EXPERIMENT NO

OBJECT
Determination of peroxide value in oil and fat by iodometric titration.

THEORY
IODOMETRY
The term “iodometry” describes the type of titration that uses a standardised sodium thiosulfate
solution as the titrant, one of the few stable reducing agents where oxidisation of air is
concerned. Iodometry is used to determine the concentration of oxidising agents through an
indirect process involving iodine as the intermediary. In the presence of iodine, the thiosulphate
ions oxidise quantitatively to the tetrathionate ions.

To determine the concentration of the oxidising agents, an unknown excess of potassium iodide
solution is added to the weakly acid solution. The iodine, which is stoichiometrically released
after reduction of the analyte, is then titrated with a standard sodium thiosulphate solution
(Na2S2O3)

INDICATOR IN IODINE/THIOSULPHATE TITRATION

Titration involving iodine commonly uses a starch suspension as indicator. This suspension is a
watery solution of starch with a few drops of bactericide added to prevent decomposition, as this
would stop the starch behaving as an indicator.

Once the bond between the iodine (I2) and the helical chain of beta-amylose is formed it turns an
intense blue.

IMPORTANT CONSIDERATIONS
Iodometric titration needs to be done in a weak acid environment which is why we need to
remember that:

1. The iodine solution used needs to be at pH < 8.5 because at a base pH iodine
disproportionates (a particular kind of oxidoreduction reaction where one substance
partly oxidises and partly reduces);
2. Sodium thiosulphate needs a neutral or weak acid environment to oxidise with
tetrathionate (in an alkaline solution we would get sulphate oxidation);
3. In a strong acid environment thiosulphate decomposes to S2;
4. In acid environments the iodide is oxidised to iodine as in the reaction below:

O2 + 4I- + 4H+ ↔2I2 + 2H2O

OXIDATIVE PROCESS
 Initiation
in the presence of a catalyst, alkyl
radicals (L·) form and these tend to
accumulate.
 Propagation
the alkyl radicals which have
formed react with atmospheric
oxygen to form a peroxyl radical
(LOO· ). This radical can react with
another available hydrogen atom
(LH) to form another free radical
(L·) and a hydroperoxide (LOOH).
 Termination
hydroperoxides, which are highly
unstable chemicals, decompose to
produce additional free radicals and /or Reaction of radicals responsible for formation of
hydroperoxides in edible fats and oils. LH is a
secondary oxidation products which monosaturated or polyunsaturated acid
accumulate and so increase the rancidity
of oil.

WHAT IS OIL OXIDATION?

Oil oxidation is an undesirable series of chemical reactions involving oxygen that degrades the
quality of an oil. Oxidation eventually produces rancidity in oil, with accompanying off flavours
and smells. All oil is in a state of oxidation - you cannot stop it completely - but there are ways to
reduce it. Attempts should therefore be made to reduce oxidation at each stage of oil
manufacture.

Oxidation is not one single reaction, but a complex series of reactions. When oil oxidizes it
produces a series of breakdown products in stages,

 starting with primary oxidation products (peroxides, dienes, free fatty acids),
 then secondary products (carbonyls, aldehydes, trienes)
 and finally tertiary products.

Oxidation progresses at different rates depending on factors such as

 temperature,
 light,
 availability of oxygen,
 and the presence of moisture and metals (such as iron).
 The type of oil also influences the rate of oxidation. Marine oils (including fish, mussel) are
highly susceptible to oxidation due to the large number of polyunsaturated fatty acids (PUFA)
they contain. These unsaturated fatty acids have reactive double bonds between their carbon
atoms, whereas saturated fats have no double bonds so they oxidise more slowly.

HOW DO WE MEASURE OXIDATION?

Measuring oxidation involves testing for the primary and secondary breakdown products. The
most common test is peroxide value (PV). However, very rancid oils can have a reduced PV
therefore the anisidine value (AV) and a Totox value are used to show the whole oxidation story.
Other measurements of oxidation are the acid value (free fatty acid FFA), thiobarbituric acid
value (TBA) and iodine value (IV). Head space volatiles (smell) can also be tested using
artificial nose technologies.

PEROXIDE VALUE (PV)

 BACKGROUND

Primary oxidation processes in oil mainly form hydroperoxides, which are measured by the PV.
In general, the lower the PV, the better the quality of the oil. However PV decreases as secondary
oxidation products appear. Most customers will require a PV of less than 10 in marine oils, but
PV may need to be as low as 2, depending on the market.

The PV test is a good way to measure the amount of primary oxidation products in fresh oils.
Oils with significant levels of peroxides may still be odourless if secondary oxidation has not
begun. If oxidation is more advanced, the PV may be relatively low but the oil will be obviously
rancid.

 DEFINITION
The peroxide value (PV) is the concentration of hydroperoxide, the primary oxidation products
present in the sample.

Peroxides are the primary oxidized products produced, which on further oxidation would
degrade to aldehydes, ketones, esters, etc., which are the secondary oxidized products.

 PRINCIPLE
The principle involves peroxides liberating iodine from potassium iodide, i.e.

ROOH+KI→ROH+KOH+I2

The amount of ROOH is then determined by measuring the amount of iodine formed, which is
done by titration with sodium thiosulfate and using a starch indicator:
 I2+starch+2Na2S2O3(blue)→2NaI+starch+Na2S4O6(colorless)

The amount of peroxides is calculated back by the amount of sodium thiosulfate (Na2S4O6)
consumed. It is expressed as peroxide value (PV) in units of milli-equivalents (meq) peroxide per
1 kg of fat extracted from the food. A general rule is that PV should not be above 10–20 meq/kg
fat to avoid rancidity flavor

IMPORTANCE OF DETERMINING PEROXIDE VALUE

The peroxide value (PV) is a very important characteristic of lipid quality. The assessment of
hydroperoxides provides an estimate of the overall oxidation status for lipids and lipid-
containing foods especially in the primary phase of oxidation, generally known as the induction
period.

From this value, the propagation step of the free radical chain mechanism and the accumulation
of hydroperoxides can be followed. However, it is not possible to use the peroxide value alone to
judge the quality of edible oils, because hydroperoxides decompose during storage. This
decomposition can take place faster than the formation of new hydroperoxides, depending on
certain storage conditions such as temperature, light or metal traces. Although the oil has already
been damaged by oxidation, and higher levels of degradation products have already formed, the
speed of hydroperoxide decomposition can result in falsely low levels of these compounds. In
this instance, the peroxide value tells us nothing about the real quality of the product. In order to
avoid misinterpreting peroxide values, it is necessary to know the history of the sample.
However, the peroxide value is a suitable parameter for measuring the deterioration of quality
over time. After the induction period, during which the peroxide value increases slowly, a steep
increase indicates that the oil has gone bad.

In general, the aim of oil production should be to produce oils with peroxide values as low as
possible, without the formation of secondary reaction products. A higher peroxide value at the
beginning of the storage period has a negative effect on the storage stability of the oil. For
refined oils, producers should aim for a peroxide value below 1, better 0.5 meq O2/kg oil, while
the peroxide value for virgin oils can be higher, up to 3 meq O2/kg oil.

MATERIALS
 Oil or lipid extract sample
 3:2 (v/v) acetic acid/ chloroform solution
 Saturated potassium iodide solution
 Sodium thiosulfate 0.1 N solution
 1%(w/v) starch indicator solution
 250mL glass-stopperedErlenmeyer flasks
 10 or 25 mL graduated glass burette `
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PROCEDURE

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OBSERVATION AND CALCULATION
SODIUM PV VALUE
STANDARD
SAMPLE WEIGHT (g) THIOSULFATE (meq active
DEVIATION
(ml) O2/kg)
Blank
Sample 1
Sample 2
Sample 3

PV= [(S-B)xNx1000]/W
Where S is the volume (ml) of sodium thiosulfate required to titrate the sample

B is the volume (ml) of sodium thiosulfate required for the blank

N is the calculated normality of the
standardized sodium thiosulfate
solution

W is the weight of the sample (g)

From this equation, the PV is

expressed as meq active oxygen/kg sample and is equal to mmol active oxygen/ 2kg sample.

DISCUSSION
 How does starch indicate iodine?
When starch is mixed with iodine in water, an intensely colored starch/iodine complex is formed.
Many of the details of the reaction are still unknown. But it seems that the iodine (in the form of
I5- ions) gets stuck in the coils of beta amylose molecules (beta amylose is a soluble starch). The
starch forces the iodine atoms into a linear arrangement in the central groove of the amylose coil.
There is some transfer of charge between the starch and the iodine. That changes the way
electrons are confined, and so, changes spacing of the energy levels. The iodine/starch complex
has energy level spacings that are just so for absorbing visible light- giving the complex its
intense blue color.

The complex is very useful for indicating redox titrations that involve iodine because the color
change is very sharp. It can also be used as a general redox indicator: when there is excess
oxidizing agent, the complex is blue; when there is excess reducing agent, the I5- breaks up into
iodine and iodide and the color disappears.

 Why do we not use starch in the beginning of iodometric titration?

The starch indicator is added to the solution near the end of the titration, at the point where dilute
iodine imparts a pale yellow color to the solution.

There are two reasons why the indicator is not added at the beginning of the titration when the
iodine concentration is high.

 First, a diffuse endpoint would result from the slow dissociation of the starch-iodine
complex if a large amount of iodine were absorbed in the starch.
 Second, iodometric titrations are carried out in strongly acid media, a situation that
promotes the reaction between oxidizing agents and iodide. Unfortunately starch has a
tendency to hydrolyze (decompose) in acidic media, destroying its indicator qualities.

 What are the advantages of using starch as an indicator in iodometric

titration?
 1. Starch give deep blue color which is helpful for detection of endpoint.
2. Starch made complex with iodide when heated with water decomposition occure ,beta-
amylose formed which combined with iodine to form starch/iodine complex of highly
intense color.
3. Freshly prepared starch is used because it is biodegraded.
4. Starch added at the end of reaction because in the beginning pale yellow color of iodine
present but when starch added it turn into dark blue color show the end point of reaction.

 Why is a conical flask place in dark in iodometric titration?

Iodine is weakly soluble in water and easily lost out of solution because it is volatile. Keeping
the solution in the dark slows down the loss of iodine from the water.

 What is the purpose of mixture of acetic acid and chloroform?

The role of chloroform and acetic acid in peroxide value is to dissolve fat or oil.

The following reaction takes place on adding acetic acid

2ROOH+2H++2I−⟶2ROH+I2+H2O

The acidic conditions (excess acetic acid) prevents formation of hypoiodite (analogous to
hypochlorite), which would interfere with the reaction
Acetic acid also has the fortunate property of being miscible with many polar and non-polar
solvents. Because this is a method for the analysis of oil samples, it is necessary to chose a non-
polar solvent capable of dissolving the sample that is also miscible with the acetic acid.
Chloroform is one solvent that fits that description nicely.