1

FST261
SEMESTER 2 MAC-OGOS 2021

GROUP MEMBERS :

NUR AYUNI NADHIRAH BINTI NASERUDIN 2020888066

SHARIFAH NUR BASHIRA SYED YUSOF 2020494778
IFFA HAZWANIE BINTI MOHD HAZRULL 2020479498
NUR HAFNI QISTINA BINTI ZAHARI 2020491254
NURUL NAZURA BINTI MOHAMMAD AZMAN SHAH 2020826824

GROUP :

AS1162C1

DATE OF SUBMISSION :

24 / 5 / 2021

LECTURER’S NAME :

MDM. NUR AISYAH BINTI ISMAIL

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Table of Contents

EXPERIMENT 1 : DETERMINATION OF PHYSICAL PROPERTIES OF FATS AND

OILS 3
A. DETERMINATION OF MELTING POINT 3
B. DETERMINATION OF SPECIFIC GRAVITY (SG) 4
C. DETERMINATION OF REFRACTIVE INDEX (RI) 6

EXPERIMENT 2 : ANALYSIS OF MEAT 11

A. MEAT PIGMENT FORMATION 11
B. EFFECT OF SALT ON BINDING CAPACITY 13

EXPERIMENT 3 : ANALYSIS OF MILK 18

A. DETERMINATION OF PH 18
B. DETERMINATION OF CASEIN FROM MILK WITH AN ACID (VINEGAR) 19

EXPERIMENT 4 : FORMATION OF GLUTEN IN DIFFERENT TYPES FLOUR 24

EXPERIMENT 5 : ENZYMATIC BROWNING OF FRUITS/VEGETABLES 30

REFERENCES 35
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EXPERIMENT 1 : DETERMINATION OF PHYSICAL PROPERTIES OF FATS AND OILS

Objective

1. To determine the melting point of fats and oils.

2. To determine the specific gravity (SG) of fats and oils at a standard temperature.
3. To determine the refractive index (RI) according to the degree and type of saturation
of fats and oils.

Introduction

Fats and oils are also called triglycerides because they are esters that contain glycerol with 3
fatty acids. If the substance is solid at the normal temperature, it tends to be fat. On the other
hand, oils tend to be liquid at the normal temperature. Next, the smoke point usually refers to
the temperature at which the fats or oils start to break down into glycerol and free fatty acids.
Fats will melt if the temperature is in a higher condition and oils are solidified at the lower
temperature. There are three physical properties that will be determined in this experiment
which are the melting point, specific gravity and refractive index.

A. DETERMINATION OF MELTING POINT

Introduction

Commercial fats or oils are heterogeneous mixtures of glycerides and therefore do not have
a sharp melting point. The slip point or also known as melting point is a temperature at which
fat or oils become to slip out of the tube. There will be two temperatures that should be
recorded which is the temperature before and after the samples of fats or oils slip out from
the open capillary tube. In this experiment, we will know whether fats or oils have a higher
melting point based on the temperature that is observed. The samples that were used in this
experiment are butter and olive oil. Each observation will be recorded in the table.
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Materials / Apparatus

Fats / Oils
Capillary tube (7.5cm long, 1.5cm internal diameter, and 0.9mm capillary bore)
Hot plate
Thermometer
Beaker, 1000mL
Tong

Procedure (Capillary Tube Method)

1. 1 cm length of fat or oil which has been melted was inserted into a capillary tube by
using a piece of filter paper.
2. Then, the capillary tube was cooled at a room temperature of <0 0C for 2 hours.
3. After that, the tube was submerged in the water so that the top end of the fat or oil is
1 cm below the level of water. Approximately 700mL water was used to contain a
1000mL beaker.
4. A thermometer was hung in the middle of the beaker. The water then was heated at
the rate of 10 0C / minute.
5. The temperature was recorded at which the fat has started to slip out of the tube and
also the final temperature when all the fat has slipped out of the tube. The
temperature or range of temperature was referred to as the slip point (melting point).

B. DETERMINATION OF SPECIFIC GRAVITY (SG)

Introduction

Specific gravity can be defined as the ratio of the density oils or fats divided with the density
of water. The determination measures the weight fats or oils in a vessel or bottle which
contains a specific volume. In this experiment, we will know whether fats or oils have a
higher specific gravity based on the calculation. The samples that are used in this
experiment are the same with the samples used to determine the melting points which are
butter and olive oil.
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Materials / Apparatus

Fats / Oils
Specific Gravity Bottle
Disposable plastic pipette/dropper
Refrigerator
Balance
Label sticker

Procedure

1. An empty SG bottle was weighed in gram (x).

2. The SG bottle was filled with oil (or melted fat). The bottle was closed with a stopper
which has a capillary bore.
3. Then, the SG bottle was cooled at 5-7 0C in chilled temperature for 24 hours.
4. After 24 hours, the SG bottle was warmed to 25 0C or room temperature until
expansion had ceased.
5. The bottle then was wiped and cleaned on the outside and was weighed in gram (y).
6. The SG of the oil or fat was calculated.

Calculation

Specific Gravity = Density of oil

Density of water

= Mass of Oil/ Volume of Oil

1 gram/mL

= (x-y) gram/ z mL
1 gram/mL

z = total volume of oil the SG bottle can hold = 50mL

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C. DETERMINATION OF REFRACTIVE INDEX (RI)

Introduction

Refractive index (RI) of fats and oils can be defined as the ratio of the speed of light in air
and in the fats or oil. Then, the refractive index will increase if the degree of unsaturation,
temperature or the length of the chain increases. Other than that, it also helps to implement
the quality control of oils and fats. In this experiment, a refractometer is used to determine
the refractive index and it is measured by using different temperatures such as 20˚C for oils,
40˚C to 60˚C for fats.

Materials / Apparatus

Fats / Oils
Disposable plastic pipette/dropper
Abbe Refractometer
Tissue paper
Dropping bottle filled with acetone

Procedure

1. For scale adjustment procedure, 2-3 drops of distilled water was placed on the main
prism surface using a syringe, covered it with a secondary prism and was looked
through the eyepiece. The refractometer was set at 1.3330 (Brix 0%) if the
thermometer scale was in 20 0C. If temperature is other than 20 0C, an adjustment
was carried out according to the chart in the refractometer manual.
2. Then, the secondary prism was opened and 2-3 drops of oil (or fat at 40 0C) was
placed at the centre of the main prism. Make sure the sample was distributed evenly
and that there were no air bubbles.
3. The secondary prism was closed with caution.
4. While looking through the eyepiece, the measurement knob was turned slowly until
the boundary line can be observed in the view. Therefore, the measurement knob
was turned until the view was changed from dark to light. This degree of lightness
can be changed by using a “high-low” switch on the thermometer box.
5. The colour compensator knob was turned to remove the colour of the boundary line.
After that, a clear boundary line was seen.
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6. Then, the measurement knob was turned again to coincide the boundary line with the
crossed line.
7. The RI then was recorded. (top scale shows the RI, bottom scale shows the brix
reading)

Data & Results

Table 1.1 : Melting points of fats/oil

Sample : butter

Temperature Trail 1 Trail 2 Trail 3 Average

Initial (OC) 20 21 20 20.33

Final (OC) 32 33 35 33.33

Sample: olive oil

Temperature Trail 1 Trail 2 Trail 3 Average

Initial (OC) -4 -5 -4 -4.3

Final (OC) -6 -6 -6 -6

Table 1.2 : Specific gravity of fats/oils

Sample Weight of empty SG Weight of SG bottle + oil Specific gravity oil

bottle (g) ( after removed from
refrigerator)(g)

Butter 23.66 55.34 0.634

Olive oil 23.66 55.61 0.639

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Table 1.3 : Refractive index of fats/ oils

Sample Refractive index

Peanut oil 1.460

Maize oil 1.465

Sunflower oil 1.461

Soybean oil 1.466

Conclusion

In conclusion, the melting point of butter and margarine, the specific gravity and refractive
index of palm oil and corn oil have been determined. Butter shows higher slip melting point
than olive oil due to the presence of saturated fats. It shows that the slip melting points will
increase when the saturation level is higher. The specific gravity of olive oil is higher than
butter because it contains longer carbon chain fatty acids. Meanwhile soybean oil has the
highest refractive index than peanut oil, maize oil and sunflower oil because the higher the
length of the carbon chain and number of double bonds the higher the refractive index.

Discussion

Experiment A shows that the average melting point of butter is 33.33°C whereas the average
melting point of olive oil is -6°C. This is because the olive oil used has more unsaturated fats
thus, it melts at lower temperatures compared to butter. Another reason olive oil has a
relatively low melting point is due to it having a long carbon chain of unsaturated fatty acids.
Although usually a high number of carbon chains would mean a high melting point, the
opposite can be said when that fatty acid is unsaturated.

Specific gravity is the ratio of the density of oil to the density of water at the same
temperature. Specific gravity can also predict whether said substance will float on the water
or sink in the water. Substances with a specific gravity that is higher than 1 will sink in the
water whereas, substances with a specific gravity that is lower than 1 will float. Moreover,
fats and oils have higher molecular weight, thus oil is not soluble in water and tends to float
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when dissolved in water. In experiment B, we get to see the specific gravity of oils used.
Based on the experiment, olive oil has a specific gravity of 0.639 and butter has a specific
gravity of 0.634. The reason olive oil has a higher specific gravity is due to its containing
unsaturated fatty acids. Unsaturated fatty acids can coil up depending on its structure thus
more fatty acids can fit in a certain volume. Olive oil contains long carbon chain fatty acids,
this may also affect the specific gravity.

Refractive index is the ratio of the speed of light at a specific wavelength in the air to the
speed of light in fats or oils. In experiment C we get to compare the refractive index of
various fats or oils to that in air. Based on the experiment, peanut oil has the lowest
refractive index with 1.460, then sunflower oil with a refractive index of 1.461. Maize oil has a
refractive index of 1.465 and the oil with the highest refractive index of 1.466 is soybean oil.
Soybean oil has the highest refractive index because it has the longest carbon chain among
the samples used. Other than that the high number of double bonds also affects the
refractive index.

Questions

1. What are the differences between fat and oil?

- Fats are usually derived from animals, whereas oils are usually derived from plants. Fat is
a substance which is solid at room temperatures, on the other hand, oil is a substance which
is liquid at room temperatures. Fats are more saturated compared to oils. Fats are melted at
higher temperatures while oils are solidified at lower temperatures. Fats are less digestible
than oils.

2. Why do different fats have different melting points?

- Different fats such as saturated and unsaturated fats have a different melting point
because how saturated the chemical bonds in the fat molecule depends on the hydrogen
atoms, the more hydrogen atoms a fatty acid has, the more “saturated” it is and the higher its
melting point will be. On the other hand, unsaturated fats have a lower melting point because
the cis- configurations will cause the fatty acid to be in a bent configuration which produces
weak van der Waals interactions between molecules.
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3. Which sample has the highest refractive index? Why?

- Soybean oil has the highest refractive index because it has the longest carbon chain and
contains a high number of double bonds. Furthermore, the more optically dense a material is,
the slower the wave will move through the material.

4. Which sample has higher specific gravity? Why?

- Olive oil has higher specific gravity because it contains more unsaturated fatty acids, has a
longer carbon chain and higher degree of unsaturation.
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EXPERIMENT 2 : ANALYSIS OF MEAT

Objective

1. To study the effect of oxygen to the colour of the meat.

2. To investigate the effect of sodium nitrate on the colour of the meat.
3. To analyse the effect of adding the salt to the meat.

A. MEAT PIGMENT FORMATION

Introduction

The color in meat shows their quality and freshness. When it comes to the red color, some
people have misconceptions. The red color in meat is not indicated as blood. However, it is
myoglobin that occurs in the muscles of animals like cows and poultry. Besides, the red color
that occurs in meat depends on the amount of myoglobin in the muscles of animals. For
example, beef has a lighter red color, while poultry has a pinkish to white color. That is
because the amount of myoglobin in beef is higher than in poultry.

In the food industry, product packaging is really important to maintain the freshness and
avoid any food spoilage. As an example, saran wraps are commonly used to ensure that the
meats retain their consistency and freshness. Other than that, for cured meat, use of nitrate
solution in meat will show a brighter red color than non-nitrate solution in meat.

I) FRESH MEAT PIGMENTS

Materials / Apparatus

Meat White paper plate

Chopping board Saran wrap
Food processor Gloves
Balance Refrigerator
Knife Label sticker
Spatula
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Procedure

1. The meat was cut into small pieces, and placed in the food processor for grinding.
2. 70 g of ground meat was placed on a chopping board after weighing.
3. A beef burger patty was formed with 1 inch thickness.
4. The burger patty was placed on a white paper plate and the colour near the edge of
the patty and near the centre are observed.
5. Colour differences were observed, which are due to the amount of oxygen that
entered the different parts of the meat.
6. Two of 1 inch thick patties were cut off.
7. One patty was wrapped using saran wrap while the other one was left unwrapped.
8. Each of the patties were placed on a white paper plate.
9. The patties were refrigerated and colour changes over the next 2 hours were
observed.

II) CURED MEAT PIGMENTS

Materials / Apparatus

Ground meat
1% sodium nitrate solution (NaNO2)
Pestle
Graduated pipette, 2mL
Beaker, 100mL
Spatula
Aluminium foil
Refrigerator
Balance

Procedure

1. 200 g of ground meat were divided into two equal batches.

2. Each batch of ground meat were put into a labelled beaker (1 and 2)
3. The meat in beaker 1 was controlled (nothing is added)
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4. 1.6 mL 1% sodium nitride solutions were added to the meat in beaker 2 and it was
thoroughly mixed for 3 minutes.
5. Each batch of ground meat was stuffed with pestle and into a labelled beaker.
6. Air was eliminated from the meat by packing it down.
7. The beakers are covered with aluminium foil and placed in a refrigerator for 1 night.
8. The beakers were removed from the refrigerator and the colour of the surface and
the interior of the meat were observed.
9. The meat was cooked by heating the beakers in a boiling water bath for 15-20
minutes.
10. The colour of the surface and the interior of the non-nitrite meat and the nitrite meat
were observed.

B. EFFECT OF SALT ON BINDING CAPACITY

Introduction

When salt is added to the ground meat, it makes the salt-soluble proteins come to the
surface of the meat. In scientific terms, it extracted the salt-soluble proteins from the cellular
structure. Then, the proteins acted like glue. The main purpose of using salt in meat is to
reduce and prevent the growth of spoilage microorganisms. Besides, it will increase the
binding of meat especially during cooking and plays an important role to make sure that the
meat products are safe to be consumed. It can also help to lower the water activity in meat
products which can inhibit the growth of microbes. Other than that, the tastes of food
products can be improved by adding some salt into it. In this experiment, we will know either
the meat can bind perfectly with water or salt.

Materials / Apparatus

Ground meat Beaker, 25mL

Salt Spatula
Glove Weighing boat
Measuring cylinder, 10mL Food processor
Mixing bowl Balance
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Procedure

1. When working with raw meat, gloves have been worn.

2. 50 g of ground meat were placed into a food processor.
3. 10ml of water is added to the meat and it’s been chopped for 15 second.
4. The meat was removed from the food processor.
5. A ball of meat was formed and shaped like a burger patty.
6. The flattened meat was put in the palm of our gloved hand (palm up).
7. Our hand was turned over. The observation is based on whether the meat sticks to
our hand or does it fall down.
8. 50 g of ground meat were placed into a food processor.
9. 4 g of salt was dissolved into 10 ml of water before adding it to the meat.
10. The salt solution is then added to the meat and chopped for 15 seconds.
11. The meat was removed from the food processor.
12. A ball of meat was formed and shaped like a burger patty.
13. The flattened meat was put in the palm of our gloved hand (palm up).
14. Our hand was turned over. The observation is based on whether the meat sticks to
our hand or does it fall down.

Data & Results

Table 3.1 : Meat pigment formation (fresh meat pigment)

Before refrigerate After refrigerate for 2 hours

part of meat colour of meat packaging colour of meat

near the centre cherry red saran wrap red

near the edge red unwrap pale red

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Table 3.2 : Meat pigment formation (cured meat pigments)

Treatment Colour of meat

uncooked cooked

surface interior surface interior

Non-nitrite meat Pale read Pale read Completely brown Brownish red

Nitrite meat Bright red Bright red pink pink

Table 3.3 : Effect of salt on binding capacity

Meat Observation (stick or fall)

Added with water stick

Added with salt solution fall

Conclusion

In conclusion, we can say that the colour and quality of the meat can indicate that the meat
is fresh. Salt can act as the preservative agent to make the meat look as fresh as before it
was stored. At the end of this experiment, we were able to understand the analysis of meat.
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Discussion

Based on the experiment, we were done the experiment of analysis of meat. First
experiment that had been observed was meat pigment formation. In this experiment, we
observed 2 meat pigments which are fresh meat pigment and cured meat pigment. In fresh
meat pigment, the fresh meat pigment was red in colour at the centre and at the edge in the
beginning. After refrigerating for 2 hours the unwrap meat is still red in colour compared to
saran wrap meat, pale brown. This is because the red colour comes from a pigment,
myoglobin. The unwrap meat was exposed to oxygen which is turning it into oxymyoglobin.
The meat with saran wrap turns pale brown because it is not exposed to oxygen, like the
meat that’s on the interior, will often lose its red colour after a few days.

Next, in the cured meat pigment experiment, the uncooked meat of non-nitrate meat is also
brown in colour but different with nitrate meat, red in colour. When the meat was cooked, we
obtained that the non-nitrate meat was brown and the interior was pinkish red. This is
because the nitrates give cured meat. Nitrates also are antioxidants and have microbial
properties, so they give significant food safety benefits, which prevent the growth of
Clostridium botulinum which causes botulism, one of the deadliest foodborne illnesses. The
nitrates are considered a preservative because their antimicrobial properties also control
spoilage bacteria, so cured meats have longer shelf-life than fresh meats.

Lastly, we ended the experiment of the effect of salt on binding capacity. 2 meats were
shaped like a burger patty, in which one was added with water while another one with salt
solution. Then, put it in the palm and turn it over. The meat with water fell down while the
meat with salt solution was stuck on the palm. This is because when we take all the moisture
out of the fresh meat, they will stick to the palm. This is also one of the ways on how to
preserve food. As salt level increases in solution, the growth of potential and survivability of
microorganisms like fungi and bacteria decreases, it is well known that proper salt
preservation prevents contamination by clostridium botulinum, the bacterium that causes the
fatal disease known as botulism.
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Questions

1. What types of reactions are responsible for the colour observed in the various
treatments? What is the name of each pigment formed? Which reactions are
desirable?

As soon as meat is cut, oxygen reacts with the myoglobin and creates the bright red colour
associated with oxymyoglobin. This will continue to develop until the iron in the myoglobin
oxidizes to the point of the metmyoglobin stage. Oxidation can also occur when iron in the
meat binds with oxygen in the muscle.

2. What is the effect of heating on the globin portion of pigment?

Effect of heating on the globin portion of pigment is the protein will be denatured and
produce hemichromes which are tan to brown in color.

3. What are the functions of salt in meat products?

The functions of salt in meat products is salt helps to bind the meat proteins together and
acts as a binding agent between meat and fat emulsions.
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EXPERIMENT 3 : ANALYSIS OF MILK

Objective

1. To determine the pH of milk

2. To determine the coagulated casein from milk with adding acid (vinegar)

Introduction

Milk is a natural source that contains high nutrients which can also help for human growth. It
also consists of important nutrients like protein, calcium, riboflavin, vitamin A, vitamin B
complex and many more. Moreover, milk is really good for bones since it provides a high
amount of calcium and vitamin D which can help to prevent the risk of osteoporosis. Milk can
be considered as consisting of three basic components namely the water, fat and non-fatty
solids or solids-non-fat. In this experiment, there are two things that should be determined
which are the pH of milk and the casein from milk with an acid (vinegar).

A. DETERMINATION OF PH

Introduction

This experiment looks at the changes happening in pH of milk as it goes sour. When the milk
goes sour, it will become more acidic and the pH becomes lower than before. The changes
occur when the milk is contaminated with bacteria which is harmful for consumption. The
activities of processing and handling dairy products may introduce further microorganisms.
Generally, there are two groups of bacteria that occur in milk which are Lactic acid and
Coliforms. Lactic acid is bacteria normally identified in milk and also dairy products while
Coliforms are bacteria that can cause spoilage in milk.
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Materials / Apparatus

Milk
Universal bottle or sample bottle with cap
pH Meter
pH Buffer 4 and 7
Wash bottle filled with distilled water
Tissue paper
Beaker, 10mL
Label sticker

Procedure

1. The pH meter calibrated with pH buffer 4 and 7.

2. The milk was allowed to reach room temperature without cooking and re-pasteurising
it.
3. The milk was poured into the universal bottle.
4. The pH Meter was inserted into the bottle of milk. The pH was recorded.
5. The pH of milk was recorded from 0 day, 1 day and 2 days.
6. A graph of pH versus day was plotted.

B. DETERMINATION OF CASEIN FROM MILK WITH AN ACID (VINEGAR)

Introduction

Milk consists of many proteins. Casein is one of the main groups. These kinds of proteins
are usually found in mammalian milk and the percentage of casein in mammalian milk such
as cows is higher than in human’s milk. Casein has a negative charge in its natural state.
This can permit the casein to disperse in milk. If an acid is added in milk, H+ concentration
will neutralize the negative charge of casein. When the milk is in the acidic condition, the pH
will reach to 4.6 which is its isoelectric point. In addition, casein is a major component for the
manufacture of cottage cheese and cream cheese. Other than using for cheese making, acid
casein is also used in the chemical industry, textile and cosmetics, and in the food
processing as a binding agent and stabilizer. In this experiment, it indicates the precipitation
of casein from milk when an acid (vinegar) is added into it.
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Materials / Apparatus

Soy milk Spatula

Pasteurized whole milk Thermometer

Rennin enzyme Cheesecloth

Hot plate Rubber band

pH meter Stirring rod

pH buffer 4 and 7 Heat Proof glove

Beaker, 250mL Weighing boat

Measuring cylinder, 250mL Wash bottle filled with distilled water

Measuring cylinder, 25mL Tissue paper

Balance

Procedure

1. The pH meter was calibrated with pH buffer 4 and 7.

2. The empty beaker was weighed and recorded.
3. 120mL of milk in the beaker was weighed and recorded. Then, weight of the milk
recorded in the data table (weight of beaker with milk - weight of beaker = weight of
milk).
4. pH of the milk was measured. The thermometer was placed in the beaker.
5. The beaker was placed with the milk on a hot plate. The milk was heated to 37℃.
The hot plate was turned off and the beaker was removed.
6. 3mL of rennin enzyme was added to the warm milk and stirred for 2 minutes, then
the milk was allowed to sit for 5 minutes. The casein was precipitated into heavy
white curds. The remaining liquid was the whey.
7. The pH of the liquid portion was measured again.
8. A (2-3 layers) of cheesecloth large enough was cut to cover the top and 2 inches
down the sides of a beaker.
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9. The cheesecloth was fastin over the top of the beaker using the rubber band.
10. The curdled milk was poured into the beaker. The curds (casein) in the cheesecloth
were collected and the vinegar and whey were allowed to drain off into the bottom of
the beaker.
11. The cheesecloth was gathered with the casein and rinse in cool water by dipping into
another beaker containing water.
12. The casein was squeezed until almost dry and then spread out the cheesecloth to let
the casein dry for 5 minutes.
13. The precipitate was weighed. (The cheesecloth did not weigh with the precipitate).
The result was recorded. Texture and colour of the curd were noted.

Data & Results

Table 6.1 : pH of milk

pH Sample 1 Sample 2 Sample 3 Average

Day 0 6.8 6.6 6.8 6.7

Day 1 6.1 5.9 6.2 6.1

Day 2 5.1 5.3 5.0 5.1

Table 6.2 : Determination of casein whole milk and soy milk

Sample Weight of beaker Weight of beaker + 120ml milk Weight of

(g) (g) precipitate

Whole milk 48.75 169.12 3.48

Soy milk 50.03 170.24 0.06

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Table 6.3 : Determination of casein whole milk and soy milk

Sample Initial pH pH after vinegar Texture and colour of

added curd

Whole milk 6.7 4.4 Clumpy solids, white

Soy milk 7.1 5.5 Smooth semi-solid, off-

white

Conclusion

In conclusion, we can analyze milk by reaction with some chemicals and undergo some
process. We can analyze the pH value and the formation of casein in fresh milk and soy milk.
At the end of this experiment, we were able to understand the analysis of milk.

Discussion

In this experiment, we already observed the pH of milk as it goes sour and casein formation
from milk with vinegar. We used whole milk and soy milk as food samples for this experiment.

Experiment A shows the pH of milk that is recorded for 3 days. The result shows the average
pH for sample 1, sample 2 and sample 3 for day 0 is 6.37. The average pH of milk for day 1
is 6.07 which is less than day 0. Then, for the last day which is day 2 the pH decreases to
5.13. After a few days,the milk becomes more acidic and the pH gets lower than before and
causes the milk to go sour. This occurs as bacteria in milk convert the sugar lactose into
lactic acid. The pH of milk accounts for the amount of lactic acid produced by microbial
activity. The more lactic acid present, the higher the acidity. So, we can say that, the longer
the milk was stored, the higher the amount of lactic acids, the lower the pH value.

For experiment B, the purpose of this experiment is to distinguish the determination of

casein between whole milk and soy milk. Casein also can be coagulated with the enzyme
rennin, which is found in rennet (an extract from the stomach of calves).The weight of
precipitate for the whole milk is lighter than soy milk which is 11.56 to 23.87. It is supposedly
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the amount of casein in milk is higher than soy milk. The texture for the whole milk after
added with vinegar is clumpy solids and the colour of the curd is white. Meanwhile, the
texture for the soy milk is smooth semi-solids and the colour of the curd is white. Casein is
present in milk as calcium salt and calcium caseinate. During the addition of acid to milk, the
negative charges on the outer surface of the micelle are neutralized (the phosphate groups
are protonated), and the neutral protein precipitates. From this milk analysis, we know that
whole milk had higher casein compared to soy milk.

Questions

1. What causes the reduction in the pH of milk?

The pH of milk changes as it goes sour. The milk is contaminated by bacteria even
before it leaves the udder. The processing and handling activities can introduce
further microorganisms that cause the reduction in the pH.

2. Compare the amount of acid casein precipitated from the whole milk with the
amount of soy protein coagulated from the soy milk. How do your results
compare with the nutrition facts label for each product ?

The amount of acid casein precipitated from the whole milk is 11.56g while the
amount of soy protein coagulated from the soy milk is 23.87g.

3. List 3 factors that influence the precipitation of casein milk ?

Factors that influence the precipitation of casein milk is rennin,an enzyme that is
found in calves’ stomach. Next, acid (vinegar) is also one of the factors to precipitate
casein from milk. Lastly, when casein loses its negative charge it will precipitate as
curds.
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4. What is the importance of casein in the food industry?

The importance of casein in the food industry is when casein is used in paste
products, it is mainly to enhance their nutritional quality. It is because the high lysine
content of the casein complements the low casein and high sulfur containing amino
acids of wheat and other vegetable protein present in such products.
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EXPERIMENT 4 : FORMATION OF GLUTEN IN DIFFERENT TYPES FLOUR

Objective

1. To observe the characteristics of the dough ball by using different types of flour.
2. To observe the gluten formation by using different types of flour.

Introduction

Gluten is a protein found in grains, including wheat, rye and barley. It is also used as an
additive for the purpose to improve texture and give moisture to the processed foods. Gluten
is a substance that forms when two main groups of protein: glutenin and gliadin come into
direct contact with water. As the proteins are worked, they become long and flexible strands.
Carbon dioxide is mostly produced in the dough by the yeast itself. The dough will be more
able to hold the gas bubbles when the flour produces more gluten and makes the dough rise
before it is baked.

Bread free of gluten would be very dense or flat shows that gluten is really important in
bread making. Other than that, rice, potato, oat and corn flours cannot form the gluten at all.
If the bread is made using those flours, it will not turn out well.

This experiment generally indicates what gluten is, the importance of gluten in bread making
and its presence in all samples of flour.

Materials / Apparatus

Normal wheat flour

High protein wheat flour
Cake flour
Rice flour
Mixing bowl
Plastic plate
Spatula
Balance
Measuring cylinder, 25mL
 26

Procedure

1. Each type of flour weighed 55 gram and was put into the mixing bowls.
2. 25mL of water was added to each bowl of flour and mixed.
3. Dough mixtures were kneaded for about 5 minutes each.
4. The texture, appearance, colour, elasticity and flexibility of each dough ball were
noted.
5. The dough ball was placed under the faucet and cool water was run over it. The
dough ball was squeezed to drain white, starchy water.
6. This under running water continued until more starch came out.
7. How the dough shrank in size, changed colour and became thread like were noted.
These were the gluten threads. The water that drained from the dough ball was white
as the starch was washed out of the dough ball.

Data & Results

Table 7.1 : Observation of dough ball

Sample Texture Appearance Colour Elasticity Flexibility

Normal Soft Can form a Creamy Elastic Flexible

wheat flour dough ball white

High protein Soft Can form a White Elastic Flexible

wheat flour dough ball

Cake flour - Cannot form White - -

a dough ball

Rice flour - Cannot form White - -

a dough ball
 27

Table 7.2 : Observation of gluten formation

Sample Gluten

Colour Size

Normal wheat flour Creamy white Small

High protein flour White Big

Cake flour - -

Rice flour - -

Conclusion

In conclusion, we can conclude that the gluten will affect the texture and overall appearance
of the dough. The higher the amount of the gluten, the higher the quality of the dough ball. At
the end of this experiment, we were able to understand the formation of gluten in different
types of flour.

Discussion

Proteins are very large molecules composed of amino acids and two naturally occurring
proteins in flour which are glutenin and gliadin. Gluten is a water-insoluble protein that forms
when water is mixed with wheat flour. These two proteins become flexible when sufficient
water is added to dry flour and this process is called hydration. As water and flour are mixed
together in the kneading process, the hydrated proteins are brought together and begin to
interact. They literally begin to stick to each other through the formation of chemical bonds.
From this process, gluten bonds form and a firm rubber substance is created providing
strength and structure. The bonds that form between the glutenin and gliadin are called
disulfide bonds.
 28

There are different results of observation of dough balls and the gluten formation because
the formation of gluten is dependent on the content of protein. In normal wheat flour it
contains about 8 to 11% of gluten. High-gluten flour which is typically used for bread making
contains 12 to 15% of gluten. Cake flour contains about 7 to 8% of gluten and there is no
gluten content in rice flour. This shows that increasing the total protein content of the flour
positively correlates to the gluten content.Generally, different types of flour have different
amounts of gluten.

In this experiment, we used different types of flour which is normal wheat flour, high protein
wheat flour, cake flour and rice flour to observe the texture, appearance, colour, elasticity
and flexibility of the dough ball. High protein wheat flour and normal wheat flour have the
softest texture because the high ratio of protein will produce softer dough. The strong gluten
matrix will provide structure in raising the dough. Secondly, the appearance of normal wheat
flour and high protein wheat flour is that there is a formation of curdle and it is easy to form
into dough balls while the appearance of rice flour and cake flour there was no formation of
curdle as it is difficult to form rice flour into dough balls. Third, the colour of normal wheat
flour was creamy white while rice flour, high protein wheat flour and cake flour showed white
color respectively. The elasticity for normal wheat flour, high protein wheat flour are more
elastic than rice flour and cake flour which shows that there is no elasticity in rice flour.

Next, we observed the gluten formation in each type of flour and the differences in their
colour and size by washing the dough ball under running water. The wet gluten content is
determined as the starch and other soluble from the dough ball. The colour form in normal
wheat flour and high protein were creamy white and white respectively. Meanwhile the
colour of gluten in cake flour and rice flour cannot be observed because the flour disappears
when washed under running water. When rinsing, the dough ball of normal wheat lost its
white colour. This is because the bleach in the flour is draining away while being washed.
Next, the size of gluten formation in high protein wheat flour is larger than normal wheat flour.
There is no gluten formation in the rice flour as it disappears when being washed under
running water.

This result can be applied to the bread making potential of flour which is mainly based on the
protein quantity and quality. Gluten plays a key role in determining the unique baking quality
of wheat by conferring water absorption capacity, cohesiveness, viscosity and elasticity on
dough. Through this experiment, it shows that different types of flour contain different
amounts of gluten. As the hypothesis states, the higher the content of protein present in flour,
the higher the gluten will be formed when dough is kneaded.
 29

Questions

1. List the major proteins present in wheat flour?

The major proteins present in wheat flour are gliadin and glutenin.

2. What is gluten?

Gluten is a protein composite found in grains, including wheat, barley and rye. Gluten
gives elasticity to dough, helping it rise and keep its shape and often gives the final product a
chewy texture.

3. Which flour has greater gluten content?

High protein wheat flour has greater gluten content among all. It contains 13 to 14 g of
protein per cup, making it 11.5 to 12.5 percent protein. It is best for yeast breads, pasta and
pizza dough.

4. Which flour do you think is suitable for bread making? Why?

High protein wheat flour is the most suitable for bread making because it has higher gluten
content which makes the bread become more fluffy and give better quality.
 30

EXPERIMENT 5 : ENZYMATIC BROWNING OF FRUITS/VEGETABLES

Objective

1. To determine the enzymatic browning of fruits or vegetables

Introduction

Browning is a process of vegetables or fruits turning their natural color into brownish due to
the chemical reactions that occur. Generally, it happens when the fruits or vegetables are
peeled or cut. In the presence of oxygen from the air, the enzyme polyphenol oxidase
catalyzes one step in the biochemical conversion of plant phenolic compounds to brown
pigments which are known as melanin. This kind of reaction is called enzymatic browning.
Other than that, the browning process may lead to a significant problem which can limit the
shelf life of fruits and vegetables. However, there are some benefits of the enzymatic
browning which produce color and flavor in coffee, cocoa, tea and also in the dried fruit such
as raisins and prunes.

The control of enzymatic browning has always been the biggest challenge for many food
industries. Several substances have been used to prevent or slow down the reaction. The
releasing of sulphite ions may prevent the formation of melanin. In this experiment, there will
be three different types of acid solution used which are citric acid, ascorbic acid and acetic
acid with two differences of percentage. In addition, the apple slices must also soak into
plain water and untreated(control) solution. It is used for the purpose of knowing the level of
browning in apple slices. The observations must be taken every 10 minutes for 2 hours and
record all the results in a data table.
 31

Materials / Apparatus

Fresh fruits/vegetables
1% ascorbic acid
1% citric acid
1% acetic acid
2% acetic acid
Beaker or cup
Paper towel
Tong
Glove
Knife
Stopwatch or timer

Procedure

1. One piece of fruit/vegetable was sliced into 6 parts in the same size.
2. An untreated fruit/vegetable slice was placed on a paper towel. The towel was
labeled as “Control”.
3. Another fruit/vegetable slice was dipped into one of the test solutions (1% ascorbic
acid) for 10 minutes using gloves or tongs, it was placed on the paper towel and
labelled with the name of the solution.
4. The glove and tong were rinsed and the same procedure was repeated for the other
three solutions (1% acetic acid,2% acetic acid).
5. One fruit/vegetable slice was soaked in water for 10 minutes, then it was placed on a
paper towel and labelled the towel “Water Soak”.
6. The time and level of browning were noted in your data table. The slices were
observed every 10 minutes for 2 hours and the observations were recorded.
 32

Data & Results

Table 9.1 : Level of browning in apple slices

Level of browning

Time (min) Untreated Ascorbic Citric acid Acetic acid Acetic acid Water
(uncontrol) acid (1%) (1%) (1%) (2%)

0 1 1 1 1 1 1

20 2 1 1 1 1 1

30 2 1 1 1 1 2

40 3 1 1 1 1 2

50 3 1 1 1 1 3

60 4 1 1 2 1 3

70 4 1 1 2 1 4

80 4 1 1 2 1 4

90 4 2 1 3 2 4

100 4 2 1 3 2 4

110 4 2 1 3 2 4

120 4 2 1 3 3 4

Conclusion

As a conclusion, several substances have been used in the food industry to prevent
browning of fruits and vegetables such as ascorbic acid, citric acid and acetic acid. The
browning is caused by the conversion of phenolic compounds to melanins catalysed by the
enzyme polyphenol oxidase. At the end of this experiment, we were able to understand the
enzymatic browning of fruits and vegetables.
 33

Discussion

Browning is a process of vegetables or fruits turning their natural color into brownish due to
the chemical reactions that occur. It usually happens when the fruits or vegetables are cut or
peeled. In this experiment, about 6 apple slices are used to obtain the level of browning
every 10 minutes for 2 hours. Each slice will be dip into 1% of ascorbic acid, 1% of citric acid,
1% of acetic acid, 2% of acetic acid and water for 10 minutes thus placing it on the paper
towel. The last slice is untreated and labeled as control. After 2 hours observations, apple
slices with untreated and water soaked show the higher level of browning among other test
solutions which is 4. This level represents that the apple slice is fully covered with light
brown. It commonly happens because the enzymes contained in the plant tissues come with
direct contact with oxygen from the air.

Other than that, Ascorbic acid (1%) has a lower level of browning compared with Acetic acid
(1%) and Acetic acid (2%) which is 2. This level shows that the apple slice that dip in 1% of
Ascorbic acid is slight or scant brown patches. In addition, browning will not proceed until all
the ascorbic acid is used in the reaction. In the food industry, ascorbic acid may help to slow
down the browning process. Furthermore, the test solution that can prevent the browning
process is citric acid (1%). It is because from the observation, the level of browning for citric
acid is the lowest among others which is 1. This level indicates that there is no browning
present in the apple slice. Citric acid is also used in the food industry as it can prevent and
avoid the browning process. Other than that, citric acid can lower the pH level of fruit tissue
and retard the action of polyphenol oxidase. As the pH is decreased below than 3, the
polyphenol oxidase will be automatically inactivated.

Questions

1. What causes browning when fresh fruits and some vegetables are peeled or cut?

The fruits and vegetables that are already peeled or cut will be exposed with the oxygen and
the enzymes in the plant tissues are released. In the presence of oxygen, polyphenol
oxidase catalyzes one step in the biochemical conversion of plant phenolic compounds to
brown pigments known as melanin.
 34

2. What conditions enhance the browning process? Why?

Bruising or any injury to the plant tissue enhances the browning process. It is because it can
disrupt the structural arrangement of constituents within the cells and allows the contents to
be exposed with oxygen.

3. How do food additives or treatment processes in use today prevent or retard

browning in fruits and vegetables?

Food additives or treatment processes can retard or prevent browning in fruits and
vegetables as they can inactivate or inhibit the polyphenol oxidase, which is an enzyme that
may lead to change the color of fruits or vegetables into brown.

4. Why do citrus juices retard browning in fresh fruits?

Citrus juices retard browning in fresh fruits because it can lower the pH level of the fruit
tissue and inhibit the action of the polyphenol oxidase.
 35

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