(Đồ án hcmute) soft cheese production using citric acid and rennet as coagulants

MINISTRY OF EDUCATION AND TRAINING HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY FOR HIGH QUALITY TRAINING CAPSTONE PROJECT FOOD TECHNOLOGY SOFT CHEESE PRODUCTION USING CITRIC ACID AND RENNET AS COAGULANTS LECTURER: PhD PHAM THI HOAN STUDENT: PHAN NGUYEN NGOC KHANH LE THI NGOC YEN SKL 0 4 Ho Chi Minh City, December, 2021 n HO CHI MINH UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY OF HIGH QUALITY TRAINING GRADUATION THESIS CODE: 2021-17116013 SOFT CHEESE PRODUCTION USING CITRIC ACID AND RENNET AS COAGULANTS Advisor: PhD PHAM THI HOAN Student: PHAN NGUYEN NGOC KHANH LE THI NGOC YEN HO CHI MINH CITY - 12/2021 i n Student ID: 17116013 17116045 ii n ACKNOWLEDGEMENTS Firstly, we would like to express our heartfelt gratitude to all of the lectures at Ho Chi Minh City University of Technology and Education in general and lecturers at the Faculty of Chemical and Food Technology especially We gained valuable knowledge and skills regarding Food Technology throughout our four years at the school We would like to express our appreciation to PhD Pham Thi Hoan - lecturer at the Department of Food Technology, Faculty of Chemical and Food Technology, Ho Chi Minh City University of Technology and Education She was always enthusiastic in her support and teaching us, mentoring and imparting knowledge and experiences from the idea to the completed graduation thesis We are truly grateful for that, and we will keep her teaching in mind We would like to show our thanks to Ms Ho Thi Thu Trang, consultant in charge of the Department of Food Technology laboratories and workshops She had been supportive and helpful with tools, machinery and equipment during our experimental research process We also would like to express our deep sense of gratitude to the instructors in charge of Food Technology workshop 1, workshop 3, Microbiology laboratory, Biochemistry laboratory and Sensory laboratory who created an ideal environment for us to complete this project We are very grateful to everyone who made my experience at Ho Chi Minh University of Technology and Education very positive and enjoyable; this experience would not have been possible without my friends, family, fellow students and mentors This project has been one of the most challenging things we have done, but also one of the most enjoyable and rewarding It is impossible to prevent faults while implementing the thesis, so we look forward to hearing your feedback on our thesis Once again, we gratefully acknowledge you and wish you all the best Best regards! iii n iv n v n vi n vii n viii n ix n 7.2 137.3333 21.19748 12.23837 84.6759 189.9908 118.00 160.00 14.1 212.5000 24.87971 14.36431 150.6954 274.3046 185.50 234.50 14.2 146.6667 25.64339 14.80522 82.9650 210.3684 128.50 176.00 21.1 290.6667 40.42689 23.34048 190.2407 391.0926 244.00 315.00 21.2 160.3333 29.24181 16.88277 87.6927 232.9740 128.50 186.00 28.1 349.3333 32.52051 18.77572 268.5479 430.1187 322.50 385.50 28.2 261.0000 40.00937 23.09942 161.6112 360.3888 221.50 301.50 Total 30 212.3000 74.57010 13.61458 184.4551 240.1449 88.50 385.50 1.1 0133 00577 00333 -.0010 0277 01 02 1.2 0300 02646 01528 -.0357 0957 01 06 7.1 0167 01155 00667 -.0120 0454 01 03 7.2 0400 03464 02000 -.0461 1261 02 08 14.1 0333 02082 01202 -.0184 0850 01 05 Adhesiveness 14.2 0467 03786 02186 -.0474 1407 02 09 21.1 0333 00577 00333 0190 0477 03 04 21.2 0367 02887 01667 -.0350 1084 02 07 28.1 0600 01000 00577 0352 0848 05 07 28.2 0467 00577 00333 0323 0610 04 05 Total 30 0357 02285 00417 0271 0442 01 09 Springiness Gumminess 1.1 2.5133 03786 02186 2.4193 2.6074 2.47 2.54 1.2 2.6867 07024 04055 2.5122 2.8611 2.62 2.76 7.1 2.6500 03000 01732 2.5755 2.7245 2.62 2.68 7.2 2.6467 08505 04910 2.4354 2.8579 2.56 2.73 14.1 2.5567 17898 10333 2.1121 3.0013 2.35 2.66 14.2 2.6233 13317 07688 2.2925 2.9541 2.51 2.77 21.1 2.8200 01732 01000 2.7770 2.8630 2.81 2.84 21.2 2.7100 03000 01732 2.6355 2.7845 2.68 2.74 28.1 2.7967 01155 00667 2.7680 2.8254 2.79 2.81 28.2 2.7300 07211 04163 2.5509 2.9091 2.65 2.79 Total 30 2.6733 11669 02130 2.6298 2.7169 2.35 2.84 1.1 139.0000 25.81240 14.90280 74.8784 203.1216 109.20 154.40 1.2 101.8300 37.54697 21.67775 8.5582 195.1018 69.69 143.10 7.1 167.4000 34.06861 19.66952 82.7689 252.0311 134.00 202.10 7.2 111.6333 17.11179 9.87950 69.1253 154.1414 94.90 129.10 14.1 164.1000 28.81996 16.63921 92.5073 235.6927 142.40 196.80 14.2 123.2000 27.81438 16.05864 54.1052 192.2948 103.40 155.00 21.1 255.5000 32.93084 19.01263 173.6953 337.3047 217.50 275.70 21.2 129.4333 24.14650 13.94099 69.4501 189.4166 103.90 151.90 28.1 303.7000 27.44431 15.84498 235.5246 371.8754 281.00 334.20 28.2 231.0667 17.41273 10.05325 187.8110 274.3223 212.30 246.70 Total 30 172.6863 69.70897 12.72706 146.6566 198.7161 69.69 334.20 n Chewiness 1.1 4.1833 37448 21620 3.2531 5.1136 3.78 4.52 1.2 2.6933 1.05548 60938 0714 5.3153 1.83 3.87 7.1 4.3533 90512 52257 2.1049 6.6018 3.44 5.25 7.2 2.9067 54049 31205 1.5640 4.2493 2.38 3.46 14.1 4.4167 85237 49212 2.2993 6.5341 3.53 5.23 14.2 2.5267 37072 21404 1.6057 3.4476 2.11 2.82 21.1 7.0667 93243 53834 4.7504 9.3830 5.99 7.61 21.2 3.4433 67826 39159 1.7584 5.1282 2.73 4.08 28.1 8.3267 74097 42780 6.4860 10.1673 7.69 9.14 28.2 5.2667 1.61630 93317 1.2515 9.2818 3.49 6.65 Total 30 4.5183 1.98020 36153 3.7789 5.2578 1.83 9.14 Test of Homogeneity of Variances Levene Statistic df1 df2 Sig L 1.114 20 398 a 2.520 20 041 b 4.658 20 002 893 20 548 Adhesiveness 4.403 20 003 Springiness 4.156 20 004 Gumminess 526 20 839 Chewiness 1.373 20 264 Hardness ANOVA Sum of Squares Between Groups L 22.899 61.094 20 3.055 267.183 29 1.760 196 358 20 018 2.118 29 Between Groups 96.447 10.716 Within Groups 22.520 20 1.126 118.967 29 141787.467 15754.163 19472.833 20 973.642 161260.300 29 Between Groups 005 001 Within Groups 010 20 000 Within Groups Between Groups Within Groups Total b Total Between Groups Hardness Within Groups Total Adhesiveness Mean Square 206.089 Total a df n F Sig 7.496 000 10.920 000 9.517 000 16.181 000 1.187 355 Springiness Total 015 29 Between Groups 253 028 Within Groups 142 20 007 Total 395 29 125205.585 13911.732 15715.309 20 785.765 140920.894 29 Between Groups 98.273 10.919 Within Groups 15.441 20 772 113.714 29 Between Groups Gumminess Within Groups Total Chewiness Total L Date N Subset for alpha = 0.05 Tukey HSDa 14.1 82.1233 7.1 82.8633 1.1 84.3600 21.1 85.6967 85.6967 1.2 86.3767 86.3767 86.3767 14.2 86.7567 86.7567 86.7567 21.2 87.1100 87.1100 87.1100 28.1 87.1433 87.1433 87.1433 28.2 89.9600 89.9600 7.2 90.8100 Sig .052 145 117 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 a Date N Subset for alpha = 0.05 Tukey HSDa 28.1 -1.5700 14.2 -1.5633 7.2 -1.5067 -1.5067 1.2 -1.3867 -1.3867 21.2 -1.3767 -1.3767 28.2 -1.3533 -1.3533 21.1 -1.2767 -1.2767 -1.2767 7.1 -1.1633 -1.1633 n -1.1633 3.975 005 17.705 000 14.143 000 1.1 14.1 -.9400 -.9400 -.8133 Sig .244 109 122 097 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 b Date N Subset for alpha = 0.05 Tukey HSDa 21.2 8.7333 1.2 9.3167 9.3167 28.2 10.3667 10.3667 10.3667 7.2 10.5300 10.5300 10.5300 14.2 11.1600 11.1600 11.1600 11.1600 14.1 11.5967 11.5967 11.5967 11.5967 11.5967 7.1 12.1200 12.1200 12.1200 12.1200 28.1 12.8167 12.8167 12.8167 1.1 14.0300 14.0300 21.1 14.4767 Sig .080 091 192 079 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 Hardness Date N Subset for alpha = 0.05 Tukey HSDa 1.2 127.3333 7.2 137.3333 137.3333 14.2 146.6667 146.6667 21.2 160.3333 160.3333 14.1 212.5000 212.5000 212.5000 1.1 214.6667 214.6667 214.6667 7.1 223.1667 223.1667 28.2 261.0000 261.0000 21.1 290.6667 290.6667 28.1 Sig 349.3333 063 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 n 071 125 058 077 Adhesiveness Date N Subset for alpha = 0.05 Tukey HSDa 1.1 0133 21.1 0133 7.1 0167 0167 21.2 0167 0167 14.1 0267 0267 1.2 0300 0300 7.2 0400 0400 14.2 1767 Sig .999 054 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 Springiness Date N Subset for alpha = 0.05 Tukey HSDa 1.1 2.5133 14.1 2.5567 2.5567 14.2 2.6233 2.6233 2.6233 7.2 2.6467 2.6467 2.6467 7.1 2.6500 2.6500 2.6500 1.2 2.6867 2.6867 2.6867 21.2 2.7100 2.7100 2.7100 28.2 2.7300 2.7300 2.7300 28.1 2.7967 2.7967 21.1 Sig 2.8200 107 Means for groups in homogeneous subsets are displayed n 055 181 a Uses Harmonic Mean Sample Size = 3.000 Descriptives N Mean Std Deviation Std Error 95% Confidence Interval for Minimum Maximum Mean Lower Bound Upper Bound 1.1 8.8700 23065 13317 8.2970 9.4430 8.61 9.05 7.1 9.7200 47508 27429 8.5398 10.9002 9.25 10.20 14.1 9.2667 25166 14530 8.6415 9.8918 9.00 9.50 21.1 8.3333 25166 14530 7.7082 8.9585 8.10 8.60 28.1 8.1700 30447 17578 7.4137 8.9263 7.90 8.50 Total 15 8.8720 65302 16861 8.5104 9.2336 7.90 10.20 1.1 2767 19425 11215 -.2059 7592 11 49 7.1 6633 39879 23024 -.3273 1.6540 30 1.09 14.1 2.0067 42852 24741 9422 3.0712 1.53 2.36 21.1 2.3567 70501 40704 6053 4.1080 1.65 3.06 28.1 3.0867 54721 31593 1.7273 4.4460 2.46 3.47 Total 15 1.6780 1.16473 30073 1.0330 2.3230 11 3.47 Acidity Syneresis ANOVA Sum of Squares Between Groups Acidity Within Groups Total Between Groups Syneresis Within Groups Total df Mean Square 4.974 1.243 997 10 100 5.970 14 16.639 4.160 2.354 10 235 18.992 14 Acidity Date N Subset for alpha = 0.05 Tukey HSDa 28.1 8.1700 21.1 8.3333 1.1 8.8700 14.1 7.1 Sig 8.8700 9.2667 9.2667 9.7200 121 Means for groups in homogeneous subsets are displayed n 563 445 F Sig 12.477 001 17.673 000 a Uses Harmonic Mean Sample Size = 3.000 Syneresis Date N Subset for alpha = 0.05 Tukey HSDa 1.1 2767 7.1 6633 14.1 2.0067 21.1 2.3567 28.1 3.0867 Sig .860 119 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 Descriptives N Mean Std Std Deviation Error 95% Confidence Interval for Minimum Maximum Mean Lower Bound Upper Bound 1.2 17.4833 24685 14252 16.8701 18.0965 17.21 17.69 7.2 18.0300 49031 28308 16.8120 19.2480 17.53 18.51 14.2 17.5333 35119 20276 16.6609 18.4057 17.20 17.90 21.2 16.7167 23629 13642 16.1297 17.3036 16.45 16.90 28.2 12.9333 30551 17638 12.1744 13.6922 12.60 13.20 Total 15 16.5393 1.93751 50026 15.4664 17.6123 12.60 18.51 Acidity 1.2 1.2900 79505 45902 -.6850 3.2650 73 2.20 7.2 1.7133 90561 52286 -.5363 3.9630 92 2.70 14.2 2.7700 85504 49366 6460 4.8940 1.91 3.62 21.2 3.9433 93168 53791 1.6289 6.2578 3.24 5.00 28.2 4.2433 86031 49670 2.1062 6.3805 3.47 5.17 Total 15 2.7920 1.41772 36605 2.0069 3.5771 73 5.17 Syneresis n Test of Homogeneity of Variances Levene Statistic df1 df2 Sig Acidity 409 10 799 Syneresis 049 10 995 Acidity Date N Subset for alpha = 0.05 28.2 12.9333 21.2 16.7167 1.2 17.4833 17.4833 14.2 17.5333 17.5333 7.2 Tukey HSDa Sig 18.0300 1.000 084 342 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 Date Tukey HSDa Syneresis N Subset for alpha = 0.05 1.2 1.2900 7.2 1.7133 1.7133 14.2 2.7700 2.7700 2.7700 21.2 3.9433 3.9433 28.2 Sig 4.2433 298 Means for groups in homogeneous subsets are displayed a Uses Harmonic Mean Sample Size = 3.000 n 064 302 APPENDIX 11: CHEMICAL COMPOSITONS DETERMINATION METHODS Total solid content The total solid content in soft cheese is determined according to TCVN 8174: 2009 (ISO 5534: 2004) Total solid content is the mass fraction of substances remaining after the end of the drying process as specified in TCVN 8174: 2009 Water in the tested moisture is allowed to evaporate in the presence of zinc oxide in an oven at 102 ℃ ± ℃ The total solid of the tested sample was conducted as follows The tested sample was brought to 20 - 25 ℃ and stirred thoroughly with a spatula The petri dish was opened and contained approximately grams of zinc oxide, with the lid and stirring rod placed on the lid in an oven at 102 ℃ for at least hour The dish was covered with a stirring rod on the top The covered dish and the rod were immediately transferred to a desiccator to avoid moisture reabsorption The prepared dish with lid and the rod were weighted and the dish was tilted to push zinc oxide to one side of it gram of the tested sample was placed in the remaining space of the dish The dish containing the sample was covered and weighed with the rod together mL of water was added to the dish and the rod was used to thoroughly mix the diluted sample and zinc oxide The mixture was evenly speeded on the dish bottom The dish was heated in a water bath for 30 minutes and the mixture was frequently stirred during the initial heating so that the liquid evaporates as much as possible After that, the dish was wiped to remove the steam in the dish The rod was placed in the dish and the dish with its side lid was dried in the oven until the mixture gained a constant mass Total solid W of the test sample, as a percentage by mass, is calculated by the following formula 𝒎 −𝒎 W = 𝒎𝟐 −𝒎𝟎 × 𝟏𝟎𝟎 + (𝟎, 𝟏 ∗ 𝒂) 𝟏 𝟎 With m0 is the weight, in grams, of the dish (including zinc oxide), with the lid and the stirring rod (g); m1 is the weight of the dish (including zinc oxide), together with the lid and the rod and the test sample (g); m2 is the weight of the dish, together with the lid, the stirring rod and dried test sample (including zinc oxide) (g); n a is the weight of titrated lactic acid obtained per 100 g of sample, determined according to ISO 11869 (g) 0.1 is the compensation value for water loss resulting from acid neutralization of cheese with zinc oxide Protein content Protein content was determined by the Kjeldahl method according to TCVN 8099- 1:2015 (ISO 8968 1:2014) The Kjeldahl method involves three steps including digestion, neutralization and titration In the Kjeldahl digestion step, protein and other organic components in a sample were digested with sulfuric acid and potassium sulfate Potassium sulfate was used to raise the boiling point of sulfuric acid to accelerate digestion Copper sulfate was also used as a catalyst to convert organic nitrogen into ammonium sulfate The digest was diluted with water Excess sodium hydroxide was added to neutralize sulfuric acid The ammonia formed was distilled into a boric acid solution Borate anion (proportional to the amount of nitrogen) was titrated with standardized hydrochloric acid The content (Carlson et al.) of total nitrogen in the sample is determined according to the following formula: 𝑵= (𝒂 − 𝒃𝑲) × 𝟎, 𝟎𝟎𝟏𝟒 × 𝑽 × 𝟏𝟎𝟎 𝒗×𝒎 With N is nitrogen content, in mass percent (Carlson et al.) a is the standard 0.1N H2SO4 volume to absorb NH3, mL b is 0.1 N NaOH volume to titrate, mL m is the tested sample weigh, g V is the total volume to digest solution, 100 mL v is digested solution volume used for distillation, 10 mL 0.0014- the amount of nitrogen (g) equivalent to mL of 0.1N H2SO4 K - correction factor for the concentration of NaOH 0.1 N The crude protein content of the test sample was determined according to the following formula: n 𝑾𝒑 = 𝑵 × 𝟔, 𝟑𝟖 With, Wp is crude protein content, in mass percentage (Carlson et al.) N is the nitrogen content of the cheese sample, in mass percentage (Carlson et al.) 6.38is the factor to convert nitrogen content to crude protein content Fat analysis • Fat content of recombined milk The lipid content was determined by the Mojonnier method according to TCVN 7084:2010 (ISO 1736:2008) The Mojonnier was developed for and applied mainly to dairy products In this method, the used organic solvents used include ammonia, alcohol 95%, diethyl ether and petroleum ether Ammonia and alcohol are used to break protein membranes, precipitate proteins and separate proteins from fatty globules Diethyl ether and petroleum ether are added to extract polar and nonpolar fats, respectively The mixture after adding solvent and stirring will separate into two phases: the light upper phase consists of solvent and fat, the lower heavy phase consists of precipitated protein, alcohol, NH3 and other components remaining in the milk The light phase is collected and dried to a constant weight, obtaining the total fat content in the milk powder sample The fat content of recombined milk was calculated as follows Milk powder was recombined with 15% of the total solid content 10 mL recombined milk was taken in an erlen-25 mL 1.5 mL ammonia and 10 mL of 95% alcohol were added and shake vigorously in 90 seconds Ammonia neutralized the acidic sample and dissolved protein, while the alcohol prevented possible gel formation After that, 25mL diethyl ether was added and shake gently for 90 seconds 25mL petroleum was subsequently added and shook gently for 90 seconds The diethyl ether dissolved the lipid, while the petroleum ether removed moisture from the diethyl ether extract and dissolved more non-polar lipid The erlen was rinsed with petroleum ether several times to extract the remaining fat on its wall The solution in the erlen was transferred to the separating funnel and the funnel was stood for 30 minutes to allow the solvents to separate After that, the light phase was collected to the petri dish The solvent in the dish was evaporated on the electric hot stove at ≤ 100 ± 1℃ until the solvent was almost completely evaporated The dish and fat were nextly dried to a constant n weight in a drying oven at 102 ± ℃ The dish was cooled to room temperature in the desiccator and weigh The experiment was triplicated Fat content was determined according to the formula: W= (𝒎𝟏 −𝒎𝟐 )−(𝒎𝟑 −𝒎𝟒 ) 𝒎𝟎 × 𝟏𝟎𝟎 With m0 is the sample weight (g) m1 is the weight flask containing lipid and extracts (g) m2 is the weight flask used to contain lipid (g) m3 is the weight flask of the blank test containing extracts (g) m4 is the weight flask used in the blank test (g) • Fat content of cheese The fat content in cheese was determined according to TCVN 8181:2009 (ISO 1735:2004) The sample is digested with hydrochloric acid followed by the addition of ethanol The acid-ethanol solution is extracted with diethyl ether and the pentane solvents are removed by distillation or evaporation The extract is weighed This principle is usually called the Schmid-Bondzynski-Ratzlaff principle The fat content of cheese was determined by the following steps grams of cheese sample (with a precision of mg) was diluted with 10 mL of hydrochloric acid in a 250-mL erlen The erlen was heated in a water bath until the sample was completely dissolved The erlen was subsequently boiled on the stove for about 10 minutes and cooled under tap water The sample mixture was transferred from the erlen to a separating funnel and the erlen was rinsed with 10 mL of ethanol and shook well for 30 seconds The erlen was continuously rinsed with diethyl ether; the top of the separating funnel was also rinsed with diethyl ether and shook well for 90 seconds The erlen was finally rinsed with 25 mL of pentane; the top of the funnel was also rinsed with pentane The funnel was stood for 30 minutes to allow the solvents to separate After that, the light phase was collected to the petri dish The solvent in the dish was evaporated on the electric hot stove at ≤ 100 ± 1℃ until the solvent was almost completely evaporated The dish and fat were subsequently dried to a constant weight in a n drying oven at 102 ± ℃ The dish was cooled to room temperature in the desiccator and weigh The experiment was triplicated The lipid content in cheese is determined W= (𝒎𝟏 −𝒎𝟐 )−(𝒎𝟑 −𝒎𝟒 ) 𝒎𝟎 × 𝟏𝟎𝟎 With, m0 is the sample weight (g) m1 is the weight flask containing lipid and extracts (g) m2 is the weight flask used to collected lipid (g) m3 is the weight flask containing extracts in the blank test (g) m4 is the weight flask used in the blank test (g) n APPENDIX 12: TESTED SAMPLE AND CONTROL SAMPLE COMPARISION Sample Cheese yield Total solid in %, w/w Moisture CSY cheese content %, w/w %, w/w %, w/w Cheese hardness, g CS 18.83 ± 0.32a 29.67 ± 0.67b 70.33 ± 0.67a 37.26 ± 1.46a 195.33 ± 2.93a RS 20.74 ± 0.20bc 36.51 ± 0.24ab 63.49 ± 0.34b 360.67 ±12.64c 50.46 ±0.46b The different letters (a-d) in each column showed significant difference (p < 0.05) *RS: rennet – coagulated cheese (tested sample) and CS: acid – coagulated cheese (control sample) n S n K L 0