DATE: 02/10/2018

LAB: 1

TITLE: Identification of biochemical in pure form and in food samples.

AIM:

1) To carry out test on pure form of reducing sugars, starch, proteins and lipids to master
techniques and become familiar with the color change.
2) To identify the biochemical

APPARATUS & MATERIAL:

 Water bath  Dilute HCl  Egg Solution

(electric heater)  Ethanol  Milk
 15 test tubes  KOH or NaOH  Fruit Juice
 Test tube holder  CuSO4 solution  Meat Pizza
 5 5ml syringes  I2/KI solution  Apple
 Teat pipette  Sucrose Solution
 Labels  Glucose Solution
 Glass rod  Starch Solution
 Benedict’s solution  Oil

METHOD:

1) Benedict’s test (reducing sugars): 2cm3 of the glucose sulotion was added to a test tube
and equal volume of the Benedict’s solution was added. The solution was shaken and
brought to a gentle boil.
2) Benedict’s test (Non-reducing sugar) 2cm3 of the sucrose solution was added to a test
tube and 1cm3 of dilute HCl was also added. The contents of the test tube were boiled
for one minute. It was then neutralized with the NaOH. Benedict’s test was then carried
out.
3) Starch Test: 2cm3 of the starch was added to a test tube. A few drops of I2/KI solution
were added.
4) Emulsion Test (lipids): 2cm3 of test substance was added to the test tube containing
absolute ethanol. The lipid was dissolved by shaking the test tube thoroughly and an
equal volume of cold water was added.
 5) Biuret test (proteins): 2cm3 of the egg albumen was added to a test tube. Potassium
hydroxide solution was added to the mix and two drops of copper sulphate solution
were added to the test tube.
6) Procedure was repeated on the meat pizza, apple, milk and juice respectively in order to
test for the presence of biochemicals (reducing sugars, starch, fats and proteins).
7) All data were tabulated.

RESULTS:

TEST SOLUTION TESTED OBSERVATIONS INFERENCE

Glucose Blue of Copper sulphate Large amount of
greenyelloworangebrick- reducing sugar
red were present.
Meat pizza Blue of Copper sulphate Small amounts of
green yellow reducing sugar
were present.
BENEDICT’S TEST Apple Blue of Copper sulphate Large amount of
greenyelloworange reducing sugars
REDUCING SUGAR brick-red were present.
TEST Milk Blue of Copper Small amounts of
sulphategreen yellow reducing sugars
were present.
Juice Blue of Copper Small amounts of
sulphategreen yellow reducing sugar
were present.
After hydrolyzing sucrose Contained non-
BENEDICT’S TEST Sucrose solution the blue of copper reducing sugars.
NON-REDUCING sulphate
SUGAR TEST greenyelloworange
brick-red
Starch Brown of iodine to blue-black. Starch was
present.
Meat pizza Brown of iodine to blue-black. Starch was
present.
STARCH TEST Apple No color change. Remained No starch was
brown of iodine. present.
Milk No color change. Remained No starch was
brown of iodine. present
Juice No color change. Remained brown No starch was
of iodine. present
 TEST SOLUTION TEST OBSERVATION INFERENCE
Oil Cloudy white emulsion formed. Lipids were
present.
Meat pizza Cloudy white emulsion formed. Lipids were
present.
EMULSION TEST- Apple No emulsion formed. Lipids were not
LIPIDS present.
Milk Cloudy white emulsion formed. Lipids were
present
Juice No emulsion formed. Lipids were not
present.

Egg Albumen Blue of copper sulphate to Proteins were

violet. present.
Meat pizza Blue of copper sulphate to Proteins were
BIURET TEST- violet. present.
PROTEIN TEST Apple No color change. Proteins were not
present.
Milk No color change. Proteins were not
present.
Juice No color change. Proteins were not
present.

DISCUSSION:

Biochemicals are chemicals such as carbohydrates, lipids and proteins which are necessary for
life to occur. Monosaccharides (glucose), disaccharides (sucrose), polysaccharides (starch) are
the three groups in which all carbohydrates are classified. Benedict’s solution is used to
identify any reducing sugars present in a solution when boiled. The polysaccharide starch, are
made up of molecules that are insoluble in nature and an iodine solution is used in the starch
test to identify them. Fatty acids and glycerol’s make up the compound known as fats
(triglycerides) and they are identified by the emulsion test. Amino acids are held together by
peptide bonds which forms the polypeptide known as proteins. Proteins make up most of the
organic molecules found in cells and are very important in both structural and metabolic
activities occurring within an organism. The biuret test contains two reagents, copper sulphate
and sodium hydroxide which are used to identify proteins present in solution.

The meat pizza and apples were crushed and cut up into small pieces before mixing with water.
The egg albumen was also mixed thoroughly with water. This was done so that the biochemical
present in the foods would have been suspended or dissolved in the water in order to react
while being tested (to indicate if the biochemical was present or not in the different foods).

After combining the glucose (reducing sugar) solution with Benedict’s solution which contained
the blue copper (II) sulphate ions (Cu2+) was reduced by the reducing sugar (glucose) to an
insoluble brick-red copper oxide which contained copper (I) ions. The brick-red precipitate
indicated large amounts of reducing sugar were present. The apple also produced a brick-red
precipitate when tested with Benedict’s solution which indicated large amounts of reducing
sugars were present. The meat pizza, milk and juice produced a yellow precipitate when tested
with Benedict’s solution which indicated small amounts of reducing sugars were present.

Sucrose which is a non-reducing sugar had to be hydrolyzed (by breaking the glyosidic bonds
between the two monosaccharides which exposed the part of the molecule needed to react
with the reagent in the Benedict’s solution) with HCl into its two reducing sugar components of
beta fructose and alpha glucose in order to react and reduce the (Cu2+) ions present in the
Benedict’s solution. After hydrolysis of sucrose sodium hydroxide was used to neutralize the
HCl acid and the Benedict’s test was carried out which produced a brick-red precipitate that
indicated large amounts of reducing sugars were present.

When potassium iodide was added to the starch, the iodine molecules got entrapped within the
molecules of starch which formed a starch iodine complex that had a blue-black color. Starch
was found to be present when the potassium iodide was added to the meat pizza; however
starch was absent when the milk, juice and apples were tested.

In water, lipids are immiscible; however they dissolve in absolute ethanol (non-polar solvent).
When absolute ethanol was added to the oil (lipids) and shaken vigorously, the emulsion of tiny
lipid droplet reflected light when water was added; therefore it gave a white, opalescent
appearance on the water. The meat pizza and milk both showed a cloudy emulsion when tested
which indicated that lipids were present in those foods. However there was no white, cloudy
emulsion present in the apples and juice which indicated that lipids were not present.

When biuret solution was added to the egg albumen solution it produced a purple color
change. This was due to the nitrogen atoms in the peptide chains reacting in the presence of
copper sulphate in alkaline solution to form a purple complex with the Cu 2+ ions. The milk and
meat pizza changed to purple when biuret solution was added to them hence indicating protein
was present in those foods. However the apple and the juice contained no proteins which were
indicated by no change in color when the biuret solution was added to them.

The meat pizza has the highest nutritional value since it contains all biochemical tested for. The
milk contained all the biochemical tested for except starch which indicates that its nutritional
content is high. The apple contained large amounts of reducing sugars only hence its nutritional
content is low. However the juice contained small amounts of reducing sugars only therefore it
was considered to have the lowest nutritional content.

The milk would be a suitable food for a baby since it has proteins, fats and small amounts of
reducing sugars that are adequate for a baby to grow healthily. The apple would be suitable for
a young boy since it contains large amounts of reducing sugars which would be necessary for
energy to fuel an active lifestyle (playing outdoors). Pizza would be suitable for an adult since it
contains fats for large amounts of energy (fats produces more energy than carbohydrates when
burnt by the body), proteins (for muscle repair due to strenuous activity), starch and small
amounts of reducing sugars.

CONCLUSION:

It can be concluded that tests on the pure form of reducing sugars, starch, proteins and lipids
were carried out in order to master the technique and become familiar with the color changes
associated with the specific test. The biochemicals present in pizza were found to be; reducing
sugars (carbohydrates), starch, lipids and proteins. The biochemical present in apple was found
to be only reducing sugars. The biochemicals present in milk were found to be; reducing sugars,
lipids and proteins. The biochemical present in juice was found to be only reducing sugars.