NOTE: Students are required to fill in the

followings clearly before submission.

Lecturer’s Name:

Dr. Pui

FACULTY OF APPLIED SCIENCES ..................................................................

DEPARTMENT OF FOOD SCIENCE & NUTRITION

Subject code & Name:

LABORATORY REPORT SUBMISSION

Experiment Title : Lab 3: Yogurt Making

Group :2

STUDENTS DECLARATION OF WORK

We declare that the work submitted is our own. We confirm that we have read and
understood the University regulations with regard to Plagiarism, Collusion and
Cheating in this work will be penalised.

No. Student ID Group Members Signature

1. 1002058607 Oo Wei Hong Wei Hong
2. 1002162512 Yong Zhi Wei Zhi Wei
3. 1002058088 Chuan Yik Shuan Yik Shuan
4. 1002162103 Shaggy Santosh Shaggy

LECTURER’S ASSESSMENT CRITERIA

Criteria Wtg. Mark

Awarded Percentage
Introduction & objective 1
Marks: Gained:
Materials & Methods 1
Results & calculations 2
Discussion & precaution steps 2
Conclusion
Questions
1
1 10 10
References 0.5
Format & cover page 0.5
Performance 1

Total 10
Lecturer’s Remarks/ Comments (if applicable):

1
Objective

To demonstrate how nutrient rich milk can be preserved by lactic acid produced by lactic
acid-producing microbial culture during fermentation.

Introduction

Yogurt is a popular dairy product that humans have enjoyed for thousands of years. Its roots
may be traced back to ancient Mesopotamia and India. Yogurt can be used in a wide variety
of ways. It is available in several forms and flavors, making it suitable for satisfying a variety
of preferences and tastes. The most common types of yogurts are plain, Greek, and flavored;
each has distinctive characteristics. Due to its mild sourness and smooth texture, plain yogurt
is a versatile ingredient for both sweet and savory dishes. Greek yoghurt, on the other hand,
undergoes a straining procedure that eliminates the whey. This results in a product that is
denser and creamier, with more protein and less lactose. Typically, flavoured yogurts are
sweetened and combined with extra ingredients, such as fruit, nuts, or other foods, in order to
create a delectable snack that is also easy to consume (Chandan, 2017).

Besides, the process of producing yogurt dates back thousands of years. There are several
physical forms of yogurt, including set, stirred or liquid (drinking yogurt), frozen, and dried
yogurt. In several nations, including the United States and Europe, yogurt culture (>10 6
cfu/mL) is required in yogurt recipes. A recent tendency has been to enhance the medicinal
benefit of yogurt by including probiotic organisms as adjunct cultures. Both cow and buffalo
milk are used to make yogurt. Milk is often prepared by standardizing it to satisfy the
minimum legal criteria established by local food regulatory bodies. The fermentation of milk
into yogurt is accomplished with lactic acid bacteria (LAB), most notably Lactobacillus
bulgaricus and Streptococcus thermophilus. These bacteria collaborate to convert lactose, the
sugar in milk, into lactic acid, responsible for yogurt's characteristic sour taste and thick
consistency (Pothuraju et al., 2018).

Moreover, yogurt has been held in high regard throughout history for the many nutritional
and physiological benefits it provides. It is an excellent source of protein, calcium, vitamins

2
B2 and B12, potassium, and magnesium, making it a meal that is high in nutritional density.
It is well knowledge that the presence of living bacterial cultures, often known as probiotics,
in yogurt helps to maintain digestive health, strengthens the immune system, and may even
improve mental well-being. Since it has such a minimal amount of lactose, yogurt is an
excellent choice for those who are lactose intolerant or sensitive. This allows these
individuals to get the health advantages of dairy products without experiencing any of the
negative side effects (Kaur et al., 2017).

Apparatus

1 500mL Beaker
2 Stainless steel pots (1 big, 1 small)
1 Schott bottle (500 ml)
Stove
Incubator at 45ºC
Ice Bath
Refrigerator
Thermometer
Glass rod (sterile)
Measuring spoon 1Tbs

Materials

Liquid milk (500ml)

Instant milk powder (50g)
Fresh yogurt (30mL)

3
Procedures

1. 50g of instant milk powder was mixed into 500mL of milk in a stainless-steel pot and
was stirred till completely dissolved. It was then placed in a water bath and heated
slowly to 85-90℃, stirred gently all the time. The temperature was maintained for 2
minutes to kill undesirable pathogens, restructure proteins to help with the viscosity as
well as denature inhibitory enzymes that retard the subsequent yoghurt fermentation.

2. The milk was cooled in an icy cold water bath to 43℃.

3. 2 tablespoons of 30mL fresh yoghurt containing live bacteria was added to the cooled
milk and mixed with a sterile glass rod.

4. The mixture was transferred to Schott bottles and sealed with a sterile cap.

5. The mixture was incubated at 45℃ for 5 hours undisturbed until the desired custard
consistency was reached, and the pH fell to 4.4-4.6. Yoghurt is considered coagulated
when it stops flowing as the beaker is tipped slowly. Fluid yoghurt results if the
mixture is stirred as the coagulum is being formed.

6. The warm yoghurt was refrigerated for 4-5 hours to stop the growth of the
thermophilic culture.

4
Results

Table 1.0 Yogurt results on Day 0 and after 1 week

Time Day 0 After 1 week

Reading pH mL of NaOH Titratable pH mL of NaOH Titratable
acidity acidity
1 6.02 4.50mL 0.45% 4.49 11.9mL 1.19%
2 5.96 4.20mL 0.42% 4.47 12.1mL 1.21%
Average 5.99 4.35mL 0.44% 4.48 12.0mL 1.20%

Calculation 1:

Final reading ( mL )−Initial reading (mL)

% lactic acid=
10

Day 0:

17.4 mL−12.9mL
1 st reading=
10

4.5 mL
¿ =0.45 %
10

21.6 mL−17.4 mL
2 nd reading=
10

4.2 mL
¿ =0.42 %
10

0.45 %+0.42 %
Average=
2

0.87 %
¿ =0.435 %
2

≈ 0.44 %

After 1 week:

25.8 mL−13.9 mL
1 st reading=
10

5
 11.9mL
¿ =1.19 %
10

37.9 mL−25.8 mL
2 nd reading=
10

12.1mL
¿ =1.21 %
10

1.19 % +1.21 %
Average=
2

2.40 %
¿ =1.20 %
2

Calculation 2:

Reading 1+ Reading 2
Average pH=
2

Day 0:

6.02+5.96
Average pH=
2

¿ 5.99

After 1 week:

4.49+ 4.47
Average pH=
2

¿ 4.48

6
Discussion

Yogurt is a type of dairy product that is fermented with specific microorganisms, such as
Streptococcus thermophilus (ST) and Lactobacillus delbrueckii subsp. bulgaricus
(LB), according to the Codex standard for fermented milk. The starter culture must contain
viable, active and abundant microorganisms in the product to the date of minimum
durability. Yoghurt production is traditionally used to preserve and extend the shelf life of
milk by reducing the pH of milk. The majority of the milk used to make yoghurt back then
was unpasteurized. Nonetheless, manufacturers must pasteurize the milk before including the
starting culture for reasons concerning the food safety (Behare et al., 2016). To kill
pathogenic microbes, milk will be heated slowly to 85–90°C in a water bath while being
gently swirled. This temperature was held for 2 minutes.

According to the results shown in Table 1.0, the first and second reading of the pH of the
mixture at day 0 were 6.02 and 5.96, respectively. Average pH of was then calculated, which
is 5.99. The pH of the mixture was slightly lower than the pH of natural milk which was
generally around 6.5 to 6.7. This is due to the milk having previously been mixed with 2
tablespoons of fresh yoghurt, which contains lactic acid that had lowered down the pH of the
mixture. Streptococcus thermophilus's production of lactic acid may have caused the pH to
initially drop. The reason is because that ST grows more quickly during the first 8 hours and
will then slows down when the pH reaches roughly about 5. However, after 1 week, the first
and second reading of the pH of the mixture had decreased, which were 4.49 and 4.47. The
average pH was then calculated, which is 4.48. Although yoghurt commonly contains ST and
LB that help in the fermentation of milk, ST usually grows slowly when the pH environment
is below 5, whereas LB can grow well at pH 4. It is unlikely that ST can grow well after the
pH has fallen to 4.48 after 1 week and undergoes the breakdown of lactose into lactic acid.

7
As a result, there is a high possibility that LB is the microbe that is responsible for the lactose
breakdown (Corrieu & Béal, 2016). As a result of lactose being broken down into lactic acid
by ST and LB, the pH of yoghurt dropped from 5.99 to 4.48 within that one week.

The titratable acidity was determined by adding 10 drops of phenolphthalein solution as

indicator into the yogurt. The yoghurt is then titrated with NaOH until the pink colour shown.
A pH range of 8.3 to 10 will allow the phenolphthalein to turn pink. Yogurt turns pink
because NaOH is titrated and neutralized by the acid until the solution has changed to a base
environment. Therefore, the amount of lactic acid contained in the yoghurt increases as the
volume of NaOH is titrated increases. The titratable acidity was calculated at day 0 and after
1 week. Based on the results shown in Table 1.0, the volume of NaOH titrated at day 0 were
4.50 mL and 4.20 mL. The average of NaOH volume was calculated, which is 4.35 mL. After
1 week, the volume of NaOH titrated were 11.9 mL and 12.1 mL. The average NaOH volume
was calculated, which is 12.0 mL. The average titratable acidity on day 0 is 0.44% whereas
the average titratable acidity after 1 week is 1.20%. The continual breakdown of lactose by
lactic acid bacteria for 1-week results in a rise in the average titratable acidity. During day 0,
the milk's lactose is not entirely converted to lactic acid. After 1 week, it shows an increase in
the titratable acidity as the lactic acid bacteria continues to turn lactose into lactic acid.
Hence, the average titratable acidity after 1 week will be higher than the average titratable
acidity at day 0 as shown. Therefore, the longer the yogurt is incubated, the higher amount of
lactic acid is produced which will leads to lower the pH decreases (Mozzi, 2016).

In this experiment, there are a few precautions steps that must be take note of. At
first, the milk should be cooled to 43°C in an icy cold water bath because that is the
temperature at which probiotics develop the fastest (Burton et al., 2017). The glass rod that
will be used to mix the fresh yoghurt and chilled milk must first be sanitized to avoid
contamination that could affect the results. Furthermore, the titration should be carried out
drop by drop to ensure that the endpoint won't be missed to obtain accurate and precise
results. Moreover, the eyes must be parallel at the meniscus level when observing the amount
of NaOH used in the titration to avoid parallax error.

8
Questions

1. Prepare two graphs showing the development of acidity in the yogurt with time
(pH and percent lactic acid)

Figure 1.0 Graph of Average pH in Yogurt against Days

9
 Fig
ure 2.0 Graph of Average Titratable Acidity (%) in Yogurt against Days

2. Explain the purpose of adding milk powder to the liquid milk.

The purpose of adding milk powder into the liquid milk is to improve the texture. This is
because milk powder contains proteins, specifically caseins, which can improve the texture of
yogurt. The proteins help to create a thicker, creamier texture by forming a firmer curd during
fermentation. The different volumes of water will affect burger patties’ sensory and physical
characteristics.

3. In the preparation of yogurt from raw milk, the milk is pasteurized at a temperature-
time combination much more severe than the normal pasteurization conditions (72 oC
and 15 seconds) before cooling and adding the starter culture. Why was this procedure
followed? What two purposes does the heating procedure fulfill?

The severe pasteurization procedure is allowed as the increase in pasteurization temperature

which is up to 85-90 oC can help to destroy more spoilage microorganisms as well as heat
resistance bacteria such as Coxiella burnetii which can survive under normal pasteurization.
The main purpose is the elimination of harmful bacteria. This is because raw milk contains

10
various types of harmful bacteria such as Salmonella and E. coli. These pathogenic bacteria
will cause foodborne illness if consumed. Hence, the main purpose of the heating procedure
is to eliminate the harmful bacteria before making yogurt. This also will allow the starter
culture to thrive without competing with other microbes. Other than that, the heating
procedure allows standardization. It helps to denature some of the whey proteins. The
denatured whey proteins then interact with other proteins and form a more stable network.
This allows the final product can be stabilized and create a consistent texture and flavour.

4. Why is the shelf life for unpasteurized yogurt longer than that for pasteurized milk?

The shelf life of unpasteurized yogurt is longer than that of pasteurized milk because
unpasteurized yogurt has a lower pH than pasteurized milk. The pH of unpasteurized yogurt
is between pH 4.0-4.5 while pH of pasteurized milk is around 6.7. The acidic environment
created by the lactic acid bacteria helps to prevent the growth of pathogenic bacteria and
spoilage microorganisms, which can extend the shelf life of the yogurt.

Then, pasteurized milk has a higher pH and is not as acidic as yogurt. Pasteurized milk has a
pH of around 6.7, which is higher than that of yogurt. The heat treatment during
pasteurization kills off any lactic acid bacteria in the that contribute to the acidic
environment. This causes the pH of the milk less acidic and more prone to spoilage as most
spoilage bacteria prefer an environment with neutral pH of around 6.5-7.5.

Conclusion

All in all, yogurt can be produced via fermentation process by introducing a small quantity of
yogurt product into milk. Once the yogurt is successfully produced, the presence of lactic
acid will affect the characteristics of the yogurt which pH value will decrease (from 5.99 to
4.48), and the titratable acidity will increase (from 0.44% to 1.20%).

11
References
Behare, P., Lule, V. K., & Patil, P. (2016). Yogurt: Dietary importance. Encyclopedia of
Food and Health, 612–616. https://doi.org/10.1016/b978-0-12-384947-2.00765-0

Burton, J. P., Chanyi, R. M., & Schultz, M. (2017). Common organisms and probiotics:
Streptococcus thermophilus (streptococcus salivarius subsp. thermophilus). The
Microbiota in Gastrointestinal Pathophysiology, 165–169.
https://doi.org/10.1016/b978-0-12-804024-9.00019-7

Chandan, R. C. (2017). An overview of yogurt production and composition. Yogurt in Health

and Disease Prevention, 31–47. https://doi.org/10.1016/b978-0-12-805134-4.00002-x

Corrieu, G., & Béal, C. (2016). Yogurt: The product and its manufacture. Encyclopedia of
Food and Health, 617–624. https://doi.org/10.1016/b978-0-12-384947-2.00766-2

Kaur, R., Kaur, G., Rima, Mishra, S. K., Panwar, H., Mishra, K. K., & Brar, G. S. (2017).
Yogurt: A nature's wonder for mankind. International Journal of Fermented Foods,
6(1), 57. https://doi.org/10.5958/2321-712x.2017.00006.0

Mozzi, F. (2016). Lactic acid bacteria. Encyclopedia of Food and Health, 501–508.
https://doi.org/10.1016/b978-0-12-384947-2.00414-1

Pothuraju, R., Yenuganti, V. R., Hussain, S. A., & Sharma, M. (2018). Fermented milk in
protection against inflammatory mechanisms in obesity. Immunity and Inflammation in
Health and Disease, 389–401. https://doi.org/10.1016/b978-0-12-805417-8.00029-9

12
13