TOPIC : Biological Molecules

AIM : Identification of Biochemicals in Pure Form

APPARATUS :

1. Ph Paper 11. Iodine

2. Test Tube 12. 1 % Starch Solution

3. Test – Tube Rack 13. 1 % Glucose Solution

4. Bunsen Burner 14. 1 % Sucrose Solution

5. Teat Pipettes 15. Olive oil or Corn oil

6. 1cm Syringe 16. Absolute Ethanol

7. Benedict’s Reagent 17. Egg Albumen

8. Sodium Hydrogen 18. 1 % Lactose Solution

Carbonate

9. 5 % Potassium 19. 20. 1 % Fructose Solution

Hydroxide Solution
10. 1 % Copper Sulphate 20. Dilute Suphuric Acid
Solution
METHOD :

CARBOHYDRATES – REDUCING SUGARS

1. 2cm3 of a solution of reducing sugar was added to a test – tube. An equal

volume of Benedict’s was added. It was brought gently to the boil and

shaked. To minimize spitting it wass shaked continuously.

2. CARBOHYDRATES – NON – REDUCING SUGARS

1. 2cm3 of sucrose solution was added to a test – tube. 1 cm3 of dilute

hydrochloric acid was added. It was boiled for one minute. With sodium

hydrogen carbonate ( check wit pH paper ) it was carefully neutralized.

Effervescence occurs care is required. Benedict’s Test was carried out.

STARCH - IODINE TEST

1. 2cm3 1 % starch solution was added to a test – tube. A few drops of I2 /

KI solution was added. Alternatively the latter was added to the solid form

of starch.
3.LIPIDS – EMULSION TEST

1. 2cm3 fat or oil was added to a test – tube containing 2 cm3 of absolute

ethanol. The lipid was dissolved by shaking vigorously. An equal volume

of cold water was added.

1. PROTEINS – BIURET TEST

1. 2cm3 of protein solution was added to a test – tube. An equal volume

of 5 % potassium hydroxide solution was added. 2 drops of 1 % copper

sulphate solution was added.

OBSERVATION

AND

RESULTS
 IMAGE 1 : Show My Lab – Partner and Me Holding our test – tubes

after the experiment. The test – tube that I am holding shows the results for

lipids – Emulsion Test that forms a cloudy white suspension and the other test –

tube shows the Starch – Iodine Test that forms a blue – black colouration. The

test – tube Ruben is holding shows the results for Carbohydrates – Reducing

Sugars that finally forms a brick – red precipitate and the other test – tube

shows the Proteins - Biuret Test that finally changes into a mauve or purple

colouration.
 IMAGE 2 : Shows the results of the Carbohydrates – Reducing

Sugars which involves monosacharides. The initial blue colouration of the mixture

turns green then yellowish and finally forms a brick – red precipitate.

IMAGE 3 : Shows the result of Lipids that is the Emulsion Test. It forms a cloudy

white suspension.
 IMAGE 4 : Shows the test – tube wit a pH Paper in it. This test –

tube shows the result for the Carbohydrates that is the Non – Reducing Sugars. A

Benedict’s Test was carried out and a brick – red precipitate was finally formed.

IMAGE 5 : Shows the results for the Biuret Test to test

proteins. A mauve or purple colouration was formed and observed.

 IMAGE 6 : Shows the three test – tubes with different colouration in it.

The test – tube with the red colouration shows the Benedict’s Test for reducing

sugars. It is used to test carbohydrates. It finally forms a brick – red precipitate.

The test tube with the blue – black colouration shows the Iodine Test to test the

presence of Starch. While, the test – tube with the white colouration shows the

Emulsion Test that is to test the presence of Lipids. It finally forms a cloudy white

suspension.
 IMAGE 7 : Shows the results of all the test that was carried out in the

laboratory. The test –tube with the blue – black colouration shows the test for

Iodine Test. It shows the presence of starch. The test – tube with the brick – red

precipitate shows the Benedict’s Test for non – reducing sugars. The initial blue

colouration of the mixture turns green then yellowish and finally form a brick – red

precipitate. The white colouration in the test – tube shows the Emulsion Test that

was carried out, a cloudy white suspension was observed. It used to test lipids. The

test – tube with the brick – red precipitate wit the pH Paper in it shows the

Benedict’s Test that was carried out on non – reducing sugars. The test tube with

the mauve colour at the left shows the Biuret Test to test the presence of

proteins.
ADDITONAL

NOTES

CARBOHYDRATES
A carbohydrate is an organic compound general formula Cm(H2O)n, containing the

elements carbon ( C ), hydrogen (H), and oxygen (O ) in the proportion 1 : 2 : 1. In

general, carbohydrates play three important roles in cells :

 The main source of energy which is a simple carbohdydrates

 Acts as a food storage

 It forms structural components in plant cells

Carbohydrates can be divided into three main groups based on their molecular

structures :

 Monosaccharides ( Simple Sugars )

 Disaccharides ( Double Sugars )

 Polysaccharides ( Complex Sugars )

MONOSACCHARIDES
1. Monosaccharides are basic subunits for the formation of more complex

carbohydrates. These molecules cannot be broken down further into smaller

units of carbohydrates. The α and β anomers of glucose. Natural saccharides

are generally built of simple carbohydrates called monosaccharides with

general formula (CH2O)n where n is three or more. A typical monosaccharide

has the structure H-(CHOH)x(C=O)-(CHOH)y-H that is an aldehyde or ketone

with many hydroxyl groups added, usually one on each carbon atom that is

not part of the aldehyde or ketone functional group. Examples of

monosaccharides are glucose, fructose, and glyceraldehyde.

2. Monosaccharides are classified according to three different

characteristics: the placement of its carbonyl group, the number of

carbon atoms it contains, and its chiral handedness. If the carbonyl

group is an aldehyde, the monosaccharide is an aldose. However, if the

carbonyl group is a ketone, the monosaccharide is a ketose. Monosaccharides

with three carbon atoms are called trioses, those with four are called

tetroses, with five are called pentoses, six are hexoses. These two systems

of classification are often combined. For example, glucose is an aldohexose

which is a six-carbon aldehyde, ribose is an aldopentose which is a five-

carbon aldehyde, and fructose is a ketohexose which is a six-carbon ketone.

3. The classification of monosaccharides is as follows :

 Based on the number of carbon atoms in their molecules,

trioses, pentoses, and hexoses

 Based on whether they carry the aldehyde or ketone functional

groups : Aldoses and ketoses.

4. Both the functional groups, either aldehyde or ketone have reducing sugars

properties. Therefore, all monosaccharides functions as a reducing sugars.

5. Benedict’s Test is used to detect the presence of reducing sugars in a

solution. Benedict’s reagent is blue in colour because it is an alkaline solution

of copper ( II ) sulphate. Reducing sugars reduce copper ( II ) ions to

copper ( I ) ions which are orange red in colur as in copper ( II ) oxide.

6. All monosaccharides have a sweet taste, dissolve easily in water and form

white crystals readily. The sweetness and water solubility of

monosaccharides are results of the hydroxyl groups attach to the carbon

atoms.
 MAJOR FUNCTIONS OF MONOSACCHARIDES

MONOSACCHARIDES FUNCTIONS
Trioses  Important Intermediates During Respiration And

Photosynthesis

Pentoses  Ribose And Deoxyribose are components of

neulotides and therefore provide structural support

for RNA and DNA which is the components of

coenzymes involves in hydrogen transport for

example NAD, NADP and FAD which is a component

of ATP. Ribose bisphospate acts as a receiver for

carbon dioxide during photosynthesis.

Hexoses  Glucose is the most important respiratory

substrate for plants and animals and is an

intermediate source of energy for cells, that is the

raw material for synthesis of disaccharides and

certain polysaccharides. All other carbohydrates

are converted to glucose.

DISACCHARIDES
1. The general formula for a disaccharide is Cm(H2O)n, . A disaccharide is

formed from two units of monosaccharides linked together through

condensation. They are composed of two monosaccharide units bound

together by a covalent bond known as a glycosidic linkage formed via a

dehydration reaction, resulting in the loss of a hydrogen atom from one

monosaccharide and a hydroxyl group from the other. The formula of

unmodified disaccharides is C12H22O11. Examples include maltose, sucrose

and lactose. Sucrose, also known as table sugar, is a common disaccharide.

It is composed of two monosaccharides : D-glucose and D-fructose.

2. Sucrose is the most abundant disaccharide, and the main form in which

carbohydrates are transported in plants. It is composed of one D-glucose

molecule and one D-fructose molecule. The systematic name for sucrose,

O-α-D-glucopyranosyl-(1→2)-D-fructofuranoside, indicates four things :

 Its monosaccharides: glucose and fructose

 Their ring types: glucose is a pyranose, and fructose is a furanose

 How they are linked together: the oxygen on carbon number 1 (C1) of α-D-

glucose is linked to the C2 of D-fructose.

 The -oside suffix indicates that the anomeric carbon of both

monosaccharides participates in the glycosidic bond.

3. Lactose, a disaccharide composed of one D-galactose molecule and one D-

glucose molecule, occurs naturally in mammalian milk. The systematic name

for lactose is O-β-D-galactopyranosyl-(1→4)-D-glucopyranose. Other

notable disaccharides include maltose (two D-glucoses linked α-1,4) and

cellulobiose (two D-glucoses linked β-1,4).

MAJOR FUNCTIONS OF DISSACHARIDES

DISACCHARIDE FUNCTIONS
MALTOSE ( MALT SUGAR )  Respiratory substrate

SUCROSE ( CANE SUGAR )  Respiratory substrate. Most

plants in carbohydrates are

transported in the form of

sucrose.

LACTOSE ( MILK SUGAR )  Respiratory substrate

POLYSACCHARIDES
 1. Polysaccharides are polymers that are formed from the condensation

of hundreds of monosaccharides bound together by glycosidic bonds.

The general formula of polysaccharides is Polysaccharides have a

general formula of Cx(H2O)y. Polysaccharides are amorphous in nature.

The important polysaccharides are starch, glycogen and cellulose.

STRUCTURE OF AN AMYLOPECTIN MOLECULE

2. Polysaccharides represent an important class of biological polymers.

Their function in living organisms is usually either structure- or

storage-related. Starch is a polymer of glucose that is used as a

storage polysaccharide in plants, being found in the form of both

amylose and the branched amylopectin. Amylose is a linear polymer of

glucose mainly linked with α(1→4) bonds. It can be made of several

thousands of glucose units. It is one of the two components of starch,

the other being amylopectin. In animals, the structurally similar glucose

polymer is the more densely branched glycogen, sometimes called

'animal starch'.

3. Cellulose is an example of structural polysaccharides. Cellulose is used

in the cell walls of plants and other organism. It is a long straight chain

polymer consisting of 2000 – 3000 units of Beta glucose linked

together through Beta 1, 4 glycosidic bonds.

BENEDICT’S TEST
 Benedict's reagent is also called Benedict's solution or Benedict's test.

Benedict's reagent is used as a test for the presence of reducing sugars. This

includes all monosaccharides and the disaccharides, lactose and maltose. All

monosaccharides are reducing sugars. They all have a free reactive carbonyl

group. Some disaccharides have exposed carbonyl groups and are also reducing

sugars.  Other disaccharides such as sucrose are non-reducing sugars and will not

react with Benedict's solution.  Starches are also non-reducing sugars, Benedict's

test will detect the presence of aldehydes and alpha-hydroxy - ketones, including

those that occur in certain ketoses.

Thus, although the ketose fructose is not strictly a reducing sugar, it is

an alpha-hydroxy-ketone, and gives a positive test because it is converted to the

aldoses glucose and mannose by the base in the reagent. Benedict's reagent

contains blue copper ( II ) ions (Cu 2+) which are reduced to copper ( I ) (Cu +). These

are precipitated as red copper ( I ) oxide which is insoluble in water. The copper

sulfate (CuSO4) present in Benedict's solution reacts with electrons from the

aldehyde or ketone group of the reducing sugar to form cuprous oxide (Cu 2O), a

red-brown precipitate.
 ++ --
CuSO4 Cu + SO4
 

2 Cu++  +  Reducing Sugar Cu+

        (electron donor)

Cu+ Cu2O (precipitate)

The final color of the solution depends on how much of this precipitate

was formed, and therefore the color gives an indication of how much

reducing sugar was present.

Increasing amounts of reducing sugar

green orange red brown

IODINE TEST
 The Iodine test is used to test for the presence of Starch. Iodine

solution is the iodine dissolved in an aqueous solution of potassium iodide. It reacts

with starch producing a blue black color. This reaction is the result of the

formation of polyiodide chains from the reaction of starch and iodine. The amylose,

or straight chain portion of starch, forms helices where iodine molecules assemble,

forming a dark blue or black color.

The amylopectin, or branched portion of starch, forms much shorter

helices and iodine molecules are unable to assemble, leading the color to be of an

orange/yellow hue. As starch is broken down or hydrolyzed into smaller

carbohydrate units, the blue-black color is not produced. Therefore, this test can

determine completion of hydrolysis when a color change does not occur. Iodine

solution will also react with glycogen, although the color produced is browner and

much less intense.

 BIURET TEST

The characteristic color of a positive Biuret Test. The Biuret Test is a

chemical test used for detecting the presence of peptide bonds. In the presence

of peptides, a copper ( II ) ion forms a violet-colored complex in an alkaline

solution. Several variants on the test have been developed. The Biuret reaction can

be used to assay the concentration of proteins because (for most proteins) peptide

bonds occur with approximately the same frequency per gram of material. In spite

of its name, the reagent does not in fact contain biuret ((H2N-CO-)2NH). The test

is so named because it also gives a positive reaction to the peptide bonds in the

biuret molecule.
 LIPIDS : OIL AND FATS

Lipids are organic compounds that contain carbon, hydrogen and

oxygen. However the ratio of oxygen atoms to hydrogen atoms in lipid molecule is

much lower than 1 : 2 ratio found in carbohydrates. Lipids are a broad group of

naturally occurring molecules which includes fats, waxes, sterols, fat-soluble

vitamins (such as vitamins A, D, E and K), monoglycerides, diglycerides,

phospholipids, and others. The main biological functions of lipids include energy

storage, as structural components of cell membranes, and as important signaling

molecules.

Lipids may be divided into groups. There are three main groups

that are commonly found in plant and animal cells are triglycerides (fats and oils),

phospolipids, and steroids. The other group of lipid consists of a agroup of

waterproof substances called waxes. Lipids may be broadly defined as hydrophobic

or amphiphilic small molecules. The amphiphilic nature of some lipids allows them to

form structures such as vesicles, liposomes, or membranes in an aqueous

environment. Biological lipids originate entirely or in part from two distinct types

of biochemical subunits or "building blocks", ketoacyl and isoprene groups. Using

this approach, lipids may be divided into eight categories :

  Fatty acyls,

 Glycerolipids

 Glycerophospholipids

 Sphingolipids

 Saccharolipids

 Polyketides

Lipids as hydrophobic organic compounds are insoluble in water but

can dissolve in organic solvents such as avetone, ether, chloroform, and alcohol.
 FATS AND OILS

The most familiar lipids are found in fats and oils. Fats consist of a wide

group of compounds that are generally soluble in organic solvents and largely

insoluble in water. Chemically, fats are generally triesters of glycerol and fatty

acids. Fats may be either solid or a liquid at room temperature, depending on their

structure and composition. Fats are animal origin. Although the words "oils", "fats",

and "lipids" are all used to refer to fats, "oils" is usually used to refer to fats that

are liquids at normal room temperature, while "fats" is usually used to refer to fats

that are solids at normal room temperature. "Lipids" is used to refer to both liquid

and solid fats, along with other related substances. Fats and oils or known as

triglycerides are esters since they they are formed from condensation of one

molecule of alcohol ( glycerol ) and three molecules of fatty acid. This reaction is

known as esterification. The bonds form between the glycerol and fatty acids are

called ester bonds.

These examples of fats can be categorized into

  Saturated Fats

 Unsaturated Fats

Saturated fat is fat that consists of triglycerides containing only

saturated fatty acid radicals. There are several kinds of naturally occurring

saturated fatty acids, which differ by the number of carbon atoms, ranging from 3

carbons (propionic acid) to 36 (Hexatriacontanoic acid). Saturated fatty acids have

no double bonds between the carbon atoms of the fatty acid chain and are thus

fully saturated with hydrogen atoms.. Examples of foods containing a high

proportion of saturated fat include dairy products for example butter.

An unsaturated fat is a fat or fatty acid in which there is at least one

double bond within the fatty acid chain. A fat molecule is monounsaturated if it

contains one double bond, and polyunsaturated if it contains more than one double

bond. Where double bonds are formed, hydrogen atoms are eliminated. Thus, a

saturated fat has no double bonds, has the maximum number of hydrogens bonded

to the carbons, and therefore is "saturated" with hydrogen atoms. In cellular

metabolism, hydrogen-carbon bonds are broken down – or oxidized – to produce

energy, thus an unsaturated fat molecule contains somewhat less energy (i.e.,
fewer calories) than an equivalent amount of saturated fat. The greater the

degree of unsaturation in a fatty acid ( the more double bonds in the fatty acid ).

PROTEINS

AMINO ACIDS

Protein consists of one or more polypeptide chains. Each polypeptide

chain is a polymer consisting of many units of amino acids linked together through a

condensation reaction. All amino acids share the basic structure but differ in their

side chains, ( -R ). Amino Acids are amphoteric. This is because each amino acid

consists of both the acidic carboxyl group ( -COOH ) and the basic amino group

( -NH2 ). Proteins (also known as polypeptides) are organic compounds made of

amino acids arranged in a linear chain and folded into a globular form.

The amino acids in a polymer are joined together by the peptide bonds

between the carboxyl and amino groups of adjacent amino acid residues. Each

polypeptide chain is a polymer consisting of many units of amino acids linked

together through a condensation reaction. The sequence of amino acids in a protein

is defined by the sequence of a gene, which is encoded in the genetic code. In

general, the genetic code specifies 20 standard amino. The amino acid is classified
into four groups based on the properties of their side chains. Amino acids have

hydrocarbons as side chains are non – polar. Protein molecules that consist of a

large amount of such amino acids are insolouble and non – reactive. Such proteins

usually form a structural protein for example, collagen. Amino acids with polar R

group produice partial charges but do not receive or donate electrons.

The presence of amino acids increases the soloubility of the protein and

enables hydrogen bonding between polypeptide chains. Basic or acidic amino acids

produce passively or negatively charged ions. These ions are strongly hydrophilic.

Proteins with a high content of these amino acids, such as globular proteins are

solouble in water.

FORMATION OF POLYPEPTIDES

1. A dipeptide consists of two amino acids linked together through a peptide

bond. This process is refereed to as condensation. The dipeptide can

undergo further reactions with other amino acids since it has a free amino

group at one end ( N – Terminal ) and free carboxyl group at the other end
 ( C – Terminal ). When one more amino acidis linked to the dipeptide, a

tripeptide is formed.

ESSENTIAL AND NON - ESSENTIAL AMINO ACIDS

Essential amino acids are "essential" not because they are more important

to life than the others, but because the body does not synthesize them, making it

essential to include them in one's diet in order to obtain them. In addition, the

amino acids arginine, cysteine, glycine, glutamine, histidine, proline, serine and

tyrosine are considered conditionally essential, meaning they are not normally

required in the diet, but must be supplied exogenously to specific populations that

do not synthesize it in adequate amounts. Examples of essential amino acids are

His, Ile, Leu, Lys, Met, Phe and Thr.

Non essential amino acids are amino acids that can be produced in our body.

Their uses and functions in our body are equally as important as the limiting amino

acids. The difference is that those kind of amino acids can be found in our food.

The functions of non-essential amino acids includes the 12 non-essential amino

acids. Included is a some of the functions and benefits and side effects (if any) of
the amino acids. For example Alanine removes toxic substances released from

breakdown of muscle protein during intensive exercise. Cysteine is a component of

protein type abundant in nails, skin and hair. It acts as an antioxidant.

PROTEIN STRUCTURES

During protein synthesis, individual individual amino acids are linked

together to form a linear polymer called polypeptide. However, a polypeptide chain

is not yet a protein. A protein is a polypeptide chain that has attained a unique,

three –dimensional shape. Some proteins consist of a single polypeptide chain that

undergoes folding and coiling. These proteins are called monomeric proteins. Many

proteins are multimeric proteins, consisting of two or more polypeptide units.

There are four levels of organization in three – dimensional structure of protein :

1. Primary Structure

2. Secondary Structure

3. Tertiary Structure
 4. Quaternary structure

PRIMARY STRUCTURE

The amino acid sequence of the peptide chains. .The primary structure is

held together by covalent or peptide bonds, which are made during the process of

protein biosynthesis or translation. These peptide bonds provide rigidity to the

protein. The two ends of the amino acid chain are referred to as the C-terminal

end or carboxyl terminus (C-terminus) and the N-terminal end or amino terminus

(N-terminus) based on the nature of the free group on each extremity.

SECONDARY STRUCTURE

Highly regular sub-structures (alpha helix and strands of beta pleated

sheet), which are locally defined, meaning that there can be many different

secondary motifs present in one single protein molecule.

 Simple Alpha Helix :

 The polypeptide chain is coiled to form a simple alpha helix. The loops

of alpha helix can be extended just like wires. The alpha helix is

maintained by hydrogen bonds.

Double or Triple Helix :

 Two or three polypeptide chains are coiled together to form fibre

 Keratin – The protein that forms hair, horns, nails, breaks

and feathers is in the form of a double helix.

 Collagen - The protein that forms tendons, ligaments

and cartilage has a triple helical structure.

Beta Pleated Sheet

 This structure is formed when some streched polypeptide chains are

arranged parallel to each other and are held together by hydrogen

bonds and folded longitudinally. It is strong but flexible.

TERTIARY STRUCTURE
 Three-dimensional structure of a single protein molecule, a spatial

arrangement of the secondary structures. It also describes the completely

folded and compacted polypeptide chain. It is the way alpha helix is folded

to form a compact protein molecule.

QUATERNARY STRUCTURE

Relative arrangement of polypeptide chains in a protein molecule that

consists of more than one polypeptide chains in a protein molecule. Multimeric

proteins such as haemoglobin have Quaternary structure.

PROPERTIES OF PROTEINS

 All globular proteins are soluble

 Large molecule of globular proteins do not dissolve completely in water

 They form a colloidal solution

 All globular proteins are amphoteric, they exhibit both base and acid.
  Proteins are easily damage by heat ( temperatures greater than 40 C ) due

to the breakage of their cross lingkages

 Chemical that brings extreme pH denatures proteins

CLASSIFICATION OF PROTEINS

Proteins are classified according to their structure :

1. Fibrous Proteins

 These proteins are involved in the formation of body structures of

animals. Most fibrous proteins have secondary structures are

insolouble in water. Examples are : collagen, myosin and fibrin .

2. Globular Proteins
  All globular proteins have tertiary structures and some also

quaternary structures. The polypeptide chains are tightly coiled and

folded to form a sphere. Examples are : Globulin, enzymes and

antibodies.

COMPARISON BETWEEN FIBROUS PROTEINS AND

GLOBULAR PROTEINS

FIBROUS PROTEIN GLOBULAR PROTEIN

 Stable Structure  Relatively Unstable

Structure

 Insoluble in water  Soluble in water

 Structural Functions  Metabolic Functions

 Polypeptide chains form  Polypeptide chains ‘ roll up ‘

long strands into spherical shape

 Examples include collagen in  Examples include all

bone and keratin in hair enzymes, antibodies and

 hormones.

Proteins can also be classified according to their composition :

1. SIMPLE PROTEINS

 Simple proteins are pure proteins that do not contain any other

substance. Examples include fibrous proteins and gloubular proteins.

2. CONJUGATED PROTEINS

 Conjugated proteins are made up of two portions attached together, a

portion attached to a non – protein known as prosthetic group.

Examples include lipoprotein and glycoprotein.

 CONCLUSION

Test was carried out to test the presence of carbohydrates in Reducing

Sugars such as glucose and fructose. Here, the Benedict’s Test was carried out.

The colour change was observed. The initial blue colouration of the mixture turns

green, then yellowish and finally form a brick – red precipitate. The Benedict’s

solution reduces soluble blue copper sulphate with copper ( II ) ions to insolouble

red – brown copper oxide containing copper ( I ). The carbohydrate test was also

carried out for Non – Reducing sugars such as sucrose. The brick - red precipitate

was formed. This is because the dissacharide can be hydrolysed to its

monosaccharides by boiling with dilute dilute hydrochloric acid. Thus, Benedict’s

Test was carried out.

The test for starch or known as the Iodine Test. Here, a blue – black

colouration was formed. The polyiodide complex is also formed with starch. Next,

the test for lipids such as oils or known as the Emulsion Test was carried out. A

cloudy white suspension was observed. This is because lipids are immiscible with
water. Thus, adding water to alcohol results in tiny lipid droplets in water which will

give a white appearance. The final test that was carried out was to the test for

the presence of proteins. Here, a purple colouration or mauve is observed.

 DISCUSSION

The carbohydrate test that was carried out for Reducing Sugars

consist of glucose and fructose. The solution of the reducing sugar initial was blue

in colour then turns green, yellowish and finally turns into a brick – red precipitate.

The Benedict’s Test was carried out during the experiment. The Benedict reagent

contains blue copper ( II ) ions which are reduced to copper ( I ) ions. It is

precipitated to insolouble red - brown copper oxide containing copper ( I ). The

carbohydrate test for Non – Reducing sugars such as sucrose was also carried out.

Here, Benedict’s Test was also carried out. The dissacharide will be hydrolysed to

its monosaccharides by boiling with hydrocholoric acid. Thus, this will finally give

the brick – red precipitate results.

The Iodine Test was carried out to test the presence of starch. A

blue – black colouration was formed. Besides, a polyiodide complex is formed with

starch. The starch becomes a paste and increases further with viscosity. Next, the
Emulsion Test was carried out to show the presence of lipids. The fat or oil was

first dissolved in alcohol. A cloudy white suspension was formed. This is because

the lipids are immiscible with water. Thus, adding water to a solution of lipid in

alcohol results in an emulsion of tiny droplets in water which will give a white,

opalescent appearance. Finally, Biuret Test was carried out to test the presence of

proteins. It gives a purple or mauve colouration. This is because of the presence

of potassium hydroxide solution which gives a purple colouration.

 TEST OBSERVATION BASIS OF TEST
Reducing Sugars :  The initial blue  Benedict’s
colouration of solution contains
 Glucose the mixture copper sulphate.
turns green, Reducing sugars
 Fructose
then yellowish reduce solouble
and may finally blue copper (II)
form a brick – ions to insolouble
red percipitate red – brown
copper oxide
containg copper
(I). The latter is
seen as a
precipitate.

Non – Reducing Sugars :  As Benedict’s  A dissacharide

can be hydrolysed
Test to
 Sucrose
monosaccharide
constituents by
boiling with dilute
hydrochloric acid.
Sucrose is
hydrolysed to
glucose and
fructose, both of
 which are
reducing sugars
and give the
reducing sugar
results with the
Benedict’s Test.

Starch :  A blue – black  A polyiodide

colouration complex is
Iodine Test
formed

Lipids :  A cloudy white  Lipids are

immiscible with
Oils, fats and waxes suspension water. Adding
water to a
solution of the
lipid in alcohol
results in an
emulsion of tiny
lipid droplets in
the water which
reflect light and
give a white,
opalescent
appearance.

Proteins :  A mauve or

purple
Biuret Test
colouration