ACTIVITY # 6

LIPIDS
GROUP #2
NUCLEOTIDE, FATTY ACID, POLYMERIZATION, MUTATION,
ALCOHOL, PROTEIN, DISACCHARIDE, AMINES, CATABOLISM
 OBJECTIVES

• To examine some physical and

chemical properties of lipids.
 MATERIALS

• Test tubes (10) • Molder • Watch glass (to cover

• Test tube rack • pH paper the150 ml. Beaker)

• Test tube holder • Large test tube • Water bath

• Test tube brush • Bunsen burner • Tripod

• Funnel • Pipette and Aspirator • Wire gauze

• Filter paper • Graduated cylinder • Stirring rod

• Spatula • Dropper • 600 ml. Beaker

• Evaporating dish • Platform balance

• Hot plate • 150 ml. Beaker
 REAGENTS
• coconut oil • egg yolk • 0.5% ferric chloride
• cottonseed oil • butter • 0.5% magnesium
• olive oil • KHSO chloride

• linseed oil • Tincture of iodine • HCI

• soybean oil • CHC13 • 6N

• mineral oil • acetic anhydride • methylene chloride

• cholesterol • concentrated HSO • NaOH pellets

• glycerol • powdered detergent • Anhydrous chloroform

• oleic acid • distilled water

• stearic acid • 0.5% calcium chloride
 PROCEDURE

NOTE: Be sure dry test tubes when performing procedure, A, B and C

I. TEST FOR FATS AND OILS
ACROLEIN TEST
 Place 2 drops of glycerol,
coconut oil, cholesterol,
olive oil in separate labeled
test tubes.
 We noticed that the
CHOLESTEROL mixed with
3mL CHLOROFORM turned
into a white colored solution
while the others are still a clear
solution.
then add a pinch of KHSO4.
Heat it gently, then
vigorously.
After heating, we noticed that
the KHSO4 didn’t dissolved
from the 4 samples.
 THIS PROCESS WILL INHIBIT THE
FORMATION OF SO 2 , FUMES WHICH
WOULD OBSCURE THE ODOR DUE TO
ACRYLIC ACID ALDEHYDES. DESCRIBED THE
ODOR PRODUCED.
 WHICH OF THE SUBSTANCES TESTED GAVE
ACROLEIN? WHY?
- WHEN THE G LYC EROL I S TREATED S TRONG LY WI TH THE P RES ENCE OF
K H S O 4 ( D E H Y D R AT I N G AG E N T ) .
- THE G LYCE ROL I S D E H YDRAT ED TO F O R M AN UN S AT UR AT E D
AL D E H YDE , W H I CH I S AC RO L E IN.

WRITE THE CHEMICAL EQUATION FOR THE

FORMATION OF ACROLEIN FROM GLYCEROL AND
POTASSIUM HYDROGEN SULFATE.
I. TEST FOR FATS AND OILS
TEST FOR UNSATURATION
 Place 1 drop of olive oil,
oleic acid, stearic acid, in dry,
separate labeled test tubes.
Add tincture of iodine drop
by drop, shaking it until a
reddish violet color persists.
After 2 drops of iodine in
the oleic acid, a reddish-
violet color persisted.
After 2 drops of iodine to
olive oil, a reddish-violet
color persisted.
After 5 drops of iodine to
the stearic acid, still no
reddish-violet color
persisted.
 BE SURE TO ELIMINATE CROSS
CONTAMINATION SAMPLES. RECORD THE #
OF DROPS CONSUMED.
I. TEST FOR FATS AND OILS
LIEBERMANN-BURCHARD
TEST FOR CHOLESTEROL
 Place a few grains of your
cholesterol, 0.5 mL egg yolk,
butter and a drop of olive
oil in separate labeled test
tubes.
Transfer 3 mL of chloroform
and 1 mL of acetic
anhydride to each test tube.
 Finally, carefully add 1 drop
of concentrated sulfuric acid
to each mixture.
Note: when we dropped
sulfuric acid inside the egg
yolk solution, we noticed that
their was a purple dot. Which
was the sulfuric acid.
MIX THE CONTENTS AND RECORD
THE COLOR CHANGES, IF ANY.
After mixing the contents,
we noticed that the white
colored solution remained
at the bottom of the
solution.
 After mixing the
contents, we noticed that
the white foam-like
substance is scattered
inside the test tube.
After mixing the content,
we noticed that the oil is
more visible. But it is still
clear.
After mixing the content, the
color of the butter scattered
so the solution turned yellow
and at the top, their was a
white foam.
WAIT 5 MINS. RECORD AGAIN THE COLOR
OF YOUR SOLUTION. RECORD YOUR
OBSERVATIONS ON YOUR DATA SHEET.
II. CHEMICAL TESTS ON SOAP
AND SYNTHETIC DETERGENT
1.Prepare a solution of 1 g
of a powdered detergent in
50 mL of distilled water.
2. Test the pH of your soap
solution and detergent
solution by dipping the pH
paper. Record your results
in the report sheet.
 3. Pour 10 mL of your soap
solution in three labeled test
tubes. Add 2 mL of 0.5%
calcium chloride solution in
test tube 1 2 mL of 0.5%
ferric chloride on the
second tube and on the
third tube place magnesium
chloride. Repeat the same
procedure this time testing
reactivity of detergent on
the three solutions.
 4. In a clean large test tube
place 10 mL of your soap
solution and 10 mL of the
detergent solution in
another large test tube. Test
it solubility in mineral oil by
adding 4 drops of mineral oil
to each tube and shake.
After shaking allow the
tubes to stand several
minutes and then record any
changes you observe.
 5. Place 4 mL of your
soap solution in a large
test tube and 4 mL of
detergent solution on
another test tube. Add
dilute HCl drop by drop on
both solution until the
solution is at pH 1. Check
the pH using pH paper.
Record the results.
6. To same test tubes used in
the previous procedure, add
4 mL of methylene chloride
to each of the test tube and
shake vigorously. Record
your observations.
DATA ANALYSIS
 ACROLEIN TEST
SAMPLE ODOR ODOR RATE
1- HIGHEST & 4- LOWEST

GLYCEROL SOUR-LIKE ODOR 2

OLIVE OIL ODORLESS 4

CHOLESTEROL ACETONE 1

COCONUT OIL FRAGRANT 3

 CONCLUSION

Acrolein test is used to detect the presence of glycerol or fat. When fat is
treated strongly in the presence of a dehydrating agent like potassium bisulphate
(KHSO4), the glycerol portion of the molecule is dehydrated to form an
unsaturated aldehyde, acrolein that has a pungent irritating odor.

It is concluded that each sample has different odors (or in some cases, odorless).
The glycerol has a sour odor. The olive oil was odorless. The cholesterol has an
odor of an acetone. The coconut oil has a fragrant odor. Each sample was rated
from lowest to highest (1 to 4). Lowest (1) has the weakest odor while highest
(4) has the strongest odor. The cholesterol sample is 4 for the reason that it has
the strongest odor whereas the olive oil sample is 1 as it was odorless which
means there was no odor or smell detected.
TEST FOR UNSATURATION
SAMPLE # of drops of IODINE
SOLUTION
STEARIC ACID
(C18H36O2) 5

OLIVE OIL
((CH3(CH2))n COOH 2

OLEIC ACID
(C18H36O2) 2
 TEST FOR UNSATURATION
SAMPLE OBSERVATION INTERPRETATION

- didn’t
changed - it’s also SATURATED FATTY
- Not SOLUBLE in iodine; doesn’t ACID because it didn’t show any
STEARIC ACID show any reaction to iodine. reaction to the iodine.
(C18H36O2) - Orange color of the iodine.

OLIVE OIL - Olive oil and the iodine solution didn’t - it’s SATURATED means it’s a single
mixed unless being shake , it’s bond and contain maximized # of
((CH3(CH2))n COOH IMMISCIBLE. hydrogen.
- Some particle of the oil didn’t mixed in the
- It’s saturated because it didn’t react
iodine.
- Light red and contains light purple color that fast to iodine takes time before
around it. we see the reaction.

OLEIC ACID - it’s also IMMISCIBLE, not unless being shake. - Saturated it also takes time to
- The iodine stays in the middle of the oleic react with iodine.
(C18H36O2) acid
- Light orange with light reddish color
around it.
 CONCLUSION

Fatty acids are important biocompounds which take part in complex

metabolic pathways, thus having major biological roles. They are obtained from
various dietary sources which determine the type of fat consumed and
consequently health outcome.
 LIEBERMANN-BURCHARD TEST
SAMPLE COLOR CHANGE WITH COLOR CHANGE (5 MINS)
OBSERVATION WITH OBSERVATION
- HAS WHITE FOAM MORE WHITE EGG WHITE LIKE
SCATTERED STARTED TO FORM AT THE TOP
EGG YOLK - IT WAS HOT ON THE TOP OF THE SOLUTION.
(ALBUMIN) PART OF THE SOLUTION.
-YELLOW
BUTTER - WARM AT THE TOP PART YELLOW WITH WHITE
OF THE SOLUTION DOTS

NO COLOR
OLIVE OIL CHANGE NO COLOR CHANGE

WHITE COLOR AT
CHOLESTEROL THE BOTTOM NO COLOR CHANGE
AFTER 5 MINS.
AFTER 5 MINS.
 CONCLUSION

The Liebermann–Burchard or acetic anhydride test is used for the detection of

cholesterol. The formation of a green or green-blue color after a few minutes is positive.
Lieberman–Burchard is a reagent used in a colorimetric test to detect cholesterol, which
gives a deep green color. This color begins as a purplish, pink color and progresses
through to a light green then very dark green color. The color is due to the hydroxyl
group (-OH) of cholesterol reacting with the reagents and increasing the conjugation
of the un-saturation in the adjacent fused ring.
It is concluded that most of the sample tests developed a white or yellow color change
whereas one sample test which is the olive oil, did not develop a color change. These mean
that all of the sample tests are not positive for cholesterol after conducting the experiment.
CHEMICAL TESTS ON SOAP AND
SYNTHETIC DETERGENT
SOLUTION OBSERVATION

DETERGENT SOLUTION

2 mL of 0.5% CaCl2 No reactivity

2 mL of 0.5% FeCl3 Dark brown settled at the

bottom.

MgCl2 There is a white substance that

settled at the bottom.
CHEMICAL TESTS ON SOAP AND
SYNTHETIC DETERGENT
SOLUTION OBSERVATION

SOAP SOLUTION

2 mL of 0.5% CaCl2 No reactivity

2 mL of 0.5% FeCl3 Yellowish-orange light brown at

the top.

MgCl2 Cloudy
 CHEMICAL TESTS ON SOAP AND
SYNTHETIC DETERGENT
10 mL soap solution with 4 drops of 10 mL detergent solution with 4
mineral oil drops of mineral oil

The mineral oil was seen at the top and at The solution became cloudy.
the bottom of the soap solution.

4 mL soap solution with dilute HCl 4 mL detergent solution with dilute

HCl

It took 5 drops for the litmus paper to It took 40 drops for the litmus paper to
react reddish. react almost reddish.
 CHEMICAL TESTS ON SOAP AND
SYNTHETIC DETERGENT

Methyl Iodine with HCl soap solution Methyl Iodine with HCl detergent
solution

The brown part feels hot while the white It feels hot, the bottom is white and it has
part of the solution doesn’t feel as hot. a strong sour odor.
 CONCLUSION
CaCl2, MgCl2 and FeCl3 are generally insoluble in detergent/soap solution creating
precipitate and it makes the solution break down and lose it effectiveness.
Mineral oil and Soap/Detergent solution
Soap and detergent acts as an emulsification that helps bring oil and water together so that
the dirt and grease can be easily rinsed away because soap and detergent have hydrophilic
head and hydrophobic tail.
4 mL Soap/Detergent solution with dilute HCl
Laundry detergents include enzymes. The enzymes would be inactive if enough acid is
added to get the pH outside the enzymes working range. The anionic surfactants would
combine with H+ ions from the acid to form non-charged poorly soluble forms of the
compounds, which Would try to separate as an oily layer. Soap solution with diluted HCl
produces a mixture of fatty acids. Fatty acids are long chain carboxylic acids which may be
saturated or unsaturated.
 CONCLUSION

Iodine reacts with chlorine in the presence of water to form iodic acid. Iodine reacts with
hot concentrated nitric acid to form iodic acid.
Iodine dissolves well in tetrachloromethane without any reaction. It dissolves to a small
extent in water. There is some reaction involved here I2 + H2O --> HI + HOI, but by no
means does all of the iodine react. Iodine reacts with many reducing agents (and seems to
dissolve).
SOUP SOLUTION
 10mL of Detergent solution
with 4 drops of mineral oil

10mL of Soap solution with 4

drops of mineral oil
4 mL of Soap solution with 4 mL of Detergent solution
dilute HCl with dilute HCl
Methyl iodine with HCl Soap Methyl iodine with HCl
solution Detergent solution