FORMAL REPORT

OF EXPERIMENT IN BIOCHEMISTRY
LIPID 1 and LIPID 2

Written by group

4:

Ira Novita Sari

Sarry Saraswaty

(K3309049)
(K3309073)
Yoga Bririan Jati

(K3309089)

Chemistry Education Study Program

Department of Mathematics and Natural Sciences Education
Faculty of Teacher Training and Education
Sebelas Maret University
SURAKARTA
2012
FORMAL REPORT
OF EXPERIMENT IN BIOCHEMISTRY

I. TITLE

: LIPID 1 and LIPID 2

II. PURPOSE :
Can isolate several types of lipids from egg yolk
Can identifying of lipis
III.BASIC THEORY
Lipids constitute a broad group of naturally occurring molecules that include fats, waxes,
sterols, fat-soluble vitamins (such as vitamins A, D, E, and K), monoglycerides, diglycerides,
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triglycerides, phospholipids, and others. The main biological functions of lipids include energy
storage, as structural components of cell membranes, and as important signaling molecules.
Lipids may be broadly defined as hydrophobic or amphiphilic small molecules; the
amphiphilic nature of some lipids allows them to form structures such as vesicles, liposomes, or
membranes in an aqueous environment. Biological lipids originate entirely or in part from two
distinct types of biochemical subunits or "building-blocks": ketoacyl and isoprene groups.[4]
Using this approach, lipids may be divided into eight categories: fatty acids, glycerolipids,
glycerophospholipids, sphingolipids, saccharolipids, and polyketides (derived from condensation
of ketoacyl subunits); and sterol lipids and prenol lipids (derived from condensation of isoprene
subunits).
Although the term lipid is sometimes used as a synonym for fats, fats are a subgroup of
lipids called triglycerides. Lipids also encompass molecules such as fatty acids and their
derivatives (including tri-, di-, monoglycerides, and phospholipids), as well as other sterolcontaining metabolites such as cholesterol. Although humans and other mammals use various
biosynthetic pathways to both break down and synthesize lipids, some essential lipids cannot be
made this way and must be obtained from the diet.

Categories of lipids
Fatty acids
Fatty acids, or fatty acid residues when they form part of a lipid, are a diverse group of
molecules synthesized by chain-elongation of an acetyl-CoA primer with malonyl-CoA or
methylmalonyl-CoA groups in a process called fatty acid synthesis. They are made of a
hydrocarbon chain that terminates with a carboxylic acid group; this arrangement confers the
molecule with a polar, hydrophilic end, and a nonpolar, hydrophobic end that is insoluble in
water. The fatty acid structure is one of the most fundamental categories of biological lipids, and
is commonly used as a building-block of more structurally complex lipids. The carbon chain,
typically between four and 24 carbons long, may be saturated or unsaturated, and may be
attached to functional groups containing oxygen, halogens, nitrogen, and sulfur. Where a double
bond exists, there is the possibility of either a cis or a trans geometric isomerism, which
significantly affects the molecule's molecular configuration. Cis-double bonds cause the fatty
acid chain to bend, an effect that is more pronounced the more double bonds there are in a chain.
This in turn plays an important role in the structure and function of cell membranes.
Glycerolipids
Glycerolipids are composed mainly of mono-, di-, and tri-substituted glycerols, the most
well-known being the fatty acid triesters of glycerol, called triglycerides. The word
triacylglycerol is sometimes used synonymously with triglyceride, however this is misleading
with respect to these compounds as they contain no hydroxyl group. In these compounds, the
three hydroxyl groups of glycerol are each esterified, typically by different fatty acids. Because
they function as an energy store, these lipids comprise the bulk of storage fat in animal tissues.
The hydrolysis of the ester bonds of triglycerides and the release of glycerol and fatty acids from
adipose tissue are the initial steps in metabolising fat.
Additional subclasses of glycerolipids are represented by glycosylglycerols, which are
characterized by the presence of one or more sugar residues attached to glycerol via a glycosidic
linkage. Examples of structures in this category are the digalactosyldiacylglycerols found in
plant membranes and seminolipid from mammalian sperm cells.
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Glycerophospholipids
Glycerophospholipids, usually referred to as phospholipids, are ubiquitous in nature and are
key components of the lipid bilayer of cells, as well as being involved in metabolism and cell
signaling. Neural tissue (including the brain) contains relatively high amounts of
glycerophospholipids, and alterations in their composition has been implicated in various
neurological disorders. Glycerophospholipids may be subdivided into distinct classes, based on
the nature of the polar headgroup at the sn-3 position of the glycerol backbone in eukaryotes and
eubacteria, or the sn-1 position in the case of archaebacteria.

Phosphatidylethanolamine
Examples of glycerophospholipids found in biological membranes are phosphatidylcholine
(also known as PC, GPCho or lecithin), phosphatidylethanolamine (PE or GPEtn) and
phosphatidylserine (PS or GPSer). In addition to serving as a primary component of cellular
membranes and binding sites for intra- and intercellular proteins, some glycerophospholipids in
eukaryotic cells, such as phosphatidylinositols and phosphatidic acids are either precursors of or,
themselves, membrane-derived second messengers. Typically, one or both of these hydroxyl
groups are acylated with long-chain fatty acids, but there are also alkyl-linked and 1Z-alkenyllinked (plasmalogen) glycerophospholipids, as well as dialkylether variants in archaebacteria.
Sphingolipids
Sphingolipids are a complicated family of compounds that share a common structural
feature, a sphingoid base backbone that is synthesized de novo from the amino acid serine and a
long-chain fatty acyl CoA, then converted into ceramides, phosphosphingolipids,
glycosphingolipids and other compounds. The major sphingoid base of mammals is commonly
referred to as sphingosine. Ceramides (N-acyl-sphingoid bases) are a major subclass of
sphingoid base derivatives with an amide-linked fatty acid. The fatty acids are typically saturated
or mono-unsaturated with chain lengths from 16 to 26 carbon atoms.

Sterol lipids
Sterol lipids, such as cholesterol and its derivatives, are an important component of
membrane lipids, along with the glycerophospholipids and sphingomyelins. The steroids, all
derived from the same fused four-ring core structure, have different biological roles as hormones
and signaling molecules. The eighteen-carbon (C18) steroids include the estrogen family
whereas the C19 steroids comprise the androgens such as testosterone and androsterone. The
C21 subclass includes the progestogens as well as the glucocorticoids and mineralocorticoids.
The secosteroids, comprising various forms of vitamin D, are characterized by cleavage of the B
ring of the core structure. Other examples of sterols are the bile acids and their conjugates, which
in mammals are oxidized derivatives of cholesterol and are synthesized in the liver. The plant
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equivalents are the phytosterols, such as -sitosterol, stigmasterol, and brassicasterol; the latter
compound is also used as a biomarker for algal growth. The predominant sterol in fungal cell
membranes is ergosterol.
Prenol lipids
Prenol lipids are synthesized from the five-carbon-unit precursors isopentenyl diphosphate
and dimethylallyl diphosphate that are produced mainly via the mevalonic acid (MVA) pathway.
The simple isoprenoids (linear alcohols, diphosphates, etc.) are formed by the successive
addition of C5 units, and are classified according to number of these terpene units. Structures
containing greater than 40 carbons are known as polyterpenes. Carotenoids are important simple
isoprenoids that function as antioxidants and as precursors of vitamin A. Another biologically
important class of molecules is exemplified by the quinones and hydroquinones, which contain
an isoprenoid tail attached to a quinonoid core of non-isoprenoid origin. Vitamin E and vitamin
K, as well as the ubiquinones, are examples of this class. Prokaryotes synthesize polyprenols
(called bactoprenols) in which the terminal isoprenoid unit attached to oxygen remains
unsaturated, whereas in animal polyprenols (dolichols) the terminal isoprenoid is reduced.
Saccharolipids
Saccharolipids describe compounds in which fatty acids are linked directly to a sugar
backbone, forming structures that are compatible with membrane bilayers. In the saccharolipids,
a monosaccharide substitutes for the glycerol backbone present in glycerolipids and
glycerophospholipids. The most familiar saccharolipids are the acylated glucosamine precursors
of the Lipid A component of the lipopolysaccharides in Gram-negative bacteria. Typical lipid A
molecules are disaccharides of glucosamine, which are derivatized with as many as seven fattyacyl chains. The minimal lipopolysaccharide required for growth in E. coli is Kdo2-Lipid A, a
hexa-acylated disaccharide of glucosamine that is glycosylated with two 3-deoxy-D-mannooctulosonic acid (Kdo) residues.[39]
Polyketides
Polyketides are synthesized by polymerization of acetyl and propionyl subunits by classic
enzymes as well as iterative and multimodular enzymes that share mechanistic features with the
fatty acid synthases. They comprise a large number of secondary metabolites and natural
products from animal, plant, bacterial, fungal and marine sources, and have great structural
diversity. Many polyketides are cyclic molecules whose backbones are often further modified by
glycosylation, methylation, hydroxylation, oxidation, and/or other processes. Many commonly
used anti-microbial, anti-parasitic, and anti-cancer agents are polyketides or polyketide
derivatives, such as erythromycins, tetracyclines, avermectins, and antitumor epothilones.

IV.

EQUIPMENTS
a. Tools
No

Name of Tool

Amount

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1
2
3
4
5
6
7
8
9
10
11
12
13
14

Measurement glass
Beaker glass
Drop pipette
Glass stirrer
Test tube
Erlenmeyer
Glass funnel
Electric stove
Exicator
Watch glass
Electric balance
Tweezers
Sentrifugace
Filter paper

1
2
2
1
4
1
1
1
1
1
1
1
1
sufficient

b. Materials
No
1
2
3
4
5
6
7
8
9

Name of Material

Amount

Chicken egg (egg yolk)

Alcohol
Ether
Acetone
Cadmium Chloride
KOH alcoholis 10%
Aquadest
Glycerine
KHSO3

3
sufficient
sufficient
sufficient
sufficient
sufficient
sufficient
sufficient
sufficient

V. PROCEDURE and OBSERVATION DATA

No

Treatment

Cracking egg, take egg yolk + 25 ml ether + 50ml

Occur coagulating, residue has

alcohol + 50 ml alcohol, stir it, let it during 10

yellow color and yellow filtrate.

2
3

minutes then filter it.

Residue + 7 ml ether + 14 ml alcohol, filter it and
its filtrate was vapored.

Observation

Yellowish filtrate
Old yellow filtrate, existence
residue
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4
Residue + ether + acetone then filtered
Dissolving sediment + alcohol + CdCl2 alcoholis,
5

6
7

stir it,
let it then filter it.
Filtrate was vapored,
+ KOH alcoholis, stir it, heated
+ ether and filter it.
Filtrate + ether, vapored until dry
Cholesterol Test
Residue + alcohol, centrifuged + alcohol

VI.

Centrifuged again
Let it about 30 minutes.
Acrolein Test
0,5 gr fat + 0,5 gr KHSO3 ,
heated
Repeat with glycerol

Turbid yellowish solution

Yellow solution, existence
clumped,
Yellow sediment more pale
Become pasta, turbid yellow
More turbid solution
Formed 2 layers :
Up
: clear yellow
Bottom : orange
Filtrate : clear yellow
Residue : orange
Yellow solution
Formed sediment
Yellow solution
Formed sediment
Mixed
More dissolve and bad smell
Bad smell very sting

ANALYZE DATA
At trial lipids, aiming to isolate several types of lipids from egg yolk lecithin that will be

generated. Where, lecithin is a

phospholipid

that becomes a major component of

fractionphosphatides to extract the yellow egg or peanut soy isolates are mechanical , and chemical
by using hexane or can be interpreted is a complex substance that is found in eggs and institutions
supply the bone marrow. In its application, the lecithin in egg yolks and most often used as an agent
emulsifier to mix oil and water , as in mayonnaise . This can happen because lecithin has a head that
is hydrophilic and the tail is hydrophobic . Lecithin in eggs is dominated by the content of
phosphatidyl kolina high, gliserolfosfolipid , long-chain fatty acids unsaturated, arachidonic acid ,
and DHA content not found in other sources of lecithin (such as beans). Lecithin is commercially
can be obtained with high purity for the additives of food or medical purposes. In addition, lecithin
is also known to help the stability of mayonnaise and mayonnaise to make it look more bold .
Lipids are compounds which are prevalent in nature. This compound can be obtained by
extracting natural materials both plants and animals with polar solvents not just as petroleum ether,
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benzene, chloroform, and others. Judging from the lipid compound structure composed of long
hydrocarbon chains, so that lipids are not soluble in water. Lipid compounds are named based on the
physical properties (solubility) than on the chemical structure. In general, lipids are divided into two
major categories, namely simple lipid and lipid complexes. Lipids are included in the class are
simple compounds that do not have the ester group and can not be hydrolyzed. This group is a
steroid. Lipid complex group of compounds composed of ester groups and may have hydrolyzed, the
fatty oils and waxes melipti.
In this experiment, will be isolated or lipid content of lecithin and cholesterol from chicken
egg yolk using alcohol and ether solvents. The first step in this experiment is to take as many as 3
eggs yolks, stir then add the alcohol and ether and then stirred again. Egg yolks into slightly
thickened or lumpy. This indicates that the solvents (alcohols and ethers) can exstracted fat or lipid
from the yolk. To be more complete extraction, the mixture allowed to stand for about 10 minutes
and the resulting yellow laruran.
Second, filter the mixture with filter paper that has been moistened with alcohol. The residue
is then washed with alcohol and ether, while the filtrate is then evaporated in a water bath until dry.
Washing the residue with ether alcohol intended to bind any residual alcohol ether in the residue
thus obtained is pure fat deposition. Thick yellow precipitate then dissolved in ether and acetone was
added. Dissolution is intended to bind the alcohol and ether remaining. While acetone has the
function to precipitate the mixture. The precipitate formed is called lecithin.
Third, filter and precipitate (lecithin) was dissolved again in the alcohol and CdCl 2, then
stirred and left for 10 minutes. After it was filtered and the residue obtained is used to test akreolin.
Lecithin can also be called phosphatidyl kolina . Where, phosphatidyl kolina is sulfuric acid
and base containing n-choline that serves to facilitate the transport and use of fatty acids by using
the enzyme lecithin cholesterol acyl transferase. The structure of lecithin:

Can be derived from egg lecithin. Lecithin has a structure like fat but contains phosphoric
acid, polar groups and nonpolar groups. Polar groups contained in the ester, phosphate is hydrophilic
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(water soluble likely), while non-polar groups contained in the fatty acid ester is lifofilik (likely to
dissolve in fat). Lecithin is a waxy solid soft white color and turns brown due to exposure to light.
Because it is hydrophilic, it can form a colloid. Lecithin is soluble in all organic solvents, except
acetone. Goal is the addition of acetone to precipitate the lecithin.
Lecithin (choline phospatidil) with the main components of choline, is an important nutrient that
is found widely in various foods and available as supplements. Lecithin contains about 13% by
weight of choline. Lecithin also zwiter ion, has a positive charge on N atom of choline and negative
charge on the O atom of the phosphate group. Lecithin can be either polar (the choline) and nonpolar (the fatty acid) making it very effective as an emulsifier.
The filtrate that was still containingacetone evaporated to residual ether-acetone to
evaporate. Then added therewith 10% KOH alcoholicwhile stirring then heated for 30 minutes. The
results obtained are orange-colored solution. This is because the solution contains lecithin, when
heated with alkali to form glycerol fatty acid choline phosphoric acid. The addition of 10% KOH
alcoholics aims to transform fatty acids into soap and glycerol.
Reaction of fatty acid with a base KOH:

Next, add 50 mL of ether into the solution and then filter it. Of ether was added, forming two
layers where the top layer is yellow and bottom layer is orange. Ether filtrate containing cholesterol
and then evaporated to dry residue thus obtained in the form of orange granules. The residue is then
dried in the exicator which will then be used to test for cholesterol.
Cholesterol is a metabolite of sterol-containing fat ( English : Waxy steroids) are found in
cell membranes and circulated in the blood plasma . Is a type of lipid which is a molecule of fat or
the like it. Cholesterol is a special type of lipid called steroids . Steroids are lipids that have a
chemical structure specific. This structure consists of four ring atomsof carbon . Other steroids,
including steroid hormones such as cortisol , estrogen , and testosterone . In fact, all steroid
hormones are made from chemical changes in the basic structure of cholesterol.
Cholesterol is a fat component and is one nutrient that is needed by the body in addition to
other nutrients such as carbohydrates, protein, vitamins, and minerals. Cholesterol has a chemical
structure as shown in the following figure:

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These elements consist of fats in the blood cholesterol, triglycerides, phospholipids and free
fatty acids. Only a quarter of the cholesterol contained in the blood are derived directly from the
digestive tract is absorbed from food, the rest is the body's own production by liver cells.
Fat contained in food will be elaborated into cholesterol, triglycerides, phospholipids and
free fatty acids when digested in the gut. The fourth element of this fat will be absorbed from the
intestine and enter the blood. Cholesterol, triglycerides, phospholipids and free fatty acids are not
soluble in blood. In order to be transported in the bloodstream, cholesterol along with other fats
(triglycerides and phospholipids) must bind to proteins to form a soluble compound and is called a
lipoprotein.
A cholesterol test performed by dissolving the residue in alcohol as much as 5 mL and then
disentrifuge. Will get the white granules on the bottom of the tube in small amounts. Discard the
solution above for the next step and add alcohol into granules and then disentrifuge again. The
addition of alcohol is intended to dissolve cholesterol, because cholesterol is fat soluble in solvents
such as ether, chloroform, benzene, and hot alcohol. Then, the mixture was let stand for 30 minutes.
The results that obtained are yellowish solution, and the precipitate formed at the bottom of the tube
in the form of yellowish granules called cholesterol.

Acrolein test
The purpose of acrolein test on lecithin and glycerin is to prove the existence of dehydrated
glycerin into aldehydes, aksilat, or akreolin, and the presence of glycerol or fat. Glycerin
usually be hydrated into an unsaturated aldehyde or akreolin and usually marked with white
smoke and the smell. Akreloin test is usually positive in the form of free glycerol, but not for
fatty acids.
Of experiments lecithin which is yellow at first and then added KHSO 4 in the form of fine
grains resulting mixture is mixed evenly. This mixture is then put into a test tube and then
heated in a water bath with a small fire in the early beginning and then becomes a large fire.
This warming resulted from strong odors and yellow. With the yellow color and pungent
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odor which indicates that the test indicated its positive acrolein. This means that the lecithin
contained in the dehydration is aldehit aksilat or akreolin and there are fat.
Then repeat the experiment with the same steps, but replace the lecithin with glycerin. And
the result is a fishy smell and stinging occur. But the odor generated no more and no fishy
smell more pungent than lecithin. But these results indicate a positive test for akreolin
glycerin.
Function of KHSO4 in this experiment is to draw water that will hydrate glycerol into the
aldehyde or akreolin aksilat characterized by white smoke and strong odors.
Reactions that occur

Lecithin + KHSO4

Glycerol + KHSO4

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VII.

CONCLUSION
1. Lipids are organic compounds that have physical properties such as fat, and can be obtained
from animal or plant by extraction with hot alcohol, ether and other fat solvent.
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2. Lecithin is one component of egg yolk contains fat and cholesterol.

3. Inside there is a chicken egg yolk cholesterol in the form of a white crystalline solid.
4. Cholesterol is a necessary intermediate in the biosynthesis of steroid hormones.
5. Akroelin test on fatty acids and glycerol are both positive. This is evidenced by the stench of
fatty acids and glycerol to be dehydrated into aldehydes, and akroelin aksilat.
6. KHSO4 additional function is to attract water so be hydrated glycerol into unsaturated
aldehydes are marked with white smoke and the smell.
7. Function of the addition of:
a. alcohol, ether: as a solvent of the remaining egg whites or a protein that may be left behind
when the separation of egg white and yolk.
b. KOH alcoholics: to provide the alkaline conditions
c. CdCl 2: to hydrolyze lecithin
d. acetone: to precipitate the lecithin
8. chemical structure of lecithin:

9. chemical structure of cholesterol:

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10. fatty acid reaction with KOH

11. Reactions that occur

Lecithin + KHSO4

Glycerol + KHSO4

VIII. BIBLIOGRAPHY
(2012, March

3).

was

taken

April

30,

2012,

from

Wikipedia:

http://id.wikipedia.org/wiki/Lesitin
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 (2012, April 12). was taken April 30, 2012, from Wikipedia:
http://id.wikipedia.org/wiki/Kolesterol
Anna, P., & Supriyanti, T. (2006). Dasar-Dasar Biokimia. Jakarta: UI Press.
Fajri , E. (2011, June 28). Archives: 2011 (81): Juni (23). was taken April 30, 2012, from
elfa Fajri: http://elfafajri.blogspot.com/2011/06/lipida.html
Fessenden dan Fessenden. (2000). Dasar-Dasar Biokimia. Jakarta: Erlangga.
muhammadpajri . (2012, January 4). Arsip Blog: Januari (18). Dipetik April 30, 2012,
from MUHAMMAD PAJRI THP 09' UNJA:
http://muhammadpajri1991.blogspot.com/2012/01/lesitin-sebagai-emulsifier.html
Sugiharto. (1989). Biokimia. Jakarta: Gramedia.
Tim Dosen. (2012). Petunjuk Praktikum Biokimia. Surakarta: Lab. P. MIPA Kimia UNS.
Winarno, F. G. (2002). Kimia Pangan dan Gizi. Jakarta: Erlangga.

IX.

ATTACHMENT
One sheet of temporary report Lipid 1 and Lipid 2

Assitant

Surakarta, 3rd may 2012

Practicant,

Group 4

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