Title : Biochemical test in food

Aim :

To identify the presence of lipid, protein and carbohydrates in food samples.

Problem Statement :

What is the nutrient that contain in the food samples given?

Theory :

Carbohydrates, proteins, lipids, and vitamins and minerals are the nutrients in food.
There are three types of carbohydrates. First is monosaccharides or simple sugar consisting
of a single sugar molecule such as glucose, fructose and galactose. Then, two
monosaccharides joined together to form disaccharides such as sucrose is a combination of
glucose and fructose. Lastly, polysaccharides, the complex carbohydrates like starch and
glycogen. They are major sources of energy for cellular work. Common sources of
carbohydrates in food are rice, breads and cereals.

Proteins are chains of amino acids. Proteins have numerous functions. They are the
basis for tissue and organ structure. Proteins also act as hormones and enzymes. Food
sources of proteins include fish, soybeans, meat and dairy products. Lipids are hydrophobic
where they are insoluble in water. Lipids include fats, oils, waxes, phospholipids, and
steroids. Concentrated sources of energy, each gram of lipid has more calories than a gram
of protein or carbohydrates. Lipids provide support for joints, tendons, and internal organs.
Food sources of lipids include nuts, meat, butter and cheese.

Simple chemical test using indicator can be used to determine the presence of
nutrients in food. A colour change of an indicator is usually a positive test for the presence of
a certain nutrient. In this experiment, we will use several indicators to test for the presence of
nutrients in food samples. To test the presence of protein, we use Millon. While to test the
presence of lipid, we use Ethanol. This test is also called as Emulsion test. Then, to test the
presence of carbohydrates, we use two solutions. We use Iodine to test the presence of
polysaccharides (starch) and Benedict to test the presence of reducing sugar which consists
of monosaccharides (glucose) and disaccharides (except sucrose).

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Materials :

Sample food A, B, C, D, E, F and G, iodine solution, benedict solution, millon, ethanol and
distilled water.

Apparatus :

Test tube, boiling tube, boiling tube holder, beaker, dropper, Bunsen burner and tripod stand.

Procedures :

1. Seven samples of unknown food, A, B, C, D, E, F and G were prepared by the lecturer.

2. All of the test tube were sterile before we use.

A. Identification of carbohydrates (monosaccharides) using Benedict solution

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1. Boiling water bath were set up using beaker filled with 4 water.

2. About 5 ml of every sample was added into test tube and labelled with A, B,
C, D, E, F and G.
3. A few drops of Benedict solution were added to each test tube and shake so
that the sample and benedict solution is well mixed.
4. All the test tubes were put into the boiling water for 5 minutes and the result
had recorded.

B. Identification of carbohydrates (polysaccharides) using Iodine solution

1. About 5 ml of every sample was added into test tube and labelled with A, B,
C, D, E, F and G.
2. A few drops of Iodine solution were added to each test tube.
3. Shake the test tube so that the sample and Iodine solution is well mixed.
4. The result was recorded.

C. Identification of proteins using Millon

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1. Boiling water bath were set up using beaker filled with 4 water.

2. About 5 ml of every sample was added into test tube labelled with A, B, C, D,
E and F.
3. A few drops of Millon were added to each test tube.
4. Shake the test tube so that the sample and Millon is well mixed.
5. All the test tubes were immersed into the boiling water bath for 5 minutes and
the result was recorded.

D. Identification of lipids using Ethanol

1. About 5 ml of every sample was added into test tube labelled with A, B, C, D,
E and F.
2. A few drops of Ethanol were added to each test tube.

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 3. Shake the test tubes so that the sample and Ethanol is well mixed
4. Add about 3 ml of distilled water..
5. Shake the test tube again.
6. The result was recorded.

Observations and Analysis :

Solution Samples Observation Analysis

A The colour turn to purple Reducing sugar absent

Benedicts
solution

B White precipitate form in a light blue Reducing sugar absent

solution

C No change Reducing sugar absent

D The colour turn to yellow Reducing sugar absent

E No change Reducing sugar absent

F The colour turn to green Reducing sugar absent

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 G Brownish-orange precipitate formed Reducing sugar
present

Iodine A The colour turn to brownish-orange Starch absent

solution

B The colour turn to dark blue Starch present

C The colour turn to dark green Starch absent

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 D No change Starch absent
E No change Starch absent
F No change Starch absent
G No change Starch absent
Millon A Brick red precipitate formed Protein present
solution

B No change Protein absent

C No change Protein absent
D No change Protein absent
E Brick red precipitate formed Protein present

F No change Protein absent

Ethanol A Jelly like precipitate formed Lipid absent

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B No change Lipid absent
C No change Lipid absent
D No change Lipid absent
E A layer of cloudy white suspension Lipid present
formed. Tiny globules of fat
suspended in the solution.

F A layer of cloudy white suspension Lipids present

formed. Tiny globules of fat
suspended in the solution.

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Discussion :

For Test 1, we use Benedicts solution to test the presence of reducing sugar.
Benedict solution will change from blue to green or orange or brown, depending on the
amount of reducing sugar present. So, from the test, we find out that only sample G contains
reducing sugar. When we add Benedict solution into the sample G and put the test tube into
the boiling water, brownish-orange precipitate was formed. So, it is proven that reducing
sugar (glucose) is present in sample G.

For test 2, we use Iodine solution to test the presence of starch. Iodine solution will
change from brown to purple or dark blue, depending on the amount of starch present. So,
based on the test, we find out that only sample B contains starch. When we add Iodine
solution into the sample B, the colour turned to dark blue. So, it is proven that starch is
present in sample B.

For test 3, we use Millon reagent to test the present of protein. If proteins are present
in the sample, red brick precipitate will form if protein is present. So, from the test, we find
out that sample A and E contain protein. When we add Millon reagent into the samples and
immersed into boiling water, brick red precipitate formed. So, it is proven that protein is
present in sample A and sample E.

For test 4, we use Ethanol to test the present of lipid. If lipids are present in the
sample, a layer of cloudy white suspension will form. Tiny globules of fat suspended in the
solution. Based on the test, we find out that sample E and sample F contain lipids. When we
add Ethanol into the sample, the lipid in the sample is dissolve and when we add water, they
form tiny dispersed droplets in the water. It is called an emulsion. So, it is proven that lipid is
present in sample E and sample F.

When we carried out the experiment, a few of precautions must be taken. Firstly, we
must sterile all the test tube before we use, to make sure that there is no impurities that will
affect the result of experiment. Then, we used rubber glove to avoid direct contact with
Millon reagent. Besides, we handled the apparatus carefully especially when we use the
Bunsen burner to avoid injuries. Lastly, we labelled all the test tube to easily recognise the
sample of A, B, C, D, E, F and G.

During the experiment, we had face some problems and weakness. However, we
managed to overcome these problems and weaknesses. The weakness is, we did not do the
revision on the procedures of the experiment. So, we did not know the steps to carry out the

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experiment. To solve this problem, we find the information on the internet. So, we manage to
carry out the experiment.

Conclusion :

Based on the experiment, we can conclude that sample A contains protein, sample B
contains starch, sample E contains protein and lipid, sample F contains lipid and sample G
contains reducing sugar (glucose).

References :

Atifah Ruzana (2015). Laporan Amali Biologi: Ujian Pemakanan. Retrieved from
http://www.slideshare.net/eepapoppins/laporan-amali-biologi-ujian-makanan

Charlie L, Jacqueline R.S & et.al. (2010). STPM Text Biology. Selangor: Oxford Fajar.

Ching, L. (2011). Pre-U Text STPM Biology Volume2. Selangor: Pearson Malaysia Sdn Bhd.