MINISTRY OF EDUCATION AND TRAINING NONG LAM UNIVERSITY-HO CHI MINH CITY FACULTY OF BIOLOGICAL SCIENCES RESEARCH ON THE PRODUCTION OF CHOCOLATE SYNBIOTIC CANDY Major : BIOTECHNOLOGY Student : DUONG YEN VY Student ID : 19126264 Academic year : 2019-2023 Thu Duc City, 08/2023 MINISTRY OF EDUCATION AND TRAINING NONG LAM UNIVERSITY-HO CHI MINH CITY FACULTY OF BIOLOGICAL SCIENCES UNDERGRADUATE THESIS RESEARCH ON THE PRODUCTION OF CHOCOLATE SYNBIOTIC CANDY Instructor Student CAO THI LOAN THANH. PHD DUONG YEN VY Thu Duc City, 08/2023 THANK YOU I would like to thank the Board of Directors, Faculty of Biological Sciences, Nong Lam University, Ho Chi Minh City, for creating conditions for me to study here. I would like to thank the Research Institute of Biotechnology and Environment for creating favorable conditions for facilities and equipment for me to carry out my graduation thesis. I would like to express my sincere gratitude to Ms.
Cao Thi Thanh Loan, who gave me direction, enthusiastically guided me, and supported me to complete my thesis. I would like to thank Mr. Vo Tran Quoc Thang, Ms. Nguyen Thi Van Anh, and Mr.
Le Nguyen Thanh Dong for always supporting and encouraging me during the completion of my graduation thesis. I would like to thank my family for always supporting, encouraging, and creating the best conditions for me to study and practice. Thank you sincerely. AFFIRMATION AND COMMITMENT My name is Duong Yen Vy, student ID: 19126264, class: DH19SHC (Phone: 0918664797, email: 19126264@st.vn), of Biotechnology, Nong Lam University Ho Chi Minh City.
This is my graduate thesis which was conducted by myself. All the data and information are completely honest and objective. I take full responsibility in front of the committee for these commitments. Thu Due City, July 31st, 2023 Student’s signature il ABSTRACT Recent studies have demonstrated the advantages of consuming probiotics and prebiotics, and many products containing probiotics and fibers have been introduced to the market.
This study's objective is to develop a synbiotic chocolate candy as a healthy promoting food. In this study, the commercial probiotic was combined with cacao powder, chocolate, and two different types of sugars (sucrose and isomalt) to make synbiotic chocolate candy. The sensory evaluation of candies was carried out. In addition, the probiotic density of the synbiotic candy was determined under different storage temperatures for 30 days.
The results revealed that the chocolate candy made with isomalt (21.95% point) was more favorable than the one with sucrose (19. The probiotic microorganisms in chocolate synbiotic candy demonstrated remarkable resilience to the challenging conditions of the artificial stomach and bile salts media which are 7. These findings are significant as they indicate the potential of Chocolate synbiotic candy to withstand harsh digestive conditions, ensuring the survival of probiotic microorganisms and the delivery of beneficial effects to consumers. Furthermore, the optimal storage temperature of 4°C is crucial for maintaining the quality and efficacy of the product throughout its shelf life.
This study contributes valuable insights into the development of synbiotic candies as functional foods, promoting digestive health and overall well- being. Keywords: synbiotic, chocolate, Isomalt, Sucrose, stomach, bile salts, probiotics, lactic acid bacteria. 1H TÓM TẮT Các nghiên cứu gần đây đã chứng minh những lợi ích của việc tiêu thụ men vi sinh và prebiotic, và nhiều sản phẩm có chứa men vi sinh và chất xơ đã được tung ra thị trường. Mục tiêu của nghiên cứu này là phát triển một loại kẹo sô cô la synbiotic như một loại thực phâm tăng cường sức khỏe.
Trong nghiên cứu này, lợi khuân thương mại được kết hợp với bột ca cao, sô cô la và hai loại đường khác nhau (sucrose và isomalt) dé tạo ra kẹo sô cô la tổng hợp. Keo được đánh giá cảm quan và khả năng sống sót của các vi sinh vật lợi khuẩn được đánh giá trong môi trường đường tiêu hóa nhân tạo sử dụng môi trường dạ dày và muối mật. Ngoài ra, mật độ lợi khuẩn của kẹo tổng hợp được xác định ở các nhiệt độ bảo quản khác nhau trong 30 ngày. Kết quả cho thấy kẹo socola làm bằng isomalt (21,95% điểm) được ưa chuộng hơn kẹo làm bằng sucrose (19,85% điểm).
Các vi sinh vật lợi khuẩn trong kẹo tổng hợp sô cô la đã chứng minh khả năng phục hồi đáng ké đối với các điều kiện khắc nghiệt của dạ dày nhân tạo và môi trường muối mật lần lượt là 7,41 (log CFU/g) và 7,66 (log CFU/g). Những phát hiện này rất có ý nghĩa vì chúng cho thấy tiềm năng của kẹo tổng hợp sô cô la có thê chịu được các điều kiện tiêu hóa khắc nghiệt, đảm bảo sự sống sót của các vi sinh vật có lợi và mang lại tác dụng có lợi cho người tiêu dùng. Hơn nữa, nhiệt độ bảo quản tối ưu là 4°C là rất quan trong dé duy tri chất lượng và hiệu quả của sản phẩm trong suốt thời hạn sử dụng. Nghiên cứu này đóng góp những hiểu biết có giá trị về sự phát triển của kẹo synbiotic đưới dạng thực phẩm chức năng, thúc đây sức khỏe tiêu hóa và sức khỏe tông thể.
Từ khóa: Synbiotic, sô cô la, Isomalt, Sucrose, da day, muối mật, lợi khuẩn, vi khuẩn lactic. 2 AFFIRMATION AND COMMITMENT. il AB LAI GG T son Si 510030250300920SS002TR011G2H-bSSHESEBS3nit8dg-glEEosibogtellies4RiiiL-0GG0018:345000E3138223)/3u8. iv G@ISWUE ES eee V LISE OF ABBRE VIA TIONS sung ngunioiniooiiddnggddcististsgbepifiobtibssdliub4biioi8ilibieriidi vill II509)v0.
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19 De) Die MATHS AVOT|binssseesosiiiistixiv1i1GES034138186850813843835186581365353X4890046XESSL4SGGI4S4580538 3836148800336 20 CHAPTER 3. MATERTALS & METHODS sát counavnnnanasonasveanavonves 22 3. time-of TP Ï6fiieHilaHOiliseeeeesseeseeinidbiietidt3i4S1i3055013131139383350398090130188004788 DD, Bids Material sessescsverosssesensasesnanesevccussenin yeeeuaraa arn eee eERREN 22 Sedo IES'PEfiHT6iil ÌSVDGEsaessssssasnnrsnndasiottbalttIGE458330503S5H-G. Determine the formula of chocolate synbiotic candy.
Determination of the total number of microorganism lactic acid bacteria in the CHOCOLATE CAH EY sueiestesortottoSBinSS0060TE59NBISG1089212J00Ợ8Đ3DĐSSESESDISSHIIES. Investigation of survival rates in the stomach and artificial bile salts. Evaluation of storage time of synbiotic candy. Implementation Methods wssecsscrsnssseesnenavsvemcananssvasessnemneresmeossenrvuecevsennanrereasessnees 26 3.
Quantification of total polyphenol confenf.42 MIRS preparation method sec 6010606156461011380/22250GE/001180020E06:G350000/0581230/8 88. Methods of sensory assessment of tastes. Survey on microbial content in artificial stomach environment and artificial bile SA tS cas ssvsessanaasescssssnaxs gE33Ä.0314054538013833853ELEESSBEKENESSL-IEIHESIGIGI2BEE363i0gi-ESSEEGEE01G1381388028033Q8300340038 2] 3. Method for determining resistance to gastric acid production (SGPF).
Determination of viability in bile salt medium (STIF). Identification of microorganisms in samples by biochemical test. Identification of microorganisms in samples by biochemical test. Determine the formula of chocolate synbiotic canidy,.
Survey on microbial content 1n artificial stomach environment and artificial bile KốI|EEbcoarsstrgEcoixgietsitiargpsptosgtoyisbsbituiiEdUeolbglS6gSg-23W2g509:80i/50.00200830:15/8881/gi0/0M8H:i3u/d9đ0cóiEagttSuiöngS/E8u8i2786E:5:0uđi80 37 4. Investigate the viability of bacteria in the artificial stomach environment. Investigate the ability of bacteria to survive 1n the bile salt environment. Evaluation of storage time of synbiotic ©añndy.
CONCLUSION AND SUGGESTIONS. -----cc-cccccereerree 48 hh®U0 Sẽ. 48 REEEREBNE scoescosszaesssezescxotEtcggtisptsgseGooogstgieibtindibrlitapidb3LfpgBsi2liSiGpisptsste28sasiszegssi 49 [35019902 .ÔỎ VI LIST OF ABBREVIATIONS FAO _ : Agriculture Organization of the United Nations LAB _ : Lactic acid bacteria SCFAs : Short-chain fatty acids SGF — : Simulated Gastric Fluid SIF : Simulated Intestinal Fluid VTEC : Verocytotoxin WAO_: World Health Organization of the United Nations vill LIST OF TABLE Table 3. Ingredients Synbiotic Chocolate Candy.
Gallic acid calibration curve results .00000cccccccceeeeeeeeeeeeeseeccceeceeeeeeeeseneees 36 1X LIST OF FIGURES THƠIIEG 2. ACC BC sere ceacenercosnmeuans Ng8660300303168 260 303835GNSGSSBSBSUS8E83i070ãb48/3i1G83BSIE2DiBS38383/30 8 Figure 2:2. Prebioties tietabOliStit GHHÊ si sccsccensaasnsanssuavasaanssaesasneaconesnwamaneaanecenessin 12 Figure 3. Candy production process.
Gram-positive bacteria, spherical (x100). Identification of microorganisms in samples by biochemical test. Enumerating yeasts and molds in the chocolate samples that contain isomalt (a) and sucrose; (b) grown on DRBC medIum.- 5555 +>5-<>ss+sesseszezseezeezsee dL Figure 4. The input microbial LAB colony (at the dilution factor 10° in the chocolate samples that contain isomalt (b) and sucrose (c) grown on MRS medium with Dr G Bitiduys (A):aS:48 COMO), cer civtencceeesnsveteererecommrrcenmicvieseeannaneremearemsmeriemennemeres2 Figure 4.
LAB count in the chocolate samples that contain isomalt (a) and sucrose (b) grown on MRS medium with Dr G Bifidus as a controlL. Percentage of scores in chocolate samples containing isomalt (a) and sucrose (b) in the three structural, color, and taste parameters .- ---- +52 +++s=+sss+sssss2 34 Figure 4. Sensory evaluation survey chart based on a total score. Gallic acid standard curve .ceccescssceeceeeceeeeeeeeseeeeeseeseeeseereeseeseeeees dO Figure 4.
LAB density of Isomalt samples media after spreading in SGF. LAB density of Sucrose samples media after spreading in SGP. Viable LAB density of Isomalt-containing products in SGF artificial gastric medium during incubation after 2 hOUTs. Surviving microbial density of Sucrose-containing products in SGF artificial gastric medium during incubation.-- 5-5-5222 <+2*++£+s+zee+zezeesxs 39 Figure 4.
LAB density of Isomalt samples media after spreading in SIF. LAB density of Sucrose samples media after spreading in SÏIF. Survival density of LAB in the products containing Isomalt in SIF arti tieial bile Salt CUTTS T6UDSLDbsssesessssoaesditsoisdoilSEisdgSgGSE4G390083. Survival density of LAB in the products containing Sucrose in SIF artificial bile salt medium during incubation.
The survival rate of LAB of chocolate products during incubation in SIF Figure 4. Investigation of LAB density under storage conditions tì HSOTHAIV SANT LOS oars cox ssssssscanseocense crass 2038 1888610L13860SH08L386EB48-. Investigation of LAB density under storage conditions IN SUCTOSE SAM PIES verscsssseessesseresreseve eave wesu amen ener EE 45 Figure 4. LAB density of chocolate products during storage at 4°C and 25°C.
Problem statement Today, we seem to be too familiar with functional products that support human health, and prevent and can cure many diseases. Among them, which have a large effect and are also mentioned quite a lot, are products with probiotics added, such as Probi yogurt, Yakult, probiotic ice cream, and probiotic cheese. However, the concept of synbiotic chocolate candy is still new, probiotics are microorganisms that have beneficial effects on humans and animals, such as benefits for the gastrointestinal tract, strengthening of the immune system, and reduction of lactose allergy, reduce cancer. Prebiotics are substances that cannot be digested, but can be fermented by the intestinal microflora to bring benefits to the host body.