FST653/663

RANCIDITY MEASUREMENT IN
EXPERIMENT 4
FATS/ OILS

STUDENT’S NURUL HUSNA BINTI ZAIDI

NAME

GROUP AS246 7Q

DATE OF
29th OKTOBER 2020
EXPERIMENT

LECTURER’S DR. RASEETHA VANI SIVA

NAME MANIKAM
 Experiment 4: Rancidity Measurement in Fats/Oils

INTRODUCTION

Edible oils are vital constituents of our daily diet, which provide energy, essential fatty acids
and serve as a carrier of fat-soluble vitamins. Rancidity is a term generally used to denote
unpleasant odours and flavours in foods resulting from deterioration in the fat or oil portion of
a food. Three different mechanisms of rancidity may occur. Lipids and triacylglycerol naturally
occur in oils and fats. Their chemical composition contains saturated and unsaturated fatty
acids and glycerides. These physicochemical parameters include iodine value (IV),
saponification value (SV), viscosity, density, and peroxide value (PV). The main cause of
rancidity is auto oxidation. It can be defined as a reaction between oxygen and the
unsaturated lipids to form a lipid hydro-peroxide, which then undergoes further reaction with
or without the participation of other compounds. This reaction is enhanced by light and metal
ions (Sun et al., 2011).

Peroxide value points out the state of the oxidation in oil. It is a measure of the extent
of glycerides constituent decomposition by lipase action, which is added by light, air, and
moisture (Mariod, Mirghani and Hussein, 2017). The amount of peroxides of palm oil indicates
the degree of primary oxidation and therefore its likeliness of becoming rancid. A lower
number of peroxides indicates a good quality of oil and a good preservation status. Unsaturated
free fatty acids react with oxygen and form peroxides, which determine a series of chain
reactions that generate the production of smelling volatile substances. Those reactions are
accelerated by high temperature and by light and oxygen exposure

Acid value of fats or oils is defined as the weight of KOH in milligram (mg) that is
needed to neutralize the organic acids present in 1 g of fat and it is a measure of the free fatty
acids (FFA) present in the fats or oils (Vitz et al., 2020). An increase in the amount of FFA in
oil or fat sample indicates hydrolysis of triglycerides. It occurs by the action of lipase enzyme
and it is an indicator of inadequate processing and storage conditions such as high temperature
and relative humidity.

Thiobarbituric acid number (TBA) is uses to measure deterioration in both extractable

and non-extractable lipids and therefore has been more frequently applied to compound fatty
foods, particularly fresh foods. The increase in the amount of red pigment formed in the
reaction between 2-thiobarbituric acid and oxidized lipids as oxidative rancidity advances has
been applied to a wide variety of fatty foods.

The objectives of the experiment are to determine acidity and peroxide value of oil
samples, and to determine thiobarbituric acid number (TBA) of fish sample.

MATERIALS AND METHODS

A. Determination of Acidity
Materials and apparatus needed are diethyl ether, alcohol, phenolphthalein, 0.1 M
NaOH, fresh palm oil, rancid palm oil, conical flask, retort stand, white tile, and burette.

Firstly, the smell of fresh palm oil and rancid palm oil were recorded. 25 mL of diethyl
ether was mixed with 25 mL of alcohol and 1 mL of 1 % phenolphthalein solution and
carefully neutralized with 0.1 M NaOH. 1 to 10 g of the oil was dissolved in the mixed
neutral solvent and titrated with 0.1 M NaOH. Constantly shake it until a pink colour
persists for 15 seconds was obtained.

B. Determination of Peroxide Value

Materials and apparatus needed are fresh palm oil, rancid palm oil, potassium iodide
powder, glacial acetic acid, chloroform, distilled water, 0.002 M sodium thiosulphate,
starch indicator, 5 % potassium iodide solution, test tubes, conical flask.

1 g of oil was weighed and added into a clean test tube. 1 g of potassium iodide powder
and 20 mL solvent mixture (Glacial Acetic Acid: Chloroform, 2:1 v/v) was added when
sample is still a liquid. The test tube was placed in boiling water so that the liquid would
boil within 30 seconds. It was let boiled vigorously for less than 30 seconds. The test
tube content was poured immediately into a conical flask that contain 20 mL of 5 %
potassium iodide solution. The test tube was rinsed twice with 25 mL water and titrated
with 0.002 M sodium thiosulphate solution using starch as an indicator. The blank was
simultaneously determined.
C. Determination of Thiobarbituric Acid Number (TBA)
Materials and apparatus needed are fatty foods, distilled water, distillation flask, 4 M
HCl, glass beads, heating mantle, TBA reagent, 90 % glacial acetic acid, distillation
flask, and glass-stoppered tube.

10 g of fish was macerated with 50 mL distilled water for 2 minutes and washed into a
distillation flask with 47.5 mL distilled water. 2.5 mL of 4 M hydrochloric acid was
added to bring pH to 1.5, followed by an antifoaming agent preparation and a few glass
beads. The flask was heated by means of an electric heating mantle so that 50 mL
distillate would be collected from the time boiling commences. 5 mL distillate pipetted
into a glass-stoppered tube, 5 mL TBA reagent (0.2883 g/100 mL of 90 % glacial acetic
acid) was added, stopper, shakes and heated in boiling water for 35 minutes. The tubes
then cooled in water for 10 minutes. The blank was prepared similarly using 5 mL
distilled water with 5 mL TBA reagent. The absorbance (D) was measured against the
blank at 538 nm using 1 cm cells.
 RESULTS

A. Determination of Acidity

Table 1.0: Acidity Determination of Fresh and Rancid Palm Oil by Titration
Final volume
Samples Initial volume (mL) Average ± Std Dev
(mL)
1 0.0 0.2
0.2 ± 0.0
Fresh palm oil 2 0.0 0.2
Blank 0.0 0.1
1 0.0 1.4
1.4 ± 0.0
Rancid palm oil 2 0.0 1.4
Blank 0.0 0.1

Calculation

For fresh palm oil,

Titre (mL)
Acid Value = 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 (𝑔) × 0.0256 𝑔 𝑝𝑎𝑙𝑚𝑖𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 × 100 %
0.2 mL
= × 0.0256 𝑔 𝑝𝑎𝑙𝑚𝑖𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 × 100 %
5𝑔

= 0.10 %

For rancid palm oil,

Titre (mL)
Acid Value = 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 (𝑔) × 0.0256 𝑔 𝑝𝑎𝑙𝑚𝑖𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 × 100 %
1.4 mL
= × 0.0256 𝑔 𝑝𝑎𝑙𝑚𝑖𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 × 100 %
5𝑔

= 0.72 %
 B. Determination of Peroxide Value

Table 1.1: Sample Volume of Fresh and Rancid Palm Oil by Titration
Weight of sample Initial volume Final volume Sample volume,
Samples
(g) (mL) (mL) Vs (mL)
Blank, Vb 0.0 8.0
Fresh palm oil 1 1.0089 0.0 26.2 26.2
2 1.0165 0.0 27.8 27.8
Blank, Vb 0.0 3.8
Rancid palm oil 1 1.0234 0.0 21.4 21.4
2 1.0630 0.0 23.5 23.5

Table 1.2: Peroxide Value of Fresh and Rancid Palm Oil

Samples Peroxide value (PV) Average ± Std Dev
1 36.0789
Fresh palm oil 37.5181 ± 2.0353
2 38.9572
1 34.3952
Rancid palm oil 35.7301 ± 1.8878
2 37.0649

Calculation

𝑉𝑠−𝑉𝑏
Peroxide Value = 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 × 𝑇 × 103

T – Molarity of sodium thiosulphate

Vs – Volume in mL titration for sample
Vb – Volume in mL titration for blank

For fresh palm oil 1,

26.2 𝑚𝐿−8.0 𝑚𝐿
Peroxide Value = × 0.002 𝑀 × 103
1.0089 𝑔

= 36.0789 mEq/kg
For fresh palm oil 2,
27.8 𝑚𝐿−8.0 𝑚𝐿
Peroxide Value = × 0.002 𝑀 × 103
1.0165 𝑔

= 38.9572 mEq/kg

Average PV of fresh palm oil = 37.5181 mEq/kg

For rancid palm oil 1,

21.4 𝑚𝐿−3.8 𝑚𝐿
Peroxide Value = × 0.002 𝑀 × 103
1.0234 𝑔

= 34.3952 mEq/kg

For rancid palm oil 2,

23.5 𝑚𝐿−3.8 𝑚𝐿
Peroxide Value = × 0.002 𝑀 × 103
1.0630 𝑔

= 37.0649 mEq/kg

Average PV of rancid palm oil = 35.7301 mEq/kg

 C. Determination of Thiobarbituric Acid Number (TBA)

Table 1.3: TBA Determination of Fish

Samples Absorbance at 538 nm TBA

Blank 0.000 0.000

1 0.134 1.045
2 0.139 1.084
3 0.142 1.108
4 0.142 1.108
Average 0.139 1.086

Calculation

Sample 1,
TBA = 7.8 × 0.134
= 1.045

Sample 2,
TBA = 7.8 × 0.139
= 1.084

Sample 3 and 4,
TBA = 7.8 × 0.142
= 1.108
DISCUSSIONS

In acidity determination, based on table 1.0, the volume of sodium hydroxide (NaOH) needed
to titrate the fresh palm oil sample is 0.2 mL which is low compared to rancid palm oil that
need 1.4 mL NaOH to obtain a pink colour that persists for 15 seconds. However, it is good
that the titre for both fresh palm oil and rancid palm oil sample is not more than 10 mL because
if the titre is more than 10 mL, two phases may not be formed. For palm oil, 1 mL of 0.1 M
NaOH contain 0.0256 g palmitic acid and this palmitic acid value is uses to calculate acid value
for both fresh palm oil and rancid palm oil. The acid value for fresh palm oil is 0.10 % indicates
that 0.10 % free fatty acid (FFA) is present in the fresh palm oil. Rancid palm oil contains 0.72
% FFA. The increase in the amount of FFA in rancid palm oil sample shows that hydrolysis of
triglycerides is high compared to in fresh palm oil. The triglyceride in rancid palm oil hydrolyse
by lipase or other reaction into FFA. Lipase is an enzyme known as hydrolase that is
responsible for the hydrolysis of triglycerides into fatty acid and glycerol. The lower the acid
value, the better the quality of the alimentary fats or oils and their state of preservatives.

Next, in peroxide value determination, the average peroxide value for fresh palm oil is
37.5181 mEq/kg which is higher than the average peroxide value for rancid palm oil with
35.7301 mEq/kg. This indicates that both sample of fresh palm oil have rancid taste since its
peroxide value is between 20 to 40 mEq/kg. Based on the peroxide value, it can be said fresh
palm oil sample use in this experiment is already rancid. However, there must be some error
that happen during the analysis that may affect the result. The peroxide value of fresh palm oil
should be lower than the peroxide value of rancid palm oil because rancid palm oil has high
level of rancidity. Peroxide value is an indicator of the lipid peroxidation or oxidative
degradation levels. It is used to assess the stability or rancidity of fats by measuring the amount
of lipid peroxide and hydroperoxides formed during the initial stage of oxidation, so it estimates
to which level the oil has rancidity.

Lastly, rancidity of frozen fish can be determined by TBA value. The principle for this
method is the reaction of 1 molecule malonaldehyde and 2 molecules TBA to form a pink
pigment malonaldehyde-TBA complex, which can be quantitated spectrophotometrically
(Tokur, Korkmaz and Ayaz, 2006). Thiobarbituric acid reactive substances (TBARS) is
considered as standard marker for the lipid peroxidation. In this TBA determination, four fish
samples were used to obtain the accurate TBA number. The average TBA number for fish
sample is 1.086.

During the experiment, there may be some errors happened that may affect the result
of the analysis. Firstly, in the peroxide value determination, the reagent used might be
contaminated or contain impurities that may affect the reading as there is no significant
difference between the peroxide value of fresh palm oil and rancid palm oil. Next, parallax
error as the observer’s eyes are not parallel to the scale when taking reading of titre.

CONCLUSION

To conclude acid value of fresh palm oil is lower than rancid palm oil because more triglyceride
hydrolyses by lipase thus produce more FFA in rancid palm oil while less FFA produce in fresh
palm oil because less glyceride hydrolyses by lipase. Fresh palm oil has better quality than
rancid palm oil because of less peroxidation.

STUDY QUESTIONS

1. Define rancidity. List two (2) types of rancidity which normally occur in food products.
Rancidity can be defined as complete or incomplete hydrolysis or oxidation of fats and
oils when exposed to air, light, moisture, and bacterial activity and this generally
occurs in food items making them undesirable for consumption (Anand et al., 2020).
Types of rancidity are hydrolytic rancidity and oxidative rancidity.

2. What is meant by Induction Period (IP)?

Induction Period is the initial slow phase of a chemical reaction which later accelerates.
Induction periods are often observed with radical reactions, but they may also occur in
other systems for example before steady-state concentration of the reactants is reached.
 3. Suggest approaches that would be effective in reducing the rancidity problems
associated with fatty foods.
The approaches that would be effective in reducing the rancidity problems are chemical
preservatives, use of low temperatures, control of humidity, storage in carbon dioxide or
oxygen are considered.

REFERENCES

Anand, J., Lunawat, R., Shah, S., Patel, P., Yusuf, S., Staughton, J., … Vig, M. (2020, February
26). Rancidity: Definition, Explanation, Examples and Types. Retrieved April 19,
2020, from https://www.scienceabc.com/pure-sciences/what-is-rancidity.html

Mariod, A. A., Mirghani, M. E. S., & Hussein, I. H. (2017). Unconventional oilseeds and oil
sources. Academic Press.

Sun, Y. E., Wang, W. D., Chen, H. W., & Li, C. (2011). Autoxidation of unsaturated lipids in
food emulsion. Critical reviews in food science and nutrition, 51(5), 453-446.

Tokur, B., Korkmaz, K., & Ayaz, D. (2006). Comparison of Two Thiobarbituric Acid (TBA)
Method for Monitoring Lipid Oxidation if Fish. Su Urunleri Dergisi, 23(3), 331-334.

Vitz, E., Moore, J. W., Shorb, J., Prat-Resina, X., Wendorff, T., & Hahn, A. (2020, February
7). Foods: Acid Value and the Quality of Fats and Oils. Retrieved April 19, 2020,
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