FST306 LABORATORY REPORT

PREPARED BY:
KHAIRUNNISA BINTI KHAIRUL AZAM 2017258506
NUR FATIHA BINTI ADNAN 2017486316
NURNADHIRAH RASYIDAH BINTI MOHD RAZANI 2017258768
NUR ATIQAH BINTI MOHD NASRI 2017205634
NUR AFIQAH BINTI ROSNAN 2017205626

GROUP: AS1163B1
NO EXPERIMENT: 2
TITLE: DETERMINATION OF FAT
LECTURER’S NAME: MADAM MARINA
SUBMISSION DATE: 14/12/2018
INTRODUCTION

Lipids are substances that are soluble in organic solvent but insoluble in water.
They may exist in liquid or solid form in the room temperature depending on their structure and
composition. Liquid triglycerides at room temperature are called oils and generally in plant
origin food such as olive oil. Solid triglycerides at room temperature are fats and generally in
animal origin food such as lard. One of the major constituents in foods are lipids. Besides
playing an important role in our diet as source of energy and providing essential fatty acids, it
also plays a major role in determining the overall physical characteristics of food such as flavor,
texture and appearance.

Fat content in food varies widely from less than 1% to almost 100%. The importance of
lipids analysis can be varied but the most important include labelling law, health, quality and
processing. A proper sampling and preservation of the sample prior to analysis affects the
validity of the results. So, the sample preparation required in lipid analysis depends on the type
of food being analyzed, the nature of lipid content and the type of method chosen. Types of
analytical methods in lipid determination are either solvent extraction, non-solvent extraction or
instrumental. Solvents used to extract lipids include ether, petroleum ether, acetone, chloroform,
benzene, alcohols and water saturated butanol.

OBJECTIVES

i. To determine the fat content of selected food samples using Sohxlet method
ii. To determine the fat content of selected food samples using Mojonnier method

PROCEDURE

SOXHLET METHOD

1. 2 g of dried sample was weighed accurately into an extraction thimble.

2. The opening was plugged of the thimble loosely with cotton the filter and was plugged
with cotton.
3. The thimble and content was placed into a soxhlet extractor.
4. A dried round bottom flask was weighed accurately and 150 ml petroleum ether was
added.
 5. The apparatus was connected to the condenser, the water was turn on and it was extracted
for a minimum of 8 hours on an elctrothernal extraction unit.

MOJONNIER METHOD

First extraction

1. The nearest 0.1 mg was weighed, 10 g milk into a Mojonnier fat extraction flask.
2. 1.5 ml NH4OH was added and shaked vigorously. 2 ml was added if the sample is sour.
3. 10 ml 95% ethanol was added and shaked 90 seconds.
4. 25 ml diethyl ether was added and shaked 90 seconds.
5. Cooled if necessary, 25 ml petroleum ether was added and shaked 90 seconds
6. Ether solution was decant from the Mojonnier flask into the previously weighed
evaporating dish.

Second extraction

1. 5 ml 95% ethanol was added and shaked vigorously 15 seconds.

2. 15 ml diethyl ether was added and shaked 60 seconds.
3. 15 ml petroleum ether was added and shaked 60 seconds
4. Solution was decant into the same evaporating dish.

Third extraction

1. 15 ml diethyl ether was added and shaked 60 seconds.

2. 15 ml petroleum ether was added and shaked 60 seconds.
3. The solution was decant into the same evaporating dish.
4. The solution was distil from the flask, the fat was dried at 105°C for 1 hour, cooled it and
weighed.
5. Any non- fatty material was appeared to be present, the fat was washed out from the flask
with light petroleum ether, dried the fat, reweighed and the result was corrected
accordingly.
CALCULATIONS &RESULTS

Result
Determination of Fat Data using Soxhlet Method
Sample Trial Weight of sample (g) Weight of Weight of
round bottom round bottom % Fat
flask (g) flask + fat (g)
1 2.0004 112.0974 112.2216 6.21 %

Biscuit 2 2.0001 101.9042 102.2258 16.08 %

Mean 2.0003 107.0008 107.2237 11.14 %
1 2.0001 112.1042 112.2258 6.08 %

Oats 2 2.0000 100.9406 101.2484 15.39 %

Mean 2.0001 106.5224 106.7371 10.73 %

Calculation for % of fat for Soxhlet Method:

% Fat in Biscuit = Weight of fat in sample x 100

Weight of dried sample
= (107.2237 – 107.0008) x 100 = 11.14 %
2.0003

% Fat in Oats = Weight of fat in sample x 100

Weight of dried sample
= (106.7371 – 106.5224) x 100 = 10.73 %
2.0001

Sample Trial Weight of sample Weight of Weight of % Fat

(g) evaporating dish evaporating
(g) dish + fat (g)
 1 10.0002 99.1222 106.0006 68.78 %

Milk 2 10.0003 99.3706 101.1812 18.11 %

Mean 10.0000 99.2464 103.5909 43.46 %

Determination of Fat Data using Mojonnier Method

Calculation for % fat for Mojonnier Method:

% Fat = [(weight of evaporating dish + fat) – (weight of evaporating dish)] x 100
Weight of sample taken

= (103.5909 – 99.2464) x 100 = 43.46 %

10.0000

DISCUSSIONS

Two experiments were conducted for determination of crude fat, which is Soxhlet
method and Mojonnier method. The Sohxlet method is the process of transferring the partially
soluble components of a solid to the liquid phase using a Soxhlet extractor. Meanwhile, for
Mojonnier method is the process where defatted solids may be determined by drying the residue
that remains in the extraction flask after the solvents are removed. Mojonnier method could
provide a very precise measurement of milk fat as it involves the chemical ether and alcohol. The
ether dissolves fats, oils and other fat soluble substances. The general fat percentage of biscuit,
oats and milk powder are 9%, 7% and 26-40% respectively.

Based on the experiment using the Sohxlet method, the sample used are biscuit and oats.
The results obtained shows different amount of fat that can be extracted by using the same
sample. As for the biscuit, the first trial is 6.21% and second trial is 16.08% of fat that can be
extracted. This gives an average of 11.14% of fat contained in the biscuit. The first trial gives a
low value of fat percentage even though the weight of sample used is 2.0004g. As for the second
trial, by using 2.0001g of biscuit sample, it gives a higher value of fat percentage which is
16.08%. This indicates that there could be an error happening in the experiment. This is probably
due to the particle size of sample. By reducing the particle size of the biscuit, the lipid extraction
would be better as the area surface is bigger. It can be concluded that by grinding the sample
properly, it will give a higher value of fat percentage.

As for the oats sample, the first trial gives a 6.08% of fat and second trial is 15.39% of
fat. The average of both trials gives a 10.73% of fat percentage. Even though the weight of
sample for the first trial and second trial is 2.0001 and 2.0000 respectively, it gives a huge
difference of fat extraction that can be obtained from the same sample. The first trial gives a
value that almost the same as the general value of fat percentage but not for the second trial, as
the value is way too big to be compared to the general value. This error could be from the
interference of other compound as it shows a huge amount of fat that was extracted. It is also
could happen due to using the same flask of doing another fat extraction, that some of the
compound may stick on the wall of the flask.

For Mojonnier method, the sample used was the milk powder. The first trial gives a high
value of fat percentage which is 68.78%. As for the second trial, the amount of percentage of fat
that can be extracted from the milk powder is only 18.11%. This shows a totally big difference of
fat percentage between these two trials. The error that occurs in this experiment could be due to
the reagent used. In this method, the extraction is repeated three times and the shaking times in
each repetition is different. Also, there is no addition of 95% ethanol in the third extraction. By
mistakenly adding ethanol, it could lead to a difference in result. Therefore, the value of fat
percentage could be lower than the actual value.

QUESTIONS
1) Draw and label the main components of Soxhlet apparatus

2) Describe the function of the various reagents used in each method.

Alcohol used to prevents possible gel formation. Ammonia used to neutralizes
acidic sample and dissolves protein. Petroleum ether uses in this experiment to
remove moisture from the ethyl ether extract and dissolves more non-polar lipid.
 3) Which is the best extraction procedure either one extraction with a large volume
of solvent OR several extractions with smaller volumes but the same total
volume? Explain.
The best extraction is by doing a several extractions with smaller volumes but the
same total volume. It is because the greater the number of small extractions, the
greater the quantity of solute removed. If one extraction can be recover 90% of the
compound, the second extraction with the same solvent may also be able to pull out
the same percent what is left. So it will double up the efficiency of pulling out the
compound compared to one extraction that only extracted 90% of the compound.

CONCLUSION

This experiment able to determine the fat content of selected food sample using Soxhlet
method which is 11.14 % in biscuit sample and 10.73% in oats sample. This experiment also
able to determine the fat content from the selecting food using Mojonier method which is milk
that contain 43.46% of fat. The preparation of a sample for solvent extraction have been done
such as drying sample, particle size reduction, acid hydrolysis and choosing the ideal solvent for
efficient lipid extraction. For this experiment, petroleum ether, diethyl ether and ethanol are
used. This step is important to get accurate and precise result.

REFERENCES

i. D. L. Luthria. (2003). Oil Extraction and Analysis: Critical Issues and Competitive
Studies. The American Oil Chemists Society.
ii. http://web2.slc.qc.ca/jmc/www/Chemweb/oldchemweb/FatLab.htm
iii. http://discoverfoodtech.com/soxhlet-extraction-method/
iv. https://www.sciencedirect.com/science/article/pii/S0022030244925737