FOOD CHEMISTRY LABORATORY

STKM 1121
SEMESTER 1 (2024/2025)

NAME NURIL FADILAH BINTI SAFII

MATRIC NUMBER A202868

GROUP NUMBER GROUP 8

TITLE PRACTICAL 2
DETERMINATION OF PEROXIDE VALUE

LECTURER’S NAME PROF. MADYA TS. DR. LIM SENG JOE

DETERMINATION OF PEROXIDE VALUE

OBJECTIVES
1. To determine the peroxide value of fresh palm oil, used palm oil, fresh sunflower oil and
used sunflower oil.

2. To compare the peroxide values of fresh palm oil, used palm oil, fresh sunflower oil, and
used sunflower oil.

INRODUCTION
The peroxide value is defined as the miliequivalents (meq.) of peroxides expressed as meq. of
active oxygen per kilogram fat which oxidize potassium iodide under the conditions of the test. A
higher peroxide value suggests a greater degree of oxidation, which can lead to rancidity, off-
flavors, and a decline in nutritional quality. In practical terms, the determination of peroxide
value is often conducted using titration methods, where the sample is treated with an acidic
solution, and the resulting peroxides are quantified, providing valuable information about the
oxidative status of the fat or oil.
In this experiment, we use iodometric method to determine the peroxide value of the oils. This
process begins by dissolving a specific quantity of the fat or oil sample in a solvent mixture of
acetic acid and chloroform. Potassium iodide is then added to the solution, which reacts with
any peroxides present, liberating iodine. After swirling the mixture for one minute, distilled water
is added. If the resulting solution turns pale yellow, we can directly add a few drops of starch
solution and titrate it with sodium thiosulfate until the blue color disappears. Conversely, if the
solution appears a deeper yellow, indicating a higher peroxide content, we first titrate with
sodium thiosulfate until the yellow color of iodine is eliminated. Following this, we add a few
drops of starch solution and continue the titration until the blue color disappears.
In this experiment, we examine both used and fresh samples of palm oil and sunflower oil to
determine their peroxide values. Fresh palm oil and sunflower oil are baseline samples that
have not experienced significant oxidation. These oils are light in color and mild in flavor, with
low peroxide values that show they are not oxidized. Used oils, however, have been cooked at
high temperatures and exposed to air, which can cause them to oxidize and form peroxides.
This means that used oils usually have higher peroxide values, indicating they are more rancid.
By comparing the peroxide values of fresh and used oils, we can learn about their quality and
safety for cooking.
RESULTS AND CALCULATIONS
Sample Replication Sample Weight (g) Titration Volume
(mL)
Fresh Palm Oil 1 5.0170 10.0
2 5.0407 10.0
Used Palm Oil 1 5.1857 6.4
2 5.1632 6.8
Fresh Sunflower Oil 1 5.0517 2.2
2 5.0155 1.9
Used Sunflower Oil 1 5.0268 5.0
2 5.0166 5.1
Blank (V2) - 0.0000 0.5
1.0 Experimental Data for Peroxide Value

Calculations:
(𝑽𝒔 −𝑽𝒃 ) × 𝑵 ×𝟏𝟎𝟎𝟎
Peroxide Value (meq. peroxide/kg fat) =
𝑾

Where,
N = the normality of sodium thiosulphate solution;
𝑉𝑆 = the volume, in mL, of the sodium thiosulphate solution used for the determination;
𝑉𝑏 = the volume, in mL, of the sodium thiosulphate solution used for the blank test;
W = the weight, in grams, of the test portion;
1000 = Conversion of unit (g/kg)

Palm Oil

Replication 1 Replication 2
Fresh Palm Peroxide Value (meq. peroxide/kg fat) Peroxide Value (meq. peroxide/kg fat)
Oil
(10.0−0.5) × 0.01 ×1000 (10.0−0.5) × 0.01 ×1000
= =
5.0170 5.0407

= 18.94 ≈ 18.9 = 18.85 ≈ 18.9

Used Palm Oil Peroxide Value (meq. peroxide/kg fat) Peroxide Value (meq. peroxide/kg fat)

(6.4−0.5) × 0.01 ×1000 (6.8−0.5) × 0.01 ×1000

= =
5.1857 5.1632

= 11.38 ≈ 11.4 = 12.20 ≈ 12.2

 18.94+18.85
Mean = = 18.90
2

(18.94−18.90)2 + (18.85−18.90)2
Standard deviation = √ = 0.0640
2−1

The peroxide value for fresh palm oil is (𝟏𝟖. 𝟗 ± 𝟎. 𝟏).

11.38+12.20
Mean = = 11.79
2

(11.38−11.79)2 + (12.20−11.79)2
Standard deviation = √ = 0.5798
2−1

The peroxide value for used palm oil is (𝟏𝟏. 𝟖 ± 𝟎. 𝟔).

Sunflower Oil
Replication 1 Replication 2
Fresh Peroxide Value (meq. peroxide/kg fat) Peroxide Value (meq. peroxide/kg fat)
Sunflower Oil
(2.2−0.5) × 0.01 ×1000 (1.9−0.5) × 0.01 ×1000
= =
5.0517 5.0155

= 3.37 ≈ 3.4 = 2.79 ≈ 2.8

Used Peroxide Value (meq. peroxide/kg fat) Peroxide Value (meq. peroxide/kg fat)
Sunflower Oil
(5.0−0.5) × 0.01 ×1000 (5.1−0.5) × 0.01 ×1000
= =
5.0268 5.0166

= 8.95 ≈ 9.0 = 9.17 ≈ 9.2

3.37+2.79
Mean = = 3.08
2

(3.37−3.08)2 + (2.79−3.08)2
Standard deviation = √ = 0.4101
2−1
The peroxide value for fresh sunflower oil is (𝟑. 𝟏 ± 𝟎. 𝟒).

8.95+9.17
Mean = = 9.06
2

(8.95−9.06)2 + (9.17−9.06)2
Standard deviation = √ = 0.1556
2−1

The peroxide value for used sunflower oil is (𝟗. 𝟏 ± 𝟎. 𝟐).

DISCUSSION
Based on the results, the sequence of peroxide values from lower to higher is fresh sunflower oil
(𝟑. 𝟏 ± 𝟎. 𝟒), used sunflower oil (𝟗. 𝟏 ± 𝟎. 𝟐), used palm oil (𝟏𝟏. 𝟖 ± 𝟎. 𝟔), and fresh palm oil
(𝟏𝟖. 𝟗 ± 𝟎. 𝟏). The results show that the peroxide value of used sunflower oil (𝟗. 𝟏 ± 𝟎. 𝟐) and
used palm oil (𝟏𝟏. 𝟖 ± 𝟎. 𝟔) are higher than fresh sunflower oil (𝟑. 𝟏 ± 𝟎. 𝟒), indicating that the
fresh sunflower oil is in a stable, unoxidized state. This lower peroxide value of fresh sunflower
oil is due to the fresh oil being stored in a sealed container, limiting its exposure to air and light
and thereby preventing reactions with oxygen that would lead to peroxide formation. Meanwhile,
both used sunflower oil and palm oil have higher peroxide values due to repeated heating and
use, which break down the molecular structure of fats, making them more reactive with oxygen.
This increased oxygen exposure further promotes oxidation, resulting in greater peroxide
formation with each use and leading to a higher peroxide value.
However, the peroxide value of fresh palm oil (𝟏𝟖. 𝟗 ± 𝟎. 𝟏) is higher than both used palm oil
(𝟏𝟏. 𝟖 ± 𝟎. 𝟔) and used sunflower oil (𝟗. 𝟏 ± 𝟎. 𝟐), indicating that the oil has undergone oxidation,
which contradicts the theory that fresh oil should have a lower peroxide value. This is because
fresh oil has not undergone repeated heating, a process that accelerates oxidation. Without
thermal degradation, the molecular structure of the fresh oil remains intact, maintaining a low
peroxide value. The incorrect results we obtained are due to errors that we make during the
experiment.
Errors in this experiment can arise from various sources, leading to inaccurate results. For
instance, during titration, the person may misinterpret the endpoint, causing either excess or
insufficient titrant to be added, which results in inaccurate iodine value measurements. Other
than that, the error can occur when incorrect amounts of reagents are added, which can lead to
unreliable results. This may happen if reagents are measured improperly or if the equipment
used for measuring (such as pipettes or burettes) is not calibrated correctly, leading to
unexpected results in the reaction. Furthermore, using reagents that are contaminated or
expired also can give incorrect results.
The determination of peroxide value is crucial in the food industry as a quality control measure
to evaluate the freshness, stability, and shelf life of fats and oils. Peroxide value specifically
measures hydroperoxides, the primary oxidation products in fats and oils. By tracking this value,
manufacturers can detect early signs of rancidity, ensuring that only fresh, high-quality oils are
delivered to consumers. Additionally, peroxide value helps indicate the potential lifespan of fats
and oils, allowing companies to set accurate expiration dates. If peroxide value rise too quickly,
this could indicate a shorter shelf life. Regular monitoring also can help manufacturers to
improve formulations and select ingredients that enhance stability.

CONCLUSION

In summary, the peroxide values for fresh and used palm oil, as well as fresh and used
sunflower oil, were determined. The results show that both used sunflower oil (𝟗. 𝟏 ± 𝟎. 𝟐) and
used palm oil (𝟏𝟏. 𝟖 ± 𝟎. 𝟔) have higher peroxide values compared to fresh sunflower oil (𝟑. 𝟏 ±
𝟎. 𝟒). This indicates that the fresh sunflower oil is not oxidized since it has a low peroxide value.
However, the peroxide value of fresh palm oil is not accurately obtained, as the peroxide value
for fresh palm oil (𝟏𝟖. 𝟗 ± 𝟎. 𝟏) should be lower than both used sunflower oil and used palm oil.
This discrepancy is likely due to experimental error.

QUESTIONS

1. Explain what are oxidative and hydrolytic rancidity?

Oxidative rancidity takes place when fats and oils react with oxygen from the air, resulting in
the formation of peroxides and other volatile compounds. These volatile compounds are
often responsible for the off-flavors and odors associated with rancid foods. This type of
rancidity is particularly common in unsaturated fats, where double bonds make them more
susceptible to oxidation. On the other hand, hydrolytic rancidity occurs when fats and oils
break down through hydrolysis, resulting in the formation of glycerol and fatty acids due to a
reaction between fats and water molecules. Hydrolytic rancidity is more likely to develop in
foods with high water content, as water serves as a medium for lipase enzyme activity.
Furthermore, heat from cooking or processing can speed up the hydrolysis reaction.

2. Describe the steps taken to overcome the rancidity problem in food.

To overcome the rancidity problem in foods, antioxidants such as Vitamin E, Vitamin C, or

synthetic options like BHA (butylated hydroxyanisole) and BHT (butylated hydroxytoluene)
can be added. These antioxidants help slow down oxidative rancidity by interrupting the
oxidation process and protecting fats and oils from reacting with oxygen. Additionally, storing
fats and oils in cool, dark, and dry conditions minimizes exposure to heat, light, and
moisture, which can speed up both oxidative and hydrolytic rancidity. Using airtight
containers also helps by reducing exposure to oxygen and moisture. In some packaged
foods, modified atmosphere packaging (MAP) can be applied, where the oxygen is replaced
 with an inert gas like nitrogen to limit oxidation by reducing the available oxygen for
reactions with fats.

3. A sample of 5.0g of food-grade oil was reacted with excess KI to determine peroxide value.
The free iodine was titrated with a standardized solution of 0.10N Na2S2O3. The amount of
titrant required was 0.60 mL (blank corrected). Calculate the peroxide value of the oil.

(𝑽𝒔 −𝑽𝒃 ) × 𝑵 ×𝟏𝟎𝟎𝟎

Peroxide Value (meq. peroxide/kg fat) =
𝑾
𝟎.𝟔 × 𝟎.𝟏 ×𝟏𝟎𝟎𝟎
= = 12 meq. peroxide/kg fat
𝟓.𝟎

REFERENCES

• Hydrolytic Rancidity vs. Oxidative Rancidity - What’s the Difference? | This vs. That. (2023).

This vs. That. https://thisvsthat.io/hydrolytic-rancidity-vs-oxidative-rancidity

• Chef Science. (2023, May 15). The Chemistry of Rancidity in Food. FoodCrumbles.

https://foodcrumbles.com/food-spoilage-rancidity-and-food-chemistry/

• Zhang, N., Li, Y., Wen, S., Sun, Y., Chen, J., Gao, Y., Altayuly Sagymbek, & Yu, X. (2021).

Analytical methods for determining the peroxide value of edible oils: A mini-review. Food

Chemistry, 358, 129834–129834. https://doi.org/10.1016/j.foodchem.2021.129834

• R.D. O'Brien. (1995). Soybean Oil Products Utilization: Shortenings. Elsevier EBooks, 363–

379. https://doi.org/10.1016/b978-0-935315-63-9.50024-3