FOOD ANALYSIS LABORATORY

(STKM2211)

TITLE :
DETERMINATION OF FAT CONTENT

NAME :
ZAINAH NAJIHAH BINTI ZAINAN

MATRIC NO. :
A190670
Objectives
 To determine the fat content of selected food samples using Soxhlet method and
Mojonnier method.

Introductions
One of the main components of food is lipids since it is one of major source of energy. As
a major source of energy and providing essential fatty acids, they are important in our
diet. It also plays a major role in deciding foods’ overall physical characteristics such as
taste, texture and feel and appearance in the mouth. Lipids are generally classified as a
component that are insoluble in water and soluble in organic solvents such as ether and hexane.
The solvent extraction method and even the non-solvent wet extraction method are the method
that can determine lipids or fat content in food samples. The method of collection depends on the
quality of the samples, the intent of the study and the available instrument. One of the most
widely used methods of isolating lipids from foods and determining the overall lipid content of
foods is solvent extraction method. The idea that lipids are soluble in organic solvents but
insoluble in water gives a simple way for the food analyst to distinguish lipid components
from water-soluble components such as proteins, carbohydrates and minerals in foods. The most
suitable sample preparation technique should be conducted in order to obtain precise and
reliable results. Typically, a variety of steps are needed to prepare a sample for solvent
extraction, such as drying sample, reducing particle size, acid hydrolysis and selecting the ideal
solvent for successful lipid extraction.
Method
Determination of crude fat content in peanut (grounded) (Soxhlet method)
1. Weight 2g pre-dried sample into pre-dried extraction thimble.
2. Place the weighed groundnuts in to the pre-dried extraction thimble.
3. Weight a pre-dried boiling flask.
4. Add 200mL hexane into the boiling flask.
5. Insert the thimble with samples into the Soxhlet apparatus.
6. Assemble the Soxhlet apparatus, boiling flask and condenser.
7. Heat up the boiling flask, and allow the Soxhlet extractor to extract fats from sample for
2 hour.
8. Remove the Soxhlet apparatus, and pour the excess solvent into the boiling flask.
9. Evaporate the solvent using a rotary evaporator.
10. Further dry boiling flask in an oven at 105⁰C for 1 h.
11. Cool flask in a dessicator and weigh.

Determination of crude fat content in minced chicken breast (Mojonnier method)

1. Weight 4g of chicken breast samples into an Erlenmeyer flask.
2. Add 20mL 4N HCl into the Erlenmeyer flask containing sample.
3. Heat in a water bath at 60℃ for 20min, stirring occasionally with a glass rod.
4. Cool and transfer samples into a Mojonnier fat extraction flask.
5. Add in 30mL of diethyl ether into the Mojonnier flask to extract the fats from the
samples.
6. Extract crude fat by shaking the Mojonnier tube 30 times.
7. Keep Mojonnier tube stationary and allow two layers to form.
8. Decent the ether layer into a pre-weighed boiling flask.
9. Repeat fat extraction 2 more times using 30mL diethyl ether each time.
10. Evaporate the solvent using a rotary evaporator.
11. Further dry boiling flask in an oven at 105⁰C for 1 h.
12. Cool flask in a dessicator and weigh.
Result

For Soxhlet method

Experimental Data
Sample: Peanut (Grounded)

Replication Sample Weight (g) Empty flask weight (g) Flask with sample oil Fat (g)
s extract weight (g)
1 2.012 220.210 221.072 0.862
2 2.108 225.808 226.863 1.055
3 2.002 217.613 218.531 0.918

Calculation:

[(Weight of flask + fat )– (Weight of flask )]

% fat = × 100
Weight of sample
0.862
Replications 1 : % fat = × 100 = 42.84%
2.012
1.055
Replications 2 : % fat = × 100 = 50.05%
2.108
0 .918
Replications 3 : % fat = × 100 = 45.85%
2.0 02
42.84+ 50.05+ 45.85
Mean = = 46.23
3

Standard deviation = √(42.84−46.23) +( 50.05−46.23 ) +(45.85−46.23)²

2 2
= 9.0634
3−1
Mean ± SD = (46.23 ± 9.0634)
 For Mojonnier method
Experimental Data
Minced Chicken
Sample: Breast

Replications Sample Weight (g) Empty flask weight Flask with sample Fat (g)
(g) oil extract weight
(g)
1 4.082 229.224 229.348 0.124
2 4.052 222.936 223.081 0.145
3 4.108 219.336 219.464 0.128

Calculation:

[(Weight of flask + fat )– (Weight of flask )]

% fat = × 100
Weight of sample
0.124
Replications 1 : % fat = × 100 = 3.038%
4.082
0.145
Replications 2 : % fat = × 100 = 3.578%
4.052
0.128
Replications 3 : % fat = × 100 = 3.116%
4.108
3.038+3.578+3.116
Mean = = 3.244
3

Standard deviation = √(3.038−3.244 ) +( 3.578−3.244 ) +( 3.116−3.244) ²

2 2
= 0.16697
3−1
Mean ± SD = (3.244 ± 0.16697)
Discussion
In experiment Soxhlet method peanuts were used as a food sample. Based on the results,
replicate 1 has 42.84% of crude fat, replicate 2 has 50.05% crude fat and replicate 3 has 45.85%
of crude fat. The average of peanuts crude fats content is 46.23 ± 9.0634. For theoretical
value fat content for potato chip is ranged within 50% (SS Arya, 2016). Thus, from the result
obtained it show that the fat content is less than the theoretical value and there is some
consideration when using Soxhlet method which while placing the thimble-containin
gsample in the Soxhlet flask, make sure that the top of the thimble is above the siphon tube. A
cotton swab mut be put on the mouth of the condenser to avoid the loss of ether vapours.

The determination of fat in food is done because of a variety reason such as for health,
nutrition, weight loss and want to know the precise amount before making nutritional decisions.
It is important for food producers to be able to report fat content in their product because of
commercial regulations (Medallion Labs, 2018). Soxhlet method used because it is fairly simple
to use and it is officially recognized method for a wide range of fat content determinations. This
method also efficient and rapid extraction method for lipids in foods.

In experiment Mojonnier method minced chicken breast were used as a food sample.
Based on the results, replicate 1 has 3.038% of crude fat, replicate 2 has 3.578% crude fat and
replicate 3 has 3.116% of crude fat. The average of minced chicken breast crude fats content
is 3.244 ± 0.16697. Mojonnier method suitable used for milk and dairy product, but we try to
used this method for minced chicken breast.
Conclusion
Concluding everything, this experiment use Soxhlet extraction method to obtain the
percentage of fat in peanuts and Mojonnier method to obtain the percentage of fat in minced
chicken breast. From the Soxhlet extraction method, we can obtain the weight of fat in the food
sample after the solvent was evaporated. Then, using the formula we can determine the
percentage of fat in peanuts. From the result, it shows that peanuts contains more fat than minced
chicken breast. In conclusion the objective of this experiment which is to determine the fat
content of selected food samples using Soxhlet method and Mojonnier method was successfully
obtained.

Questions
1. Why is the Mojonnier extraction method more suitable for extracting fat from meat
compared to the Soxhlet method?
Mojonnier Method is a discontinuous solvent extraction method and does not require
removal of moisture from the sample. Soxhlet method required dried and grounded
sample prior analysis.

2. What role does 4N HCl play in the Mojonnier fat extraction method?
Mojonnier extraction is typically used to extract fatty acids after acid hydrolysis. Thus,
hydrochloric acid is used to extract fatty acid of the samples.

3. Why is it necessary to predry your samples if they are moist? What drying method would
you employ?
Fats cannot be efficiently extracted from moist foods with ethyl ether; ether saturated
with water is inefficient for fat extractions; dried samples are easier to grind. To extract
the fat from a food sample, you have the choice of using ethyl ether or petroleum ether
as the solvent, and you can use a Soxhlet apparatus.