DETERMINATION OF FATS AND OILS CONSTANTS

Fats and oils are recognized as essential nutrients in both human and animal diets. Nutritionally,
they are concentrated sources of energy( 9 Kcal/gram); provide essential fatty acids which are
the building blocks for the hormones needed to regulate bodily systems; and are a carrier for the
oil soluble vitamins A, D, E, and K. They also enhance the foods we eat by providing texture and
mouth feel, imparting flavor, and may contribute to the feeling of satiety after eating. Fats and
oils are also important functionally in the preparation of many food products. They act as
tenderizing agents, facilitate aeration, carry flavors and colors, and provide a heating medium for
food preparation. Fats and oils are present naturally in many foods, such as meats, dairy
products, poultry, fish, nuts and in prepared foods, such as baked goods, margarines, dressings
and sauces. To understand the nutritional and functional importance of fats and oils, it is
necessary to understand some constants used in their description:

Acid value (Acid Number)

The acid value (AV) is the number that expresses, in milligrams the quantity of potassium
hydroxide required to neutralize the free acids present in 1 g of the substance. The acid value
may be overestimated if other acid components are present in the system, e.g. amino acids or
acid phosphates. The acid value is often a good measure of the breakdown of the triacylglycerol
into free fatty acids, which has an adverse effect on the quality of many lipids.

ml of KOH x N x 56
AV = = mg of KOH
Weight of Sample

N = Normality of KOH
% Free Fatty Acid (FFA) = AV x 0.503

Material
 Fat or Oil
 Absolute ethanol alcohol
 Phenolphthalein
 0.1 N KOH

Procedure

1. Place 5.0 g of fat or oil in a dried conical flask.

2. Add 25 ml of absolute ethanol alcohol and add ( 2-3) drops of phenolphthalein
3. Heat with shaking in water bath (65%) for 10 minutes ,then cool Titrate the solution
1
 against 0.1 N KOH until pink color appears (end point).
4. Record your observations.
5. Calculate the acid value (AV) and free fatty acid (%FFA) using above laws.

[2] Saponification Number

The saponification value is the number of mg of potassium hydroxide required to neutralize

the free acids and to saponify the esters in 1 g of the substance. The saponification number is a
measure of the average molecular weight of the triacylglycerol in a sample. Saponification is the
process of breaking down a neutral fat into glycerol and fatty acids by treatment with alkali. The
smaller the saponification number the larger the average molecular weight of the triacylglycerol
present i.e. Saponification value is inversely proportional to the mean molecular weight of fatty
acids (or chain length).
O
H2 C OH
H2 C O C R O
O
+ 3 KOH HC OH + 3R C OK
HC O C R
O
H2 C OH
H2 C O C R

Saponification Value of Fats and Oils

Fat or oil Saponification Value

Milk fat 210-233
Coconut oil 250-264
Cotton seed oil 189-198
Soybean oil 189-195
lard 190-202
Butter fat and vegetable fats ∼ 220 – 250

Material
 Fat or Oil
 0.5 N alcoholic potassium hydroxide ( alcoholic KOH) ( prepared by dissolving 30 g
potassium hydroxide in 20 mL of water and make the final volume to 1 L using 95 %
ethanol. Leave the solution to stand for 24 h before decanting and filtering the solution.
 0.5 N Hydrochloric acid
 Phenolphthalein.

2
Procedure
1. Weigh approximately 2 g of the fat or oil into a 250 mL conical flask.
2. Add 25 mL of alcoholic potassium hydroxide solution (0.5 N).
3. Attach a reflux condenser and heat the flask contents on a boiling water bath for 1 hour
with occasional shaking.
4. While the solution is still hot, add 3 drops of phenolphthalein indicator and titrate the
excess potassium hydroxide with the 0.5 N hydrochloric acid (Vml of hydrochloric acid
at end point represents S).
5. Repeat the procedure but without sample (Vml of hydrochloric acid at end point
represents B).
6. calculate the saponification number by using the following law:

B: ml of HCl required by Blank.

56.1(B -S) x N of HCl
SP# =
Gram of Sample S: ml of HCl required by Sample.

[3] Ester Value

The ester value is defined as the mg of KOH required to react with glycerin (glycerol / or
glycerin) after saponify one gram of fat. It is calculated from the saponification
Value (SV) and the acid Value (AV):

Ester Value (EV) = Saponification Value (SV) – Acid Value (AV)

% glycerin Ester Value 0.054664

[4] Iodine Value (I.V)

The iodine value (IV) gives a measure of the average degree of unsaturation of a lipid: the
higher the iodine value, the greater the number of C=C double bonds. By definition the iodine
value is expressed as the grams of iodine absorbed per 100g of lipid. Iodine value (I.V.) is
directly proportional to the degree of unsaturation (No of double bonds.) and inversely
proportional to the melting point (M.P.) of lipid. An increase in I.V indicates high susceptibility
of lipid to oxidative rancidity due to high degree of unsaturation.
One of the most commonly used methods for determining the iodine value of lipids is
"Hanus method". The lipid to be analyzed is weighed and dissolved in a suitable organic solvent,

3
to which a known excess of iodine chloride is added. Some of the IBr reacts with the double
bonds in the unsaturated lipids, while the rest remains:

R-CH=CH-R + IBr excess  R-CHI-CHBr-R + IBr remaining

The amount of IBr that has reacted is determined by measuring the amount of IBr remaining
after the reaction has gone to completion (IBr reacted =IBr excess – Ibr remaining). The amount of IBr
remaining is determined by adding excess potassium iodide to the solution to liberate iodine, and
then titrating with a sodium thiosulfate (Na2S2O3) solution in the presence of starch to determine
the concentration of iodine released:

IBr remaining + 2KI  KBr + KI + I2

I2 + starch + 2Na2S2O3 (blue)  2NaI + starch + Na2S4O6 (colorless)

TYPICAL IODINE NUMBERS

Coconut oil 8 - 10

Butter 25 - 40

Beef tallow 30 - 45

Palm oil 37 - 54

Lard 45 - 70

Olive oil 75 - 95

Peanut oil 85 - 100

Cottonseed oil 100 - 117

Corn oil 115 - 130

Fish oils 120 - 180

Soybean oil 125 - 140

Safflower oil 130 - 140

Sunflower oil 130 - 145

Linseed oil 170 - 205

Material
 Oil or fat
 Hanus solution ( it’s prepared by dissolving 18.2 g of iodine in 1L of glacial acetic acid
and then add 3 ml of bromine water for increasing the halogen content.

4
  15% potassium iodide solution
 1% starch solution
 0.1 N Sodium thiosulfate solution.

Procedure
1. Weigh approximately 0.25 g of the fat or oil into a 250 mL conical flask.
2. Add 10 ml of chloroform.
3. Add 30 ml of Hanus solution and close the flask completely by Para film, then
leave the solution for 30 minutes with shaking continuously.
4. Add 10 ml of 15% potassium iodide solution and then shake.
5. Add 100 ml of distilled water (DW).
6. Titrate the iodine solution against 0.1 N Sodium thiosulfate solution till yellow
color formed , then add 2-3 drops of starch solution where blue solution formed
and then continue with titration till the blue color is disappeared (Volume (ml) of
Na2S2O3 at end point represents S)
7. Do same above procedure but without sample (Volume (ml) of Na 2S2O3 at end
point represents B).
8. Calculate the iodine number by using the following law:

( B - S )  N of Na2S2O3  0.127g/meq  100

Iodine Value =
Weight of Sample (g)

B: V ml of Na2S2O3 volume for blank

S: V ml of Na2S2O3 volume for sample

Questions

1. What is the difference between fats and oils?

2. a. What is fatty acid?
b. What are the classes of fatty acid?
c. Give three examples of each class of fatty acid.
d. what factors will increase the free fatty acid in an oil/fat?
3. What is the significance/importance of the following
a. acid value
b. saponification value
c. ester value
d. free fatty acid

Significance ( acid value)

5
Acid value is the measure of hydrolytic rancidity. In general, it gives an indication about edibility
of the lipid.
- Edible oil contain > 1%
- Pharmaceutical oil must not have any acidity.