INSTITUTE OF MEDICAL SCIENCE TECHNOLOGY

LABORATORY REPORT
HDB 10603
Estimation of Iodine Value of Fats and Oils

 NAME : NIDA’UL ‘ADNIE BINTI AHMAD RIDZUAN

 PROGRAMME: BACHELOR OF BIOMEDICAL SCIENCES
 ID NUMBER : 12213119087
 LAB GROUP : L01-B01
INTRODUCTION:
The main constituents of natural fats and oils are triglycerides. Triglycerides are formed from three
fatty acids which are linked to glycerol by fatty acyl esters. Fatty acids are long aliphatic chains with
carboxyl groups. These are grouped into saturated or unsaturated based on the number of double
bonds present in fatty acid. Saturated fatty acids consist of only single bonds between the carbon
atoms and are likely to exist as solids at room temperature. Unsaturated fatty acids consist of single
bonds as well as double bonds and will be liquids at room temperature. Naturally occurring
unsaturated fats contain double bonds in the cis form. Trans fatty acids are linked with cardiovascular
diseases and health problems. Hydrogenation leads to the conversion of unsaturated fatty acids to
saturated fatty acids. The fatty acids combine with oxygen or halogens depending upon the extent of
unsaturation to form saturated fatty acids. So, it is significant to know the degree of unsaturation.
There are different methods to know the level of unsaturation in fatty acids, one among them is by
determining the iodine number of fats.

OBJECTIVES:
To determine whether the fats are saturated or unsaturated by determining the iodine value

HYPOTHESIS:
Iodine value is the measure of the degree of unsaturation in fats and oils. The iodine number or value
is defined as the number of grams of iodine consumed by 100g of fats. The principle behind the
reaction is the addition of halogens like iodine to the double bonds present in the unsaturated fats. The
iodine monochloride reagent allowed to react with fat in the dark. The amount of iodine consumed is
then determined by titrating the iodine released on adding potassium iodide with standardized sodium
thiosulphate solution and comparing it against the contents in the iodination flask labelled ‘Blank’.
The iodine value can be calculated using the formula:

iodine value=¿
equivalent weight of iodine x volume of sodium thiosulphate used x normality of sodiumthiosulphate x 100
weight of fat sample used for analysis
where equivalent weight of iodine is 127 and normality of sodium thiosulphate is 0.1.

MATERIALS:
1. Iodine Monochloride Reagent
2. Potassium Iodide
3. Standardized 0.1 N Sodium thiosulphate
4. 1% Starch indicator solution
5. Reagent bottle
6. Chloroform
7. Fat sample in chloroform
8. Iodination flask
9. Burette and burette stand with magnetic stirrer
10. Glass pipette
11. Measuring cylinder
12. Distilled water
METHODS:
1. All the reagent solutions were prepared and the requirements were arranged on the table.
2. 10ml of fat sample dissolved in chloroform were pipetted out to an iodination flask labelled as
“TEST".
3. 20ml of Iodine Monochloride reagent were added into the flask. The contents were mixed in
the flask thoroughly.
4. Then, the flask was allowed to stand for a half an hour incubation in dark.
5. A BLANK was set up in another iodination flask by adding 10ml Chloroform to the flask.
6. 20ml of Iodine Monochloride reagent were added to the BLANK and the contents were mix
in the flask thoroughly.
7. The BLANK was incubated in dark for 30 minutes.
8. Meanwhile, the TEST are taken out from incubation after 30 minutes and 10 ml of potassium
iodide solution were added into the flask.
9. The stopper and the sides of the flask were rinsed using 50 ml distilled water.
10. The “TEST” was titrated against standardized sodium thiosulphate solution until a pale straw
colour is observed.
11. About 1ml starch indicator were added into the contents in the flask, a purple colour is
observed.
12. The titration were continued until the colour of the solution in the flask turned colourless.
13. The disappearance of the blue colour was recorded as the end point of the titration.
14. Similarly, the procedure was repeated for the flask labelled ‘Blank'.
15. The endpoint values of the BLANK were recorded.
16. The iodine number were calculated using the equation below:

equivalent weight of iodine x volume of Na2 S 2O 3 x 100 x 10−3

iodine number of fat=
weight of fat sample used for analysis(g)
Equivalent Weight of Iodine = 127
Normality of sodium thiosulphate (Na2S2O3) = 0.1

RESULTS:

Initial burette Final burette reading Volume of the sodium

reading thiosulphate run down (ml)
Blank (without fat) 0 65 65
Test (with fat) 0 64 64
CALCULATION:
Volume of sodium thiosulphate used = [blank - test] ml
= 65 – 64
= 1 ml

equivalent weight of iodine x ( volume of sodium thiosulphate used ) x normalityof sodium

Iodine number of fat=
weight of fat sample used for analysis ( g)
= 6.35

Volume of Volume of Blank - Volume of Normality Equivalent Iodine

blank test solution test sodium of sodium weight value
(mL) (mL) (mL) thiosulphate (mL) thiosulphate (mg)
5 2 3 0.3 0.1 127 19.05
10 4 6 0.6 0.1 127 38.1
15 6 9 0.9 0.1 127 57.15
20 8 12 1.2 0.1 127 76.2
20 10 15 1.5 0.1 127 95.25

DISCUSSION:
Iodine value is the gram (g) of iodine consumed by 100g of fat.
Iodine value indicates: a. The degree of unsaturation in an oil
b. The presence of double bonds
c. Degree of saturation of an oil
d. None of the above
Answer: None of the above
Equivalent weight of Iodine is 127.
The relationship between iodine value and unsaturation of oil is they are directly proportional.
6.35 will be the iodine number of a fat which consumed 1 ml of sodium thiosulphate solution.
The indicator used for iodine estimation analysis is starch.
ASSIGNMENTS:
1. A fat has an iodine value of 250, what can you conclude about the nature of the fat? Can this fat be
provided to a patient having cardiovascular disease?
Iodine esteem is the mass of iodine that has been devoured by 100 grams of substance. So, it
tends to be utilized to decide the measure of unsaturation in unsaturated fats. The more the
iodine value of a compound the higher the number of c=c bonds therefore there is a higher
unsaturation.

2. If a 0.2g of oil consumed 1ml of sodium thiosulphate, calculate its Iodine value and classify the oil?

equivalent weight of iodine x ( volume of thiosulphate blank−volume of thiosulphate test ) x 100 x

Iodine value=
weight of fat sample used for analysis(g)

127 x (K −1)x 100 x 10−3

=
0.2
= 63.5 (K-1)
Iodine value is a measure of the degree of unsaturation in fats and oils. It is essentially the
number of grams of iodine consumed by 100 g of fat. If the iodine number is in the range of 0-70
then it is a fat, any value above 70 is considered an oil.

3. What is the role of starch in the titration?

Starch is a sustainable indicator in the titration process because it will turn purple when there is
iodine present in the iodine is added. The colour change should be a purple colour which
appears only in the case if positive results which are that the starch is present in the solution.
When the starch is not present, no colour change appears in the solution. The result could be
negative. It also could be negative if the experiment was not performed properly.

4. What is the practical use of the iodine number?

The iodine number is the measure of the unsaturation of fats and oils. It is expressed as the
number of grams of iodine absorbed by 100 g of the fat under the test conditions used.

5. What can be concluded about the solvents used for in this experiment?
Iodine number is used to calculate the Degree of the unsaturation of the Faty acids. Fats contain
the lower Iodine number and Oils rich in unsaturated fatty acids have high iodine numbers. The
iodine number equals the number of mg of iodine required to saturate the fatty acids present in
100 mg of the oil or fats.
6. Calculate the number of double bonds in the fat?
The double bonds present in unsaturated fatty acids react with the iodine. The more the iodine
number, the more the number of C=C bonds. There are different methods to know the level of
unsaturation in fatty acids, one among them is by determining the iodine number of fats. By
using the formula:

equivalent weight of iodine x volume of x 100 x 10−3

iodine number of fats=
weight of fat sample used for analysis( g)

7. Why starch indicator is added during the titration rather adding it before the titration?
Starch indicator is added to titration mixtures that involve iodine because the colour change is
highly visible. Titration is the process of adding a solution with a known concentration to a
solution with an unknown concentration until the mixture neutralizes. To achieve measurable
results, you must know the volume of the unknown solution. A colour change typically occurs
once the solution neutralizes. Starch is a viable indicator in the titration process because it turns
deep dark blue when iodine is present in a solution. When starch is heated in water,
decomposition occurs and β-amylose is produced. β-amylose combines with iodine, resulting in a
dark blue colour change. For example, a starch-iodine titration can be used to determine how
much vitamin C is in a solution. Vitamin C converts iodine to iodide, so the starch mixture
would not turn blue until all the vitamin C is used up. Another example of the usefulness of
starch in the titration process is determining how much copper is in brass.

REFERENCES:
1. Amrita Vishwa Vidyapeetham Virtual Lab. (n.d.). Retrieved from https://vlab.amrita.edu/
2. Unsaturated Fatty Acid. (n.d.). Retrieved from
https://www.sciencedirect.com/topics/neuroscience/unsaturated-fatty-acid
3. Metrohm. (n.d.). Determination of iodine value (IV) in fats and oils. Retrieved from
https://www.metrohm.com/en-my/applications/AN-H-076