Biochemistry Lab Manual
Biochemistry Lab Manual
EXPERIMENT 13:
Saponification Reaction
I. OBJECTIVES
At the end of the experiment, you shall be able to:
II. DISCUSSION
A soap is a salt of a fatty acid. Fatty acids are carboxylic acids with a long
unbranched hydrocarbon (aliphatic) chains.
The ionic (highly polar, charged) nature of salts typically makes them water
soluble. The non-polar hydrocarbon (aliphatic) tail of soap molecules allows them to be
miscible with non-polar (greasy) substances and ‘help’ these substances dissolve in
water. Successful bathing is all about improving the water solubility of ‘dirt’.
Triglycerides are the main constituents of vegetable oil and animal fats. A
triglyceride is a tri-ester with three long fatty-acid-like hydrocarbon chains. When a
triglyceride reacts with three equivalents of a strong base such as NaOH, the three
ester bonds are hydrolysed to yield three fatty acid salts (soap molecules) and one
molecule of glycerol as shown below. This ‘saponification’ can also be referred to as
base hydrolysis.
In this experiment you will make soap out from olive oil. The chemical structure
of the major constituent of olive oil is as follows (C52H96O6).
III. MATERIALS
Part A.
Part B.
500 ml beaker 500 ml molder
Stirring rod
IV. REAGENTS
Part A and B.
V. PROCEDURE
2. After drying them, cut them into smaller pieces using a scissor or grind it into powder
using a blender or a coffee grinder.
3. Prepare 232 ml olive oil, 27 ml coconut oil, and 14 ml castor oil at room temperature
each in a separate beaker. Measure them using a graduated cylinder.
4. Prepare your lye solution in a 100 ml beaker by adding 52 ml water and 38 grams of
Sodium Hydroxide. That will constitute 42% lye concentration.
1. Mix the 232 ml olive oil, 27 ml coconut oil, and 14 ml castor oil in a 500 ml beaker
using a stirring rod.
2. After mixing the oils, you can now add the lye solution in the mixture.
3. Add the orange peels (in powdered form). You can also add citrus scent.
4. Continue stirring until the solution reach a light to medium trace (color).
Name: Date: _
PRE-LABORATORY TASK 1
Measuring
accurately
using a
measuring
cylinder
Making 42%
concentration
lye solution
Handling
reagents
properly
Using of
appropriate
laboratory
materials
PRE-LABORATORY TASK 2
Name: Date: _
DOCUMENTATION SHEET
PREPARATION
OF MATERIALS
THE OLIVE OIL WAS BOUGHT
FROM THE GROCERY
TOGETHER WITH THE
ORANGES THAT WERE USED
TO OBTAIN THE ORANGE
PEELS. THE COCONUT AND
CASTOR OIL CAN BE
BROUGHT FROM WATSONS.
MAKING OF
SOAP
THE ORANGE PEELS WERE
DRIED USING AN OVEN IN
LOW HEAT SINCE WE DON’T
HAVE A DEHYDRATOR. WE
MADE SURE THAT IT IS
ALREADY DRIED BEFORE
BRINGING IT TO THE LAB.
THEY WERE NOT YET IN
POWDERED FORM SO WE
CUT IT IN SMALL PIECES
USING A SCISSOR.
FORMATION OF
SOAP
THE SOAP MIXTURE WAS
LEFT UNDISTURBED FOR
ABOUT 12-24 HOURS TO
HARDEN AND DRY.
WE EXAMINED IT AFTER
REMOVING IT FROM THE
CONTAINER.
Post-laboratory questions:
Most soaps contain glycerol because it draws water to skin, effectively balancing it`s
moisture levels. Its presence in skin has been shown to prevent signs of aging by
helping skin cells mature properly.
Glycerin is a thick, clear fluid that is used in a wide variety of bath ad beauty products.
It is humectant which means it has the ability to attract moisture in the skin. It draws
moisture up through the skin layers and slows or prevents excessive drying and
evaporation.
Triglyceride- an ester formed from glycerol and three fatty acid groups. Triglycerides
are the main constituents of natural fats and oils. Soap is produced by the
saponification (hydrolysis) of a triglyceride (fat or oil).
Surfactant- a substance which tends to reduce the surface tension of a liquid in which
it is dissolved. Surfactants function by breaking down the interface between water and
oils and/or dirt
When triglycerides in fat/oil react with aqueous NaOH or KOH, they are converted into
soap and glycerol. This is called alkaline hydrolysis of esters. Since this reaction leads
to the formation of soap, it is called the Saponification process.Thus, the soap
molecule has a polar head and a non-polar hydrocarbon tail.