(Đồ án hcmute) phytochemical, antioxidant capacity and antifungal property screening of passiflora edulis seed ethanolic extract

HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION KING MONGKUT’S UNIVERSITY OF TECHNOLOGY THONBURI A THESE MAJORS CHEMISTRY AND FOOD TECHNOLOGY PHYTOCHEMICAL, ANTIOXIDANT CAPACITY AND ANTIFUNGAL PROPERTY SCREENING OF PASSIFLORA EDULIS SEED ETHANOLIC EXTRACT ADVISOR: SONGSIN PHOTCHANACHAI CANDIDATE: NGUYEN HUYNH TIEN STUDENT NO: 15116051 SKL 0 2019 an i Phytochemical, antioxidant capacity and antifungal property screening of Passiflora edulis Seed Ethanolic Extract MISS NGUYEN HUYNH TIEN STUDENT NO 15116051 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE DEGREE OF BACHELOR OF SCIENCE (FOOD TECHNOLOGY) SCHOOL OF BIORESOURCES AND TCHNOLOGY KING MONGKUT‘S UNIVERSITY OF TECHNOLOGY THONBURI HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION 2019 an ii Phytochemical, antioxidant capacity and antifungal property screening of Passiflora edulis Seed Ethanolic Extract Miss Nguyen Huynh Tien (Intership student) A Thesis Submitted in Partial Fulfilment of the Requirement for the Degree of Bachelor of Science (Food Technology) Ho Chi Minh City University of Technology and Education 2019 Thesis Committee ………………………………………………… Chairman of Thesis Commitee (Asst Prof Songsin Photchanachai, Dr Agr Sci.) ………………………………………………… Member and Mentor (Ms Wissanee Pola, Ph.D student) Coppyright Reserved an iii Thesis Title Phytochemical, antioxidant capacity and antifungal property screening of Passiflora edulis Seed Ethanolic Extract Candidate Miss Nguyen Huynh Tien Thesis Advisor Assoc Prof Songsin Photchanachai Field of Study Chemistry and Food Technology/ Postharvest Technology Faculty Faculty for high quality training/ School of Bioresources and Technology Academic year 2019 Abstract Passion fruit seeds are byproduct from juice production and abundant in nutritional and health benefits The objective of this study was to evaluate total phenolic content (TPC), antioxidant capacity and antifungal activities of ethanolic extract of passion fruit seeds The seeds were grounded and extracted with 50% and 70% ethanol The TPC was evaluated by Folin-Ciocalteau assay Antioxidant capacities were measured by free radical scavenging activity (DPPH; 2,2-diphenyl-1-picrylhydrazyl), ferric reducing antioxidant power (FRAP) and antifungal acvity against Aspergillus niger using the radial growth technique Results showed that the TPC and antioxidant capacities of the crude extraction using 70% ethanol are significantly higher than that of the 50% ethanol Moreover, the ethanolic extract of P.edulis seed cake showed antifungal activities against important fungal foodborne pathogen ( Aspergillus niger) Conversely, the concentration of the solvent did not influence the crude yield These findings demonstrated that the 70% ethanol would be an appropriate concentration for extraction of passion fruit seeds However, further investigation is recommended using higher concentration of ethanol and other solvents to maximize the nutritional value of passion fruit seed Keywords:Antioxidant capacity, Passion fruit seed, Total phenolic content, Aspergillus Niger an iv ACKNOWLEDGEMENTS This work would not have been possible without the financial support of the King Mongkut‘s University Technology Thonburi, Thailand I am especially indebted to Asst Prof Dr Songsin Photchanachai of the Seed and Grain Laboratory, who have been supportive of my thesis and encouraging me throughout the study, the chairman of my committee I am grateful to all of those with whom I give so much heling to work during this time Each of the members of the Seed amd Grain Laboratory has provided me extensive personal and professional guidance and taught me a great deal about both scientific research and life in general I would especially like to thank Ph.D Wissanee Pola, as my teacher and mentor, she has taught me a lot of things and trained me to be better in the work A special thanks to my lectures in Ho Chi Minh City University of Technology and Education who have given me the opportunity to exchange student programs at King Mongkut‘s University Technology Thonburi, Thailand They always support me during this time Especially, Dr Trinh Khanh Son who gives me good opinions and take care of me Lastly, I would like to thank my family members and my boyfriend for their kind understanding and wholehearted support throughout my working thesis time here at KMUTT an v CONTENTS PAGE Abstract iii ACKNOWLEDGEMENTS iv CONTENTS v LIST OF TABLES vii LIST OF FIGURES viii CHAPTER 1: INTRODUCTION 1 Research background Objectives Scope of study 4 Expected benefits CHAPTER 2: LITERATURE REVIEW Passion fruit seed Benefits of passion fruit seed Antioxidants 3.1 Introduction antioxidants 3.2 Phenolic compounds 3.3 Mechanism action of antioxidant 11 Extraction method 15 5.1 Effect of solvent type 16 5.2 Effect of solvent concentration 18 5.3 Effect of DMSO 19 Assessment methods of antioxidant activity 19 Aspergillus niger 20 an vi CONTENTS PAGE CHAPTER 3: MATERIALS AND METHODS 23 Preparation of materials 23 Extraction procedure 23 Determination of total phenolic content 23 Free radical scavenging activity by DPPH assay 24 Ferric reducing antioxidant power (FRAP) assay 25 Evaluation of crude extract from Passiflora edulis seed to Aspergillus niger 25 6.1 Preparation of crude extract 25 6.2 Preparation of the inoculum 25 6.3 Mycelial growth inhibition 26 CHAPTER 4: RESULTS 27 Extraction yield 27 Evaluation of antioxidant capacity 27 The influence of crude extract on Aspergillus niger growthing 29 CHAPTER 5: DISCUSSION 31 Experiment 31 Experiment 33 CHATER 6: CONCLUSION AND SUGGESTION 35 REFERENCES 36 APPENDIX A 48 an vii LIST OF TABLES TABLE PAGE Table Chemical composition of passion fruit seed meal Table Conditions for vegetative growth of Aspergillus niger 21 Table Extraction yield of crude is extracted using ethanol 50% and ethanol 70% 27 Table Total phenolic content of crude extracts using 50% and 70% ethanol 48 Table Antioxidant activity of crude is extracted using ethanol 50% and ethanol 70% determined by DPPH, FRAP assays 48 Table Percentage inhibition of mycelial growth of crude extract from Passion fruit seed using 70% and 50% ethanol 48 Table The growthing of fungi in four different mediums 49 an viii LIST OF FIGURES FIGURE PAGE Figure (A) Purple passion fruit (Passiflora.edulis Sims) and (B) Yellow passion fruit (Passiflora edulis f.flavicarpa) Figure The seed from Passion fruit Figure Chemical structure of (a) Scirpusin B, (b) resveratrol and (c) piceatannol 10 Figure Illustration mechanism of primary and secondary antioxidants 12 Figure Free radical-scavenging mechanism of plant phenolic compounds 13 Figure Molecular structures of trans-resveratrol and piceatannol The conversion of trans-resveratrol to piceatannol is catalyzed by P450 enzyme in the presence of NADPH 14 Figure Microscopic structure of Aspergillus niger 21 Figure Standard curve of garlic acid solution at 765nm absorbance 24 Figure Standard cruve of Trolox solution at 593 nm absorbance 25 Figure 10 The total phenolic content of crude is extracted using ethanol 50% and ethanol 70% 27 Figure 11 Antioxidant activity of crude is extracted using ethanol 50% and ethanol 70% determined by DPPH assay 28 Figure 12 Figure Antioxidant activity of crude is extracted using ethanol 50% and ethanol 70% determined by FRAP essay 28 Figure 13 The effect of DMSO on A niger growth at different concentrations 29 Figure 14 The growthing of fungi in four different mediums 30 an CHAPTER 1: INTRODUCTION Research background The passion fruit, also named as egg-shaped fruit which originated from Brazil, has been cultivated in tropical, subtropical and temperate regions on a large scale (Liu, S., et al., 2009) Passion fruit belongs to the Passifloraceae family and originally from tropical America, featuring over 500 species worldwide Among these, the fruits of only about 20 varieties are edible including the most cultivated species in the world namely the yellow passion fruit (Passiflora edulis f flavicarpa), purple passion fruit (P edulis) and sweet passion fruit (P alata) (Malacrida, C R., & Jorge, N., 2012) A B Figure (A) Purple passion fruit (Passiflora.edulis Sims) and (B) Yellow passion fruit (Passiflora edulis f.flavicarpa)  Classification Domain Eukaryote Subclass Dileniidae Kingdom Plantae (Plants) Order Violales Tracheobionta (Vascular plants) Family Superdivision Spermatophyta (Seed plants) Genus Subkingdom an Passifloraceae Passiflora 36 REFERENCES AACC., 2000, Approved Methods of the AACC Method 44-15A, 10thed., The Association, St Paul, Minnesota,USA Abarca, M L., Bragulat, M R., Castella, G., & Cabanes, F J., 1994 Mycoflora and aflatoxin-producing strains in animal mixed feeds Journal of Food Protection, 57(3), pp 256-258 Abarca, M.L., M.R., Bragulat, G., Sastella and F.J Cabanes, 1994 Ochratoxin a production by strains of Aspergillus flavus var niger Applied Environ Microbiol., 60 pp: 2650-2652 Agizzio AP, Carvalho AO, Ribeiro 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SD (n=4) 2) Significant differences between samples are presented as * for p≤0.05 Table Antioxidant activity of crude is extracted using ethanol 50% and ethanol 70% determined by DPPH, FRAP assays Treatments DPPH (%) Ethanol 50% Ethanol 70% p 36.74 ± 1.481) 48.97 ± 3.151) **2) FRAP (mmol Trolox/100 g FW) 56.95 ± 8.9921) 116.99 ± 9.161) **2) 1) Values are means ± SD (n=4) 2) Significant differences between samples are presented as ** for p≤0.05 Table Percentage inhibition of mycelial growth of crude extract from Passion fruit seed using 70% and 50% ethanol TRT Day Day Day Day Day 70% Ethanol 0.5% 37.1429 3.93939 -4.7228 -5.6669 -6.5565 50% Ethanol 0.5% 34.2857 5.75758 -1.232 -5.6669 -7.7367 Possivetive 13.6857 6.06061 2.95688 5.47902 7.39575 an 49 Table The growthing of fungi in four different mediums Treatments 1) Diameter (cm) Day Day Day Day Day Ethanol 50% Ethanol 70% DMSO 1.38c1) 3.11a1) 4.96a1) 6.75a1) 8.24a1) 1.32c1) 3.17a1) 5.1a1) 6.75a1) 8.12a1) 1.65b 3.24a 4.67b 6.34ab 7.60b Positive 1.81a 3.10a 4.72b 6.04b 7.06c Values are means ± SD (n=5) The same letter are not significantly different an an ... CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION 2019 an ii Phytochemical, antioxidant capacity and antifungal property screening of Passiflora edulis Seed Ethanolic Extract Miss Nguyen Huynh Tien...i Phytochemical, antioxidant capacity and antifungal property screening of Passiflora edulis Seed Ethanolic Extract MISS NGUYEN HUYNH TIEN STUDENT NO 15116051... total phenolic content (TPC), antioxidant capacity and antifungal activities of ethanolic extract of passion fruit seeds The seeds were grounded and extracted with 50% and 70% ethanol The TPC was