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EXPERIMENT NO. 3
LIPIDS A. SPOTTING EFFECT
GROUP 1
INUR-1
Alfonso, Dorothy Angela O.
Antero, Ma. Layla Y.
Avila, Alyanna Ainah A.
Arceno, Anthony C.
Anes, Daniela Louisse B.

A. Spotting Effect A. Spotting Effect

• Samples
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A. Spotting Effect A. Spotting Effect

Samples With (+) or without (-) translucent spot • Lotion
1. Lotion +
2. Hair wax +
3. Vegetable oil +
4. Cream +
5. Vitress +

A. Spotting Effect A. Spotting Effect

• Hair wax • Vegetable oil
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A. Spotting Effect A. Spotting Effect

• Cream • Vitress (Hair oil)

B. Solubility
Vegetable oil

B. SOLUBILITY
•
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B. Solubility B. Solubility
• Vegetable oil

Sample Water Methylene Ether Toluene

chloride Oleic acid

Vegetable oil Immiscible Miscible Miscible Miscible

B. Solubility B. Solubility
• Vegetable oil and ether • Lecithin
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B. Solubility B. Solubility
Sample Water Methylene Ether Toluene
chloride
Lecithin Immiscible Miscible Miscible Miscible
Soybean oil (33-35% of Lecithin)

C. Test for Unsaturation

C. TEST FOR Test for: determining the presence of double bonds in unsaturated lipids

UNSATURATION
•

• Principle: Halogenation via addition

• Positive visible result: decolorization of the iodine solution
• Reagents: methylene chloride (CH2Cl2), iodine solution
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C. Test for Unsaturation C. Test for Unsaturation

• Color of iodine solution: orange Sample Color Change of Iodine Solution
Vegetable oil Turned yellow
Lecithin No change

Iodine
solution in Iodine
vegetable oil solution in
lecithin

C. Test for Unsaturation C. Test for Unsaturation

• Fatty acids in animal fats are usually saturated, whereas those in vegetable oils are
generally unsaturated.
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D. Acrolein Test
Test for: determining the presence of fats or glycerin (glycerol)

D. ACROLEIN TEST
•

• Principle: dehydration
• Results: burnt fat odor
• Reagents: glycerol, potassium bisulfate (KHSO4)

D. Acrolein Test D. Acrolein Test

Sample Odor of the Vapor
Glycerol Slightly burnt smell
Cooking oil Fried
Lecithin Burnt fat

After heating
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D. Acrolein Test
Purpose of potassium bisulfate
E. EXTRACTION OF
BRAIN LIPIDS
•
• Dehydrating agent

• Compound responsible for the odor

• Acrolein
• smell of burnt fat is caused by glycerol in the burning fat breaking down into acrolein
• toxic and is a strong irritant for the skin, eyes, and nasal passages

E. Extraction of Brain Lipids E. Extraction of Brain Lipids

Methods: • To the Decantate:
• In an Erlenmeyer or Florence Flask, place the homogenized brain and add enough • Add acetone gradually until precipitation is complete
ether to completely immerse the brain sample.
• Filter the Mixture
• Cover Tightly and set aside
• The Residue (Residue B) is to be divided into 3 portions
• Decant the incubated brain sample
• These portions will be used in the Ninhydrin, Soda Lime, and Ammonium
Molybdate Tests.
• Evaporate the filtrate until it dries using a steam bath
• Label the residue obtained as Residue C to be used in the Leibermann-Burchard
Test.
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E. Extraction of Brain Lipids E. Extraction of Brain Lipids

• To the Residue: • Done in order to obtain Residue B to be used in the Ninhydrin, Soda Lime, and
Ammonium Molybdate Tests.
• Get a small portion of the incubated brain sample and add 10ml of hot 95%
ethanol • Residue C to be used in the Leibermann-Burchard Test
• Mix thoroughly and decant • And Decantate A to be used in the Molisch Test
• Use the decantate and label it as Decantate A to be used in the Molisch Test

E. Extraction of Brain Lipids E. Extraction of Brain Lipids

• Done in order to obtain Residue B to be used in the Ninhydrin, Soda Lime, and
Ammonium Molybdate Tests.
• Residue C to be used in the Leibermann-Burchard Test
• And Decantate A to be used in the Molisch Test
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F. 1. Molisch Test
F. DETECTION OF BRAIN LIPIDS
1. MOLISCH TEST

F. 1. Molisch Test F. 1. Molisch Test

• Molisch Test is a sensitive chemical test for the presence of carbohydrates, but for • To 2.0 ml of decantate A, add 5 drops of Molisch’s Reagent. Mix Thoroughly.
the case of lipids. There are types of lipids that have carbohydrates attached to
them (glycolipids). When the concentrated sulfuric acid is added to the mixture of • Tilt the tube and carefully add 1.0 ml of concentrated sulfuric acid, drop by drop.
decantate A and the molisch reagent, A purple ring is formed. • Note the color of the ring formed at the junction of the lipids.
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F. 1. Molisch Test
F. DETECTION OF BRAIN LIPIDS
2. NINHYDRIN TEST

F. 2. Ninhydrin Test F. 2. Ninhydrin Test

• Objective • Materials
• To detect the presence of an alpha amino group 1. Residue B
2. Water
3. Ninhydrin reagent
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F. 2. Ninhydrin Test F. 2. Ninhydrin Test

• Procedures • Results
1. Dissolve a portion of residue B in 2.0 ml of water • Formation of a blue violet solution or
2. Add 1.0 ml of 1% ninhydrin reagent. Mix thoroughly and heat in a boiling water bath for 2 Ruhemann’s purple
minutes.
3. Note the color of the resulting solution. Record observations

F. 2. Ninhydrin Test F. 2. Ninhydrin Test

Test Purpose Reagent Compound Tested Principle + VR Compound
responsible
for +VR

Ninhydrin Detect the Ninhydrin Phosphatidylserine Decarboxylation Ruhemann’s

test presence of Deamination purple
an a-nh2 or a- Condensation
nh group
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F. 3. Soda Lime Test

F. DETECTION OF BRAIN LIPIDS
Objective

3. SODA LIME TEST

•
• To detect the presence of an amino (NH2) or an imino (NH) group

F. 3. Soda Lime Test F. 3. Soda Lime Test

• Procedures • Results
1. Grind a pinch of soda lime and portion • Red to blue color of litmus paper
of residue B
2. Transfer the mixture in a dry test tube
and heat gently over a small flame.
3. After a min of heating, test the vapor
by placing pieces of red and blue
litmus paper at the end of the stirring
rod.
4. Record observations
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F. 3. Soda Lime Test F. 3. Soda Lime Test

Test Purpose Reagent Compound Tested Principle + VR Compound
responsible for +VR

Soda lime To detect the NaOH, CaO Phosphatidyl- Deamination Red to

test presence of Ethanolamine blue
amino (nh2) litmus
and imino(nh) Phosphatidyl- paper
group Serine

F. DETECTION OF BRAIN LIPIDS

F. 4. Ammonium Molybdate Test
4. AMMONIUM • Ammonium molybdate test is a test for the presence of phosphate group.

MOLYBDATE TEST
• When added to phosphate ions, warmed with nitric acid it forms a yellow
precipitate of ammonium phosphomolybdate complex. A Confirmatory test for
presence of phosphates.
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F. 4. Ammonium Molybdate Test F. 4. Ammonium Molybdate Test

• PO43- + 12(NH4)2MoO4 + 21HNO3 + 3H+ -> (NH4)3PO4·12MoO3↓ + 21HN4NO3 +
12H2O

• the lipid hydrolyses, producing free phosphate.

F. 4. Ammonium Molybdate Test F. 4. Ammonium Molybdate Test

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F. DETECTION OF BRAIN LIPIDS

F. 5. Leibermann-Burchard Test
5. LEIBERMANN- • Liebermann – Burchard is a test for the presence of sterol.

BURCHARD TEST
• Cholesterol produces a characteristic green color when it is mixed with the
Liebermann–Burchard reagent, a mixture of acetic anhydride and sulfuric acid.
The change in color may be gradual, initially pink, then blue-purple, and finally
deep green.

F. 5. Leibermann-Burchard Test F. 5. Leibermann-Burchard Test

• Acetic anhydride is used as solvent and dehydrating agents and the sulfuric acid is
used as dehydrating and oxidizing agent.
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F. 5. Leibermann-Burchard Test F. 5. Leibermann-Burchard Test