksii using response surface methodology. J Appl Phycol 29(6): 3161-3173. doi.org/10.1007/s10811-017-1162-y. 4. Paper IV: Dang TT, Bowyer MC, Van Altena IA & Scarlett CJ. (2017). Optimum conditions of microwave assisted extraction for phenolic compounds and antioxidant capacity of the brown alga Sargassum vestitum. Sep Sci Technol (In Press). doi.org/10.1080/01496395.2017.1414845. 5. Paper V: Dang TT, Vuong QV, Bowyer MC & Scarlett CJ. Chemical profile and antioxidant activities of the crude extract and different fractions prepared from the brown alga Hormosira banksii (Turner) Decaisne. Submitted to Journal of Botanica Marina. 6. Paper VI: Dang TT, Bhuyan DJ, Bond DR, Bowyer MC, Van Altena IA & Scarlett CJ. iv Fucoxanthin content, isolation and cytotoxic activity against pancreatic cancer from brown alga Hormosira banksii (Turner) Decaisne. Submitted to Journal of Biotechnology. 7. Paper VII: Dang TT, Sakoff JA, Bowyer MC, Van Altena IA & Scarlett CJ. Antioxidant and cytotoxic activity (in vitro) of phlorotannin-enriched fractions from the brown alga Hormosira banksii (Turner) Decaisne. Submitted to Journal of MarineBiotechnology. 8. Paper VIII: Dang TT & Scarlett CJ. Extraction and cytotoxic activity of the sulfated polysaccharides (fucoidans) against pancreatic cancer in vitro from brown alga Hormosira banksii (Turner) Decaisne. Submitted to Journal of Biomedicine andPharmacotherapy. v STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, We, Quan V. Vuong, Maria J. Schreider, Christopher J. Scarlett, Michael C. Bowyer and Ian A. Van Altena as co-authors with contribution of planning, giving the suggestions and editing the paper, attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the published research paper entitled “The Effects of Drying on Physico‐Chemical Properties and Antioxidant Capacity of the Brown Alga (Hormosira banksii (Turner) Decaisne)”. J. Food Process Pres 41(4): e13025. doi.org/10.1111/jfpp.13025. Dr. Ian A. Van Altena Date: 27/4/2018 Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: ix STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, We, Christopher J. Scarlett, Michael C. Bowyer and Ian A. Van Altena as co-authors with contribution of planning, giving the giving the suggestions and editing the paper, attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the published research paper entitled “Comparisonof chemical profileandantioxidantproperties of the brown algae”, Inter. Food Sci. Technol 51(1): 174-181. doi:10.1111/ijfs.13571. Dr. Ian A. Van Altena Date: 27/4/2018 Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: x STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, We, Quan V. Vuong, Maria J. Schreider, Christopher J. Scarlett, Michael C. Bowyer and Ian A. Van Altena as co-authors with contribution of planning, giving the giving the suggestions and editing the paper, attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the published research paper entitled “Optimisation of ultrasound-assisted extraction conditions for phenolic content and antioxidant activities of the alga Hormosira banksii using response surface methodology”. J Appl Phycol 29(6): 31613173. doi.org/10.1007/s10811-017-1162-y. Dr. Ian A. Van Altena Date: 27/4/2018 Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: xi STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, We, Christopher J. Scarlett, Michael C. Bowyer and Ian A. Van Altena as co-authors with contribution of planning, giving the feedbacks and editing the paper, attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the published research paper entitled “Optimum conditions of microwave assisted extraction for phenolic compounds and antioxidant capacity of the brown alga Sargassum vestitum” Sep Sci Technol (In Press). doi.org/10.1080/01496395.2017.1414845. Dr. Ian A. Van Altena Date: 27/4/2018 Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: xii STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, We, Quan V. Vuong, Christopher J. Scarlett, Michael C. Bowyer as co-authors with contribution of planning, giving the suggestions and editing the paper, attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the published research paper entitled “Chemical profile and antioxidant activities of the crude extract and different fractions prepared from the brown alga Hormosira banksii (Turner) Decaisne”. Submitted to Journal of Botanica Marina. Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: xiii STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, We, Deep J. Bhuyan, Danielle R. Bond, Christopher J. Scarlett, Michael C. Bowyer and Ian A. Van Altena as co-authors with contribution of planning, giving the giving the suggestions and editing the paper, attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the published research paper entitled “Fucoxanthin content, isolation and cytotoxic activity against pancreatic cancer from brown alga Hormosira banksii (Turner) Decaisne”. Submitted to Journal Biotechnology. Dr. Ian A. Van Altena Date: 27/4/2018 Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: xiv STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, We, Christopher J. Scarlett, Jennette Sakoff, Michael C. Bowyer and Ian A. Van Altena as co-authors with contribution of planning, giving the suggestions and editing the paper and Jennette Sakoff (conducting experiments of cancer cells), attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the research paper entitled “Antioxidant and cytotoxic activity (in vitro) of phlorotannin-enriched fractions from the brown alga Hormosira banksii (Turner) Decaisne”. Submitted to Journal of Marine Biotechnology. Dr. Ian A. Van Altena Date: 27/4/2018 Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: xv STATEMENT OF AUTHORS' CONTRIBUTION TO THE PAPERS To whom it may concern, Christopher J. Scarlett as co-authors with contribution of planning, giving the suggestions and editing the paper, attest that research higher degree candidate, Thanh Trung Dang, was the principle contributor to the planning, execution, analyses of the experiments and the writing of the research paper entitled “Extraction and cytotoxic activity of the sulfated polysaccharides (fucoidans) against pancreatic cancer in vitro from brown alga Hormosira banksii (Turner) Decaisne. Submitted to Journal of Biomedicine and Pharmacotherapy. Thanh Trung Dang Date: 26/4/2018 I have seen this paper and I agree with the signatories above that it represents a substantial amount of research work and can be part of Thanh Trung Dang’ s PhD thesis. Professor Frances Martin Assistant Dean Research Training (ADRT) Date: xvi CONFERENCE PRESENTATIONS 1. Thanh T. Dang, Quan V. Vuong, Micheal C. Bowyer, Ian A. Van Altena, Christopher J. Scarlett. Phytochemical and antioxidant properties of crude and fractionated extracts from the brown alga Hormosira banksii (Turner) Decaisne. International scientific conference “Sustainable Agriculture and Environment” December, 13-14, 2016, Ho Chi Minh City, Vietnam. Oral presentation. 2. Thanh T. Dang, Quan V. Vuong, Maria J. Schreider, Micheal C. Bowyer, Ian A. Van Altena, Christopher J. Scarlett. Effect of drying methods on chemical properties and antioxidant capacity of the brown alga (Hormosira banksii). 3rd International Conference Sustainable Agriculture, Food and Energy November, 17-20, 2015, Ho Chi Minh City, Vietnam. Oral presentation. xiv LIST OF ABBREVIATIONS ° C µL ANOVA AQ BuOH DCM EA et al. g Hx h MAE min mL Mw nM RP-HPLC RSM sec TFC TLC TPC UAE UV-Vis v/v w/v w/w W Degree Celsius Microliter (s) Analysis of Variance Aqueous fraction Butanol fraction Dichloromethane fraction Ethyl acetate fraction and others gram (s) Hexane fraction hour( s) Microwave Assisted Extraction Minute (s) Millilitre (s) Molecular weight Nano-moles Reversed-phase High Performance Liquid Chromatography Response Surface Methodology Second (s) Total Flavonoid Content Thin Layer Chromatography Total Phenolic Content Ultrasonic Assisted Extraction Ultraviolet- Visible Volume by Volume Weight by Volume Weight by Weight Watt (s) xv FIGURES FOR THESIS Figure 1: Stran-fucoxanthin Figure 2: Some phlorotannins with phloroglucinol (1,3,5-trihydroxybenzene) as a basic unit xvi Figure 2: Some phlorotannins with phloroglucinol as a basic unit (continue) Structure of fucoidan unit Figure 3: Sulfated polysaccharides (Fucoidans) xvii TABLE OF CONTENTS STATEMENT OF ORIGINALITY --------------------------------------------------------------- i DECLATATION OF AUTHORSHIP ------------------------------------------------------------ ii ACKNOWLEDGEMENTS------------------------------------------------------------------------ iii LIST OF PUBLICATIONS INCLUDED AS PART OFTHE THESIS ---------------------- iv STATEMENTS OF AUTHOR S’ CONTRIBUTION TOTHE PAPERS ------------------- vi CONFERENCE PRESENTATIONS ------------------------------------------------------------ xiv LIST OF ABBREVIATIONS -------------------------------------------------------------------- xv FIGURES FOR THE THESIS-------------------------------------------------------------------- xvi TABLE OF CONTENTS ----------------------------------------------------------------------- xviii ABSTRACT ----------------------------------------------------------------------------------------- xx 1 PART 1: LITERATURE REVIEW 1.1 1.1.1 a) b) 1.1.2 a) b) c) d) 1.1.3 a) b) 1.1.4 a) b) 1.1.5 1.1.6 a) b) Background 1 Algae and health benefits Overview of marine algae Health benefits Bioactive compounds from brown algae Fucoxanthin Pholorotannins Polysaccharides (fucoidans) Other components Extraction of algal compounds Microwave Assisted Extraction Ultrasound Assisted Extraction Isolation and identification of bioactive compounds Chromatographic separation and isolation ofbioactive compounds Identification of bioactive compounds Activities of algal compounds against several cancer cell lines Algal components against pancreatic cancer cell lines Problems with pancreatic cancer Algal compounds against pancreatic cancer 1 1 2 4 4 5 6 7 8 9 10 10 10 15 16 18 18 19 1.2 Research content 20 1.3 Research Aims and Expected Outcomes 21 1.4 Experimental Rationale 21 1.5 Hypothesis, aims and objectives 23 26 PART 2: RESULTS 2.1 Synopsis of research papers published from results 26 2.2 Research papers published from results 32 xviii PART 3: GENERAL DISCUSSION AND CONCLUSIONS 3.1 3.1.1 3.1.2 3.1.3 3.2 3.2.1 3.2.2 General discussion 34 34 Preparation of the samples (dryingand extraction) Bioactive compounds and isolation processes frombrown algae Activity of algal compounds against pancreatic cancercell lines Conclusions and recommendations 35 38 41 43 Conclusions Recommendations 43 45 46 BIBLIOGRAPHY xix