HAWASSA UNIVERSITY

School of Nutrition, Food Science and Technology

Food Chemistry Laboratory Manual
(Food Science and Post-Harvest Technology)

Dec, 2012

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Experiment 1: Effect of Emulsifiers on Emulsion Stability

Introduction
Emulsion is a property where two liquids are evenly spread out in each other, yet not dissolved in
each other. Oil and water form the most common emulsions; band milk is an emulsion of
butterfat in water. Emulsions are important in the production of foods that contain water and fat,
such as mayonnaise or margarine. These products require the addition of an emulsifier, such as
the food lipid lecithin, proteins (egg yolk), gum to stabilize food emulsions.
For each test, you will use:

 90 milliliters colored water

 10 milliliters vegetable oil
 Sealable container that will hold at least 200 milliliters (empty soft drink bottle, plastic
Glad Ware with sealable lid, etc.)
 1 clear 100 milliliter graduated cylinder for water and emulsion, clean and dry
 Stopwatch

Emulsion Trial Water (mL) Oil (mL) Other Ingredient

1 90 10 None - Control

2 90 10 2.0 grams sucrose

3 90 10 2.0 grams whey protein concentrates?

4 90 10 0.25 grams xanthan gum?

5 90 10 3 milliliters Polysorbate 20 (Tween 20)?

6 90 10 3 milliliters egg yolk (fresh)

*separate egg white and yolk into two dishes or sample containers. Keep refrigerated.

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Procedure:

1. Measure 90 milliliters of room temperature water and pour into sealable container.

2. Add “Other Ingredient (stabilizer)” and shake or use whisk to stir until incorporated.

3. Measure 10 milliliters of oil and add to sealable container.

4. Seal container and shake vigorously for 20 seconds.

5. Immediately pour the emulsion into a clean, dry 100 milliliter graduated cylinder. If emulsion
is over 103 milliliters, the team will receive 0 points for the emulsion stability score.

6. Timing will begin when the team as soon as the team finishes pouring and sets down the
container.

7. Observe the emulsion stability (degree of phase separation as shown by a growing oil phase on
the top) at 0, 2, 5, 10 and 15 minutes by recording the volume of the lipid phase in a data table.

8. Wash all containers with soap and warm water to remove all residues.

Record Data

1. Qualitative observations during the experiment

2. Quantitative observations during the experiment

Results

1. Prepare a data table showing the change in volume of oil phase (milliliters of oil on top of
emulsion) as time increases for each emulsion trial.

2. Make a graph from your data tables that show all 6 emulsion trials. Plot Volume of Oil Phase
(y) vs. Time (x).

3. Calculate the rate of phase separation as (Δ milliliters/ Δ Time)

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Biochemical test on food products

The foods you eat are made of organic compounds such as carbohydrates, lipids, proteins, and
vitamins.

Carbohydrate is a compound of carbon and water with the basic formula CnH2On. [Note:
condensation products, such as sucrose, have one less H2O and a formula CnH2O (n-1)].

Carbohydrates are the most abundant of all carbon-containing compounds, composing nearly
three-fourths of the dry mass of all plant life on earth. Examples of carbohydrates include
glucose, sucrose (table sugar), starch, and cellulose. Lipids are compounds of fatty acids and
glycerol. Food lipids are divided into fats, which come from animal sources and are solid at room
temperature, and oils, which come from plant sources and are liquid at room temperature.
Another type of lipid is cholesterol. Cholesterol is a sterol compound made by animals and is
used to make certain steroid hormones in the body. It is not found in plants. Proteins are complex
polymers composed of amino acids. Amino acids contain carbon, hydrogen, nitrogen, and
sometimes sulfur and serve as the monomers for making peptides and proteins.

You can perform chemical tests to learn what foods contain carbohydrates, lipids, and/or
proteins.

Iodine solution is used to test the presence of starch in food. It has black color. In the presence
of starch, it will give blue black color. Benedict's solution is used to test for simple sugars, such
as glucose. It is a clear blue solution of sodium and copper salts. In the presence of simple
sugars, the blue solution changes color to green, yellow, and brick-red, depending on the amount
of sugar.

Biuret solution is used to identify the presence of protein. It is a blue solution that, when it
reacts with protein, will change color to pink-purple.

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Materials

8 test tubes, test-tube rack, lab apron, stirring rod, masking tape, plastic gloves, test-tube holder,
safety goggles, newsprint paper

Chemicals

Benedict's solution, Biuret reagent, Iodine solution and distilled water

Food Substances

Maize flour/ wheat flour, raw egg white, table sugar, vegetable oil

General Direction

Read all the directions for this activity before you begin your work.

Put on your safety goggles, lab apron, and plastic gloves.

Put 8 test tubes in your test-tube rack. Label each test tube by putting masking tape near the top
edge of the test tube. Use a pencil to write one of the seven food substances on each label. Mark
the eighth label water. The water is your control.

Experiment 2.1: Test for Carbohydrates

A number of color tests and other tests have been developed for the detection of the different
types of sugars. Tests are not available, however, for all such compounds.

Fehling's Solution Test.

This test is based on the principle of the reduction of the cupric ion to the cuprous. Solutions of
cupric sulfate and alkaline sodium potassium tartrate are mixed together and then with the
reducing sugar solution and heated. A positive reaction shows the red precipitate of cuprous
oxide, or with lesser amounts of reducing sugar, a green or red, or a reddish yellow color. This
test works for any reducing sugar, monosaccharide or oligosaccharide. Although the reaction that
occurs is rather complex, it can be illustrated as follows (AOAC 1980; Fehling 1849).

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Iodine solution test

Use a medicine dropper to put ~10 drops of each food in the test tube with the matching label.
Add 3-4 drops iodine to each test tube.

Starch is one form of carbohydrate. If the substance in your test tube contains starch, it will turn
a blue-black color when it mixes with the iodine solution.

Observe the contents of your test tubes and Record the amount of starch present (0, +, ++, +++, +
+++) in your data chart. The food which contains the most starch should be recorded as ++++.

Empty and wash each test tube and return it to your test tube rack.

Benedicts solution test

Use a medicine dropper to put ~10 drops of each food into the test tube with the matching label.
Add 10 drops of Benedict's solution to each test tube. CAUTION: Benedict’s solution is
poisonous. Do not get any in your mouth and do not swallow any!

Use a test-tube holder to carefully place the test tubes in the hot water bath. Heat the test tubes
for 2 to 3 minutes. CAUTION: Use a test-tube holder to handle hot test tubes. Point the open
end of a test tube away from yourself and others.

Use a test -tube holder to return the hot test tubes to the test-tube rack. If the substance in your
test tube contains sugar, Benedict's solution will change color. See Table 1 below:

Appearance of Substance after Adding Benedict's Solution

Amount of 0 + ++ +++ ++++

Sugar in Food none trace little sugar some sugar much sugar

Color blue blue/green green Yellow orange/red

Observe your test tubes (using white paper as a background). Record the amount of sugar
present, in your data table.

Empty your test tubes, clean them thoroughly, and return them to the test tube rack.

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Tests for Proteins

Biuret Test

The Biuret Reaction. This test for the presence of the peptide linkage is obtained when solutions
of proteins or polypeptides are treated with a dilute solution of cupric sulfate followed by
addition of strong alkali (20% NaOH). The resulting color varies from bluish violet to pinkish
violet. It is obtained with all native proteins and the bulk of their split products. The peptide bond
forms a coordination compound with the Cu2+, which is the cause of the color. Since two or more
peptide linkages are necessary to produce the color, a dipeptide will not give the result.

Use a medicine dropper to put ~10 drops of each food on the test tube with the matching label.
Use a medicine dropper to carefully add 10 drops of Biuret reagent to each test tube.
CAUTION: Biuret reagent can burn your skin. Wash off spills and splashes immediately with
plenty of water while calling to your teacher.

Observe the contents of each test tube (using white paper as a background). If the food contains
proteins, it will turn a pinkish purple. Record the amount (0, +, ++, +++, ++++) of protein for
each food substance in your data table. The food which contains the most protein should be
recorded as ++++.

Empty the test tubes and clean them thoroughly. Before leaving the laboratory, clean up all
materials and wash your hands thoroughly.

Exercise

Sugar and Starch - Glucose Test

1. Crush half a saltine cracker into fine powder with the back of a spoon,
then mix it with a teaspoon of water in a cup
2. Chew the other half of the saltine for at least one full minute, and then
spit it out into another cup. (Yes, it's gross. Oh, the things we must do
for the sake of science! Don't worry, it's worth it.)

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3. Fill two test tubes with an inch of Benedict's solution each. To one add 15 drops of the
saltine-and-water solution, to the other add the saltine-and-saliva solution. (Use two
different pipets and label the test tubes so you can tell them apart.)
4. Fill a glass with boiling water and then set the test tubes in it and wait for three minutes.
Remove the test tubes from the hot water and allow them to cool. Swirl the contents and
observe the color of the liquid. Is there a difference between the two? If there is
difference, explain why that difference observed

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Liquid milk adulteration test

Specific gravity is the relation between the mass of a given volume of any substance (milk) and
that of an equal volume of water at the same temperature. Its test determines level of solids, i.e.,
it is used as main indicator for fraudulent behavior by the milk suppliers (added water, or
removal of fat). The specific gravity lies in the range of 1.030-1.035 with an average of 1.032.

Requirements: Digital Gibertini-DensiMat and Alcomat-2, 97% alcohol, 100ml graduated

cylinder, distilled water, liquid milk, thermometer,

Procedures
1. Calibrate Digital Gibertini by transferring 70 ml of 97% alcohol into 100 ml graduated
cylinder
2. Insert the electrode and the temperature analyzer into the cylinder
3. Check the Gibertini for calibration with 97% alcohol at room temperature
4. Rinse temperature analyzer and graduated cylinder with distilled water
5. Ensure the milk sample temperature to about 20°C
6. Place 70 ml of the sample into graduated cylinder
7. Insert the electrodes of Gibertini and temperature analyzer into the sample
8. Record the reading of specific gravity
9. Rinse the electrode, temperature analyzer and cylinder with distilled water between tests
10. Put the electrode in electrode box after use.
Calculate the specific gravity and determine whether the sample adulterated or not.

Apparatus
Lactometer: this is a hydrometer (a device for measuring specific gravity) adapted to the
normal range of the specific gravity of milk. It is usually calibrated to read in lactometer degrees
(L) rather than specific gravity per se.

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 A lactometer

L
The relationship between the two is: +1 = specific gravity (sp. gr.)
1000
Thus, if L = 31, specific gravity = 1.031
 A tall, wide glass or plastic cylinder
 A thermometer (the lactometer may have a thermometer incorporated).

Procedure
1. Heat the sample of milk to 40°C and hold for 5 minutes. This is to get all the fat into a liquid
state since crystalline fat has a very different density to liquid fat, and fat crystallizes or melts
slowly. After 5 minutes, cool the milk to 20°C.
2. Mix the milk sample thoroughly but gently. Do not shake vigorously or air bubbles will be
incorporated and will affect the result.
3. Place the milk in the cylinder. Fill sufficiently so that the milk will overflow when the
lactometer is inserted.
4. Holding the lactometer by the tip, lower it gently into the milk. Do not let go until it is almost
at rest.
5. Allow the lactometer to float freely until it is at rest. Read the lactometer at the top of the
meniscus. Immediately, read the temperature of the milk; this should be 20°C. If the temperature
of the milk is between 17 and 24°C, the following correction factors are used to determine L:

Temp oc 17 18 19 20 21 22 23 24
Correction -0.7 -0.5 -0.3 0 +0.3 +0.5 +0.8 +1.1

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For example, if the lactometer reading is 30.5 and the temperature is 23°C:
Corrected lactometer = Lc = 30.5 + 0.8 = 31.3
Calculations
Calculations always use Lc, the corrected lactometer reading. To calculate the specific gravity,
divide the corrected lactometer reading by 1000 and add 1.
31.1
In our example: Sp. gr. = +1 = 1.0313
1000

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Condensed tannin content determination

Objective: After the successful accomplishment of this practice:

 Students will able be to check the tannin level on the selected grains by using qualitative
analysis.

Background
Diets rich in phytochemicals are beneficial to human health because of their significant
antioxidant properties. Tannins account for about 19% of total dietary antioxidant capacity.
Tannins, also known as condensed tannins or proanthocyanidins (PAs), are oligomers and
polymers of flavan-3-ols. Tannins are widespread throughout the plant kingdom, with diverse
biological and biochemical functions, such as protection against predation from herbivorous
animals and pathogenic attack from bacteria and fungi. Tannins in fruits, vegetables, and certain
beverages contribute the bitter flavor and astringency.

Materials and Consumables

 Grains (barley or sorghum)
Equipments and chemicals
 NaOH (caustic soda) and detergent (Dietol), Spatula, Sieves, Glass beakers

Procedure
1. Measure caustic soda ( 5g/100ml bleach)
2. Add bleach and mix
3. Count out sound kernels of selected grains
4. Add the mixed bleaching reagent
5. Shake, leave for 10 minutes
6. Wash the bleached kernels of selected sound grains with clean water
7. Blot dry

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Discussion question
1. What is the importance of tannin analysis in cereal grains
2. How can tannin affect the enzyme level of grains

Determination of total soluble solids

@ Materials and Consumables
 Fruits/vegetables
 Blender (food processor), distilled water, Chopping-board, Knife, small size beakers,
Refractometr

Procedures
1. calibrate the refractometer with known 0Brix
2. Take two fruits which have different ripening stage
3. Extract the juice.
4. place 2 to 4 drops of clear juice on the prism
5. Record Refractometer reading

Question: what do you observe about TSS of ripe and unripe fruits?

1.4. Determination of Acidity

@ Materials and Consumables

 Fruits/vegetables
 Blender (food processor), Chopping-board, Knife, beakers, conical flask, pH
meter, burette, NaOH, phenolphytaline indicator, distilled water

Procedures

1. Take fruits randomly from each treatment and macerate for juice
2. Thoroughly mix the Juice
3. Filter using muslin cloth
4. Place filtered juice in the beaker
5. Place pH meter in the beaker and record the reading
Note: the pH meter should be calibrated with known standard buffer or distilled water and
the tips shouldn't touch the edges of the container during measurement

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Determination of Titrable acidity

Materials and Consumables

 Fresh fruits

 0.1N NaOH

 Distilled water

 Flasks, Beakers, Test tubs, Muslin cloth

Procedure

1. Extract juice from each fruits and mix the beverage thoroughly and filter using muslin

cloth.

2. Take ten milliliters of filtrate and titrated with 0.1N NaOH using phenolphthalein

solutions as indicator until persistent pink color of indicator will be appeared

3. Indentify the dominant acid in the fruit/vegetable and Calculate titratable acidity using

the formula as follows and compare the result

for citrus fruits conversion factor = 0.0064

V l NaOH×0 . 064×NNaOH
% Acidity= ×100
Samplevolume

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Determination of texture (firmness) of fruits

The main reasons for measuring these properties

1. Tactile properties affect the consumer’s sensory perception and acceptance of a food
product. Once the range of sensory acceptability is determined, quality analysis test can
be constructed to distinguish between an acceptable and unacceptable product.
2. Physical properties of food affect the design of processing equipment. Quantifying these
physical properties is helpful in selecting and adjusting the equipment used to mix,
transport and package products.

Materials and Consumables

Fruits/vegetables
Texture analyzer

Procedures

1. Randomly take fruits with different ripening stage

2. Fit the appropriate probe on the Texture Analyzer
3. Place the fruit at the center of penetrator and allow the machine to penetrate
4. Record force required to penetrate the sample in N

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