Exercise no.

4
LIPID EXTRACTION AND LIPID TESTS
I. INTRODUCTION

Lipid, any of a diverse group of organic compounds including fats, oils, hormones, and certain components of membranes that
are grouped together because they do not interact appreciably with water. One type of lipid, the triglycerides, is sequestered as fat in
adipose cells, which serve as the energy-storage depot for organisms and also provide thermal insulation. Some lipids such as steroid
hormones serve as chemical messengers between cells, tissues, and organs, and others communicate signals between biochemical
systems within a single cell. The membranes of cells and organelles (structures within cells) are microscopically thin structures
formed from two layers of phospholipid molecules. Membranes function to separate individual cells from their environments and to
compartmentalize the cell interior into structures that carry out special functions. So important is this compartmentalizing function that
membranes, and the lipids that form them, must have been essential to the origin of life itself.
In this experiment, the acetone is used to extract invisible fats, since lipids are relatively insoluble in water but soluble in
organic solvents. When the extraction is complete, the students will be able to see, touch and smell the lipids in the petri dishes and be
able to determine if it saturated or unsaturated fatty acids. The cocoa butter found in the chocolate chips is a saturated fat and will be
solid at room temperature. The oils used to fry the potato chips and peanuts are unsaturated and will be liquid at room temperature.

II. OBJECTIVES:
At the end of the experiment, each student will be able to:

1. Conduct an experiment on their own related to the extraction of lipids.

2. Determine the outcome of the related experiment.

This study
This study source
source was
was downloaded
downloaded by
by 100000851111728
100000831778338 from
from CourseHero.com
CourseHero.com on
on 09-03-2022
11-14-2021 07:58:05
09:41:03 GMT
GMT -06:00
-05:00
 III. MATERIALS:
3-petri dish, 15-test tube, test tube brush, test tube holder, test tube rack, Bunsen burner/alcohol lamp, triple beam
balance, 1-25ml graduated cylinder 50ml acetone, 5ml ethyl alcohol, 1ml CCl4, 1ml ether, 1g KHSO4, oleic acid, Sudan
IV dye
Chocolate chips, potato chips, cashew nuts, coconut oil, olive oil, vegetable oil

IV. PROCEDURE
Lipids extraction:

1. Weigh 50ml beaker in 0.01g sensitivity using analytical balance.

2. Place the chocolate chips, potato chips, and cashew nuts in different beaker. Add acetone just enough to submerge the food to
be tested. And weigh.
3. Add a small amount of acetone to each dish (enough to submerge the foods to be tested).
4. Place each beaker in a well-ventilated area to hasten its evaporation rate for 10-15minutes.
5. Test the extractions in the paper towel (or brown paper) if they are lipids. If the paper towel (brown paper) will become greasy
to touch and transparent, it is lipids.
6. Try to smell your food sample when the acetone already dried up.
7. Record your answer in the Data table for extraction of lipids.

Food Weight (g) Weight (g) Weight (g) Weight (g) Weight (g) % lipid
of petri dish of petri dish of of petri with lost from extraction
with raw Raw food dry food food
food
Chocolate 200 205.3 5.3 204.9073 0.3927 7.8%
chips

This study source was downloaded by 100000851111728 from CourseHero.com on 09-03-2022 09:41:03 GMT -05:00
 Potato chip 200 205.12 5.12 204.204 0.916 17.89%
Cashew nuts 200 205.02 5.02 203.4849 1.2351 24.6%
(Weight of beaker with raw food) - (weight of beaker) = weight of raw food
(Weight of beaker with the raw food) – (weight of beaker with the dry food) = weight lost from the food

Weight loss from food

Weight of raw food x100 = % lipid extracted

FOOD Describe what you see in the paper towel

Chocolate chips There is a dark wet spot on the paper towel.
Potato chip There are a translucent spot like the potato shape on the paper
towel.
Cashew nuts The paper has no change.

TESTING FOR THE PRESENCE OF LIPIDS:

A. Solubility and Density of oil samples

1. Note the color of coconut oil, olive oil, and vegetable oil.
2. Test the solubility of each oil in the following solvents: water, ethyl alcohol, carbon tetrachloride and ether. Shake well.
3. Note their relative densities using hygrometer (the liquid with lesser density stays above the liquid with greater
density). Which solvent is oil soluble? Which solvent is less dense? More dense?

color density SOLUBILITY

water Ethyl CCl4 ether
alcohol
coconut oil Light 0.908- Insoluble Soluble Soluble Soluble
yellow 0.921

This study source was downloaded by 100000851111728 from CourseHero.com on 09-03-2022 09:41:03 GMT -05:00
 Olive oil Pale 0.913- Insoluble Soluble Soluble Soluble
yellow 0.917
Vegetable oil Golden 0.91-0.93 Insoluble Soluble Soluble Soluble
yellow

B. Acrolein Test

1. Put a pinch of KHSO4 and two drops of coconut oil in a small tube.
2. Heat gently at first and then vigorously.
3. Note the odor and name of the compound responsible for the odor.
4. Repeat the above procedure using oleic acid, instead of coconut oil.
5. Explain the results.

In the Acrolein test, whether the fats or oils are heated with the potassium in the test tube, Crystal of bisulfate. The presence of the fat
or oils is suggested by Acrolein’s unpleasant taste or scent.

C. Grease Spot Test

Place a small amount of vegetable oil on a clean sheet of bond paper. Observe.
The part of the clean bond paper where I put the vegetable oil started to diffract the light.

D. Dye Test

1. To 3 ml of water in a test tube, add a minute amount of Sudan IV dye.

2. Shake thoroughly then observed.

_The solution becomes clear.

This study source was downloaded by 100000851111728 from CourseHero.com on 09-03-2022 09:41:03 GMT -05:00
 3. Add 1 ml of vegetable oil.
4. Shake again and let it stand for 2 minutes. Observed.

I can see a separation of vegetable oil and the other substance

V. RESULTS AND OBSERVATIONS:

1. How can you tell that the dark wet spot on the paper is fats not water?

We can tell that the dark wet spot is fat by considering the characteristic of the spot. If it is fat, the dark wet spot will be translucent and it will still
appear even the paper towel dries. Nevertheless, if it is water, the spot will be opaque and wrinkled. Even though the paper towel dries, there will
also be wrinkle.

2. Rank from the most to least the percentage of lipid extracted from all three foods. Look at the nutrition facts label on three
food rank them. Did your ranking agree with the ranking of the product label?
The most percentage of lipid extracted of the food is the Cashew nuts 24.6% followed by Potato chips 17.89% and the least of them is
the Chocolate cookies 7.8%.. The ranking agreed with the ranking of the product labels. The first rank is cashew nuts due to the most
percentage of lipid extracted from all three foods. The second rank is potato chips and the last one is chocolate cookies.

3. Determine which lipids contained and unsaturated fatty acids in this experiment, based on your descriptions of the fats in the
Petri dish.

The extracted oil from the chocolate cookies had coarse texture, solid viscosity and changing in odor so I knew that it contained unsaturated fatty
acids. Unlike the first one, the extracted oil from the potato chips had smooth texture, medium viscosity and no changing in odor and color so I
knew that it was unsaturated fat. And the last, the extracted oil from the cashew nuts had smooth texture, solid viscosity, no changing in color, but
changing in odor so I think of it and presume that the cashew nuts contain both unsaturated and saturated fats.

VI. CONCLUSION

This study source was downloaded by 100000851111728 from CourseHero.com on 09-03-2022 09:41:03 GMT -05:00
 According to the results, I used the paper towel to detect the lipids in the three foods; chocolate cookies, cashew nuts, and potato
chips. When the foods were heated, fats came out from the foods. If fats were present, there is a dark wet spot and translucence on the
paper towel. I also extracted lipids from the food to find that which food contained the most and the least amount of lipids. I then
found that cashew nuts contained lipids the most, the second was potato chips, and the least was chocolate cookies. Cashew nuts had
the highest percentage of lipid extraction. Moreover, we can also know that which food contained saturated fatty acids and which one
contained unsaturated fatty acids by observing the description (the texture, color, odor and viscosity) of the fats, which was extracted
from each food. The extracted oil from the chocolate cookies had coarse texture, solid viscosity and changing in odor so we knew that
it contained unsaturated fatty acids. Unlike the first one, the extracted oil from the potato chips had smooth texture, medium viscosity
and no changing in odor and color so we knew that it was unsaturated fat. And the last, the extracted oil from the cashew nut had
smooth texture, solid viscosity, no changing in color, but changing in odor so I think of it and presume that the cashew nuts contain
both unsaturated and saturated fats.

LIBRARY WORK
 In an A4 bond paper research at least two (2) journals / articles on lipids (landscape, Times New Roman 13).

Author(s) Year Title Methods Results

publishe
d

This study source was downloaded by 100000851111728 from CourseHero.com on 09-03-2022 09:41:03 GMT -05:00
 JULIAN B. MARSH and 1966 Simple charring Reagent-grade concentrated sulfuric acid, tested for the Our results agree with the data of Blank et al. (8), which show that
DAVID B. WEINSTE method for presence of organic residues by heating at 2OOOC for 5 under constant charring conditions the extent of conversion of
determination of min, was used. The lipids used as standards were purified lipids to carbon is strongly influenced by structure and molecular
lipids by preparative thin-layer chromatography on Silica Gel G
weight and that the charring of compounds within the same lipid
plates which had been washed in the solvents used for the
lipid separations. The standard solutions were prepared in class does not vary to any considerable extent. The present method
chloroform at a concentration of 30 pg/ml and aliquots of is more rapid and sensitive than many of the methods currently in
1-5 ml were generally employed in the assay. Solvents use for the determination of lipids separated by column or thin-
were removed from the lipid samples under a flow of layer chromatography. In addition, the linearity of response of a
nitrogen in test tubes placed in an aluminum heating block natural and synthetic mixture of lipids indicates the possibility of
at 80-100OC. Solvent blanks were run routinely. After the using such mixtures as standards for the measurement of total lipid
tubes had been cooled, 2 ml of concentrated sulfuric acid in tissue or plasma extracts, providing that the approximate lipid
was added to each tube. At 15 sec intervals the tubes were
composition is known and that this does not change significantly
placed in an aluminum heating block at 2OOOC for 15
min. The temperature within the tubes in the heating block under the conditions of the experiment performed.
should be controlled to within *2OC during the charring
period. The tubes were placed in water at room
temperature for 15 sec and then were transferred to an ice
bath for 5 min; 3 ml of water was added to each tube, the
contents were mixed thoroughly, and the tubes were
replaced in the ice. When cool, the tubes were removed
from the ice and left standing for 10 min or until all
bubbles had disappeared. The optical density was
measured with a Spectronic 20 spectrophotometer (Bausch
& Lomb Incorporated, Rochester, N.Y.) at 375 nip
Damila Rodrigues de 2010 Evaluation of lipid A new technique, the alternative method, to extract total The total lipids extracted from human plasma varied between 0.19% and
MoraisI; Jeane Eliete extraction and lipids using a microwave was proposed and evaluated. 0.41%; the highest total lipid extracted were obtained by the Folch, Lees
Laguila VisentainerII; fatty acid and Stanley (0.41%), alternative method (0.37%) and Rose-Gottlieb
Leandra Pereira dos composition of (0.36%) methods. The Gerber method was ineffective to extract total lipids
SantosI; Makoto human plasma from human plasma. A total of 24 fatty-acid species were quantified by gas
MatsushitaIII; Nilson chromatography. Of the methods studied, the highest concentrations were
Evelázio de SouzaIII; Jesuí found using the Folch, Lees and Stanley method (p < 0.05).
Vergílio
VisentainerIII

This study source was downloaded by 100000851111728 from CourseHero.com on 09-03-2022 09:41:03 GMT -05:00
Powered by TCPDF (www.tcpdf.org)