PRACTICAL 3: DETERMINATION OF FAT

INTRODUCTION

Fat is a major constituent of foods. It is important in our diet for the source of energy,
essential nutrients, flavor, texture, mouth, feel and appearance. By the other hand, if too much
consumption certain lipids can be detrimental to health (cholesterols and saturated fats). Fats
is a food components that are soluble in organic solvents but insoluble in water. Lipid exist as
free fatty acid, phospholipids, sterols, monoacylglycerol, diacylglycerol and triacylglycerol,
waxes and fat soluble vitamins and pigment. It is important to run lipid analysis especially for
nutritional labelling and to conform to standards. Other than that it also important to determine
the food quality because food properties depends on total lipid content.

There are a few method for extraction fat in fat analysis. The method that been used
in this experiment are discontinuous extraction method (Werner-Schmid method) and semi-
continuous extraction method (Soxhlet method). For discontinuous extraction method, it based
on mixing sample and solvent in suitable flask. Then, the flask were shaken vigorously and
allow solvent and aqueous phase to separate. Aqueous phase decanted off. Concentration of
lipid in solvent can be determined by evaporating solvent and measure of the remaining lipid
after drying. While semi-continuous extraction method are more efficient and rapid extraction
method for lipid in foods. A semi continuous process is one that allows a solvent to build up
and surround a sample. The soxhlet methods allows a solvent to run through a sample to
extract and collect lipid in a flask. (Hahm, 2009).

OBJECTIVE

To determine the crude fat content in peanut and unsweetened evaporated milk.

MATERIALS

Petroleum ether, analytical balance, desiccator, Soxhlet apparatus, filter paper, round bottom
flask, heating mantle, water bath, graduated cylinder, beaker, mortar and pestle, spatula,
oven, concentrated hydrochloric acid, ethanol, diethyl ether, test tube, tongs, boiling water
bath, separatory flask, evaporating dish and pipette.

METHODS

1. Soxhlet extraction method

Dried round bottom flask were obtained and clearly labelled. Two gram of dried
sample were placed into a piece of filter paper. Then the paper been transferred into
Soxhlet extractor. The empty labeled round bottom flask were weighed and 180 ml
 petroleum ether was added. The apparatus then was connected to the condenser, and the
water been turn on and was extracted for 8 hours on an electrothermal extraction unit. The
flask that contained petroleum ether was removed when the extraction done. The round
bottom flask was placed on a steam bath to remove residual petroleum ether. After all
solvent dried the flask were placed in the oven with 1050C for 1 hour of drying. The flask
was immediately transferred into a desiccator to cool and weigh.

2. Werner Schmid method

Ten gram of sample was placed into the test tube. Ten millimeter of concentrated
hydrochloric acid were added, then been immersed in the boiling water bath and
continuously stirred with glass rod until the mixture turn violet. The tube then been cool it
down in running water to room temperature and all of the content been transferred into
separatory flask. The fat been extracted by shaking with 20 ml diethyl ether for a minute
and an equal amount of petroleum ether were added and shake for another minute and a
few drop of ethanol was added. The solution were decanted into evaporating dish and
dried at 1000C, cooled and weighed.

RESULTS

Table 1.1 crude fat analysis using Soxhlet method (Peanut)

F1 F2 Mean Standard
deviation
Weight of sample 2.11 2.01
(g)
Weight of empty 116.87 127.17
round bottom flask
(g)
Weight of round 118.37 128.64
bottom flask +
crude fat (g)
% crude fat 71.09 % 73.13 % 72.11 % 1.02
content
 Table 1.2 crude fat analysis using Werner-Schmid method (unsweetened evaporated milk)

F1 F2 Mean Standard
deviation
Weight of sample 10.04 10.01
(g)
Weight of empty 62.41 61.81
evaporating dish
(g)
Weight of 62.95 62.30
evaporating dish +
crude fat (g)
% crude fat 5.38 % 4.90 % 5.14 % 0.24
content

DISCUSSION

From the analysis of fat content of peanuts, Soxhlet method extraction has been used.
Solvent extraction of solid samples, is commonly known as solid-liquid extraction, which is one
of the oldest methods for solid sample pretreatment. (Castro, 2000). The analysis shows that
the percentage of the crude fat content of peanut is 72.11 %. By the other hand a research
been done regarding this fat content of peanuts by (Arnarson, 2019) which the analysis state
that, the fat content of peanuts are in between 44-56 % only which much lower. This shows a
contradiction with the obtained result in the analysis. This probably happen due to residual
solvent that not totally evaporated and cause overestimation to happen. This probably can
improvise in future by lengthen the drying process in the oven so that the solvent will totally
evaporated. Nevertheless, this Soxhlet extraction method is very effective because it can be
extracted multiple times. Compared with general soaking method, it has the advantage of
small solvent dosage, high efficiency and complete extraction.

Next, for unsweetened evaporated milk fat analysis, Werner-Schmid method of

extraction is used. In this method, hydrochloric acid is added to break covalently and ionically
bound lipids into free lipid form since lipids complexes with protein (lipoprotein) and
polysaccharides such as glycolipids. The solvent been used is diethyl ether and petroleum
ether. Addition of petroleum ether is important because petroleum ether will removes moisture
from ethyl ether extract and dissolves more non polar lipid. During this analysis a few drop
ethanol is needed to get a better separation of solvent and sample. If bad separation happen,
overestimation might occur where the sample already mix up with the solvent that consist fat
when decant in evaporating dish. From the analysis the crude fat content of unsweetened
evaporated milk is 5.14 %. From a study been done the minimum standard of evaporated milk
contain at least 7.9 % milk fat. (Fillipone, 2018). From the analysis, the fat content is much
lower. This probably due to incomplete extraction occur during the analysis. Other than that,
it might happen due to not homogenized sample is obtained for the analysis, which can be
improve in future by shaking the sample first, before the analysis.

QUESTIONS

1. Comment the suitability of the method with the supplied samples.

It is suitable to run fat analysis with the supplied samples, for the Soxhlet method a
solid sample is preferred, while for the Werner-Schmid method, less sugar content of
sample is required as example in this analysis unsweetened evaporated milk. It is
because in Werner-Schmid method used hydrochloric acid. If the milk sample high in
sugar, it can cause the milk to churning.
2. Difference between one extraction with large volume of solvent and use the same
amount of solvent total volume with several extractions in smaller volumes.
Several extractions with small volumes of solvent is more effective than a single
extraction with a large volume of solvent. It is because, when you only extract once
there must be a residual left, with a multiple extraction the residual can be reduced and
better extraction will be obtained.

CONCLUSION

In conclusion, the fat content in peanuts is 72.11 % while in unsweetened evaporated milk the
fat content is 5.14 %.

References
Arnarson, A. (2019, May 7). Peanuts 101: Nutrition Facts and Health Benefits. Retrieved from
healthline.com: https://www.healthline.com/nutrition/foods/peanuts

Castro, L. d. (2000). Environmental application: Soxhlet extraction. Soxhlet extration, 88-92.

Fillipone, P. (2018, January 11). Evaporated and Sweetened Condensed Milk Differences. Retrieved
from the spruce eats: https://www.thespruceeats.com/evaporated-vs-sweetened-
condensed-milk-1807001

Hahm, J. (2009, February 3). Fat Extraction. Retrieved from NFSC: Nutrient analysis in action:
https://nfsc450groupd2015.weebly.com/about.html
APPENDIX
Calculation

Soxhlet method

% Fat in sample = Weight of fat in sample (g)/ Weight of sample taken (g) x 100 %

= (118.37 g -116.87 g) / (2.11 g) x 100%

=71.09 %

Werner Schmid method

% Fat in sample = Weight of fat in sample (g)/ Weight of sample taken (g) x 100 %

= (62.95 g -62.41 g)/ (10.04 g) x100 %

= 5.38 %
School of Industrial
Technology
Faculty of Applied
Sciences
__________________________________________________
______

FST 556
Laboratory Title: Determination of fat
Date of experiment: 7.10.2019
Laboratory Group: AS2464C

Name : Muhammad Shafiq Bin Mustafa

Student ID : 2018401924
Group : AS2464C

MARK: