SBU 3013

BIOLOGY I

Lab Report

Practical 2

Molecules of Life

LECTURER’S NAME : Azi Azeyanty binti Jamaludin

NAME MATRIC NO.

NUR SYAFIQAH SYAHIRAH BINTI D20171078222
CHE ZULKIFLI
FARAH FARIZA BINTI ESHA @ ISA D20171078227

NUR HAZWANI BINTI A. D20171078242

SHIRAJZUDEEN
NIK DIANA SAHIRA BINTI NIK D20171078251
NORAINUDIN
MUHAMMAD ZUNAWANIS IQBAL BIN D20172080339
ZAIDI
 1. Introduction

A carbohydrate is an organic compound with the general formula Cm (H2O)n, that

is, consists only of carbon, hydrogen and oxygen, with the last two in the 2:1 atom
ratio. Carbohydrates make up the bulk of organic substances on earth and perform
numerous roles in living things. The carbohydrates (saccharides) are divided into
four chemical groups: monosaccharides, disaccharides, oligosaccharides and
polysaccharides. Polysaccharides serve for the storage of energy (e.g., starch in
plants and glycogen in animals) and as structural components (e.g., cellulose in
plants and chitin in arthropods).
Structural polysaccharides are frequently found in combination with
proteins (glycoproteins or mucoproteins) or lipids (lipopolysaccharides). The 5-
carbon monosaccharide ribose is an important component of coenzymes (e.g., ATP,
FAD and NAD) and the backbone of the genetic molecule known as RNA. The
related deoxyribose is a component of DNA. Saccharides and their derivatives
include many other important biomolecules that play key roles in the immune
system, fertilization, preventing pathogenesis, blood clotting and development.
2. Objective
Several basic tests frequently used to identify or test for certain carbohydrates are
performed.
3. Methodology Charts
a) Molisch Test (General carbohydrate test)

Add 5 drops of
alfa naftol
Slowly place
solution 5% to Observe the
1ml
2ml sucrose 2%, colour changes
concentrated
glucose 2%, and record the
sulphuric acid
maltose 2% and results of your
into each of
starch 5% in 4 experiment.
these test tubes
different test
tubes.

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 b) Barfoed Test (Monosaccharide)

Add 2ml of
sucrose 2%,
Place 5ml
glucose 2%,
Barfoed tester Soak them for
maltose 2% Record the
into 4 30 seconds in
and starch 5% result.
different test the boil water.
into each test
tubes.
tube
separately.

c) Benedict Test (reducing sugar)

Place 1ml of sucrose 2%,

Cooling them down before you
glucose 2%, maltose 2% and
observe any changes and
starch 5% into each test tube
record the result.
separately

Add 2ml Benedict solution into

Then, heat briefly in boiling
each test tube. Mix by gently
water for two minutes.
shaking

d) Polysaccharide

Place 5 drops of sucrose 2%, Add one to two drops of

glucose 2%, maltose 2% and Iodine-potassium iodide
starch 5% onto tile patches. solution and observe

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4. Results

2.1 Carbohydrate
2.1.1 Molish Test

Discussion:

Carbohydrates undergo dehydration reactions and loss of water because the presence of
concentrated sulfuric acid. Pentoses and hexoses form five member oxygen containing rings
on dehydration. The five member ring, known as furfural, further reacts with Molisch
reagent to form colored compounds. Pentoses are then dehydrated to furfural, while hexoses
are dehydrated to 5-hydroxymethylfurfural. Either of these aldehydes, if present, will
condense with two molecules of naphthol to form a purple-colored product. A positive
reaction is indicated by appearance of a purple ring at the interface between the acid and test
layers.

Monosaccharides give a rapid positive test. Glucose are monosaccharide. Disaccharides and
polysaccharides react more slowly than monosaccharide. Sucrose and Maltose are
disaccharide which also gave purple color ring. Starch gave slightly purple color because
they are polysaccharides.

Conclusion:

Sucrose, glucose, maltose and starch all are carbohydrates which give positive test for
Molisch test.

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2.1.2 Barfoed Test

Result:

Formation of red precipitate

Sucrose 2% - No
Glucose 2% - Yes
Maltose 2% - No
Starch 5% - No

Discussion:

Barfoed’s test distinguishes monosaccharides from disaccharides. Positive test for

monosaccharides is the appearance of red precipitate (Cu2O) within 1-2 minutes. If no
precipitate formed, indicates the presence of disaccharide. The red precipitate come from the
reaction between the reduction of copper (II) acetate to copper(I) oxide (Cu2O). RCHO +
2Cu2+ + 2H2O > RCOOH + Cu2Ov + 4H+ The aldehyde group of the monosaccharide
which normally forms a cyclic hemiacetal is oxidized to the carboxylate. Glucose are
monosaccharides show positive result in this test. Reducing disaccharides undergo the same
reaction, but do so at a slower rate. So, the timing to heat the sample is set to 3.5 minutes.
However, the samples are heated no more than 3.5 minutes to prevent the disaccharide
breaking down to monosaccharide. Sucrose, maltose, and starch showed negative result in
this test.

Conclusion:

Only monosaccharide will give an immediate red precipitate in Barfoed’s test that is glucose,
the other remaining solutions which are maltose, sucrose, and starch do not show any
changes.

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2.1.3 Benerdict test
Result:

Result of colour of the solution

Sucrose 2% - Green precipitate

Glucose 2% - Brick red precipitate are formed
Maltose 2% - Orange red precipitate formed
Starch 5% - Light blue precipitate

Discussion:

The Benedict’s test is used to detect the presence of reducing sugars (sugars with a free
aldehyde or ketone group) such as glucose. All monosaccharides are reducing sugars; they
all have a free reactive carbonyl group. Some disaccharides have exposed carbonyl groups
and are also reducing sugars. Maltose which is disaccharides also called reducing sugar as it
has the exposed carbonyl groups. Other disaccharides such as sucrose and starch are non-
reducing sugars and will not react with Benedict’s solution. Benedict’s reagent is a mild
oxidant with CuSO4, Cu (II) sulfate, as one of the reagents. In the presence of a reducing
sugar, the blue solution of Cu (II) or Cu+2, is changed to a brick red/brown precipitate of
Copper (I) or Cu+1 oxide,Cu2O. If there a small or large amount of the reducing sugar
present, the color would range from green to brick red respectively. RCHO + 2Cu2+ +
4OH- > RCOOH + Cu2O + 2H2O Sucrose indirectly produces a positive result with
Benedict’s reagent if heated with dilute hydrochloric acid prior to the test, although after
this treatment it is no longer sucrose. The addition of HCl hydrolysed the non-reducing
sugar, as it split it up into its component monomers.

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The monomers are reducing sugars which gave the positive result on the second reducing
sugar test. The acidic conditions and heat break the glycosidic bond in sucrose through
hydrolysis. The products of sucrose decomposition are glucose and fructose, both of which
can be detected by Benedict’s reagent, as described above. This same goes for starch. But
since starch has larger component compare to sucrose so it took a longer time to hydrolyse.
That the purpose of heat it in longer time compare to sucrose. Without the addition of acid
to sucrose solution, starch solution, the test given is negative. The solutions remain clear
blue after the addition of Benedict’s reagent and heating.

Conclusion:

Benedict’s test is the common test which is used to determine the existence of reducing
sugar. Maltose and glucose are reducing sugars which give positive test. Starch and sucrose
are non reducing sugars which give positive results after adding hydrochloric acid.

2.1.4 Polysaccharide test

Result :

Discussion:

Starch gives positive result in Iodine test as the color of solution change from yellow to dark
blue. The immediate formation of a vivid blue color indicates amylose. Vivid blue
coloration forms due to the polyiodide complex formed. Cellulose is derived from D-
glucose units, which condensed through beta(1->4)-glycosidic bond. This give cellulose to

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be a straight polymer therefore, it can’t coil around iodine to produce blue colour as starch
does. Only starch gives the color of vivid blue, this is because it contains amylase. The
iodine molecules slip inside of the amylase coil. The amylose, or straight chain portion of
starch, forms helices where iodine molecules assemble, forming a dark blue color.

Conclusion:

The polysaccharide test is used to test for the presence of starch. Starch is a type of
polysaccharide carbohydrate which is made up of amylose and amylopectin. It is one of the
main sources of carbohydrate and present naturally in plant. Amylose in starch form dark
blue complex with iodine.

5. Reference

a) Chemistry LibreTexts. (2019). Introduction to Carbohydrates. [online] Available at:

https://chem.libretexts.org/Bookshelves/Biological_Chemistry/Supplemental_Modul
es_(Biological_Chemistry)/Carbohydrates/Carbohydrates_Fundamentals/Introductio
n_to_Carbohydrates

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 1. Introduction

Proteins are polymers of amino acids. Twenty different types of amino acids occur
naturally in proteins. Proteins differ from each other according to the type, number
and sequence of amino acids that make up the polypeptide backbone. As a result
they have different molecular structures, nutritional attributes and physiochemical
properties. Proteins are important constituents of foods for a number of different
reasons. They are a major source of energy, as well as containing essential amino-
acids, such as lysine, tryptophan, methionine, leucine, isoleucine and valine, which
are essential to human health, but which the body cannot synthesize.

Proteins are also the major structural components of many natural foods,
often determining their overall texture, e.g., tenderness of meat or fish products.
Isolated proteins are often used in foods as ingredients because of their unique
functional properties, i.e., their ability to provide desirable appearance, texture or
stability. Typically, proteins are used as gelling agents, emulsifiers, foaming agents
and thickeners. Many food proteins are enzymes which are capable of enhancing the
rate of certain biochemical reactions. These reactions can have either a favorable or
detrimental effect on the overall properties of foods. Food analysts are interested in
knowing the total concentration, type, molecular structure and functional properties
of the proteins in foods.

2. Objective
Several basic tests frequently used to identify or test for certain protein are
performed by students.

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3. Methodology Charts
a) Biuret test

Add four
Place 3ml
(4) drops
albumin Add 3ml
cuprum
solution Sodium
sulphate Record
5%, and Hydroxide
0.5% and the result
milk into 3 2% and
observe
different mix gently.
the colour
test tubes.
changes.

b) Millon test

Place 3ml
albumin
Add three (3) Observe and
solution 5%,
drop of Millon record the
and milk into
solution result.
3 different
test tubes

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 4. Results

a) Biuret test

Before

After

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 OBSERVATION INTERPRETATION
Milk ( slightly change to light purple) Have protein but cannot analysed by
biuret test
Albumin ( change from chalky to Proteins are present
purple colour)

b) Millon test

Before

After

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OBSERVATION INTERPRETATION

Milk ( milky white) Presence of protein

Albumin ( cloudy solution) Presence of protein

5. Discussion

From this experiment we can see that protein are important constituents of foods for
a number of different reasons. They are a major source of energy, as well as
containing essential amino-acids, such as lysine, tryptophan, methionine, leucine,
isoleucine and valine, which are essential to human health, but which the body
cannot synthesize. To test the presence of the protein, in this experiment, we do
biuret test and Millon test

a) Biuret test
Biuret test is used biuret reagent which is (copper sulfate in a strong base) reacts
with peptide bonds in proteins to form a blue to violet complex known as the “biuret
complex”. From this experiment, the result of milk is from chalky colour turn to
light purple. This is because, the milk already had a process so that the cassien
protein will mix the milk serum. So the level of protein will be low and milk cannot
be test the accurate protein milk. So mixture of milk cannot be analysed by doing
biuret test. For albumin, we can see that from chalky colour turn to purple colour and
purple colour indicates the presence of protein in this experiment.

b) Millon test
Millon’s test is to detect amino acid containing phenol group ( hydroxyl group
attached to benzene ring). Millon's reagent (Hg/HNO3) gives positive results with
proteins containing the phenolic amino acid “tyrosine”. From this experiment, we
can see that albumin in the Millon test, the white precipitate is formed when millon
reagent is added into a protein. It will turn red brick precipitate when you heat it but

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in this experiment we just see the white precipitate. The white precipitate indicates
the high presence of protein. For milk in the Millon test, the clear cloudy is form and
indicate the presence of the protein.

6. Conclusion

For the conclusion, protein plays a very vital role in our body by having different
function in depending upon the direction of each. Protein, which are basically a
polymers of amino acids give us an insight on how the different R groups in each
amino acids dictate the proteins structure into primary, secondary, tertionary and
quaternary structure of protiens. This experiment which is Biuret test and Millon’s
test is example to test the presence the protein in a given sample. Moreover, the
experiment gives a closer look to what is the eefect of the different denaturing agents
to a protiens.

7. Reference

a) People.umass.edu. (2019). 6. ANALYSIS OF PROTEINS. [online] Available at:

https://people.umass.edu/~mcclemen/581Proteins.html
b) Biuret Test for Protein. (2019). Retrieved from
http://brilliantbiologystudent.weebly.com/biuret-test-for-protein.html
c) (2019). Retrieved from,
http://drugresearch.in/pdf/2019/jan-march/IJOD-5-Anita%20Maurya.pdf

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 1. Introduction

Lipids are one of the major constituents of foods, and are important in our diet for a
number of reasons. They are a major source of energy and provide essential lipid nutrients.
Nevertheless, over-consumption of certain lipid components can be detrimental to our
health, e.g. cholesterol and saturated fats. In many foods the lipid component plays a major
role in determining the overall physical characteristics, such as flavor, texture, mouthfeel
and appearance. For this reason, it is difficult to develop low-fat alternatives of
many foods, because once the fat is removed some of the most important physical
characteristics are lost. Finally, many fats are prone to lipid oxidation, which leads to the
formation of off-flavors and potentially harmful products. Some of the most important
properties of concern to the food analyst are:

 Total lipid concentration

 Type of lipids present
 Physicochemical properties of lipids, e.g., crystallization, melting point, smoke point,
rheology, density and color
 Structural organization of lipids within a food

2. Methodology Charts
a) Acrolein Test

Place five (5)

drops of
Add 1g Sodium
coconut oil, Indicate the
bisulphate and
palm oil and smell presence
heat slowly
vegetable oil and record the
using Bunsen
into three result
burner.
different test
tubes.

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 b) Colouring Test

Add a few
Place three
drops of
(3) drops of
Sudan IV.
coconut oil, Add 5ml distil
Carefully mix
palm oil and water and mix
by hard
vegetable oil by gently
shaking. Leave
into three shaking.
them for 1
different test
minute and
tubes
observe

3. Results
a) acroline test

b) colouring test

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sample Acroline test Colouring test
Coconut oil Burnt grease Reddish-orange
Palm oil Pungent smell Light-orange
Vegetable oil Pungent smell Reddish-orange

4. Discussion

Acrolein Test is the simplest unsaturated aldehyde. It is a relatively electrophilic

compound and a reactive one, hence its high toxicity. This reaction is to
determine the presence of glycerin in a fat. By heating the fat sample in the
presence of sodium bisulfate (NaHSO4), which acts as a dehydrating agent,
acrolein is formed and can easily be detected by its odor. Whenever fat is heated
in the presence of a dehydrating agent, the fat molecule will shed its glycerol in
the form of the unsaturated aldehyde–acrolein. From the experiment, we can see
that, the smell presence of coconut oil is likely burnt grease which is tolerable,
smell presence for palm oil is pungent smell and for vegetable oil , the smell
presence is pungent smell.

In colouring test, we can detect them by colouring after adding Sudan IV into the
sample. Sudan IV is a soluble in water and still soluble in lipid. Sudan IV will
stain them reddish-orange or light-orange which is the result get is a positive
result.

5. Conclusion

As the conclusion, during the experiment about the reaction of lipids, the group
was able to determine the and observe the different characteristics of the test
used to know the lipid’s unsaturation and the solubelity. Different test has
varying appearance and odor that distinguishes them from the rest. The group
was able to achieve the objective of knowing the effect of test used for the lipid’s
unsaturation. By use different test, we can get variety of result.

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6. Reference
a) People.umass.edu. (2019). ANALYSIS OF LIPIDS. [online] Available at:
https://people.umass.edu/~mcclemen/581Lipids.html

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