Evaluation of tyrosinase and antioxidant activities of brominated compounds from vietnamese red seaweeds

MINISTRY OF EDUCATION AND TRAINING NHA TRANG UNIVERSITY PATRICK A BLAMO, JR EVALUATION OF TYROSINASE AND ANTIOXIDANT ACTIVITIES OF BROMINATED COMPOUNDS FROM VIETNAMESE RED SEAWEEDS MASTER THESIS KHANH HOA - 2020 MINISTRY OF EDUCATION AND TRAINING NHA TRANG UNIVERSITY PATRICK A BLAMO, JR EVALUATION OF TYROSINASE AND ANTIOXIDANT ACTIVITIES OF BROMINATED COMPOUNDS FROM VIETNAMESE RED SEAWEEDS MASTER THESIS Food Technology Major: Topic allocation Decision Decision on establishing the Committee: September 19, 2020 Defense date: Supervisors: Dr The Han Nguyen Dr Hong Ngoc Thuy Pham Chairman: Faculty of Graduate Studies: KHANH HOA - 2020 UNDERTAKING I declare that the thesis entitled: ―Evaluation of Tyrosinase and Antioxidant Activities of Brominated Compounds from Vietnamese Red Seaweeds‖ is my own work The work has not been presented elsewhere for assessment until the time this thesis is submitted 09 2020 Patrick A Blamo, Jr (Author) iii ACKNOWLEDGMENT My health, strength and the opportunity to be part of this amazing family, I owe it to God I would like to appreciate my professors and the entire staff in the Department of Food Technology, Nha Trang University Your sacrifices, knowledge impacted and the amicable environment created to study and successfully complete this master's thesis, cannot go unappreciated My profound gratitude to Dr The Han Nguyen and Dr Hong Ngoc Thuy Pham, supervisor and co-supervisor, respectively Your guidance, support, patience and continuous courage fueled my enthusiasm Your vast knowledge and experience, created a confident roadmap that has led to the success of this thesis Finally, and importantly, my family: my mother and partner whose prayers never fainted, my uncle whose support never quenched, you are all the reason this is possible…thank you! 09 2020 Patrick A Blamo, Jr iv CONTENT UNDERTAKING iii ACKNOWLEDGMENT iv CONTENT .v LIST OF SYMBOLS ix LIST OF ABBREVIATIONS x LIST OF TABLES xi LIST OF FIGURES xii ABSTRACT xiv CHAPTER INTRODUCTION 1.1 Introduction 1.2 Background 1.3 Statement of Problem 1.4 Justification of the Study .6 1.5 Research Question 1.6 Hypothesis .7 1.7 Objective 1.7.1 General Objective 1.7.2 Specific Objective 1.8 Scope of Study .8 1.9 Limitation of the study CHAPTER LITERATURE REVIEW .10 2.1 Introduction 10 2.2 Scope 10 2.3 Overview of Seaweeds 11 2.4 Significance of Investigating Seaweeds .12 2.5 Biological Potential of Seaweeds (Laurencia) 13 v 2.5.1 Total Phenolic Profile 13 2.5.2 Tyrosinase Inhibitory Property 13 2.5.2.1 Influence of Enzymatic discoloration .13 2.5.2.2 Role of Tyrosinase in Melanin Biosynthesis 14 2.5.3 Oxidative Damages and Control 16 2.5.3.1 Effect of Reactive Oxygen Species 16 2.5.3.2 Antioxidant 17 2.5.3.3 Antioxidant Properties 18 2.5.4 Brominated Compounds 19 2.5.4.1 Definition .19 2.5.4.2 Brominated compounds from Laurencia .19 2.6 Health Effect of Synthetic Compound 20 2.7 Effect of extraction methods and parameters on the biological activity of seaweeds 21 2.7.1 Extraction method 21 2.7.2 Extraction solvent .22 2.7.3 Extraction time .23 2.7.4 Extraction temperature 23 2.7.5 Solute-solvent (volume/mass) ratio .24 2.8 Mechanism of Ultrasound-Assisted Extraction 24 2.9 Response Surface Methodology (RSM) .26 2.10 Modern Application of Seaweeds .26 CHAPTER MATERIALS AND METHODS 29 3.1 Materials .29 3.1.1 Seaweed samples 29 3.1.2 Chemicals and Reagent 29 3.2 Methods 29 3.2.1 Experimental Procedure .29 vi 3.2.2 Screening test to select the starting sample 30 3.2.3 Single factor test 31 3.2.4 Experimental optimization Design .31 3.2.5 Ultrasound Assisted Extraction (UAE) 31 3.2.6 Fractionation, Isolation and characterization of compounds from L intermedia Yamada 32 3.2.6.1 General experimental procedures 32 3.2.6.2 Extraction, fractionation and isolation 32 3.2.6.3 Characterization of isolated compounds 33 3.2.7 Determination of total phenolic content, antioxidant and tyrosinase inhibitory activities .34 3.2.7.1 Total phenolic content (TPC) 34 3.2.7.2 DPPH radical scavenging assay (DRSC) 34 3.2.7.3 Ferric reducing antioxidant power (FRAP) 34 3.2.7.4 Tyrosinase Inhibition Assay (TIA) 35 3.2.8 Statistical analysis 35 CHAPTER RESULTS AND DISCUSSION 36 4.1 Screening 36 4.2 Single factor optimization 36 4.3 Optimization .40 4.3.1 Fitting the models 40 4.3.2 Influence of extraction parameters on TPC and antioxidant capacity 45 4.3.3 Optimization and validation of the models 53 4.4 Correlation between the total phenolic content and antioxidant activities 54 4.5.Total phenolic content and biological activities of the fractions and isolated compounds 55 4.6 Purification and structural identification of aplysistatin and palisadin b 56 CHAPTER CONCLUSION recommendation AND FUTURE PERSPECTIVE .63 vii 5.1 Conclusion 63 5.2 Recommendation 63 5.3 Future Perspective 64 REFERENCE 65 APPENDICES I Appendices A I Appendices B II viii LIST OF SYMBOLS % : percent g : gram g : milligram GAE : Gallic Acid Equivalent AAE : Ascorbic Acid Equivalent m : mass M : molar nm : Nanometer °C : degree Celsius TE : Trolox Equivalent ix LIST OF ABBREVIATIONS ANOVA : Analyzes of Variance BBD : Box-Behnken Design DPPH : 1,1-Diphenyl-2-picrylhydrazyl DRSC : DPPH Radical scavenging capacity EtOAc : Ethyl Acetate EtOH : Ethanol FRAP : Ferric Reducing Antioxidant Power HCl : Hydrochloric Acid HPLC : High-Performance Liquid Chromatography MeOH : Methanol Na2CO3 : Sodium carbonate NMR : Nuclear Magnetic Resonance R2 : R-square (determined coefficient) RMSE : Root Mean Square Error ROS : Reactive Oxygen Species RSM : Response Surface Methodology TIA : Tyrosinase Inhibitory Activity TLC : Thin layer chromatography TPC : Total Phenolic Content TPTZ : 2,4,6-tris(2-pyridyl)-s-triazine UAE : Ultrasound-assisted extraction UV : Ultraviolet Radiation x 98 Mandrekar VK, Gawas UB, Majik MS Brominated molecules from marine algae and their pharmacological importance Studies in Natural Products Chemistry 2018 Jan;6:461–490 DOI: 10.1016/B978-0-444-64183-0.00013-0 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