THESIS-DEFENSE-TRỊNH-HOÀNG-HUY-final.pptx

Thesis defense with food
engineering

1. Vietnam National University International University - Department of Biotechnology Food Technology Name: Trịnh Hoàng Huy Student ID: BTFTIU18039 Advisor: M.Sc. Nguyễn Thị Hương Giang Date: 30 / 8 /2023 THESIS DEFENSE CHANGES IN PHYSICOCHEMICAL, MICROBIOLOGICAL AND SENSORY PROPERTIES OF ACEROLA ( Malpighia punicifolia L.) CIDER AFTER FERMENTATION AT VARIOUS DURATION AND TEMPERATURES 1

2. OUTLINE 2 1 2 3 4

3. Project general information Grown mostly in many provinces of the Mekong Delta, especially at Tien Giang. Natural sources of Vitamin C , powerful antioxidants. High in phytochemical components such as anthocyanins , phenolic, and carotenoids. Originated in Central America 3 Nutrients Unit Acerola Cherry Calories Kcal 32 Water g 90.6-92.4 Total Carbohydrates g 7-7.8 Protein g 0.21-1.2 Fat g 0.23 – 0.8 Vitamin C mg 470-920 Anthocyanins mg 2.70-5.2 TPC mg 1028-1131 Table 1. Macronutrient and micronutrients composition of acerola (for 100g fresh fruit) (Vendramini and Trugo, 2000) Figure 1. Acerola cherry (Vendramini and Trugo, 2000) Figure 2 . Vitamin C  ≈ 0.1%   >0.1%

4. Sources TPC (mg GAE/100g FW) Vitamin C (mg AAE/100g FW) β-carotene (μg/100g FW) ACN (mg/100g FW) TEAC DPPH (mg/100g FW) Acerola fruit 1028-1131 470- 920 536.55 2.70-5.2 101-251 Apple 197- 202 4.6 19 2.3- 3.2 52.9-54 Cherry 70.2- 73 3.2 - 122- 140 242-260 Orange 75.7- 78 59.1 110-320 - 125-128 Passion- fruit 292- 301 - 360-780 - 1125-1160 Table 2. The bioactive compounds and antioxidant activity of Acerola and common fruits (Modified from Hoang, Q. B et al, 2022) Project general information Vitamin C > 100 times apple, 20 times citrus Higher antioxidant capacity(DPPH) compared to apples and oranges Total Phenolic Content >10 times other fruits 4 Thin skin that makes them highly perishable Fast-ripening rate, and succulent structure HOWEVER, short shelf life Acerola cider fermentation Fermentation is known to increase the phenolic content of berry juice products by increasing extraction from the skins, including anthocyanins ( Heinonen et al., 1998; Su and Chien, 2007)and therefore may increase the antioxidant capacity (Martin and Matar, 2005) and potential for health benefits.

5. Project general information “Cider fermentation’’ Alcoholic fermented beverage with low alcohol content in the range 3.0–8.5% with a weakly acidic and slightly sour taste. By-products(esters, organic acids, glycerols,..) affect cider sensory quality Cider fermentation condition(time and temperature) 5 Source Condition References Rose apple wax 10 days, 25 o C Chukwan Techakanon et al, 2018 Apple(Golden Delicious) 8 days, 20 o C Lorenzini, M et al, 2018 Apple(UK) 14 days, 18 o C J. Rosend et al, 2015 Fuji Apple 10 days, 20 o C Mengqi Ye et al, 2016 Mango cider 10 days, 25 o C Reddy and Reddy (2011) Temperature had important effect on yeast growth and on the level of volatile compounds. ( According to Beltran et al, 2018) Figure 3 . Factors affect alcoholic fermentation

6. Objectives Find out the best condition to retain the antioxidant as well as vitamin C content of Acerola Cider Create a new cider product with good quality in terms of physicochemical and sensory 6 Hypothesis Different fermentation temperatures (15 o C, 20 o C, and 25 o C) may have significant effects on physicochemical properties (pH, TSS, Alcohol content, Total phenolic, Vitamin C, Antioxidant DPPH), microbiological and sensory qualities of Acerola cider Different fermentation duration (7, 10 days ) may have significant effects on physicochemical properties (pH, TSS, Alcohol content, Total phenolic, Vitamin C, Antioxidant DPPH), microbiological and sensory qualities of Acerola cider

7. 2 . Materials and Methods 7 Materials The raw materials including sour Acerola fruit, harvested from Tien Giang farm, active dry yeast Saf-instant (produced by Lesaffre), and white sugar (produced by Bien Hoa Sugar) were obtained from Mega Market An Phu, An Phu ward, district 2, HCMC Chemicals : All reagents, solvents, and chemicals used during the project were of analytical grades and the highest purity provided from various commercial suppliers. These are potassium iodate, potassium iodide concentrated sulfuric acid , DPPH reagent , pectinase enzyme, 96% ethanol, Folin-Ciocalteu's reagent, di-sodium carbonate, gallic acid, L-ascorbic acid

8. 2. Materials and Methods Cider fermentation Code Temperature ( o C) Duration (days) AT1 15 10 AT2 20 AT3 25 Table 3. Fermentation at various temperature in 10 days Table 4. Fermentation in various duration at 20 C Code Temperature ( o C) Duration (days) AD 1 20 7 AD2 10 8

9. 2 . Materials and Methods 9 Physical Analysis Methods Determination of pH: pH will be determined by using a pH meter Determination of Total Soluble Solid : Brix meter was used to determine total soluble solids Determination of Alcohol content : The alcohol content was quantified by using distillation

10. 2 . Materials and Methods 10 Chemical Analy tical Methods Determination of Ascorbic acid: The ascorbic acid was quantified according to Suganyadevi P. et al. (2010)Iodine solution was prepared by mixing KIO 3 , KI, and 3M H 2 SO 4 with the ratio 1:0.054:6 (w:w:v), respectively. L-ascorbic acid was used as a calibration and the result will be expressed as mg of ascorbic acid per 100 ml of sample (mg AA/ 100 ml) Determination of Total Phenolic contents: The determination of total phenolic contents will be done by using a spectrophotometer at 760 nm wavelength (Ainsworth et al., 2007). Gallic acid will be used to build up a calibration curve and the results will be expressed as mg of gallic acid per 1 L of sample (mg GAE/L) Determination of Total Antioxidant Capacity (DPPH assay): The supernatant was prepared, which followed the method described by Mezadri et al . (2008). 0.1 ml of supernatant was mixed with 4.9 ml of 60 µM DPPH reagent in absolute ethanol. The tube was incubated at room temperature in the dark for 30 min before being measured by spectrophotometer at 517 nm wavelength. The 80% ethanol was used as a blank

11. 11 Microbiological analysis 2 . Materials and Methods Determination of total aerobic bacteria: Based on the method of Montville T.J. et al. (1978) with some modifications, acerola cider will be used to make serial dilution to 10 -4 by using buffer peptone water. After that, dilute solutions will be inoculated to prepare plate count agar which is autoclaved and poured into Petri dishes before by using the spreading technique. The inoculated dishes will be incubated at 37 o C for 24h. Then the incubated dishes will be screened first and only dishes with the number of colonies between 25 to 250 will be counted and analyzed Determination of yeast and molds: The procedure of determining the amount of yeast and mold was described by Thomas J.M. and others (2012) . The samples will be diluted with peptone water from dilution 10 -1 to 10 -4 . 1 ml of each diluted suspension will be spread on the autoclaved Dichloramphenicol Rose Bengal Agar (DRBC) dishes. These dishes will be incubated at 25 o C for 5 days. After that, the colonies of yeast and mold will be counted separately and the data will be used for analysis

12. 12 2 . Materials and Methods Sensory Evaluation Before being sensory evaluated, acerola cider samples were microbiological tested The investigated samples should meet the satisfactory level of microbiological standards for alcohol drink in QCVN 6-3:2010/BYT, which is less than 10 3 CFU/mL of sample The sensory evaluation was carried out to know the acceptability of 30 untrained panelists using a nine-point Hedonic scale . 15ml of each qualified sample was served in and labeled with a 3-digit code with Latin square order. Each panelist did the sensory evaluation in an individual booth under controlled humidity and temperature (25 C) to prevent error The hedonic scale is from extremely dislike to extremely like in terms of appearance, color, aroma, taste, overall acceptability

13. Table 5 . Changes in the physical parameters of Acerola fermented cider under different fermentation conditions. 3. Results and Discussion 13 Data are expressed as mean ± SD (n = 3). a, b, c: Same letters in the same column express that values in different fermentation temperature are not significantly different (Tukey comparison test, p >0.05). x, y: Same letters in the same column express that values in different fermentation duration are not significantly different (2 samples T-test, p >0.05). Effect of fermentation on physical properties

14. 14 3. Results and Discussion Effect of fermentation on bioactive compounds of acelora cider Table 6. Changes in bioactive compounds of acerola fermented cider under different fermentation temperature in 10 days

15. 15 3. Results and Discussion Effect of fermentation on bioactive compounds of acelora cider Table 7. Changes in ascorbic acid content of acerola fermented cider in 7 days and 10 days at 200C Data are expressed as mean ± standard deviation (n=3). x, y: The results within the same row with the same letter is not different significantly (2 samples T tests, p < 0.05).

16. 16 3. Results and Discussion Microbial analysis of acerola cider Table 8. Microbiological quality of the acerola cider at different fermentation temperature and duration

17. 17 3. Results and Discussion Effect of fermentation on sensory properties of acerola cider Figure 4 . Sensory evaluation scores of acerola cider in term of appearance, aroma, color, taste, and overall acceptability in 9‑point scale under different fermentation temperatures a,b:Bars with same letter are not significantly different (Tukey comparison test, p>0.05).

18. 18 3. Results and Discussion Effect of fermentation on sensory properties of acerola cider Figure 5 . Sensory evaluation scores of acerola cider in term of appearance, aroma, color, taste, and overall acceptability in 9 point scale under different fermentation durations a,b: Bars with the same letter are not significantly different. (Tukey comparison test, p>0.05)

19. 4. Conclusion 19 Different temperatures resulted in similar pH values and significantly affected the soluble solids and alcohol content. More over , lower temperatures led to high alcohol content and higher in vitamin C and total phenolic compounds, probably due to growth rate of the yeasts . Longer fermentation (10days) may lead to the decreases of Vitamin C and TPC. Fermentation temperature s and durations may not significantly affect the sensory attributes such as taste or aroma hence the overall acceptability of ciders did not vary . 20°C and 7 days was considered to be the most suitable for acerola cider fermentation .

20. References Alonso-Salces, R.M .; Barranco, A.; Abad , B.; Berrueta , L.A .; Gallo, B.; Vicente, F. Polyphenolic profiles of basque cider apples cultivars and their technological properties . J. Agric. Food Chem. 2004, 52, 2938–2952 and liquors. Journal of Agricultural and Food Chemistry, 46, 25–31. Babushahi-Kouhestani , M.; Salehi , M.; Mazloomi , S.M .; Almasi-Hashyani , A. Quantitative evaluation of vitamin C in industrial lemon juice by titration method. J. Biol. Today’s World 2014, 3 , 139–141. Bangzhu Peng, Fuling Li, Lu Cui, and Yaodong Guo , 2015. Effects of Fermentation Temperature on Key Aroma Compounds and Sensory Properties of Apple Wine . Journal of food science, 2015. Beltran G, Novo M, Guillamon JM, Mas A, Roses N. 2008. Effect of fermentation temperature and culture media on the yeast lipid composition and wine volatile compounds . Int J Food Microbiol 121:169-77. Czyzowska , A.; Pogorzelski , E. Changes to polyphenols in the process of production of must and wines from blackcurrants and cherries . Part, I. Total polyphenols and phenolic acids. Eur. Food Res. Technol. 2002. Davidović , S. M., Veljović , M. S., Pantelić , M. M., Baošić , R. M., Natić , M. M., Dabić , D. Č., Pecić , S. P., & Vukosavljević , P. V. (2013). Physicochemical, Antioxidant and Sensory Properties of Peach Wine Made from Redhaven Cultivar. Journal of Agricultural and Food Chemistry, 61(6), 1357–1363. De Rosso, V.V.; Mercadante , A.Z . The High ascorbic acid content is the main cause of the low stability of anthocyanin extracts from acerola . Food Chem. 2007, 103, 935–943. Ginjom , I.; D’Arcy, B.; Caffin , N.; Gidley, M. Phenolic compound profiles in selected Queensland red wines at all stages of the wine-making process. Food Chem. 2011, 125, 823–834.

21. References Grosso, G., Bei , R., Mistretta , A., Marventano , S., Calabrese, G., Masuelli , L., ... & Gazzolo , D. (2013). Effects of vitamin C on health: A review of evidence . Front Biosci (Landmark Ed), 18(3), 1017-1029. Guyot , S.; Marnet , N.; Sanoner , P.; Drilleau , J.F . Variability of the polyphenolic composition of cider apple ( Malus domestica ) fruits and juices. J. Agric. Food Chem. 2003, 51, 6240–6247. Heinonen , Lehtonen , & Hopia . (1998). Antioxidant activity of berry and fruit wine J. Frias, L.M . Miranda, R. Doblado , C. Vidal- Valverde . Effect of germination and fermentation on the antioxidant vitamin content and antioxidant capacity of Lupinus albus L.var . Food Chemistry, 92 (2005), pp. 211-220. Jian-Ping Yuan and Feng Chen. Degradation of Ascorbic Acid in Aqueous Solution J. Agric. Food Chem. 1998, 46, 12, 5078–5082. Karthikeyan Venkatachalam , Chukwan Techakanon , and Suraphon Thitithanakul , Impact of the Ripening Stage of Wax Apples on Chemical Profiles of Juice and Cider ACS Omega 2018 3 (6), 6710-6718. Kiczorowski , P., Kiczorowska , B., Samolińska , W. et al. Effect of fermentation of chosen vegetables on the nutrient, mineral, and biocomponent profile in human and animal nutrition . Sci Rep 12, 13422 (2022). Lamuela‐Raventós , R. M. (2018). Folin-Ciocalteu method for the measurement of total phenolic content and antioxidant capacity. Measurement of Antioxidant Activity & Capacity: Recent Trends and Applications, 1st edition , 107–115. DOI : 10.1002/9781119135388. Le, T. T., & Kha , C. T. (2018). A study on processing conditions of fermented acerola juice. The Journal of Agriculture and Development , 17(5), 123-126 (in Vietnamese). Lima, V.L.A.G ., Me´lo , E.A ., Maciel , M.I.S ., Prazeres , F.G ., Musser, R.S ., Lima, D.E.S,2005 . Total phenolic and carotenoid contents in acerola genotypes harvested at three ripening stages . Food Chemistry 90, 565–568. Marques, L.G ., Ferreira, M.C., Freire, J.T ., 2007. Freeze-drying of acerola ( Malpighia glabra L.). Chemical Engineering and Processing 46, 451–457.

22. References Mezadri , T., Ferna´ndez-Pacho ´ n, M.S., Villano , D., Garcı´a-Parrilla , M.C., Troncoso , A.M., 2006. The acerola frut composition, productive characteristics and economic importance. Archivos Latinoamericanos de Nutricio ´ n 56, 101–109. Mezadri , T., Perez-Galvez, A., Hornero -Mendez, D., 2005. Carotenoid pigments in acerola fruits ( Malpighia emarginata DC.) and derived products. European FoodResearch Technology 220, 63–69. Mezadri , T., Villaño , D., Fernández-Pachón , M. S., García-Parrilla , M. C., & Troncoso , A. M.13 (2008). Antioxidant compounds and antioxidant activity in acerola ( Malpighia emarginata DC.) fruits and derivatives. Journal of Food Composition and analysis, 21(4), 282-290. Minatel IO, Borges, C. V. , Ferreira, M. I. , Gomez, H. A. G. , Lima, C. O. C. a. P. P. . Phenolic Compounds: Functional Properties, Impact of Processing and Bioavailability. Ministry of Health (2010) National technical regulation for alcoholic beverages. Link:(last entry: 06/02/2022) Montville T.J., Matthews K.R ., and Kniel K.E . (2012). Yeast-Based and Other Fermentations. Food Microbiology – an introduction (Third edition), 20, 282 – 297. Peng, Bangzhu ; Li, Fuling ; Cui, Lu; Guo , Yaodong (2015). Effects of Fermentation Temperature on Key Aroma Compounds and Sensory Properties of Apple Wine. Journal of Food Science, 80(12), S2937 – S2943 . Doi:10.1111 /1750-3841.13111 R. Doblado , H. Zielinski, M. Piskula , H. Kozlowska , R. Muñoz, J. Frías , C. Vidal- Valverde . Effect of processing on the antioxidant capacity of Vigna sinensis var. carilla . Journal of Agricultural and Food Chemistry, 53 (2005), pp. 1215-1222. Rasouli , H., Farzaei , M. H., & Khodarahmi , R. (2017). Polyphenols and their benefits: A review. International Journal of Food Properties, 20, 1700-1741. Ribeiro, B. S., & de Freitas, S. T. (2020). Maturity stage at harvest and storage temperature to maintain postharvest quality of acerola fruit. Scientia Horticulturae , 260.

23. References Rufino , M. S. M.; Alves, R. E.; Brito, E. S.; Pérez-Jiménez, J.; Saura-Calixto , F.; Mancini- Filho , J.; Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil. Food Chemistry 2010, 121, 996. Ruiz- Torralba , A., Guerra-Hernández, E. J., & García -Villanova, B. (2018). Antioxidant capacity, polyphenol content and contribution to dietary intake of 52 fruits sold in Spain. CyTA -Journal of Food, 16(1), 1131-1138. S.B . Hande , K. Nuray , K. Feryal . Degradation of vitamin C in citrus juice concentrates during storage. J. Food Eng. (2006). Safti´c Martinovi´c , L.; Birkic , N.; Mileti´c , V.; Antolovi´c , R.; Štanfel , D.; Wittine , K. Antioxidant Activity, Stability in Aqueous Medium and Molecular Docking/Dynamics Study of 6-Amino- and N -Methyl-6-amino-L-ascorbic Acid. Int. J. Mol. Sci. 2023 , 24 , 1410. Steinkraus KH . 2002. Fermented foods, feeds, and beverages. Biotech Adv. 4:219-243. Ulrich, R. Organic Acids. In The Biochemistry of Fruits and their Products; Hulme , A.C ., Ed.; Academic Press: New York, 1970; pp 89–118. Vendramini , A. L., & Trugo , L. C. (2000). Chemical composition of acerola fruit ( Malpighia Punicifolia L. ) at three stages of maturity. Food chemistry, 71(2), 195-198.

24. Thank you Thank you