FST 306

STUDENTS NAME & MATRIX NUMBER:

1.
2.
3.
3.

Students name
ASMAA SYUHADA BINTI AMINUDDIN
NUR AIN AYUNI BINTI AMAT SUJANGI
SITI NUR RUQAYYAH BINTI MOHD SAMSHUL
SITI NOOR SYAFIZA BINTI YUSRI

EXPERIMENT NO: 2
EXPERIMENT TITLE: DETERMINATION OF FAT
DATE OF EXPERIMENT: 27 JULY 2016
DATE OF SUBMISSION: 3 AUGUST 2016
LECTURERS NAME: MDM. NUR AISYAH ISMAIL

INTRODUCTION

Matrix number
2015867272
2015845854
2015874022
2015838698

Fat is one of the three main macronutrients, along with carbohydrate

and protein. Fats, also known as triglycerides, are esters of three fatty acid
chains and the alcohol glycerol. Lipids are one of the major constituents of
foods, and are important in our diet for a number of reasons. They are a
major source of energy and provide essential lipid nutrients. Nevertheless,
over-consumption of certain lipid components can be detrimental to our
health, e.g. cholesterol and saturated fats.
In many foods the lipid component plays a major role in determining
the overall physical characteristics, such as flavor, texture, mouth feel and
appearance. For this reason, it is difficult to develop low-fat alternatives of
many foods, because once the fat is removed some of the most important
physical characteristics are lost.
Lipids are usually defined as those components that are soluble in
organic solvents (such as ether, hexane or chloroform), but are insoluble in
water. This group of substances includes triacylglycerol, diacylglycercols,
monoacylglycercols, free fatty acids, phospholipids, sterols, carotenoids and
vitamins A and D. The lipid fraction of a fatty food therefore contains a
complex mixture of different types of molecule.

Although sometimes the

term fat is used to describe those lipids that are solid at the specified
temperature, whereas the term oil is used to describe those lipids that are
liquid at the specified temperature.
In order to get accurate and precise result, most appropriate sample
preparation procedure should be done. The preparation of a sample for
solvent extraction usually involves a number of steps such as drying sample,
particle size reduction, acid hydrolysis and choosing the ideal solvent for
efficient lipid extraction.

SOXHLET EXTRACTION METHOD

Objective:
To determine the fat content of selected food samples using Soxhlet method
Apparatus:
Analytical balance, dessicator, Soxhlet apparatus, thimbles, round bottom flask 250 ml, heating
mantle, water bath, graduated cylinder 500 ml, beaker 250 ml, mortar and pestle, spatula and
oven
Chemical:
Petroleum ether (boiling point: 40-60C)
Procedure:
1. About 2g of dried sample was weighed accurately into extraction thimble or a piece of filter
paper.
2. The opening of the thimble was plugged loosely with cotton or the filter was folded and was
plugged with cotton.
3. The thimble or paper and content were placed into a Soxhlet extractor.
4. A dried round bottom flask was weighed accurately and 150 ml petroleum ether was added.
5. Apparatus was connected to the condenser, turned on the water and was extracted for a
minimum of 8 hours on an electrothermal extraction unit.
6. Flask was removed containing petroleum ether extract after the extraction complete.
7. The petroleum ether was evaporated off on a boiling water bath.
8. The flask was transferred into an oven at 105C for one hour to dry the extract.
9. The flask was transferred immediately into a dessicator to cool and weigh

Calculations:
1. Butter cracker
First Trial: Weight of fat in sample

= (Weight of Flask + Fat) (Weight of Flask)

= 120.9116 120.3531
= 0.5585

% Fat in sample

= Weight of Fat in sample (g) 100

Weight of sample taken (g)
= 0.5585 100
2.0109
= 27.7736%

Second Trial: Weight of fat in sample = (Weight of Flask + Fat) (Weight of Flask)
= 126.2985 124.9456
= 1.3529
% Fat in sample

= Weight of Fat in sample (g) 100

Weight of sample taken (g)
= 1.3529 100
2.0482
= 66.0531%

2. Hup Seng
First Trial: Weight of fat in sample

= (Weight of Flask + Fat) (Weight of Flask)

= 125.2923 123.8412
= 1.4511

% Fat in sample

= Weight of Fat in sample (g) 100

Weight of sample taken (g)
= 1.4511 100
2.0951
= 69.2616%

Second Trial: Weight of fat in sample = (Weight of Flask + Fat) (Weight of Flask)
= 105.3451 104.6572
= 0.6879
% Fat in sample

= Weight of Fat in sample (g) 100

Weight of sample taken (g)
= 0.6879 100
2.0283
= 33.9151%

Third Trial: Weight of fat in sample = (Weight of Flask + Fat) (Weight of Flask)
=101.2320 99.9408
= 1.2912
% Fat in sample

= Weight of Fat in sample (g) 100

Weight of sample taken (g)
= 1.2912 100
2.0736
= 62.2685%

Results:
Table 2.1: Determination of Fat Data Using Soxhlet Method

Sample

Trial

Weight of sample Weight of round Weight of round % Fat

bottom flask (g)
bottom flask + fat
(g)
(g)

Butter
Cracker

2.0109

120.3531

120.9116

27.7736%

2.0482

124.9456

126.2985

66.0531%

Mean

46.9134%

Hup Seng 1

2.0951

123.8412

125.2923

69.2616%

2.0283

104.6572

105.3451

33.9151%

2.0736

99.9408

101.2320

62.2685%

Mean

55.1484%

MOJONNIER METHOD
Objective:
To determine the fat content of selected food samples using Mojonnier
method.
Principle:
Fat is extracted with a mixture of ethyl ether and petroleum ether, and the
extracted fat is dried to constant weight and expressed as percent fat by
weight. The assay uses not only diethyl ether and petroleum ether but also
ammonia and ethanol.
Apparatus:
Mojonnier extraction flask with stopper, evaporating dish, analytical balance,
oven, desiccator and graduated cylinder 25ml.
Chemicals:
Ammonium hydroxide, ethanol, diethyl ether, petroleum ether (boiling point:
40-60C)

Discussion:
Fat is important to all aspects of meat production and processing.
Crude fat content is determined by extracting the fat from the sample using
a solvent, then determining the weight of the fat recovered. As lipids/fats are
relatively non-polar molecules, they can be pulled out of a sample using
relatively non-polar solvents. With a non-polar solvent, only non-polar
molecules in the sample dissolve while polar ones do not. Problems arise
however, in cases where lipids are bound in animal or plant cell membranes.
Animal and plant cell membranes are made up of molecules that have both
polar and non-polar regions such as triglycerides (molecule with polar
glyceride heads and non-polar fatty acid tails) and phospholipids (similar to
triglycerides, but a phosphate group replaces the fatty acid tail). These
molecules end up grouping together with their polar heads sticking outwards
and non-polar tails inwards making it difficult for non-polar solvents to
interact with the non-polar tails and extract them
Based on this experiment, be able learn how to determine the fat
content of selecting food samples using Soxhlet method. Food samples that
using in this experiment which had butter cracker and Hup Seng biscuit. By
using Soxhlet method it will know the content of fat in both of food samples.
It had weighed accurately into an extraction thimble or a piece of filter paper.
The thimble or paper content had placed into Soxhlet extractor. All apparatus
had connected to condenser and the water had turned on to extract for
minimum 8 hours on an electro thermal extraction unit. Then, the flask
containing the petroleum extract had removed after extraction complete.
The flask had transferred into an oven at 105C for one hour to dry the
extract. After that, the flask had transferred immediately into a desiccator to
cool and weigh. The weight had recorded on Data Sheet. The Soxhlet

extraction method, semi continuous extraction method, used for determine

crude fat content in this food samples. The Soxhlet allows one to extract fats
from a solid sample into a liquid, leaving behind any impurities. It also used
small volume of solvent over and over again.
Furthermore, the result had obtained the percentage of fat in Hup Seng
biscuits had 55.1484% and for butter cracker had 46.9134%. Hup Seng
biscuits have higher percentage of fat content than butter cracker. It had
slightly different percentage fat for both of food samples because the
different amount of fats used by manufacture to produce this biscuits.
Due to the error during the experiment that the sample had not been
extracted properly by using Soxhlet method as the method only give the free
fatty acid amount of the sample. For the recommendation, the total fat
extraction method result can be used for the result in both free and bound
fat for the higher value in total fat extraction.
Conclusion:
At the end of the experiment, the fat content of selected food samples,
biscuits which came from two different companies that were used in Soxhlet
Extraction method were successfully determined. As for Mojonnier method
which used two different types of milk, the fat content could not be
determined because of some faults throughout the procedures but the
mistakes had been identified and corrected so that it would not happen in
the future.

Questions:
1. Draw and label the main components of Soxhlet apparatus.

condenser

Extraction
chamber
Siphon arm

thimble

Round bottom flask

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

Petroleum ether Removes moisture from the ethyl ether extract and
dissolve non-polar lipid (reduce proportion of water and non-fatty soluble

substances)
Ethanol Precipitates protein and prevents possible gel formation.
Diethyl ether Dissolved the lipids
Ammonia To neutralizes acidic samples and dissolve protein

3. Which is the best extraction procedure:

a. One extraction with large volume of solvent OR
b. Several extractions with smaller volumes but the same total volume? Explain.
The best extraction method is several extractions with smaller volumes but the same
total volume because the steps are simple and reading is more accurate compared with
large volume of solvent.

References:
1. INTRODUCTION OF FOOD ANALYSIS, S. Suzanne Nielsen, 2010 Edition,
Klew Academic Plenum Publishers.
2. FOOD CHEMISTRY, Fennema, Owen. R., 1996, 3rd Ed, Marcel Dekker, Inc. New York.
3. http://web2.slc.qc.ca/jmc/www/Chemweb/oldchemweb/milk.htm