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TITLE OF EXPERIMENT
Introduction of Reaction of Carbohydrate, Lipids, and Protein.
B. GOAL OF EXPERIMENT
To learn the general properties and reaction of carbohydrate, lipid, and
protein compounds.
C. LIETERATURE REVIEW
Carbohydrates are important constituents of all living organisms and have
a variety of different functions. Some are important structural components of cells;
others act as recognition sites on cell surfaces. For example, the first event in our
lives was a sperm recognizing a carbohydrate on the outer surface of an egg.
Other c arbohydrates serve as a major source of metabolic energy. The leaves,
fruits, seeds, stems, and roots of plants, for instance, contain carbohydrates that
plants use for their own metabolic needs and these also serve the metabolic needs
of the animals that eat them (Bruice, 2014: 1017).
The word carbohydrate originally described glucose, the simplest and most
readily available sugar. Because glucose has the formula C6H12O6, it was once
thought to be a “hydrate of carbon,” C61H2O26. Although this view has been
abandoned, the name “carbohydrate” persisted, and we now use it to refer to a
large class of biomolecules with similar structures. Carbohydrates have in
common many hydroxyl groups on adjacent carbons together with either an
aldehyde or ketone group. Glucose, for example, has five hydroxyl OH2 groups
and one aldehyde (-CHO) group
H H H OH H O
HO C C C C C C H
H OH OH H OH
OH OR'
H+
'
R C OR' + R' OH R C OR' + H - OH
H H
a hemiketal a ketal
The cyclic hemiacetal and hemiketal forms of monosaccharides also react with
alcohols to form acetals and ketals. These acetals and ketals are called glycosides,
and the carbon-oxygen bond formed is called a glycosidic bond. The group
bonded to the anomeric carbon atom of a glycoside is an aglycone. In aglycones,
an oxygen atom from an alcohol or phenol is linked to the anomeric carbon atom.
Glycosides are named by citing the aglycone group first and then replacing the -
ose ending of the carbohydrate with -oside (Oulette and J, 2015: 356).
. Their functional groups are involved in reactions with alcohols, lipids, or
proteins to form biomolecules with specialized functions. nosaccharides undergo
a variety of structural changes and chemical reactions. They react with each other
to form disaccharides and polysaccharides. Monosaccharides are chiral molecules
and Monosaccharides exist mainly in cyclic forms rather than the straight-chain
forms ( McMurry et al, 2013: 658).
Simple carbohydrates are monosaccharides (single sugars); complex
carbohydrates contain two or more monosaccharides linked together.
Disaccharides have two monosaccharides linked together, oligosaccharides have
3 to 10 ( oligos is Greek for “few”), and polysaccharides have 10 or more.
Disaccharides, oligosaccharides, and polysaccharides can be broken down to
monosaccharides by hydrolysis.
ℎ𝑦𝑑𝑟𝑜𝑙𝑖𝑠𝑖𝑠
−M − M − M − M − M − M − M − M − M − Xm
Polysacaharide monosacahride
In maltose the glycosidic oxygen atom of one glucose is a and is bonded to
the C-4 atom of another glucose unit that is the aglycone. Therefore, maltose is an
a-1,4′-glycoside. Maltose is produced by the enzymatic hydrolysis of starch (a
homopolysaccharide) catalyzed by the enzyme amylase. Maltose is further
hydrolyzed by the enzyme maltase. Lactose, a disaccharide found in diff erent
concentrations in the milk of mammals, including both humans and cows, is often
called milk sugar. As in maltose and cellobiose, the aglycone of lactose is a
hemiacetal, and it can be either 𝛼 𝑜𝑟 𝛽 (Oulette and J, 2015: 358,360).
Glucose is the most important simple carbohydrate in human metabolism.
It is the final product of complex carbohydrate digestion and provides acetyl
groups for entry into the citric acid cycle as acetyl-CoA. d-Galactose is widely
distributed in plant gums and pectins, the sticky polysaccharides present in plant
cells. d-Fructose, often called levulose or fruit sugar, occurs in honey and many
fruits. It is one of the two monosaccharides combined in the disaccharide sucrose.
Fructose is produced commercially in large quantities by hydrolysis of cornstarch
to make highfructose corn syrup. (McMurry at al,2013: 670,671,677,678).
Disaccharides are compounds that consist of two monosaccharide
subunits hooked together by a g lycosidic linkage. actose is a disaccharide found
in milk. It constitutes 4.5% of cow’s milk and 6.5% of human milk by weight.
The subunits of lactose are d -galactose and d -glucose. The d -galactose subunit
is an acetal (Bruice, 2014: 1038).
Lipids are less well known than carbohydrates and proteins, yet lipids are
just as essential to our diet and well-being. They have three major roles in human
biochemistry: (1) Within fat cells (adipocytes), they store energy from metabolism
of food. (2) As part of all cell membranes, they keep separate the different
chemical environments inside and outside the cells. (3) In the endocrine system
and elsewhere, they serve as chemical messengers. Chemically, lipids are defined
as naturally occurring organic molecules that are nonpolar and therefore dissolve
in nonpolar organic solvents but not in water (McMurri et al,2013:721).
The classification of blood groups relies on differences in the structures of
oligosaccharides bonded to a protein called glycophorin, which is embedded in
the membrane of red blood cells. This chain is bonded to a protein in a variety of
possible ways including a glycosidic bond to the hydroxyl group of a serine
residue in the protein (Oulette and J, 2015: 363).
Protein content determined using visible spectrophotometry in the five
samples show that protein content in green peas and peanuts are not significantly
different, but their protein contents are lower than in ginkgo nuts, kidney beans
and flageolet beans. Compared to protein contents obtained using visible
spectrophotometry, protein contents obtained using Kjeldahl are higher. This is
caused by the different principle of both methods (Sinaga, et al, 2018: 262).
Molisch test is a general test used to detect the presence of carbohydrates.
If a negative test result is obtained the presence of sugars in the sample is omitted.
It is a useful test for identifying any compound which can be dehydrated to
furfural or hydroxymethylfurfural in the presence of H2SO4. Alpha naphthol reacts
with the cyclic aldehyde to form purple colored condensation products Iodine
reagent (KI) is used to distinguish starch and glycogen from other
monosaccharides and polysaccharide ( Elzagheid, 2018: 84).
With an estimated worldwide production of 1.5 × 1012 tons annually,
cellulose is almost an unlimited raw material source and the most abundant
carbohydrate polymer available at present. Cellulose has found significant
application in the paper, film, textile, wood product and fibre industry and most
recently, it is being employed in the production of biofuel. Starch is a giant
carbohydrate molecule made up of large amount of glucose units held together by
glycosidic bonds. Starch is formed from a combination of two polysaccharides:
amylose and amylopectin (Azeez and Olekan, 2018: 69).
Glucose, fructose, mannose, xylose, maltose, sucrose and lactose were
purchased from Sangon Biotech. Co., Ltd. (Shanghai, China). GI (HR7-100,
Hampton) from Streptomyces rubiginosus was obtained from Seebio (Shanghai,
China), which was used as received. All other reagents are of the AR grade. FBA
stock solution was prepared with phosphate-buffered saline solution containing
1% . All aqueous solutions were prepared with a Millipore system. The glass
carbon electrode with a diameter of 3 mm was polished with alumina power, and
then sonicated in ethanol and water (Liu et al, 2014: 9164-9165).
D. APPARATUS AND CHEMICALS
1. Apparatus
a. Test tube (12 units)
b. Rack tube (2 units)
c. Greduated cylinder 10 ml (4 units)
d. Beaker glass 1000 ml (1 unit)
e. Beaker glass 100 ml (1 unit)
f. Beaker glass 25 ml (1 unit)
g. Funnel (1 unit)
h. Stir bar (1 unit)
i. Pavorizer cup (1 units)
j. Spray bottle (1 unit)
k. Spiritus burner (1 units)
l. Tripod and gauze (1 units)
m. Wood clamp (1 units)
n. Rough cloth (2 units)
o. Soft cloth (2 units)
p. Drop pipettle (8 unit)
q. Tube brush (1 unit)
r. Matches (1 unit)
s. Erlenmeyer flask 100 ml (1 unit)
2. Chemicals
a. Glucose 0,02 M (C6H12O6)
b. Sucrose 0,02 M (C12H22O11)
c. Fructose 0,02 M (C6H12O6)
d. Univeresal indicator paper
e. Sodium hydrocide 40% and 6 M (NaOH)
f. Sodium chloride saturated (NaCl)
g. Zinc sulphate 1% (ZnSO4)
h. Hydrochloric Acid strong (HCl)
i. Sulphuris Acid strong (H2SO4)
j. Strach solution 0,7% (C6H10𝑂5 )
k. Alpha Naphtol 5% (In alcohol)
l. Ethanol 95% (C2H5OH)
m. Calcium chloride (CaCl2)
n. Resorsinol 0,5% (C6H6O2)
o. Egg white
p. Coconut oil
q. Pb-Acetat ((C6H3O2)2)
r. Luft solution
s. Lacmus
t. Filter paper
u. Copper Sulfate 1% (CuSO4)
v. Aquadest (H2O)
E. WORK PROCEDURE
1. General reaction of Carbohydrate
a. Prepared 4 reaction tubes and filled each reaction tubes with 1ml of glucose
0,02 M, 1 ml of fructosee 0,02 M, 1 ml of sucrose 0,02 M, and 1 ml starch
solution
b. Luff Test (to examine the reduction). In each tubes and filled 1 ml of dilute
luff solution, mixed and heated until 15 minutes. Observe the change
c. Molisch Test (to test the effect of acid/dehydration) In each tubes filled 2
drops of ∝-naphtol 5% mixed and added H2SO4 1 ml. observe the change
d. Seliwanof Test (to test hydrolisis result) In each tube filled 2 ml of
concentrated HCl and boiled it. Cooled it and added 0,5 ml of resorsinol 0,5%.
Observed thet color
2. General Reaction of Lipids
Saponification Reaction
a. Filled 5 ml of NaOH 40% in pavorizer cup
b. Filled 5 ml of coconut oil and 5 ml of ethanol
c. Heated until 15 minutes. shaked
d. If the cup has been solid, cooled and addedd 40 ml of saturated solution of
NaCl
e. Filtered and washed with cold water
Made a solution with dissolvve a half of the soap that you have made
a. Added 100 ml of aquadest
b. Checked the pH of soap solution with indicator universal
c. Added 5 grams of CaCl2 solution in 10 ml of soap solution
d. Shaked and noticed the observation
e. Tried to wash your hand with a recidue of soap that you have made
3. General Reaction of Protein
a. Diluted 2 ml of egg white with 12 ml of aquadest and shaked it
b. If the solution is colorless, added a little bit of salt (NaCl)
c. Prepared 4 reaction tubes and filled 2 ml of egg white solution in each tubes
d. Precipitation reaction, in first tube added a drops of silute ZnSO4 solution and
observed it
e. Biuret test, in second tube added 1 ml of NaOH 6 M then added drop of
CuSO4 % solution, mixed it and observed its color
f. In the third test tube, added 1 ml of NaOH 6 M, heated and checked the vapor
with lacmus paper that is wet red
g. In the fourth test tube, added drops of Pb-acetat solution than added 1 ml of
NaOH 6 M and heated it
F. OBSERVATION RESULT
1. General reaction of Carbohydrates
2. Reaction of Lipid
No. Activities Observation
1. 5 ml of NaOH, 5 ml of coconut Solid solution after heated
oil, 5 ml of ethanol (heated).
2. Solid solution + saturated The solution cloudt, there are
sediment
3. pH The solution is base with pH = 12
4. Solution of pH The solution is base
5. Solid solution + H2O + CaCl2 It has beem colorless solution and
there are white sediment
HO C H 2Cu2++5OH- H C OH
Cu2O +3H2O
H C OH H C OH
HO CH H H C OH
H2C OH
CH2O CH2OH
3) Sucrose
CH2OH
OH
C O O
OH C C OH OHC + Cu2++OH-
C C O C C CH2OH
OH
4) Starch
CH2OH CH2OH
OH C O
C O C
OH C C OH + Cu2++OH-
C
C C O C C OH
OH n
OH
b. Molisch test
Molisch test is qualitative in order to know the effect of acid on
carbohydrates. This test is based on the dehydration record of carbohydrates by
dukfat asm which forms a purple furfural ring. Positive reaction with the
appearance of a purple ring on the surface between the acid layer and the bottom
layer.
In this experiment there were 4 tubes and filled with glucose, fructose,
sucrose, and starch solution. Then the tube is added to a solution of α-naphthol
which functions to form deposits. and approximately 1 ml of concentrated sulfuric
acid serves to hydrolyze the saccharide bonds to produce furfural which then
reacts with the metabolic reagent. Glucose tube, sucrose, fructose produces a
colorless solution and does not form purple rings, while the starch solution
produces a purple solution but does not form a ring. This failure is possible when
the addition of H2SO4 has an acidic reaction with air. This is not in accordance
with the theory which states that starch solutions are carbohydrate compounds
which all produce a purple ring on their reaction with the metabolic reaction. As
for the reaction:
OH OH
HO
H3O+
HO [O], H3O+ HO
O
HO
O
+2 -H2O -H+, -2e-
OH OH
Cl
CH CH OH CH OH O OH OH
H
C C C C
+ CH2 O C
HOCH2 C O C
O
OH
b. Second tube, biuret test (color reaction). In this tube add 2 ml of the mixture
and CuSO4 produces a brown mixture. The function of addition is as
deanaturation of protein. Positive test in this experiment is to produce a color
change to blue, this indicates that there is a peptide in the protein solution and
the peptide bond can be said to be strong. The reaction that occur :
c. Third tube, steam test: addition of 1 ml which is then made to produce a gray
solution and produce a smell. The reaction of amino acids and amino acids is a
protein denaturation reaction. The addition of the mixture and steam of the
heated solution were tested using lacmus paper which produced an acidic
solution characterized by the color of the lacmus paper. The reaction that occur:
O O H O H O
H H2C H CH2OH
OH glisin serin
gliserin
d. Fourth tube, addition of PB acetate; plus a few drops of PB acetate and make
sure. Function as a protein deanaturation. After that it is heated to produce a
black solution. This is not in accordance with the theory, where solution should
not be colored. The function of the PB acetate test is to identify whether or not
there is an element of sulfur. If there is an element S then the solution that has
been added to PB acetate will appear black. So in this experiment the solution
that has been added to PB acetate still contains sulfur. The reaction
Pb2+ + 4OH − → PbO2− + 2H2 O
Bruice, Paula Yurkanis. 2014. Organic Chemistry Seventh Edition. New York:
Pearson.
Liu, lin, Ning Xia, Jimin Du, Qige He, Yun Qin, Huihui Li, and Caicai Li. 2014.
A Simple and Rapid Method for Probing of Isomerization of Glucose to
Fructose with Ferroceneboronic Acid. International Journal of
Electrochemical Science. Vol. 8.
Sinaga, Siti Morin, Maria Intan and Jansen Silalahi. 2015. Protein Analysis of
Canned Legumes by using Visible Spectrophotometry and Kjeldahl
Method. International Journal of PharmTech Research. ISSN: 0974-
4304 Vol.8, No.6.