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Experiment 5b Pre Post Lab

The document summarizes an experiment involving lipid extraction from egg yolk and qualitative tests to identify various lipids. It includes pre-lab and post-lab questions. For the pre-lab, students drew structures of phospholipids, sphingoglycolipids, and cholesterol, and described the purposes and positive indicators of various tests. In the post-lab questions, students explained the extraction process used to isolate lipids from egg yolk and the principles and reactions of tests for ninhydrin, Molisch's, phosphate, Salkowski, and acrolein. The summary compared the results of each test to expected results, finding several tests were unsuccessful in properly identifying the expected lipids.

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Rue Cheng Ma
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0% found this document useful (0 votes)
205 views9 pages

Experiment 5b Pre Post Lab

The document summarizes an experiment involving lipid extraction from egg yolk and qualitative tests to identify various lipids. It includes pre-lab and post-lab questions. For the pre-lab, students drew structures of phospholipids, sphingoglycolipids, and cholesterol, and described the purposes and positive indicators of various tests. In the post-lab questions, students explained the extraction process used to isolate lipids from egg yolk and the principles and reactions of tests for ninhydrin, Molisch's, phosphate, Salkowski, and acrolein. The summary compared the results of each test to expected results, finding several tests were unsuccessful in properly identifying the expected lipids.

Uploaded by

Rue Cheng Ma
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Experiment 5B: Pre-Post Lab BSN 1H

Members: Arabis, Aranas, Bada, Idong, Peñaloga, Sumiguin

PRE-LAB QUESTIONS
1. Draw the general structure of phospholipids, sphingoglycolipids, and cholesterol.

Phospholipids

Sphingoglycolipids

Cholesterol

2. Give the purposes of the tests presented in this experiment and identify its positive
indicators.

Ninhydrin Test
Ninhydrin test is a substance test performed to distinguish the presence of ammonia,
essential/optional amines, or amino acids. This test includes the expansion of ninhydrin reagent
to the test that outcomes in the arrangement of dark blue color, regularly named Ruhemann's
purple, are within sight of an amino group. (Aryal, 2020).

Molisch's Test
Molisch's test is an overall test for all starches. In this test, starches, when responded
with concentrated H2SO4, get dehydrated to shape furfural and its subordinates. When
monosaccharide is treated with concentrated H2SO4 or concentrated HCl, the -OH gathering of
sugar is taken out as water. As a result, a furfural is shaped if from pentose sugar while a
hydroxymethylfurfural is formed if from hexose sugar. These items respond with sulphonated α-
naphthol to give a purple (violet-red) hued complex (Karki, 2018).

Test for Phosphate


Phosphate test is generally used to measure or detect phosphate ions in a given
sample. This test is used in different areas like industrial processes, scientific research, and
environmental water monitoring. In medical professionals, they use this to measure the level of
phosphate in the blood. This kind of test will produce a yellow crystalline as it confirms
phosphate's presence in a given solution.
(Wiredchemist, n.d.)

Salkowski Test
Salkowski test is a test for cholesterol. When concentrated sulfuric corrosive is added to
a chloroform arrangement of cholesterol, the chloroform layer shows a red to blue tone, and the
corrosive layer shows. (Oxford Reference n.d.).

Acrolein test
Acrolein test is a test performed to identify if there is a presence of glycerol in a solution
using potassium bisulfate as a dehydrating agent. The dehydrating agent will dehydrate the
molecule's glycerol portion to form unsaturated aldehyde that will result in a pungent irritating
odor. (OLABS, 2015)

POST-LAB GUIDE QUESTIONS


1. What types of lipids are found in egg yolk? Explain how the extraction process
isolates lipids from egg yolk?

The lipids found in egg yolk are 68% low-density lipoprotein (lipovitellin), 12% high-
density lipoprotein, and 12% livetin (which contains 0.7% IgY), and 7% phosvitin are the
proteins in egg yolk. With multiple studies showing lipid-free egg yolk protein (EYP) to have a
perfect amino acid score of 100, egg yolk's protein profile makes it an outstanding source of
nutrients for the human body. Studies have also shown that egg yolk protein (EYP) is digested
faster than milk casein and that egg protein has a high nutritional value than milk casein. ("Yolk
Proteins | Ecovatec", 2020).

The process of lipid extraction of egg yolk was to crack the eggshells carefully and
separate the egg yolk and egg white. Keep the egg yolk for the qualitative analysis. Get 1 ml of
egg yolk and add 2ml of 1m of sodium chloride. After that, add 3 ml of cyclohexane and 95%
ethanol, mix thoroughly. The mixture was then transferred into the test tube and centrifuged at
1000 mph for 5 minutes. After centrifugation, the mixture separated into three phases: 1. a clear
yellow oil layer on top, 2. yellow sediment at the middle 3. an aqueous layer at the bottom
(Larsen, JE. & Frowning, JW. , 2019).

2. Explain the principle in each test. Give corresponding chemical reactions.

Ninhydrin Test
Principle: This is a test utilized for the detection of proteins or amino acids with free amino
groups. These free amino groups react with ninhydrin, leading to corresponding chemical
reactions that would yield into or release CO2, ammonia, an aldehyde, and a hydrindantin
(reduced form of ninhydrin) (Karki, 2018). The hydrindantin will undergo condensation with
ammonia and another ninhydrin molecule, which brings to the creation of a purple or blue
complex, also known as the Ruhemann’s purple (Basnet, 2020). However, instead of a blue-
purple complex for the amino acid proline, the product would form a yellow pigment.
Reaction:

BYJUS. (2020, September 28). Ninhydrin Test Result Interpretation [Illustration].


BYJU’S. https://byjus.com/chemistry/ninhydrin-test/
Molisch’s Test
Principle: This is a test used on various samples to identify the presence of carbohydrates. The
sample is added with Molisch's reagent and concentrated acid. The addition of your
concentrated HCl causes the present pentoses or hexoses to undergo dehydration, which then
yields into furfural if it was a pentose or a hydroxymethylfurfural if from a hexose. Either of the
two can now bind and undergo a condensation reaction with an α-naphthol from the Molisch’s
reagent. This process will now form the test's positive indicator, a purple ring layer between the
interface of concentrated acid and the test sample (Sapkota, 2020).

Reaction:

Karki, G. (2018, December 18). Molisch’s Test: Objectives, Principle, Reagents, Procedure and Result [Illustration]. Online Biology Notes.
https://www.onlinebiologynotes.com/molischs-test-objectives-principle-reagents-procedure-and-result/

Test for Phosphate


Principle: This test merely focuses on the oxidative response wherein the natural structure's
transformation to inorganic phosphate occurs. There is also precipitation in response to the
continuing process. The combination or the mixture of the potassium and sodium carbonate is
utilized to eliminate the hints of natural structure in the arrangement. By warming the solution
helps the reaction to boost the response of change. In using the acidification together with the
nitric corrosive produces hydrogen phosphate, which ammonium molybdate promptly
recognizes to create yellow precipitation (Biological Zoo301, n.d)

Reaction:

Gopal, J. (n.d). [Illustration] Organic Chemistry-Purification of Organic Compound. Retrieved from:


https://www.learnpick.in/prime/documents/ppts/details/107/organic-chemistry-purification-of-organic-compound

Salkowski Test
Principle: This type of test is commonly used for determining the cholesterol in a specific
mixture (Bangash, 2017). By using this test as a pivotal procedure, it also indicates different
results, either clear or visible colors, with the help of the concentrated sulfuric acid, which
eliminates two atoms of water from two particles of cholesterol. Thus it speeds up the reaction
rate. Moreover, in observing this test's outcomes, in the layer where there is chloroform, it
shows a bluish-red to cherry-red color like feature. In contrast, a yellow with green fluorescence
feature is observed in the sulfuric acid layer (JezKagandahan, n.d).

Reaction:

JezKagandahan. (n.d). [Illustration] Salkowski Test & Conclusion. Retrieved from: https://www.scribd.com/document/435129885/Salkowski-Test-
Conclusion

Acrolein test
Principle: This is a qualitative test for glycerols. To indicate glycerols' presence, when the
crucible containing a lipid sample is heated, there is a release of a pungent smell, which gives
an irritating or sharp sensation. This is for the reason that the addition of potassium bisulfate
(KHSO₄) to the lipid sample will dehydrate the present glycerol component, forming acrolein
(OLabs, 2015). In addition, further heating of the sample contributes to the polymerization of
acrolein, which is again indicated for the release of a pungent smell.

Reaction:
BYJU’S. (2020, August 11). Tests of Oils and Fats - Chemistry Practicals Class 12 [Illustration]. BYJU’S. https://byjus.com/chemistry/tests-of-oils-and-
fats/

3. Explain the results of each test and compare it to the expected results.

A. Molisch test.

The first extracted lipids using ethanol came out positive as expected when tested on
molisch's test for carbohydrates. Each test tube contained sulfuric acid that led the carbohydrate
that is covalently linked to a lipid molecule undergo dehydration reaction and further reacts with
molisch reagent to form colored compounds or the purple ring in the sample, which can be
shown in the A1 test tube (Biology online, 2020).

B. Ninhydrin Test

The result of the ninhydrin test shows a confusing indicator for both A2 and A3, a purple-
colored solution, but A2 shows deep purple color while a3 shows pale purple and will
theoretically be negative when subjected to ninhydrin test since it is not a protein. The extracted
sample in a2 has the presence of lipoproteins as it came out positive for the test (BYJU classes,
n.d.).

C. Phosphate Test
The result of this test wasn't successfully justified as the experiment result did not give a
yellow precipitate layer of ammonium phosphomolybdate as a positive indicator for this test for
A3, which is the lecithin that has a phosphate group attached to it, and the other two shows a
yellow solution, and a yellow precipitate that is theoretically will turn out negative, but the yellow
precipitate of A1 might not be ammonium but other molecule (Wiredchemist, n.d.).

D. Salkowski Test

The experiment was again unsuccessful as the result did not show the A2 cholesterol
containing sample positive for the test and showed a clear solution instead of showing red and
yellow layers, the upper layer and lower layer as the expected result for this test. The other two
test tubes just gave a yellow solution but still negative (Bangash, 2017).

E. Acrolein Test

Since the result for this test is through a pungent irritating odor, by smelling them, we
can identify if the sample contains glycerol. We can assume that it tested positive in the A1
triglycerides since they are tri-esters consisting of glycerol and will test positive in A2 and A3
(OLABS, n.d).

References:
amrita.olabs.edu.in,. (2015). Qualitative Analysis of Oils and Fats. amrita.olabs.edu.in/?
sub=73&brch=8&sim=210&cnt=1

Aryal, S. (2020). Ninhydrin Test- Definition, Principle, Procedure, Result, Uses.


https://microbenotes.com/ninhydrin-test/.

Bangash, A. (2017). Salkowski test for cholesterol - Its principle and procedure.
https://medicalstudyzone.com/salkowski-test/

Basnet, A. (2020, April 3). Ninhydrin test: Principle, Reaction, Reagents, Procedure and Result
Interpretation. Online Biochemistry Notes. http://biocheminfo.com/2020/04/03/ninhydrin-
test-principle-reaction-reagents-procedure-and-result-interpretation/

Biology online, (2020). dictionar; Glycolipid. https://www.biologyonline.com/dictionary/glycolipid

Biological Zoo301. (n.d). University of Santo Tomas.


https://www.coursehero.com/file/p6iiq7k/Test-for-phosphate-as-the-name-suggests-
tests-for-phosphate-The-positive-result/

BYJU classes, (n.d). Chemistry. retrieved from: https://byjus.com/chemistry/ninhydrin-test/

Ecovatec Solutions INC. (2020). Yolk Proteins. http://ecovatec.com/yolk-proteins/.

JezKagandahan. (n.d). Salkowski test & conclusion.


https://www.scribd.com/document/435129885/Salkowski-Test-Conclusion

Karki, G. (2018). Molisch’s Test: Objectives, Principle, Reagents, Procedure and Result.
https://www.onlinebiologynotes.com/molischs-test-objectives-principle-reagents-
procedure-and-result/.

Karki, G. (2018, December 18). Ninhydrin Test: Principle, Requirements, Procedure and Result.
Online Biology Notes. https://www.onlinebiologynotes.com/ninhydrin-test-principle-
requirements-procedure-and-result/

Larsen, JE. & Frowning, JW. (2019). Poultry Science| Extraction and Processing of Various
Components from Egg Yolk. Retrieved from:
https://www.sciencedirect.com/science/article/pii/S0032579119357165.

OLABS,( n.d.) Chemistry: Qualitative analysis of oils and fats. https://amrita.olabs.edu.in/?


brch=8&cnt=1&sim=210&sub=73

OLabs. (2015). Qualitative Analysis of Oils and Fats (Theory) : Class 12 : Chemistry : Amrita
Online Lab. http://amrita.olabs.edu.in/?sub=73&brch=8&sim=210&cnt=1

Oxford Reference. (n.d.). Salkowski test.


https://www.oxfordreference.com/view/10.1093/oi/authority.20110803100438192.
Sapkota, A. (2020, November 19). Molisch Test- Definition, Principle, Procedure, Result, Uses.
Microbe Notes. https://microbenotes.com/molisch-test/

Stadelman, WJ. (2003). Encyclopedia of Food Sciences and Nutrition (Second Edition). EGGS:
Structure and Composition.
https://www.sciencedirect.com/science/article/pii/B012227055X003874.

Wiredchemist, (n.d.) Qualitative analysis.


http://www.wiredchemist.com/chemistry/instructional/laboratory-tutorials/qualitative-
analysis

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