IBG112 - BIOANALYSIS I

LAB REPORT 3

DETERMINATION OF IODINE VALUE - WIJS METHOD

PREPARED BY:
WONG KAR JIN

MATRIC NUMBER:
161937

GROUP NUMBER:
GROUP 8
1.0 INTRODUCTION
The iodine value (IV) is a measure of the unsaturated fats and oils. This unsaturation is in the form
of double bonds, which react with iodine compounds. Higher iodine number represents more presence
of C=C bonds in the fat. It is expressed as the number of grams of iodine absorbed by 100 g of the fats
under the test conditions used. In addition, iodine value is responsible to identify and classify the fats.
However, the iodine value may be combined with other determinations, such as the thiocyanogen
value or the separation of fatty acids by the lead salt-alcohol or lead salt-ether methods, and it thus
serves as a means of calculating the proportions of the various fatty acids which are present in the
sample(1).

There are indeed several methods to analyse iodine value: Wijs method, Hanus method, Kaufmann
method and the like. In this experiment, we would like to determine iodine value of 2 oil samples
which are sunflower and palm oil using Wijs method. This is the official method of the American Oil
Chemists' Society (AOCS), the American Chemical Society, one of the official measures of the
Association of Official Agricultural Chemists, and recommended by the 9th International Union of
Pure and Applied Chemistry (IUPAC)(1). Wijs reagent is comprised of solution of iodine
monochloride in a mixture of acetic acid and carbon tetrachloride, CCl4 to a test portion. By looking
on the adverse effect of Wijs method, Wijs solution causes severe burns, and the vapours can render
lung and eye damage. Hence, the use a fume hood is highly recommended. Furthermore, All Wijs
solutions are sensitive to temperature, moisture, and light. Store in a cool and dark place, and never
allow to come to a temperature above 25 - 30°C. Due to advancement of technology, Wijs method has
now been modified by using cyclohexane as a solvent instead of carbon tetrachloride. In this
procedure, fats are dissolved in the solvent and mixed with Wijs solution. Iodine in the Wijs solution
reacts with the fats. The solution is titrated with sodium thiosulphate solution of known molarity. The
concentration of C=C in the original sample can therefore be calculated by measuring the amount of
sodium thiosulphate needed to complete the titration. Starch is normally used as an indicator because
it forms a molecular complex with the iodine that has a deep blue colour.

This method is applicable to all normal fats and oils not containing conjugated double bonds as its
presence complicates the determination iodine value(2).

2.0 OBJECTIVE
The objective to this experiment is:

➢ To determine the iodine value of sunflower oil and palm oil

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3.0 REAGENT
There are few reagents which will be used for the experiment which are potassium iodide, KI 15 %
(w/v) aqueous solution free of iodine or iodate. It is prepared by dissolving 15 g of reagent grade KI
in 100 ml of distilled water. Then, freshly boiled for 3 minutes and allowed to cool starch solution 1 %
(w/v) aqueous dispersion. The subsequent reagents are sodium thiosulphate, Na2S2O3.5H2O, 0.1 N as
well as potassium dichromate solution, K2Cr2O7, 0.1 N by dissolving 24.8 g sodium thiosulphate
pentahydrate in distilled water and dilute to 1 litre and dissolving 14.709 g of dried K2Cr2O7 in
distilled water and make up to 1 litre in a volumetric flask respectively. Wjis reagent (Iodine
monochloride solution in acetic acid carbon tetrachloride mixture) is also required which is available
commercially. Furthermore, cyclohexane in reagent grade is needed to complete the experiment. Last
but not least, standardised sodium thiosulphate. Na2S2O3.5H2O solution is prepared. It is prepared by
adding 0.037 g of K2Cr2O7 powder into a conical flask and adding 25 mL of distilled water. 5 ml
concentrated hydrochloric acid, HCl is added and followed by 10 ml KI solution. Mix the solution.
The solution is allowed to stand for 5 minutes and 100 mL of distilled water is added. Titrate this
solution with Na2S2O3.5H2O solution. Na2S2O3.5H2O solution is swirled until the yellow colour
almost disappear. The strength of Na2S2O3.5H2O solution is expressed in terms of its normality.

Normality of Na2S2O3.5H2O solution can be expressed as formula below:

25 × W
Normality =
V

Where
W is the weight of K2Cr2O7 powder
V is the volume of Na2S2O3.5H2O solution used

3.1 APPARATUS
There are plenty of apparatus entailed to finish this experiment such as wide-necked glass bottles or
flasks with ground glass stoppers (e.g. Iodine flasks), capacity 250 or 500 mL, burettes, 50 mL
graduated in 0.1 mL and pipettes, 20 and 25 mL.

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4.0 PROCEDURE
Firstly, 1 gram of palm oil and 1 gram of sunflower oil were weighed accurately to the nearest 0.01 g
in respective 500 mL conical flasks. Then, 15 mL of cyclohexane was measured and poured to
dissolve the fat. 25 ml of Wijs solution was added and stopper was inserted and mixed gently. The
flask was then placed in the dark for over 20 minutes. After standing for 20 minutes, added 20 mL of
KI solution and 150 mL water. The solution was titrated with Na2S2O3.5H2O solution until the yellow
colour was noticed due to the iodine almost vanished. Later, 1 to 2 mL of the starch indicator solution
was added and the titration was proceeded until the blue colour just disappeared after vigorous
shaking. The procedure above was repeated to obtain three results for each sample. Prepared a blank
test (without oil) simultaneously under the same condition.
Note that the weight of the sample to be used varies according to its expected iodine value. Table
below indicates the sample weight and expected iodine value for respective products.

Product Expected iodine value Sample weight

Hydrogenated palm kernel oil, olein, stearin. <5 3.00
Palm kernel oil, olein, stearin 5 - 25 1.00
Palm oil, olein, stearin 25 - 60 0.40
Special high IV olein 60 - 100 0.30
Liquid oils, soya bean oil, etc. > 100 0.15

Where
N is the exact normality of Na2S2O3.5H2O solution used
V is the volume, in mL, of Na2S2O3.5H2O solution used for the blank test
V1 is the volume, in mL, Na2S2O3.5H2O solution used for the determination
W is the weight, in grams, the test portion of oil

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5.0 RESULT AND TABULATION

Type of oil Weight of sample (g) Volume of (mL)

1st 2nd 3rd Average 1st 2nd 3rd Average
Sunflower 1.15 1.24 1.09 1.16 3.40 3.80 3.90 3.70
oil
Palm oil 1.00 1.07 1.05 1.04 19.10 19.40 18.90 19.13

The volume of Na2S2O3.5H2O for blank test is 23.7 mL.

➢ From the titration, the volume of Na2S2O3.5H2O used is 7.40 mL during standardisation of
Na2S2O3.5H2O solution prior to the iodine value determination.
Based on the data obtained from the experiment:

Normality of Na2S2O3.5H2O:
25 × W
Normality =
V

Where
W is the weight of K2Cr2O7 powder
V is the volume of Na2S2O3.5H2O solution used
25 ×0.037
Normality =
7.40
= 0.125 N

There are 2 iodine values to be calculated which are sunflower oil and palm oil:
Iodine value:

Where
N is the exact normality of Na2S2O3.5H2O solution used
V2 is the volume, in mL, of Na2S2O3.5H2O solution used for the blank test
V1 is the volume, in mL, Na2S2O3.5H2O solution used for the determination
W is the weight, in grams, the test portion of oil

12.69 (0.125)(23.7 − 3.7)

Iodine value of sunflower oil =
1.16
= 27.349

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 12.69 (0.125)(23.7 − 19.13)
Iodine value of palm oil =
1.04
= 6.970

6.0 DISCUSSION
Based on the objectives of this experiment, we would like to calculate and identify the iodine values
for sunflower oil and palm oil. Iodine value determination is a measure of the number of double
bonds in the unsaturated fatty acids in one gram of oil(3).

Now, we will discuss issues regarding the method used to conduct this experiment. Firstly, we
poured both sunflower and palm oils and cyclohexane was added into both respective oils followed by
addition of Wijs solution with gentle shake. The purpose of adding cyclohexane is to act as a solvent
to dissolve fat in both oils. Note that the precision of the iodine value determination is not affected by
the different solvent options(4). Wijs solution contains iodine which will react with the fat and is
highly dependent of the double bond C=C in the unsaturated fatty acids in one gram of oil. Then, the
solutions were left to react in the dark. This method was required to be run in the dark for accurate
results. The entire system can be placed in an enclosure for instance a fume hood or darkened beakers
can be used along with the covering or disclose system whereby lids from beakers can be added or
removed automatically(5). After standing over some period of time, KI solution was added to the
mixture to liberate any unreacted iodine(6). Titration of the mixture with Na2S2O3.5H2O began prior to
the addition of distilled water. The colour of the mixture in the conical flask was brown and titration
was proceeded until yellow colour was observed due to the iodine almost vanished. Yet, there was still
presence of iodine for successive titration. Then, few mL of colourless starch indicator solution was
added into the flask which caused the mixture to be blue-black. The rational of adding starch indicator
is to test the presence of iodine in the mixture. Since there was remnant of iodine present in the
mixture (in conical flask), blue-black was noticed. Titration was continued until blue colour just
vanished and milky solution emerged after vigorous shaking which marked the endpoint of the
experiment.

Based on the data collected throughout the conduct of the experiment, the iodine value obtained
from sunflower oil is 27.349 while the iodine value of palm oil is 6.970. Iodine value of sunflower oil
is higher as opposed to palm oil. Iodine value is an indicator of the relative degree of unsaturation in
oil components, as determined by the uptake of halogen (Iodine). The melting point and oxidative
stability are highly correlated to the degree of unsaturation and hence, iodine value provides an
estimation of these quality factors. The relationship can be defined such that iodine value is dependent
of unsaturation and stability. The greater the iodine value, the more unsaturation and higher the
susceptibility to oxidation(7). Therefore, sunflower oil contains less C=C double bonds (more
unsaturated fat), less stable and highly susceptible to oxidation as compared to that of palm oil.

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 Repeated frying is prevalent in household and different oils may possess different properties and
downsides. According to the American Oil Chemists’ Society (AOCS), the maximum permittable
peroxide value for edible oils is 10 meq/kg oil. A study has shown that after heating five times, the
peroxide value of sunflower oil (8.42 meq/kg oil) is higher than palm oil (6.01 meq/kg oil). Larger
peroxide value indicates lower chemical stability of the oil during frying. Hence, sunflower oil is less
chemically stable during frying. In addition, sunflower oil has more presence of lipid hydroperoxides
which in the body, can decompose into free radicals that can oxidise other unsaturated fatty acids and
form compounds that are potentially toxic to one’s health. Thus, sunflower oil emerges as less
chemically stable oil for deep-frying and is less safer for repeated frying with higher potential to
produce toxic substances in mankind(8).

7.0 CONCLUSION

The objective of the experiment, we would like to calculate and measure iodine values for 2 samples
which are sunflower oil and palm oil. Based on the data obtained from the conduct of the experiment,
the iodine the iodine value obtained from sunflower oil is 27.349 while the iodine value of palm oil is
6.970 in which sunflower oil has greater iodine value than palm oil.

ATTACHMENT OF PICTURES

Photo 1 Photo 2
Photos 1 and 2: Endpoint of iodine value determination using Wijs method (Cloudy and milky)
Photo credit: Determination of Iodine Value _A Complete Procedure (AOAC 920.159), MicroChem’s
Experiments

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8.0 REFERENCE
1. Barbour, A. D. (1934). A comparison of various methods of determining iodine values, and their
effect on the calculated results of fat analysis. Oil & Soap, 11(1), 7–10.
https://doi.org/10.1007/bf02543607
2. Forbes, W. C., & Neville, H. A. (1940). Wijs iodine numbers for conjugated double bonds.
Industrial & Engineering Chemistry Analytical Edition, 12(2), 72–74.
https://doi.org/10.1021/ac50142a003
3. Kyriakidis, N. B., & Katsiloulis, T. (2000). Calculation of iodine value from measurements of fatty
acid methyl esters of some oils: comparison with the relevant American oil chemists society
method. Journal of the American Oil Chemists' Society, 77, 1235-1238.
4. Pocklington, W. D. (1990). Determination of the iodine value of oils and fats: results of a
collaborative study. Pure and applied chemistry, 62(12), 2339-2343.
5. Aocs. (2008). Chapter 41 Iodine Values of Fats and Oils. In 2007-2008 additions and revisions to
the official methods and recommended practices of the AOCS. essay, Amer Oil Chemist Society.
6. Beverage, W., Gibb, A., & Hawley, D. (2014). Chapter 2 Nature's Chemistry. In CFE Higher
Chemistry. essay, BrightRED Publishing.
7. Caballero, B., Trugo, L. C., & Finglas, P. M. (2003). Ground Nut Oil. In Encyclopedia of Food
Sciences and nutrition (pp. 2967–2974). essay, Academic Press.
8. Taufik, A. (2021, June 15). The Frying Oil Showdown: Which is the best option? Malaysian
Palm Oil Council. Retrieved May 7, 2023, from https://rb.gy/be4oz
9. British Standards BS 684: Section 2.13: 1976.
10. Goswami, G., Bora, R., and Rathore, M.. (2015). International Journal of Science Technology and
Management, 4(1):495-501.
11. Kumar, C. R. (2012). Research methodology. APH Publishing Corporation.
12. Lister, T., & Renshaw, J. (2015). Aqa Chemistry. Oxford University Press.
13. MicroChem's Experiments. (2020, September 20). Determination of iodine value _a complete
procedure (AOAC 920.159). YouTube. Retrieved May 7, 2023, from https://tinyurl.com/hr68c4bv
14. Nielsen, S. S. (2003). Instructor's Manual for Food Analysis, third edition: Answers to study
questions. Kluwer Academic/Plenum Publishers

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9.0 FLOWCHART