Name: Delda, Jovan Paul G.

CHEM 104A Lab Schedule: A 12:00 – 04:00 PM M-F

Course & Year: BSN 1 - A Name of Lab Instructor: Jemimah Joy Guarin

ANALYSIS OF FATS AND OILS

Activity No. 5

I. PURPOSE: To be able to know some common properties of fats and oils

II. APPARATUS: Bunsen burner, 50 ml beaker, 250 ml beaker, stirring rod, test
tubes, watch glass, vials, test tube holder, nichrome wire

III. MATERIALS: coconut oil, olive oil, linseed oil, rancid oil , oleic acid, stearic acid,
ethanol, carbon tetrachloride, congo red, blue and red litmus paper,
solid KHSO4, glycerol, 5% aqueous solution of glycerol. Hubl’s iodine
solution, dilute HCl, dilute NaOH, 10% NaOH chloroform, PbO, soap
solution, 1% albumin , Na2CO3 solution ,
0.01M of glucose solution, conc HNO3, K2Cr2O4 solution, borax
powder

Hubl’s reagent is prepared by dissolving 26 grams of iodine and 30 grams of

Hgl2 in 1 liter of 95% alcohol

IV. PROCEDURE:

1. Spot Test for Fat:

Place a drop of coconut oil upon a piece of white paper. Compare the spot
produced by the fat with that produced by water.
OBSERVATION:

Steps Carried Out Comparison

A drop of coconut oil on paper
A drop of water on paper
The oil droplet patch on the paper
became translucent.
After resting for a while, the water that
was dropped into the paper was
absorbed, leaving the paper just a little
damp. Additionally, unlike coconut oil,
which is slightly distributed on paper,
water adheres to it.

2. Color, Solubility, and Density:

 Note the colors of coconut oil and olive oil and glycerol. The color is due to
impurities in the oils since the glyceryl esters are colorless. Biuret test the solubility of
each by adding 1 drop of it to about 5 drops of each of the following: water, ethanol,
boiling ethanol, and carbon tetrachloride. Note the relative density of each oil with
respect to water.
OBSERVATION:

Solubility
in
Oil samples Color Density with Water Ethanol Hot ethanol
respect to
water
Coconut oil Opaque/ Very dense insoluble soluble soluble
cloudy white
with faint
yellow
Olive oil Yellow/ gold Very dense insoluble soluble soluble
Glycerol Clear/ Slightly dense soluble soluble soluble
colorless

 What can be said about the solubility of oils towards water and other
organic solvents? Explain.
Answer:

The various oil samples are soluble in ethanol, cold ethanol, and CCl4 but
insoluble in water. Oils with a non-polar character should dissolve in non-polar solvents
according to the adage "like dissolves like." Oil and water don't mix well because oil is
nonpolar and water is polar, which explains why. The oil samples, however, are soluble in
nonpolar organic solvents like ethanol and CCl4. Oil samples were not soluble since NaOH
is an inorganic substance.

3. Reaction towards indicators:

Prepare 3 separate test tubes of 1 ml of fresh coconut oil. To the first add red
litmus paper, to the second blue litmus paper and to the third add 3 drops of congo
red. Repeat the test using a rancid oil.

OBSERVATION:

Oil Samples Red litmus paper Blue litmus Congo Red

paper
Coconut oil red blue orange
Rancid oil red red blue

 Differentiate the reaction of fresh coconut oil and rancid oil towards the
indicators.

ANSWER:
 The two litmus sheets' colors remained unchanged after being exposed to
fresh coconut oil, indicating that it is neutral. On the other hand, rancid oil caused the
blue litmus paper to turn red, indicating that it is acidic.

4. Acrolein Test:

Mix three drops of coconut oil with 0.5 g KHSO 4 in a test tube. Heat the mixture
and cautiously note the odor. Repeat the test with glycerol in place of the oil. Write
the chemical equation.

OBSERVATION:

Oil Samples Steps carried out Odor

Coconut oil 3 drops of coconut oil Pungent odor
were added with 0.5
grams of potassium
bisulfate and were
heated. The odor was
carefully noted.
Glycerol Mixed 3 drops of glycerol A pungent irritating burnt
with potassium bisulfate odor (acrolein)
in a test tube. The
mixture was heated and
the odor was cautiously
noted.

Chemical Equation:

Oil or fat ---→ glycerol + fatty acid

5. Test for Unsaturation:

Place 5 ml of each sample in 4 separate test tubes; coconut oil, stearic acid,
linseed oil and oleic acid. Add 3 ml of chloroform to each tube and add Hubl’s iodine
solution drop by drop by shaking in between addition.

Make a control set up by shaking a mixture of chloroform and iodine without the
sample. Account for the result obtained.
 OBSERVATION:

Test tube: Control CHCl3 + I2 solution Flaming red (no change)

5.1 Coco oil + CHCl3 + I2 Orange to dark orange;
solution violet color in iodine does
not fade away (saturated)
5.2 Stearic acid +CHCl3 + I2 Yellow/ dark orange color;
solution violet color in iodine did not
fade away (saturated)
5.3 Linseed oil + CHCl3 + I2 Dark orange color; violet in
solution iodine fades away
(polyunsaturated; positive)
5.4 Oleic acid + CHCl3 + I2 Dark orange color; violet
solution color in iodine fades away.
(monounsaturated; positive)

Explanation of Results:

The iodine lost its violet color in test tubes 5.3 and 5.4. The oleic acid and
linseed oil are unsaturated and it denotes a favorable response. While test tubes 5.2 and 5.1
showed no fading of the iodine's violet color, this showed that the stearic acid and coco oil
were saturated. Due to their unsaturated nature, lipids have the ability to absorb iodine and
are reactive to iodine.

• Which of the oil absorb more iodine, oleic or stearic?

Because oleic acid has double bonds in its carbon chain and is an unsaturated molecule, it
absorbs more iodine. As a result of saturation, the carbon chain of stearic acid only has single
bonds. When compared to saturated oils, lipids with double bonds absorb more iodine
because they are more iodine reactive.

• Which oil contains more unsaturated fatty acids, coco oil or linseed oil?
Unsaturated fatty acids are more prevalent in linseed oil. Linoleic acid is a component of
linseed oil. This explains why there is more unsaturation in the fat the greater the iodine
levels. Linseed oil is extremely unsaturated and has an iodine value of 136–178 compared to
coconut oil's 7–10 value.

• Structurally, how does an unsaturated fat differ from saturated fat?

Unsaturated fat is made up of unsaturated fatty acids with one or more double carbon-carbon
bonds; it can either be monounsaturated (just one double bond) or polyunsaturated (two or
more double bonds). All of the carbon-carbon bonds in saturated fat are single bonds. While
unsaturated fat is liquid at normal temperature, saturated fat is solid.

• What is the relationship between iodine number and unsaturation of fat?

The level of fat or oil unsaturation is indicated by the iodine value. It is defined as the quantity
of iodine that 100 grams of fat may absorb. The amount of unsaturations present in the fat or
oil increases with the iodine levels. Unsaturated fatty acids have double bonds that interact
with iodine; the higher the iodine number, the more C=C bonds are present.

6. Drying Oil:

In a 3-inch test tube place 1 ml of linseed oil and an estimated 30 mg of litharge

(PbO). Gently warm the mixture for 10 minutes in a water bath, then pour it into a watch
glass. Set the watch glass aside and observe after 30 minutes. Repeat the test with
coconut oil.

OBSERVATION:

Oil sample Steps carried out Results

Linseed oil After adding lye (PbO) and It dried faster
soaking in a water bath for
a while, linseed oil was
added. After that, it was
given some time to rest in
a watch glass.
Coconut oil Litharge (PbO) was added Does not dry up readily
to the coconut oil, and it
was then submerged in
water for a while. It was
then exposed for a while to
a watch glass.

• What reason may be given to account for the difference in the behavior
of the two oils (linseed oil and coconut oil) towards litharge?
Answer:
The absence of water did not cause the oils to dry out; rather, it was due to
to the development of a tough coating that sticks to the watch glass's surface. Compared to
coconut oil, which has a larger percentage of saturated fat, linseed oil contains more
unsaturated fats. Additionally, it's due to the methods used to acquire these oils. Coconut oil
is produced by crushing and pressing copra to extract the oils, whereas linseed oil is
prepared from the dried, ripened seeds of the flax plant and is obtained by pressing,
occasionally followed by solvent extraction.

7. Emulsification:

Prepare 4 test tubes with the following contents:

Tube 1: 5 ml water
Tube 2: 5 ml water + 3 ml soap solution
Tube 3: 5 ml water + 1 ml of 1 % albumin solution
Tube 4: 5 ml water + 0.5ml Na2CO3 solution
To each test tube above add 5 drops of coconut oil, shake and observe the results.

OBSERVATION:
 Test tubes Mixture Classification
1 5 ml water +5 drops of Temporary emulsion
coconut oil
2 5 ml water +5 drops of Permanent emulsion
coconut oil + 3ml of soap
solution
3 5 ml water +5 drops of Permanent emulsion
coconut oil + 1% albumin

4 5 ml water +5 drops of Permanent emulsion

coconut oil + .5ml Na2CO3

• What is an emulsion?

When two liquids that are often incompatible are combined, an emulsion is
created in which one liquid contains a dispersion of the other liquid. Emulsions fall
within the more broad category of colloids, which are two-phase systems of matter.
Two types of emulsions are distinguished: temporary and permanent.

• Which tube or tubes formed a permanent emulsion?

As albumin, soap solution, and Na2CO3 are recognized emulsifiers, test tubes
2, 3, and 4 were found to permanently emulsify. Both the hydrophilic and hydrophobic
components of these emulsifiers work together to stabilize the emulsion. Test tube 1
only produced a transient emulsion. Since Test Tube 1 was devoid of an emulsifier,
they both separated after resting for a while.

8.Glycerol:

a. Fuse a drop of glycerol in a nichrome wire with powdered borax. Note the green
flame produced. This is due to the glycerol ester of boric acid.

OBSERVATION:

Glycerol + powder borax --------------------🡪 color of flame: green

glycerol is a non-sugar compound with OH groups- the green-colored flame is due
to the glycerol ester of boric acid.

b. Perform the nitro-chromic acid test on a 5% aqueous solution of glycerol.

To 5ml of 0.01M of glucose solution add 3ml of conc. HNO 3 and 5 drops K2Cr2O4
solution. A blue solution appears after a minute if its contains sugar with a
concentration of more than 1%.This test is due to the free - CHOH in the sugar
molecule. Non sugar compounds which contains this group may also give a
positive reaction.

OBSERVATION:
 Glucose solution + conc HNO3 + 5 drops K2Cr2O4 solution --------
🡪 Blue solution due to the presence of OH group in glucose solution.

• What is the color formed?

Glycerol produced a green flame in the flame test, but glucose

produced a blue solution in the nitro chromic test..

• This indicates the presence of what group in glycerol?

The presence of CH OH groups was indicated by the green flame in glycerol and the blue
color in the glucose solution. Carbon atoms are linked to the OH group in the chemical
structure of glycerol, which has the formula C3H8O3. Additionally, the carbon atoms of
glucose, C6H12O6, are linked by OH groups. Positive results could also come from non-
sugar molecules with CH OH groups like glycerol.
9. Basic Hydrolysis or Saponification:

Warm 30 ml of coconut oil in 600 ml beaker and add in small proportion, 50 ml

of a 10% NaOH solution. Heat the reacting mixture gently and stir continuously. The
reaction is complete when the mixture becomes stick and clings to the stirring rod.
Pour the mixture into a cardboard mold and allow to set.

• What are the basic chemical components of fats and oil?

Fatty acids and glycerol make up the bulk of fats and oils. The majority of dietary
oils and fats are made up primarily of triglycerides. Minor components include mono-
and diglycerides, free fatty acids, phosphatides, sterols, fatty alcohols, fat-soluble
vitamins, and other substances. A triglyceride is made up of three fatty acids and
glycerol.
• What is saponification? What is its product?
Saponification is the process of heating fats or oils with a potent base (NaOH) to
create glycerol and fatty acid salts (Soap). According to the definition of saponification,
it is a "hydration reaction where free hydroxide breaks the ester bonds between the
fatty acids and glycerol of a triglyceride, resulting in free fatty acids and glycerol," both
of which are soluble in water. By using heat and an aqueous alkali solution, the
process of saponification transforms fat, oil, and lipid into soap and alcohol. Fatty acids
are monomers that have long carbon chains, and soaps are salts of fatty acids like
sodium palmitate.

• Give an equation for the reaction using glyceryl laurate to represent

the oil.

Glyceryl Laurate —hydrolysis —> glycerol + lauric

acid
Lauric acid + sodium hydroxide = sodium laurate +
 water
P.S. (Camera scanner used)