SCES3013 Physiology and epidemiology

Title :

Biochemistry tests of organic molecule-carbohydrates, lipid and protein.

Aim :

To test for the presence of carbohydrates, protein and lipid in burger

Objectives :

1. To test the presence of carbohydrates, protein and lipid in the food sample.
2. To construct experiment and get accurate results
3. To apply scientific skills during the experiment.

Problem statement :

Do carbohydrates, protein and lipid present in the burger?

Hypothesis :

The burger contains carbohydrates, protein and lipid.

Materials / apparatus :

Burger, water, Benedict’s solution, Iodine solution, hydrochloric acid, sodium

bicarbonate powder, potassium hydroxide solution, copper (II) sulphate solution,
Bunsen burner, test tube, boiling tube, tripod stand, filter paper, beaker, dropper, wire
gauze, test tube rack, measuring cylinder, test tube holder, pH paper, glass rod
Procedure :

A. Iodine test for starch

Apparatus set-up:

Figure 1

Procedure :

1. The burger is crushed into crumbles and inserted into a test tube.
2. A few drops of Iodine solution is added into the crumbs.
3. The test tube is placed within the test tube rack.
4. The colour change of iodine solution is observed and the result is recorded
in Table 1.
Result :

Types of Reagent Observation Inference

biochemistry
tests
Iodine test Potassium The colour of iodine Starch is present.
triiodide solution changes from
solution brown colour to blue
black.

Table 1
 B. Benedict’s solution test for reducing sugar
Apparatus set-up:

Figure 2

Procedure :

1. The burger is crushed into crumbles and inserted into the boiling tube.
2. A few drops of Benedict’s solution is added into the boiling tube.
3. The mixture is shaken and then heated in a water bath for a few minutes until
the colour of Benedict’s solution changes.
4. The boiling tube is placed in the test tube rack.
5. The colour of the precipitate is observed and recorded in Table 2.
Result :

Types of Reagent Observation Inference

biochemistry
tests
Benedict’s test Copper(II) The colour of Benedict’s Burger contains
sulphate, solution changes from moderate
sodium blue to the formation of amount of
citrate and
orange precipitate. reducing sugar.
sodium
carbonate

Table 2
 C. Benedict’s solution for non-reducing sugar
Apparatus set-up :

Figure 3

Procedure :

1. The burger is crushed into crumbles and inserted into boiling tube.
2. A few drops of Benedict’s solution is added into the boiling tube.
3. 1 ml of hydrochloric acid is added into the boiling tube.
4. The mixture is shaken and heated in a water bath.
5. The mixture is left to cool.
6. The sodium bicarbonate powder is added to neutralise the acid.
7. The pH paper is dipped into the boiling tube.
8. A few drops of Benedict’s solution is added.
9. The mixture is slowly shaken
10. The boiling tube is heated in the water bath.
11. The boiling tube is placed within the test tube rack.
12. The colour change of Benedict’s solution is observed.
13. The result is recorded in Table 3.
Result :

Types of Reagent Observation Inference

biochemistry test
Benedict’s test 1. Hydrochloric An orange precipitate Reducing sugar
acid is formed. and non-
2. Sodium reducing sugar
bicarbonate
are present.
powder
3. Copper(II)
sulphate,
sodium
citrate and
sodium
carbonate

Table 3
 D. Biuret test for protein

Apparatus set-up

Figure 4

Procedure :

1. The burger is crushed into crumbles and inserted into the test tube.
2. 2 ml potassium hydroxide is measured using the measuring cylinder.
3. The mixture is shaken.
4. 2 ml copper (II) sulphate is slowly added drops by drops into the mixture.
5. The mixture is stirred using the glass rod.
6. The test tube is placed in the test tube rack.
7. The colour change is observed and the result is recorded in Table 4.
Result :

Types of Reagent Observation Inference

biochemistry test
Biuret test 4. Copper The purple colour is Burger contains
(II) only shown in a little less amount of
sulphate amount. However, protein.
5. Potassium
there is more yellow
hydroxide
residue.

Table 4
 E. Grease spot test for the presence of lipid
Apparatus set-up

Figure 5

Procedure :

1. The burger is crushed into crumbles and placed on the litmus paper.
2. The filter paper is left under the fan in order to become dried faster.
3. The oil patch can be seen on the filter paper under the sunlight.
4. The result is recorded in Table 5.
Result :

Types of Material Observation Inference

biochemistry
test
Grease spot test Filter A translucent spot is Burger contains lipid.
paper formed on the filter paper.

Table 5
Discussions :

Our group uses Mc Chicken Deluxe as our food sample. The burger contains
two pieces of breads, tomato, chicken, vegetables, some mayonnaise and a slice of
cheese. We carried out several food tests in order to test for the presence of
carbohydrates, protein and lipid.

We carried out a few precaution steps while doing this experiment. In the
Benedict’s test for non-reducing sugar, we use pH paper to ensure neutralisation
process occurred between the chemical reaction of hydrochloric acid and sodium
bicarbonate. We also use water bath when heating the boiling tube to avoid direct
heating and high temperature that may lead to the cracking of test tube. Water bath is
also used to ensure the content in the test tube does not spill out. We also handle
poisonous chemical such as hydrochloric acid and Iodine solution with care in the
experiment. Besides, we also faced problem such as malfunctioning of Bunsen burner.
We replaced it with a new Bunsen burner.

Iodine test is carried out to test the presence of starch in burger. The potassium
triiodide is used as a reagent in the test which then dissociates to form iodine
molecules, which gives the colour of brown apart from colourless triiodide pentaiodide
ions and iodide. The tri-iodide and penta-iodide ions formed are linear. Starch in the
food sample contains the structure of helical amylose where the ions slip inside the
helix of the amylose to form the starch-iodide complex in the form of charge. Electrons
are transferred between the starch and iodide ions. The electrons are excited and
promote the energy levels between amylose and iodide ion. Light is absorbed at a
different wavelength and turn the iodine solution into blue-black. This indicates that
starch is present in the burger. The presence of starch might be due to the bread
crumbles in the burger. Starch is actually polysaccharide or complex carbohydrate
which results from the condensation of glucose unit. Hence, the intake of carbohydrate
can act as an energy source for physical activities such as running and climbing.

We also carried out food test for the presence of reducing sugar by using
Benedict’s solution. The Benedict’s solution is originally blue and made up of copper
(II) sulphate, sodium carbonate and sodium citrate. The copper (II) sulphate acts as a
reducing agent and determines the colour change of Benedict’s solution. The sodium
carbonate acts as an alkali medium which are required for the redox reaction. While,
sodium citrate acts as a stabilizer so that copper (II) ions are not reduced to copper (I)
during storage. Under the alkaline solution, the reducing sugar and form enediols
which act as reducing agents. The blue copper (II) sulphate in Benedict’s solution is
reduced to copper (I) oxide which causes the formation of orange precipitate. The
positive result may be due to monosaccharide or simple carbohydrate in glucose form
from the bread crumbles. The orange colour indication also shows the approximate
glucose content in the burger is about 1500 to 2000 mg/dl.

Color Approximate glucose mg/dl Indication

Blue solution Nil

Green solution <500 mg/dl Trace

Green ppt 500-1000 mg/dl +

Yellow ppt 1000-1500 mg/dl ++

Orange ppt 1500-2000 mg/dl +++

Red to Brick red ppt >2000 mg/dl ++++

Source: Dhurba Girl.(2016,Jun 19)

While, in Benedict’s test for non-reducing sugar, we added some hydrochloric

acid and sodium bicarbonate powder. The hydrochloric acid is used to break the
glycosidic bond in sucrose through hydrolysis process into monosaccharide. The
monosaccharide has the aldehyde functional group and can reduce blue copper (II)
ions to red copper (II) ions in the presence of alkali. While, the sodium bicarbonate
powder acted as an alkali to neutralise the hydrochloric acid. It is added before the
Benedict’s solution. An orange precipitate is formed. The result shows the presence
of reducing and non-reducing sugar. The reducing sugar is present due to the bread
crumbles, while the non-reducing sugar is present due to the vegetables and tomato.
Both reducing and non-reducing sugar are actually simple carbohydrates.

Biuret test was also carried out to test for the presence of protein in the burger.
The reagents used in Biuret test are copper (II) sulphate and potassium hydroxide.
Potassium hydroxide provides an alkaline medium for the presence of peptide bonds,
while copper (II) sulphate solution produces copper (II) ions. The copper (II) ions form
coordination complex with 4 nitrogen atoms from peptide bonds. The complex of
copper (II) ions and nitrogen atoms lead to the colour change from blue to purple. For
our food sample, the content of protein might be very little as a little purple colour is
only shown. The content of protein should be increased as it is needed for growth and
repair of body tissues. The low amount of protein might be due to a few reasons. Firstly,
even the burger contained chicken, the protein might be denatured earlier on due to
the high temperature when processed in the factory. Secondly, we might not be adding
sufficient amount of potassium hydroxide solution until it covers the surface of the
mixture. The mixture might not be shaken well until a visible purple colour was shown.
The presence of peptide bond might be insufficient for the colour change. The quantity
of chicken might also not be enough when we crushed into burger crumbles.

Apart from that, the grease spot test is carried out to test for the presence of
lipid in the burger. Lipid is a derivative from glycerol and contains fat. It has a higher
boiling point and are does not evaporate easily. We have dropped some water on the
burger crumbles. The spot of water absorbs enough water and evaporate. However,
the heat is not enough for the grease to evaporate. Thus, a translucent spot is
observed under the sunlight when the water absorbed in the filter paper diffracts light.
Light can pass through the paper and form translucent spot. The result might be due
to the cheese, chicken and mayonnaise which contain lipid. The intake of lipid serves
as an insulator in our body. It also functions as energy storage molecules, structural
component of phospholipid bilayer in the membrane and cell signalling molecule.

In conclusion, Mc Chicken Deluxe contains carbohydrates, protein and lipid.

However, burger contains high amount of carbohydrates and less amount of protein.
Excess intake of carbohydrate might lead to health problem, such as diabetes mellitus
II. While, excess intake of lipid might lead to obesity. The accumulation of fat might
block the arteries and cause cardiovascular diseases. Thus, burger is considered not
a healthy food and does not provide a balanced diet.

Conclusion :

Mc Chicken Deluxe contains carbohydrate, protein and lipid.

References :

Brilliant biology student.(2015).What is the iodine test for starch. Retrieved from
http://brilliantbiologystudent.weebly.com/iodine-test-for-starch.html

Brilliant biology student.(2015).What is Benedict’s test for non-reducing

sugar.Retrieved from http://brilliantbiologystudent.weebly.com/benedicts-test-for-n
on-reducing-sugars.html.

Dhurba Girl.(2016,Jun 19).Benedict’s test principle, reagent preparation, procedure

and interpretation. Retrieved from http://laboratoryinfo.com/benedicts-test-
principle-reagent-preparation-procedure-interpretation/

Gan Wat Yeat.(2016).Success Biologi SPM.Selangor Darul Ehsan:Oxford Fajar Sdn.

Bhd..

Yeap Tok Kheng.(2011).Longman essential science PMR.Selangor Darul

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