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 0 6 1 7 4 2019 do 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 do 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. Songsin Photchanachai, Dr.) ………………………………………………… Member and Mentor (Ms. Wissanee Pola, Ph.
student) Coppyright Reserved do 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. 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.
do 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. 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.
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. do an v CONTENTS PAGE Abstract. v LIST OF TABLES.
vii LIST OF FIGURES. viii CHAPTER 1: INTRODUCTION. Scope of study. 4 CHAPTER 2: LITERATURE REVIEW.
Passion fruit seed. Benefits of passion fruit seed. Mechanism action of antioxidant. Effect of solvent type.
Effect of solvent concentration. Effect of DMSO. Assessment methods of antioxidant activity. 20 do an vi CONTENTS PAGE CHAPTER 3: MATERIALS AND METHODS.
Preparation of materials. Determination of total phenolic content. Free radical scavenging activity by DPPH assay. Ferric reducing antioxidant power (FRAP) assay.
Evaluation of crude extract from Passiflora edulis seed to Aspergillus niger. Preparation of crude extract. Preparation of the inoculum. Mycelial growth inhibition.
Evaluation of antioxidant capacity. The influence of crude extract on Aspergillus niger growthing. 33 CHATER 6: CONCLUSION AND SUGGESTION. 48 do an vii LIST OF TABLES TABLE PAGE Table 1.
Chemical composition of passion fruit seed meal. Conditions for vegetative growth of Aspergillus niger. Extraction yield of crude is extracted using ethanol 50% and ethanol 70%. Total phenolic content of crude extracts using 50% and 70% ethanol.
Antioxidant activity of crude is extracted using ethanol 50% and ethanol 70% determined by DPPH, FRAP assays. Percentage inhibition of mycelial growth of crude extract from Passion fruit seed using 70% and 50% ethanol. The growthing of fungi in four different mediums. 49 do an viii LIST OF FIGURES FIGURE PAGE Figure 1.
(A) Purple passion fruit (Passiflora.edulis Sims) and (B) Yellow passion fruit (Passiflora edulis f. The seed from Passion fruit. Chemical structure of (a) Scirpusin B, (b) resveratrol and (c) piceatannol. Illustration mechanism of primary and secondary antioxidants.
Free radical-scavenging mechanism of plant phenolic compounds. Molecular structures of trans-resveratrol and piceatannol. The conversion of trans-resveratrol to piceatannol is catalyzed by P450 enzyme in the presence of NADPH. Microscopic structure of Aspergillus niger.
Standard curve of garlic acid solution at 765nm absorbance. Standard cruve of Trolox solution at 593 nm absorbance. The total phenolic content of crude is extracted using ethanol 50% and ethanol 70%. Antioxidant activity of crude is extracted using ethanol 50% and ethanol 70% determined by DPPH assay.
Antioxidant activity of crude is extracted using ethanol 50% and ethanol 70% determined by FRAP essay. The effect of DMSO on A. niger growth at 4 different concentrations. The growthing of fungi in four different mediums.
30 do an 1 CHAPTER 1: INTRODUCTION 1. 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. 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. (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 Subkingdom Tracheobionta (Vascular plants) Family Passifloraceae Superdivision Spermatophyta (Seed plants) Genus Passiflora do an 2 Division Magnoliophyta (Flowering Species edulis plants) Class Magnoliopsida (Dicotyledons) One of the favorite fruit, passion fruit is usually consumed in its natural state, with the seeds, or processed as tropical juice. They contain many phytochemicals, such as polyphenolic compounds and carotenoid families, and vitamin C.
About structure, passion fruit is divided into three parts, rind (PF-R), the pulp (PF-P), and seed (PF- S) (Matsui, Y. Normally, the passion fruit seeds are discarded as a byproduct after extraction of the juice from the fruit (Liu, S. The fruits are used to make both fresh juice and concentrated juice. These processes generate rinds and seeds as by-products, and make up approximately 40% and 12% of the starting materials, respectively (Matsui, Y.
Hence, thousands of tons of passion fruit seeds will be discharged each year by food factories. To avoid wasting natural plant resources, many people have begun to study passion fruit seeds. Researchers have discovered that the seed is rich in crude lipid and fiber (Baker, 1997). Moreover studies conducted with passion fruit seeds showed that the bagasse is sources of several compounds of nutritional importance, particularly the capacity to oxidize.
Growing knowledge about the health-promoting impact of antioxidants in everyday foods, combined with the assumption that several common synthetic preservatives may have hazardous effects (Krishnakumar, V., & Gordon, I, 1996), has led to multiple investigations in the field of natural antioxidants (Peschel et al, 2006). The interest in the search for natural antioxidants for use in foods has increased in recent years aiming to decrease the lipid oxidation rate, or for pharmaceutical applications to prevent chronic diseases related to the production of free radicals. The abundant content of phenolic compounds in Passiflora edulis seed may serve as an excellent dietary source of natural antioxidants for disease prevention and health promotion (Chau, C. Natural antioxidants discovered can prevent food spoilage caused by oxidizing agents.
do an 3 Beside the damage losses due to the deterioration due to foods caused by oxidation, many of the phytopathogenic fungi can also cause heavy losses of food spoilage. The growth of fungi result to the reduction in the quality and quantity of food (Abarca, M. Some Aspergillus species are xerophilic fungi and Aspergillus niger is commonly involved in fruit spoilage (Frazier, W. The presence of secondary metabolites, such as mycotoxins could have harmful effects on consumers‘ health.
Pizzolitto et al, 2015). Aspergillus niger was selected to study the impact of crude extracts on fungi-causing food spoilage. It is also one of the most commonly encountered fungi contaminating foods and fruits, and occurring in soil and indoor environments. Natural plant extracts may provide an alternative protect into food and feed from fungal contamination.
Also, there is an increasing interest in the antifungal activities of some natural compounds of crude extracts may inhibit the growth of fungi (Moghaddam, M. The solvent extraction has been used widely to extract bioactive components from plants. The selection of solvents for extracting antioxidant compounds is an important factor to consider. It is noted that an extraction solvent system is based according to the purpose of extraction such as the nature of interesting components, the availability of reagents and equipment, cost and safety concerns (Yu, L.
50% and 70% ethanol are used to obtain the crude extract from Passiflora edulis seeds. A comparison of effect of using two concentrations of ethanol for extraction from P.edulis seed to antioxidant capacity and inhibition of fungal growth is reported below. Objectives The objective of this study was to evaluate the total phenolic content, antioxidant capacity and antifungal property from the crude extract of passion fruit seed using two different concentrations of ethanol solvents for extraction. Scope of study 3.
The effect of two ethanol concentrations (50% and 70%) used to extract crude extract to antioxidant capacity. The effect of two ethanol concentrations used to obtain crude extract on phenolic content. The effect of two ethanol concentrations (50% and 70%) used to obtain crude extract to the growth of fungi causes food spoilage (Aspergillus niger) 4. The use safe solvents, less expensive methods for crude extraction.
Effect of crude extract on fungal as a baseline study for studies of extracting components with anti-fungi activity. Antioxidants, anti-bacteria, anti-mildew are applied in food and pharmaceutical industries. Antioxidants, anti-mold are applied in food and pharmaceutical industries. do an 5 CHAPTER 2: LITERATURE REVIEW 1.
Passion fruit seed The soft edible pulp contains tiny seed up to 25％ of the pulp fresh weight (Chau, C. The seeds of this fruit are rich in fiber, minerals and lipids, with good amount of proteins (Oliveira et al. The high fiber inside the seed, rendered it to be unfit for stock feeding. However, the seed contain compounds such as organic acids, polyphenols, pigments, fractionation of nitrogen, moisture, starch, ash and free amino acids (Pruthi, 1963.