ACTIVITY NO.

5
REACTION OF
LIPIDS
Group 4:
 Aquino, Karenza
 Baluarte, Rachel
 Fabon, Eriane Diana
INTRODUCTIO
N
 Lipids
 Lipids are heterogeneous class of bioorganic compounds
 Lipids are nonpolar hydrocarbons.
Alcohol ------------ Ester linkage ---------------
Fatty Acids
Fats
Oils Limited solubility in
Waxes
water & widely
distributed in
Cholesterol and It’s derivatives
nature both in
Some Vitamins plants and in
Prostaglandins animals
 Fats Oils
saturat Unsatura
ed fatty ted fatty
acids *Most acids
abundant
lipids
*Tricyglycer
ols
mixtures
 Chemical properties of triacylglycerol (simple
lipids) are typical with that of esters and
alkanes.
 Important reactions of these compounds are
hydrolysis, saponification, hydrogenation and
rancidity and emulsification.
Chemical
Principles
Involves
 SOLUBILITY
 Lipids are non-polar organic compounds. The physical
properties of fatty acids, and of compounds that
contain them are largely determined by the length and
degree of unsaturation of the hydro carbon chain. The
nonpolar hydrocarbon chain accounts for the poor
solubility of fatty acids in water. Solubility of a
substance depends on a simple rule of thumb “like
dissolves like” this statement indicates that a solute will
dissolve best in a solvent that has a similar chemical
structure to itself. The overall solvation capacity of a
solvent depends primarily on its polarity.
 TRANSLUSCENT SPOTS
 Lipids have a characteristic greasy feel
 When brought in contact with a substance
like paper, it penetrate through it
producing a translucent spot
 Fats are non-volatile
 The spot of grease can never absorb
enough heat to vaporize. When the liquid
is inside the sheet of paper, it diffracts
 ACROLEIN TEST
 The Acrolein test is a general test for the
presence of glycerol in a molecule.
 When a fat is heated strongly in the
presence of a dehydrating agent such as
potassium bisulfate (KHSO). the glycerol
portion of the molecule is dehydrated to
form the unsaturated aldehyde, acrolein
(CH2=CH–CHO),
Glycer
ol

Acrolei
n
 TEST FOR
UNSATURATION
 Identify if the lipid sample have a double
bond
 All neutral fats contain glycerides of
some unsaturated fatty acids.
 These unsaturated fatty acids become
saturated by taking up iodine.
 If the fat contains more unsaturated fatty
acids, it will take up more iodine.
 TEST FOR RANCIDITY
 It is a process for deterioration or degradation of fats
 Rancidity, is the natural process of decomposition of
fats or oils by either hydrolysis or oxidation, or both.
 The process of degradation converts fatty acid
esters of oils into free fatty acids,  by reaction with
air, moisture and/or other materials.
 This gives rise to an unpleasant odour and taste in
food.
 These lipids degrade to the point of becoming either
unpalatable or unhealthy to ingest.
 Factors Causing Rancidity Catalysts:
 trace metal ions & inorganic salts
 Temperature
 Amount of PUFA
 Time
 Light
 Water
 EMULSIFICATION
 Emulsification is a process by which you
mix two liquids that are ordinarily
immiscible. Emulsification of fats to mix
them with water-based substances, which
has important implications for cooking
and for digestion. To emulsify fats, you
need substances such as bile salts or
other compounds that help connect the
fat and water. Droplets of the dispersed
 LIEBERMANN-BURCHARD
TEST
 The cholesterol is react as a typical alcohol with a strong
concentrated acids and the product are colored substances.
 Acetic anhydride are used as solvent and dehydrating agents,
and the sulfuric acid is used as dehydrating and oxidizing
agent.
 The acetic anhydride in the Leibermann-Burchard test can
react with the C3 hydroxyl group of cholesterol and related
steroid in the presence of a strong acid such as concentrated
sulphuric acid.
 A positive result is observed when the solution becomes:
(red  blue, and finally bluish –green color)
 SALKOWSKI TEST
 Salkowski Test is a test for cholesterol. It was said
that the reaction principle of this test indicates
the presence of double bond in one cholesterol
ring is responsible for its ability to form colored
products in the presence of concentrated
inorganic acids and it should yield a layering of
colors. When concentrated sulfuric acid is added
to a chloroform solution of cholesterol, the
chloroform layer shows a red to blue color and
the acid layer shows a green fluorescence. The
OBJECTIVES &
SUMMARY OF
PROCEDURES
1

• To list some chemical properties of lipids

2
• To identify the best solvents for lipids
3
• To test the different components of lipids
4
• To differentiate rancid from fresh oil using
different indicators
SOLUBILITY

 Coconut Oil
Distilled
Water
5% HCl
5% NaOH
Observe solubility of Benzene
Ether
oil in the different
solvents Ethyl
TRANSLUCENT SPOTS

Note the formation of a semi-

transparent spot. Allow to
evaporate spontaneously and
observe what happens to the
*REPEAT using GLYCEROL
transparent spot
 ACROLEIN TEST

Observe change
 
Coconut in colour and
Oil odour evolved
*REPEAT using GLYCEROL and
TEST FOR UNSATURATION  Coconut Oil
Oleic Acid
Crystals of
Palmitic
Acid

 
Shake and
allow to
stand for a
few
minutes
w/occasion
ally
shaking.
Observe
TEST FOR RANCIDITYCoconut Oil
Phenolphth
alein
Methyl
Orange
Blue & Red
Litmus

*REPEAT using RANCID

EMULSIFICATION Coconut
Oil
Bile
Solution
Albumin
Soap
Solution

Shake and compare

results
LIEBERMANN-BURCHARD
TEST
-

Place in the dark

(10 min.),
observe color
change
(0.5g) Cholesterol + 2ml
Chloroform
Acetic Anhydride
Conc. Sulfuric Acid
SALKOWSKI TEST

Note colour
produces both in
the chloroform
and sulphuric
acid layers

(0.5g) Cholesterol + 2ml

Chloroform
RESULT
S
 Treatme
Test Result
nt
Solubility of Immiscible and forms bubbles
a. Distilled water
Coconut oil in: on top of the solution
Immiscible and forms bubbles
b. 5% HCl
on top of the solution
Immiscible and forms bubbles
c. 5% NaOH
on top of the solution
d. Ether Miscible and Colorless
e. Benzene Miscible
Immiscible and forms bubbles
f. Ethyl alcohol
on top of the solution
g. Chloroform Miscible
 C D
B

H
A E G

SOLUBILITY
 Treatme
Test Result
nt
Translucent spot Coconut oil Easily dry and widely spread
Formed very slow and small spread
Glycerol
compared to coconut oil
Brown colour w/ strong pungent
Acrolein Test Coconut oil
burnt fat odor
Light brown colour w/ strong
Glycerol
pungent burnt fat odor
Dark Brown colour w/ strong
Oleic acid
pungent burnt fat odor
Formation of lighter brown layer on
Unsaturation Coconut oil
the top of the dark brown layer
Oleic acid Brown color; thick substance formed
Violet color on top of light brown
Palmitic acid
layer
TRANSLUCENT
SPOT
Coconut Oil
Palmitic acid

UNSATURATION
Oleic acid
 Test Treatment Result
Rancidity Fresh oil w/ phenolphthalein Pink color
Orange color at the
w/ methyl orange
bottom of the tube
w/ Blue litmus remained
a. Red litmus the same and Red
b. Blue litmus turned blue
Rancid Oil w/ Color of solution is light
phenolphthalein brown
w/ methyl orange Cloudy red color
w/ Red litmus remained
a. Red litmus red, while the blue
b. Blue litmus litmus turned red
 Test Treatment Result
Brown colour w/ formation
Emulsification Coconut oil w/ bile
of suspended bubbles
Light yellow colour w/
w/ albumin formation of suspended
bubbles
Cloudy white colour
w/ soap solution
mixture
Leibermann –
Dark green colour is formed
Burchard
Chloroform layer is red and
Salkowski sulfuric acid layer is yellow
w/ green fluorescence
Salkowski
ANSWERS TO
QUESTIONS
 1. Give a generalization of solubility
of lipids
 Lipids is a non-polar organic compounds and
we know that when it comes to solubility
non-polar will only dissolve in a non-polar
solvents wherein it follows the rule “like
dissolve like”. This idea was proven when
the activity was conducted, the coconut oil
is insoluble in polar solvents such as the
distilled water, hydrochloric acid, sodium
hydroxide and ethyl alcohol, but it is highly
miscible in a non-polar solvents like ether,
 2. Define:
 EMULSIFICATION is the breakdown of large
fat globules into smaller, uniformly
distributed particles. It is accomplished
mainly by bile acids in the small intestine.
 RANCIDITY is the complete or incomplete
oxidation or hydrolysis of fats and oils when
exposed to air, light, moisture or by bacterial
action, resulting in unpleasant taste or odor.
 3. What is the importance of the
acrolein test? Write a balanced
 equation.
Acrolein test is important in
detecting the presence of glycerol or
fats in such solutions. If fat is
treated with a dehydrating agent
the glycerol portion of the molecules
is dehydrated to form an
unsaturated aldehyde, acrolein that
has a pungent irritating odour.
Glycer
ol

Acrolei
n
 4. Differentiate the two types of
rancidity.
 Oxidative and hydrolytic are the two
types of rancidity. Oxidative
rancidity (autoxidation) occurs when
oxygen is absorbed from the
environment. In the presence of
oxygen and/or ultraviolet radiation,
most lipids will break down and
degrade forming several other
 On the other hand, hydrolytic
rancidity (hydrolysis or enzymatic
oxidation) occurs in the absence of
air, but with moisture present. This
normally is accomplished through
enzymatic peroxidation, where
enzymes found naturally in plant
oils and animal fats can catalyse
reactions between water and oil.
 5. What is the reaction of
phenolphthalein and methyl orange in
acid-base solution?

Acid Base
Phenolphthalei
Colorless Pink Colour
n

Methyl Orange Pink Colour Yellow Colour

 6. What is the importance of
emulsification in the digestion of lipid?
 Emulsification is important in the
breaking down of triacylglycerol fats in
digestion. Fats repel water because they
are hydrophobic and remain insoluble in
the small intestine. The enzymes that is
responsible in breaking down fats into
fatty acids and monoglycerols for easier
absorption is the lipase which is
produce by the pancreas, but lipase can
 This is where emulsification process comes
in, emulsification breaks down the fats into
smaller manageable droplets and free
floating bile salts and phospholipids are
recruited and surround each droplet. Bile
salts and phospholipids are amphipathic so
have both hydrophobic and hydrophilic
surfaces and can ensure that large fat
globules cannot reform. With the help of
emulsification process it reduces surface area
for the lipase to efficiently digest the fats.
 7. What is the positive result and the
importance of Liebermann-Burchard and
Salkowski?
Positive Importan
Test
Result ce
Important in
Change of color
Liebermann- detecting the
into a green or
Burchard green-blue
presence of
cholesterol
Color become Important in
bluish red and detecting the
Salkowski