MINISTRY OF EDUCATION AND TRAINING NONG LAM UNIVERSITY HO CHI MINH CITY FACULTY OF BIOLOGICAL SCIENCES GRADUATION THESIS ANALYSIS OF PHYTOCHEMICAL COMPOSITION, ANTIOXIDANT AND ANTIBACTERIAL ACTIVITIES OF SEED KERNEL EXTRACTS FROM TU QUY MANGO (MANGIFERA INDICA L.) Major : BIOTECHNOLOGY Student implementation : PHAN THI BICH TRAM Student code : 19126188 Academic year : 2019 - 2023 Thu Duc City, August 2023 MINISTRY OF EDUCATION AND TRAINING NONG LAM UNIVERSITY HO CHI MINH CITY FACULTY OF BIOLOGICAL SCIENCES UNDERGRADUATE THESIS ANALYSIS OF PHYTOCHEMICAL COMPOSITION, ANTIOXIDANT AND ANTIBACTERIAL ACTIVITIES OF SEED KERNEL EXTRACTS FROM TU QUY MANGO (MANGIFERA INDICA L.) Advisor Student Dr. NGUYEN THANH VIET PHAN THI BICH TRAM Thu Duc City, August 2023 ACKNOWLEDGEMENTS First of all, I would like to express my sincere thanks and deep gratitude to the teachers of Nong Lam Ho Chi Minh City University, especially the teachers of the Department of Biological Sciences who have dedicatedly taught and imparting invaluable knowledge to me during my study at school. To complete this graduation thesis, I would like to thank the Institute of Technology Application and Sustainable Development, Nguyen Tat Thanh University for providing facilities, helping me to access more equipment. I sincerely thank PhD.
Dinh Xuan Phat, a lecturer at the Faculty of Biological Sciences, has created the best conditions for me to carry out this graduation thesis. I would like to express my deep gratitude to Mr. Nguyen Thanh Viet of Nguyen Tat Thanh University in Ho Chi Minh City, who directly guided the thesis, wholeheartedly guided and helped me find a research direction to help me complete this thesis. And finally, I would like to thank my family, brothers, sisters and friends who have always been by my side to share, encourage and cheer me on during my graduation thesis.
Due to the limited amount of knowledge and reasoning ability of oneself, there will be many errors and limitations. I look forward to receiving your guidance and contributions to make my thesis more complete. Thank you from the bottom of my heart. CONEIRMATION COMMITMENT My name: Phan Thi Bich Tram, MSSV: 19126188, class: DH19SHA in Biotechnology, Nong Lam Ho Chi Minh City University.
I hereby declare that: This is my own graduation thesis with the data and information in the research process is completely honest and objective, I am fully responsible before the committee for the commitments. Ho Chi Minh City, August 2023 Students perform Phan Thi Bich Tram ABSTRACT The objective of this study is to construct a database on the antioxidant and antibacterial activities of mango kernel extracts, obtained via different methods. These methods include maceration (M), hot extraction (HRE), and ultrasonic-assisted extraction (UAE). The results showed that the extract obtained through the UAE method had the highest content of polyphenols, as well as superior antioxidant and antibacterial activities.
Phytochemical analyses detected the presence of phenolics, tannins, alkaloids, flavonoids, and saponins in the mango kernel samples, which may contribute to their biological activity. The assessment of antioxidant activity in the three tested extracts demonstrated free radical scavenging efficiency in two experiments: DPPH and ABTS. ICso values for DPPH and ABTS ranged from 121.71 ug/mL and 162.35 ug/mL, respectively. Antibacterial activity was determined by the agar well diffusion method.
The minimum inhibitory concentration (MIC) was ascertained based on the resazurin staining mechanism using 96-well plates, and the minimum bactericidal concentration (MBC) was also identified. The results demonstrated that mango kernel extracts exhibited antibacterial activity against the tested bacterial and fungal strains, namely Escherichia coli ATCC 25922, Pseudomonas aeruginosa NRRL B-14781, Salmonella typhimurium YS1646, Listeria monocytogenes ATCC 13932, Staphylococcus aureus ATCC 6738, Bacillus cereus ATCC 10876, and Candida albicans 10231. The diameters of the inhibition zones ranged from 11. The MIC of the tested samples ranged from 0.50 mg/mL, while the MBC ranged from 1.
Variations in extraction methods, temperature, and humidity are also considered factors influencing the differences in biological activity observed among the various types of mango kernel extracts. Keywords: mango seed kernel, TPC, DPPH, ABTS, MIC, MBC TÓM TẮT Phân tích thành phần hóa thực vật, hoạt tính kháng oxy hóa và kháng khuẩn từ cao chiết nhân hạt từ giống xoài Tứ Quý (Mangifera indica L.) Nghiên cứu này được tiến hành nhằm xây dựng cơ sở đữ liệu về hoạt tính kháng oxy hóa và hoạt tính kháng khuẩn của nhân hạt xoài thu được bằng các phương pháp khác nhau. Các phương pháp này bao gồm ngâm (M), chiết xuất nóng (HRE) và chiết xuất có hỗ trợ siêu 4m (UAE). Kết quả cho thấy chiết xuất thu được thông qua phương pháp UAE có hàm lượng polyphenol cao nhất, cũng như các hoạt tính chống oxy hóa và kháng khuẩn vượt trội.
Các phân tích hóa thực vật đã phát hiện ra sự hiện diện của phenolics, tanin, alkaloid, flavonoid và saponin trong các mẫu nhân xoài, những chất nay có thé góp phan vào hoạt động sinh học của chúng. Việc đánh giá hoạt tính chống oxy hóa trong ba chiết xuất được thử nghiệm đã chứng minh hiệu quả loại bỏ gốc tự do trong hai thí nghiệm: DPPH và ABTS. Giá trị ICso đối với DPPH và ABTS lần lượt nằm trong khoảng từ 121,76 đến 170,71 ug/mL và 162,67 đến 235,35 ug/mL. Hoạt tính kháng khuẩn được xác định bằng phương pháp khuếch tán giếng thạch.
Nồng độ ức chế tối thiêu (MIC) được xác định dựa trên cơ chế nhuộm resazurin sử dụng đĩa 96 giếng và nồng độ diệt khuẩn tối thiêu (MBC) cũng được xác định. Kết qua đã chứng minh rang chất chiết xuất từ hạt xoài thé hiện hoạt tính kháng khuẩn chống lại các chủng vi khuẩn và nam được thử nghiệm, cu thé là Escherichia coli ATCC 25922, Pseudomonas aeruginosa NRRL B-14781, Salmonella typhimurium YS1646, Listeria monocytogenes ATCC 13932, Staphylococcus aureus ATCC 6738, Bacillus cereus ATCC 10876, and Candida albicans 10231. Đường kính của vùng ức chế nằm trong khoảng từ 11,00 mm đến 20,33 mm. MIC của các mẫu thử nghiệm nam trong khoảng từ 0,63 đến 2,50 mg/mL, trong khi MBC nam trong khoảng từ 1,32 đến 5,00 mg/mL.
Sự khác biệt trong phương pháp chiết xuất, nhiệt độ và độ âm cũng được coi là những yếu tố ảnh hưởng đến sự khác biệt về hoạt tính sinh học được quan sát giữa các loại chiết xuất từ hạt xoài khác nhau. Từ khóa: nhân hạt xoài, TPC, DPPH, ABTS, MIC, MBC TABLE OF CONTENTS ABS TRACT wersessncensecovnnrenieeeeerr sien uremanin rn enema nwaiesi nny oy eemoemiemineeiorsetes 11 OT HN. INTRODUCTION ves cmessc<massincmaneanetamenmeaawaesauas stem 1 i(ch eee ee ee 1 Leds OB] CCV 6 vec ccesnomannerameereecenwercemeeansuaaeeeanasereneueser eum uaET Heer EEE RES 2 15a-RIESGanchGGITEGHESsssssssssssbsssosseseadipgoasTa.enlseSineiiliEgidgHBi44idl6kif,A/48s20610/207A400ãng03025008gbig. LITERATURE RE VIEW) cuc go nhàn 1401100311611501431601614184010032044 3 01a; IGEVHGNWI G-THIATTDOsssssetionestioktzetebbidsgHiqg2gBipdS38SDBSĐSGc0SSGHSSDSESHBHGDRGESSHERGHEHB.
bOIIE/TTBTĐDI.ccx-scscc2022-5-522228n6512 30326 instvecsdinnssved des scene saeearscoasues lasses insaseasitiness 3 Pu con v00 v1. 3 20:28: ÏMfELHOIAI:TIBTEOHGH án cxccsmesecenemmenceerammemcian mena eae 4 2. Mango production SIfUAfIOTI. Overview of bacteria and ÍUñnØ1.
PESCHEFICNIG, COME DACUGTIA, sxswessanscoreannnesessn sus tousnanevnsina hiS0Ag0L30838. Pseudomonas aeruginosa DaC(©TI14. c5 cv ng HH Ỹ 2. Research status of mango activity in Vietnam and in the worÌd.
Research in the (0n. MATERIALS AND METHODS. Time and place of perÍOrImaCG. Materials, chemicals and research equ1pIen(.
--- - 55+ +++*s+*++es+zexxerseerres 12 8 Dre | a AR CSCALCHy MI ALETI A 8 ssaseseoasntiniibidtDotittuinNSESENG13GGIRGDS1S. Research equipment and tools. 13 Bie Ee RIPAOUIOT TSUN OO Ss veneseserecranecaxen ser we neiaunces ee er ameiasump cans eausioran wenapaeeeamnwemanamncene 14 3. Method of maceration beams.
Hot extraction methOd.-- ---- 5 2+2 * + 33x +3 vn ng nrnn nrnrkc 15 3.3, Ultrasonic assisted extraction rii6tlO‹. Research methods on mango kernels .-- ----- ¿+ +2 ++++++s£++++tE+zE+ztesrerserseerke 16 3. Evaluation of physicochemical properties. ¿55 5552 s+ssssrerrrerres 16 321:14Ì: Define C01 OF son gzgsnngy E165 014 DA T3IQS5IRDEEBSSSSEĐHES.
IDetermingfion:of humidity 1 ssesacssbieti401806330/4N8G158204380655GSSHGS20400G0S81<gH@ids8 16 3. Method for determining water aC{TVIẨY.- -- 2 2c 22x 2222 211 2 re rệt 17 3⁄4:1:1: Deter pH saseseszbsseoiasiEESE0138101481S8EISESNSESLVBSSESISEBSAGHGSESSRSS0M9343383405381978 0008 17 3. Method for determination of vitamin C. ---- 55+ + ++c++ec+eeeeeererreerxee 18 BAKO, ASW essmscmemre meneame eerie tee EEE 19 3.
Analysis method of phytochemical components in mango kernel extract. 19 3:6; EXalữatfoi Of aiiHöGXIHaTIE RGHTY <cocecessexseuesecsssmeememernam acerca 20 3. DPPH free radical scavenging method. ABTS free radical scavenging abIÏIfV.
Evaluation of polyphenol content from mango seed kernel extract. Investigation of antibacterial and antifungal activities of mango seed kernel CXIPA C8 csnsnsnssestkis0i19056938554563655853008058005105S04SE9109860/5895319uS0GDGGH2380. Agar plate diffusion method. Methods for evaluating the minimum inhibitory concentration and the minimum bactericidal concentration 0.
Statistical processing methO.-- -- - + 5+ + *+*E+*E+tE£vEekExrExErrekrekrrerrrkrrerrkre 24 Chapter 4. RESULTS AND DISCUSSION. Results of evaluation of physicochemical componens. Obtaining mango seed €XÍTAC.
Analysis results of phytochemical compOSIHIOH. Evaluation results of polyphenol content from mango seed extract. Evaluation results of antioxidant aCfIVIẨY. Evaluation results of antibacterial and antifungal activity «0.
Determination of antibacterial and antifungal rIngs. The results of the determination of the minimum inhibitory concentration and the minimum bactericidal cOC€TitrAf1OTI. CONCLUSIONS AND RECOMMENDATIONS.- 35 Sa, "© ome Wd 2x5225615174G50155010G308SE36138E55ESEESEESBER0801024JEEĐRIEEREEEAEEIGEISGEGEEEDXRDEEHUGSEIGHREASSEES.EA88088 35 SWUM [LIAS cE (6) Wee eee eee ee et ee ee ee 35 APPENDIX vi LIST OF ABBREVIATIONS ABTS : [2,2’-azisbonis(3-ethylbenzothiazolin-6-sulfonicacid) diammonium| ANOVA : Analysis of variance B. cereus - Bacillus cereus C.
albicans : Candida albicans CFU : Colony Forming Units DCPIP : 2,6-diclophenol-indophenol DPPH : 2,2-diphenyl-1-picrylhydrazy E. coli : Escherichia coli E. faecalis : Enterococcus faecalis FAO : Food and Agriculture Organization of the United Nations GAE : Gallic acid equivalent HRE : Hot reflux extraction ICao : Inhibitory Concentration 50% L. monocytogenes: Listeria monocytogenes M : Maceration MBC : Minimum Bactericidal Concentration MIC : Minimum Inhibitory Concentration P.
acnes : Propionibacterium acnes P. aeruginosa : Pseudomonas aeruginosa S. aureus : Staphylococcus aureus S. typhimurium : Salmonella typhimurium TAA : Total Ascorbic Acid TPC : Total polyphenol content UAE : Ultrasound-assisted extraction UV - Vis : Ultraviolet - visible vii LIST OF TABLES Table 3.
Extraction parameters of the methods. Table of high dilutions in 96-well pÏaf€S. Physicochemical composition of mango seed kernel powder. Table of high recovery efficiency of mango seed kernel extract.
Quantitative results of TPC compounds. -- -- 5+ +s+++£+ses+exssessx2 28 Table 4. Evaluation results of antioxidant activity .-- -- 5c s++cs+ccsseeserrrerres 29 Table 4. Table of determination of antibacterial and antifungal rings.
Table of determination of MIC and MBC of mango seed extract. 33 viii LIST OF FIGURES Figure 3. Flowchart of the extraction process of mango kernel extract. (OLE LAB COlGE SPACE esac cs seazcconence neseuinscstens 2HGSRB3hMBSSRAGGIG.
The basis of the DPPH free radical scavenging method. ABTS free radical scavenging ÍaC1Ì1fy. Mango seed kernel powder after drying. Samples of mango seed kernel extract after reCOV€YY.
Gallic Acid CONCENIT ANON svssccsessssnescsstsvessesnveveunsoenanssnesesvivonsuaoasvavereunasenentes 28 Figure 4. Test results of agar plate diffusion test on E. MIC plate spreading results of FE. Overview The practice of supporting the treatment of diseases with plant extracts has been prevalent for centuries, playing a pivotal role in fulfilling healthcare needs, particularly in developing countries.
Antibiotic resistance has emerged as a significant global issue. Many plants are considered useful in supporting the treatment of infections. Consequently, the search for novel antioxidants and antibacterial agents with no side effects has become one of the trending topics in current scientific research. Several plant extracts have demonstrated capabilities to suppress the activity of multidrug-resistant microorganisms.
This highlights the potential application of plant extracts in fields such as food, cosmetics and pharmaceuticals. Mango (Mangifera indica L.), a common fruit tree in tropical countries, including Vietnam, ranks second in the world in terms of fruit production, after the banana. India leads globally in mango producer and acreage. In 2019, global mango production reached 51 million tons, generating about 210,000 tons of waste per year (El-Kady et al., 2017; Lebaka et al.
Mangos are typically consumed fresh, used In juices, dried, or powdered. However, businesses or processing units only focus primarily on the flesh of the mango, neglecting nutrient-rich byproducts like mango kernels. According to research by Garcia-Mahecha et al.