Definition Classification

Biological Biochemical
importance Test
Definition

Organic substances relatively insoluble

in water but soluble in organic solvents
like chloroform, ether and benzene
 Lipid compounds including:

• Fats and Oils (triglyceride)

• Waxes
• Steroids
• Fat- soluble vitamins
• Phospholipid
• Mono- di glycerides
Simple Complex Derived
lipid lipid lipids
Neutral fats or oils Waxes
Alcohol is
Alcohol is
other than
GLYCEROL
glycerol
 Fats and oils are
 also called triglycerides.

 esters of glycerol+ 3 FA
When hydrolysis of waxes gives : one fatty acid + alcohol

EXAMPLES
 Beeswax
 Subclassified according to the type of prosthetic group

Phospholipids Glycolipids Lipoproteins

FA + ALCOHOL + PHOSPHORIC ACID

They
frequently
have
nitrogen
containing
bases
Phospholipids
may be
classified on the
basis of the
type of alcohol
present
A. Glycerophospholipids B. Spingophospholipids

 Examples  Examples
 Plasmalogens  Spingomyelins
 Cardiolipins
FA + ALCOHOL[SPINGOSINE] +CARBOHYDRATE
WITH NITROGEN BASE

Example
Cerebrosides
Gangliosides
 Chylomicrons
Lipid with Very low density lipoprotein
prosthetic (VLDL)
group PROTEIN Low density lipoprotein (LDL)
High density lipoprotein (HDL)
 Example

Vitamin A
Fatty acids Steroids Cholesterol
and D
 Structural Precursor of
Storage form component of many steroid Protection of
of energy cell hormones, internal organs
membrane. vitamin D
 Fatty acids are
 long-chain carboxylic acids.

 typically 12-18 carbon atoms.

 insoluble in water.

 saturated or unsaturated.
Fatty acids are
saturated with all single C–C bonds.
are solids at room temperature

unsaturated with one or more double C=C bonds.

are liquids at room temperature.
Chemical tests for:
1. Solubility in polar and nonpolar solvent
2. Emulsification
3. Oxidation of unsaturation FA
4. Qualitative analysis of detection in natural products ( salkowski reaction)
1. Solubility in polar and nonpolar solvent
• Reagents
• Olive oil (vegetable oil), butter (animal fat),
• stearic acid (saturated fatty acid), oleic acid (unsaturated fatty acid),
• Solvents: dilute acid and alkali solutions, cold alcohol, hot alcohol,
benzene, chloroform, ether and carbon tetrachloride.

Procedure
1. Add a 2drops of the liquid fat or 0.1 g of the solid fat in labeled test
tubes.
2. To each test tube add a 1 ml of solvent and write down your
observations.
3. Repeat the experiment with a different solvent and make your
observations.
2- Emulsification

Reagents
• Neutral olive oil, 0.5 % Na2C03,

Procedure
1. Shake up a drop of neutral olive oil with a little water in a test tube, the fat
becomes finally divided forming an emulsion, upon standing fat separates and
rises to the top.

2. To 5 ml water in a test tube add 2 to 3 drops of 0.5 percent Na2CO3. Introduce a

drop of neutral olive oil and shake, the emulsion is not permanent and is not so
transitory.

3. Repeat step 2 using rancid olive oil. In this case the alkali combines with the
free fatty acids to form soap and the soap being an emulsifying agent, it
emulsifies the fat.
2- Emulsification

• An emulsion is a mixture of two or more liquids that

are normally immiscible (no mixable).

• Bile Salts Synthesized from cholesterol and stored in

the gallbladder When you eat fat, the bile salts act as
soap and help emulsify the large globules of fat.

A. Two immiscible liquids, not yet emulsified

B. An emulsion of Phase II dispersed in Phase I
C. The unstable emulsion progressively separates
D. The surfactant stabilizing the emulsion
1. Oxidation of unsaturation FA

• Into test tube place one drop of oil and 3 ml of Na2CO3

• Warm it slightly and drop solution KMnO4 .after each drop the
violet color disappear
• The end of the reaction recognized by ppt brown solid MnO2
4-Qualitative analysis of detection in natural
products ( salkowski reaction)

• Reagent:
o.s , Chloroform, sulphuric acid
Procedure:
Take 1 ml O.S
Then add 2 ml chloroform
2ml sulphuric Acid and mix it
4-Qualitative analysis of detection in natural
products ( salkowski reaction)

• Observation:
Upper layer is red and lower layer is yellow with
green floresence

Cholesterol is present.
5-Qualitative analysis. Sudan III test

• Reagent:
o.s , Chloroform, Sudan III reagent
Procedure:
Then add 0.5 ml chloroform
Add O.S drop by drop
Then add one drop of Sudan reagent
5-Qualitative analysis. Sudan III test

• Observation
Red color formed
Test is positive

Lipid present
7-Qualitative analysis. Saponification

• Reagent:
O.S, 20% ethanolic sodium hydroxide NaOH, Distilled
water, HCL
Procedure:
Take original solution then add 20% NaOH then boil and
cool. Then add 10 ml water and then boil and cool
Then add few drop HCL
Observation: A solid fatty acid mass is formed
Lipid is present
 O
O
R
C OH R OH
R C O R

Carboxylic Acid Alcohol Ester

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