12

EXPERIMENT 2
RANCIDITY MEASUREMENT IN FATS/OILS
Introduction.
Fats and oils play an important role in the flavor, aroma, texture, and nutritional quality of
foods, pet foods, and feeds. Fats and oils may be added during manufacturing or they may be
inherent to the product or ingredient. The product may be pure oil or it may be part of a complex
mixture with proteins, carbohydrates, minerals, and vitamins. The product may contain almost no
fat or it may contain a considerable amount. Regardless of the source of fat, the amount of fat, or
the product composition, predicting and monitoring fat and oil quality is an important component
of developing and manufacturing high quality products.
A. Determination of Acid Value or Free Fatty Acid.
Introduction.
Acid value is a measure of the free fatty acids in oil. Normally, fatty acids are found in the
triglyceride form, however, during processing the fatty acids may get hydrolyzed into free fatty
acid. The higher the acid value found, the higher the level of free fatty acids which translates into
decreased oil quality. Acceptable levels for all oil samples should be below 0.6 mg KOH/g
(measured in potassium hydroxide per gram) (AOCS Official Method Cd 8-53, 2003).
Objectives.
To determine the acid value or free fatty acid of new palm oil and old palm oil.
Materials/Apparatus.
Oils/fats
1% phenolphthalein solution
Diethyl ether
0.1 M NaOH
Burette
Measuring cylinder, 25 mL
Measuring cylinder, 1 mL
Disposable plastic pipetter/dropper

Test tube rack

Balance
Stopwatch
Spatula
Conical flask
Tong
Retort stand
Label sticker

Procedure.
1. 25 mL diethyl ether was mixed with 25 mL alcohol and 1 mL of 1% phenolphthalein solution
in a conical flask.
2. This solution was neutralized with 0.1 M NaOH.
3. 1 10 gram of the oil or melted fat was dissolved in the mixed neutral solvent.
4. It was titrated with 0.1 M NaOH until a pink colour persists for 15 seconds is obtained.
(Caution: Titre should not exceed 10 mL, if not two phases may be formed). However, this will
not happen if hot alcohol is used as a solvent.
Calculation.
Acid value = Titre (mL) x 5.61
Weigh of sample
Free fatty acid value is usually calculated as oleic acid (1 mL 0.1M NaOH = 0.0282 gram oleic
acid. In which case the acid value = 2 x FFA.
However, BS 684 recommended that the FFA in the following oils should be calculated as the
acid which is more appropriate to that present (major fatty acid):
FFA
For olive oil; 1 mL of 0.1 M NaOH = 0.0282 gram oleic acid
For palm oil; 1 mL of 0.1 M NaOH = 0.0256 gram palmitic acid
For palm kernel oil, coconot oil; 1 mL of 0.1 M NaOH = 0.0200 gram lauric acid
With most oils, rancidity begins to be noticeable (on the palate) when the FFA calculated as oleic
acid is about 0.5-1.5%. However, some quite rancid oils show minimal acidities.
Palm oil (new)
= 0.77 mL x 5.61
5 gram
= 0.8639
Palm oil (old)
= 1.20 mL x 5.61
5 gram
= 1.3464

B. Determination of Peroxide Value.

Introduction.
Measuring the number of peroxide is also commonly performed on pure fats and oils and shows
the stage of oxidation, or how fat oxidation has progressed already as peroxides are the first
components formed during the oxidation of fat. The peroxide value is helpful in order to
determine the quality of saturated fat but is not that helpful in assessing unsaturated fats.
Objectives.
To determine the peroxide value of fats/oils.
Materials/Apparatus.
Oils/fats
1% starch solution
Potassium iodide powder
Solvent mixture (Glacial acetic acid + Chloroform 2:1)
5% potassium iodide solution
0.002 M sodium thiosulphate
Measuring cylinder, 25 mL
Water bath, 100C
Disposable plastic pipette/dropper
Beaker, 50 mL

Test tube rack

Test tube
Stopwatch
Spatula
Burette
Conical flask
Tong
Retort stand
Balance
Label sticker

Procedure.
1. 1 gram oil or fat was weighed into dry, clean test tube. (It is best to do in the dark).
2. While sample is still a liquid, 1 gram potassium iodide powder and 20 mL solvent mixture
(Glacial Acetic Acid : Chloroform, 2 : 1 v/v) was added.
3. This test tube was placed in boiling water so that the liquid is boiled within 30 seconds. Let is
boiled vigorously for not more than 30 seconds. (This experiment had been operated in fume
cupboard).
4. The test tube content was immediately poured into a conical flask containing 20 mL 5%
potassium iodide.

5. The test tube was rinsed twice with 25 mL distilled water and was poured back into the conical
flask.
6. It was titrated with 0.002 M sodium thiosulfate solution using starch solution as an indicator.
Make sure the flask was well shaken.
7. A blank determination was carried out at the same time.
Calculation.
Peroxide value is usually reported as the mL of 0.002 M sodium thiosulphate per gram of
sample. If this value is multiplied by 2, the figure then equals miliequivalents of peroxide oxygen
per kilogram of sample (mEq/kg), which has greater international recognition.
Fresh oils usually have peroxide values below 10 mEq/kg. A rancid taste often begins to be
noticeable when the peroxide value is between 20 40 mEq/kg.
Peroxide Value =

Vs Vb
Weight of sample

x T x 103

T = Molarity of sodium thiosulphate

Vs = Volume in mL titration for sample
Vb = Volume in mL titration for blank
Palm oil (new)
= 9.56 ml 4.87 ml x 0.002 M x 103
1 gram
= 4.69 x 2
= 9.38
Palm oil (old)
= 21.80 ml 4.87 ml x 0.002 M x 103
1 gram
= 16.93 x 2
= 33.86

Results.
Table 2.1: Acid Value or FFA of Fats/Oils
Sample
Palm oil
(new)
Palm oil
(old)

Trail 1
1.0
1.1

Volume of titration (ml)

Trail 2
Trail 3
0.6
0.7
1.4

1.1

Acid Value

FFA

Average
0.77

0.8639

0.4320

1.20

1.3464

0.6732

Peroxide
Value (PV)

(PV x 2)
mEq/kg

Table 2.2: Peroxide Value of Fats/Oils

Sample /
Blank
Blank
Palm oil
(new)
Palm oil
(old)

Volume of titration (ml)

Trail 1
4.00

Trail 2
5.20

Trail 3
5.40

Average
4.87

0.00

0.00

20.00

10.00

6.50

9.56

9.38

18.76

24.50

24.50

16.40

21.80

33.86

67.72

Discussions.
Based on the results of this experiment from the table above, its shows that the acid value of
palm oil (new) is lower than palm oil (old) which is 0.8639 and 1.3464 respectively. The free
fatty acids (FFA) of palm oil (new) is lower than palm oil (old) too which is 0.4320 and 0.6732
respectively. Acid value or free fatty acids is related to hydrolytic rancidity.Hydrolytic rancidity
is caused by fat hydrolysis. Hydrolytic rancidity refers to the odor that develops when
triglycerides are hydrolyzed and free fatty acids are released. This reaction of lipid with water
sometimes requires a catalyst, but results in the formation free fatty acids and salts from free
fatty acids (soaps). In particular, short chain fatty acids, such as common butter fats, are
odorous. From this experiments result, the acid value of palm oil (old) is higher than palm oil
(new) because it has more free fatty acids and thus become more susceptible to become rancid.
The peroxide value of palm oil (new) based on the above table is lower than palm oil (old)
which is 9.38 and 33.86 respectively. The peroxide value is related to oxidative rancidit.
Oxidative rancidity is a condition cause by fat oxidation. Peroxide value is a measure of
oxidation during storage and the freshness of lipid matrix. In addition, it is a useful indicator of
the early stages of rancidity occurring under mild condition and it is a measure of the primary
lipid oxidation products. One of the most important parameters that influence lipid oxidation is
the degree of unsaturation of its fatty acids. When double bonds of unsaturated fats are oxidized,

peroxides are among the oxidation products formed. High peroxide value is an indicator of
oxidation level and the greater the peroxide value, the more oxidized the palm oil. The peroxides
decompose to yield a complex of mixtures, including aldehydes, ketones, and other volatile
products. These products are responsible for "rancid" odors and flavors. So, when the palm oil
has higher peroxide value, the higher it is susceptible to become rancid. From this experiement,
the result is valide because the palm oil (new) have peroxide value is below 10 mEq/kg since it is
fresh oil, while the peroxide value for palm oil (old) is ranging between 20 40 mEq/kg and its
shows that this old palm oil has begins a rancid taste and undergo rancidty.
Conclusions.
The conclusion is, palm oil (new) has lower acid value and peroxide value than palm oil (old),
and thus it proves that it is more healthy to consume fresh oil than the palm oil (old) because it is
not rancid or have lower chances to become rancid.
Questions.
1. What is rancidity?
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. These are oxidative, hydrolytic, and ketonic.
2. List 2 types of rancidity which normally occur in food products.
- Oxidative rancidity
- Hydrolytic rancidity
3. What are the factors that encourage the rancidity of fats and oils?
The factors that encourage the rancidity of fats and oils are too much exposure to air, heat and
light, and chemical composition (saturated or unsaturated fat) of fats and oils.

References.
- https://www.npal.com/docs/npal_document_71.pdf
-https://www.australianolives.com.au/assets/files/pdfs/Media/Health%20Effects%20of%
20Rancid%20Oils.pdf
- http://www.gcca.org/wp-content/uploads/2012/09/RancidityAntioxidants.pdf
- http://whfoods.org/genpage.php?tname=dailytip&dbid=357 (haein)
- Rancidity and Antioxidants, Revised 2008, WFLO Commodity Storage Manual.
- Estimation of total free fatty acid and cholesterol content in some commercial edible oils in
Ethiopia, Bahir DAR Dimberu G. Atinafu* and Belete Bedemo, November 2011, Journal of
Cereals and Oil seeds Vol. 2(6), pp. 71-76.