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Lipids Chemical Test
Lipids Chemical Test
CHAPTER III
General Tests
- Tests for Lipids
Reagents:
Distilled water
Olive oil
Procedure:
1. Add 1mL (20 drops) of the sample in each pair of liquids indicated.
2. Mix the contents of each tube by agitating.
3. Wait for 2 minutes.
4. Examine the tube carefully. Has the sample dissolved in the solvent or do you
see two separate layers in the tube?
5. Record your observations.
6. Save the tube for the Sudan Red test described below.
Positive Result:
Two separate layers in water and dissolved in olive oil
Principle:
Solubility, Polarity
Procedure:
1. Put a drop of the sample on a piece of unglazed paper.
2. Draw a circle around the spot with a soft pencil.
3. Allow the spot to dry thoroughly.
4. Hold the paper in front of a light source and observe the spot.
Positive Result:
Translucent
Principle:
Opacity
C. Physical Properties
Color= palest yellow to greenish yellow
Appearance= mobile liquid
Odor= typical grapefruit
Positive Result:
Within the range of 1.472 – 1.479
Principle:
Determination of concentration of volatile oils. Determination of oil whether it falls
at the expected range.
Procedure:
- Place a drop of the sample on a filter paper and let it dry.
Positive Result:
No stain in volatile oil
Principle: Volatile oils do not produce permanent spot unlike fixed oils.
Specific tests
-Test for Aldehyde
A. Baeyer’s Test
-Test for unsaturation
-Test for Multiple Bonds
Reagent/s:
potassium permanganate
unknown in 0.5 mL reagent grade acetone
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Procedure:
1. Dissolve 1 drop or 0.02 grams of the unknown in 0.5 mL reagent grade
acetone.
2. Add a 1% aqueous solution of potassium permanganate dropwise with
shaking.
3. If more than one drop of reagent is required to give a purple color to the
solution, unsaturation or an easily oxidized functional group is present.
Positive Result:
The disappearance of the KMnO4's purple color and the appearance of a brown
suspension of MnO2 is a positive test.
Principle: Oxidation
B. 2, 4- Dinitrophenylhydrazone Test
Test for aldehydes & sterically unhinedered ketone
Regent/s :
2,4-DNP reagent
95% ethanol
Procedure:
1. Dissolve 2-3 drops of your test compound in 2 ml 95% ethanol in a small test
tube.
2. Mix this solution with 2 ml of the 2,4-DNP reagent.
3. Look for the formation of an orange-yellow precipitate to indicate the presence of
an aldehyde or ketone.
Positive Result:
Orange hydrazone ppt. indicates the presence of a ketone (conjugated) , yellow
hydrazone ppt. indicates an aldehyde (non-conjugated)
Principle:
ANE
C. Fehling’s Test
Reagent/s:
Fehling’s A- 7 g copper II sulfate dissolved in 100 ml water
Fehling’s B- 35 g potassium sodium tartrate and 10 g NaOH in 100 ml water
Procedure:
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1. With equal amounts of Fehling’s A and Fehling’s B to make 1 ml, add 3 drops of
sample.
2. Water bath for 10-15 minutes.
Positive Result:
Brick red precipitate
Principle:
Oxidation of strong alkaline medium
D. Benedict’s Test
Reagent/s:
Benedict’s solution
Procedure:
1. Add 2 ml Benedict’s solution to the test tubes
2. Add 2-3 drops of test compound to each test tube.
3. Heat the test tubes in boiling water for 10-15 mins.
4. Look for the formation first of a pale green color followed by the formation of
the reddish precipitate of cuprous oxide.
Positive Result:
Pale green color followed by the formation of the reddish precipitate
Principle:
Oxidation in less basic medium
E. Bromine Test
Test for unsaturation
Reagents:
0.5% Br2 in CCl4
Procedure:
1. Place 5 drops of the sample in a dry test tube. Add 10 drops 0.5% Br2 in CCl4
reagent. Shake the test tube vigorously and observe the rate and extent
(colour change) by which the reagent is decolorized; the bromine color is
discharged. OPTIONAL: Place moistened blue litmus paper on a glass rod
across the mouth of the test tube and note any color change.
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2. Compare with water as a negative control. If the reagent failed to decolorize
within 1 minute, expose the reaction mixture to sunlight. Note down the
results.
Positive Result:
Colorless - Discharging of the bromine color without the evolution of hydrogen
bromide gas
Principle:
Radical substitution reaction
F. Nitration Test
Test for aromaticity
Reagent/s:
HNO3,
H2SO4
Nitrating Mixture
Procedure:
1. Place 2 mL of conc. HNO3 in an Erlenmeyer flask. Immerse the flask in a water
bath and gradually add 2 mL conc. H2SO4. Cool the resulting mixture to room
temperature. This will serve as the nitrating mixture.
2. Place 5 drops of the sample in a dry test tube. Add 8 drops of the nitrating
mixture and shake the test tube to ensure complete mixing. Note the formation of
a yellow oily layer or droplet. Dilute with 20 drops of water.
3. If there is no apparent reaction observed within a minute, place the test tube in a
water bath (~50°C) for 10 minutes. Dilute with 20 drops water. Note down the
result.
Positive Result:
Yellow oily layer
Principle:
Oxidation
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Experimental Result
General Tests
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Experimental Result
Specific Test
Name of the Test (+) Result Experimental Actual Figure
Result
1. Solubility in Two separate Two separate
Polar And Non layers in layers in
Polar Solvents water and water and
dissolved in dissolved in
olive oil olive oil
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Name of the (+) Result Experimental Actual Figure
Test Result
1. Bayer’s Test Disappearance of Disappearance of
purple color and purple color and
appearance of appearance of
brown suspension brown suspension
of MnO2 of MnO2
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6. Nitration Yellow oily layer Yellow oily layer
Test