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A.

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

Carbohydrates are synthesized by plants and stored as starch, a polymers of


glucose (McMurry et al, 2013: 657).
Acid catalyzes the reaction, shifting the equilibrium to the right if there is
excess alcohol or the water that forms is removed. In this substitution reaction, an
–OR group replaces the –OH group.
OH OR'
H+
'
R C OR' + R OH R C OR' + H - OH
H H
a hemiacetal an acetal

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

No. Solution Luff test Molisch test Seliwanoff test


1. Glucose Colorless and Colorless and no ring Colorless and
there is no there is no
sediment sediment
2. Fructose Blue tosca and Colorless and no Orange and there
there is no forming a ring is no sediment
sediment
3. Sucrose Blue tosca and Colorless and no Orange and there
there is no forming a ring is no sediment
sediment
4. Strach Blue tosca and Purple and no forming Colorless and
solution there is no a ring there is no
sediment sediment

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

3. General Reaction of Protein


No. Activities Observation
1. 2 ml of white egg solution + drops of ZnO Cloudy to white, there are
sediment
2. 2 ml of white egg solution + 1 ml of NaOH The solution becomes
+ 5 drops of CuSO4 (Blue solution) brown
3. 2 ml of white egg solution + 1 ml of NaOH The color is grey, and the
( heated) solution is acid
4. 2 ml of white egg solution + 5 drops of Pb- The color is black
acetic + 1 ml of NaOH (Heated it)
G. DISCUSSION
1. The general reaction of Carbohydrate
Carbohydrate or commonly called saccharide is defined as polyhydroxy
aldehyde or polyhydroxy ketone which can be hydrolyzed to produce compounds
on the basis of the number of constituent units.
a. Luff Test
The luff test is performed to test the power of reduction by using glucose,
fructose, sucrose and starch solution. Glucose, fructose, sucrose and starch
solution are put into different tubes, after which a dilute luff solution is added to
each tube. The function of adding luff as a tester, is in the solution there is sugar.
This aims to determine which solution includes reducing sugar. The solution is
then heated for 15 minutes. The function of heating is to speed up the reaction.
The results we obtained were first glucose which produced a colorless solution in
this glucose experiment which is a carbohydrate derivative (monosaccharide)
which has a reduction power faster than other solutions, and also has a free
reduction group which can reduce so that it can form Cu2O (deposits red),
fructose produces Blue toska-colored solutions and no deposits, ii does not fit the
theory that fructose can be formed from hydrolysis of a disaccharide called
sucrose and fructose is one of the reducers (Budhiman, 2009). Furthermore,
sucrose produces turquoise colors that are not much different from the initial color
before heating. This indicates that there is no reaction. This is in accordance with
the theory which states that sucrose is a reducing sugar (Budiman, 2009). The
starch solution that produces turquoise colors is not much different from the color
before heating. This indicates that there is no reaction due to the weak compound
both which consists of disaccharides and polycaraides which must first become
monosalca which is a saturated hydrocarbon. There are reactions that occur:
1) . Glucose
O
H2C H
C OH
H C OH HO C H

HO C H 2Cu2++5OH- H C OH
Cu2O +3H2O
H C OH H C OH

HO CH H H C OH
H2C OH

D-glukosa Asam D-glukonat


2) Fructose
OH
heksodiol glukosa asam glukomat
O
O OHOH COOH
OH/H2O 2+ -
2Cu +SOH
HO H C OH H OH H C OH + Cu2O+3H2O
H OH H OH
OH C H OH C H
HO H
H OH
H C OH H OH H C OH
OH C OH H C
H OH 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

5-(hidrosimetil) furfural 𝛼 − 𝑛𝑎𝑝ℎ𝑡ℎ𝑜𝑙 senyawa berwarma


c. Seliwanof Test
Seliwanof test is used to distinguish or to test the results of hydrolysis.
This experiment was conducted to test the results of hydrolysis and can also be
used to distinguish the presence of ketoses in the sample, judging by the color
change.
First mixing on each tube with HCl which serves to hydrolyze the
polysaccharide into its constituent monosaccharide and change each solution to a
colorless solution after it is heated. The heating function is to dehydrate fructose,
starch, sucrose, and glucose into hydroxy methyl furfural. After that glucose is
added resorcinol and produces a colorless solution, starch also produces a
colorless solution while sucrose and fructose produce an orange solution. This
identifies the ketose content in carbohydrates. This thing which is anchored in
seliwanof reagent dehydrates fructose and sucrose to produce hydroxy furfural so
that it forms an orange solution. The reaction :
O O
R C + HCl HCl C C C O atau
H
CH CH CH H

Cl
CH CH OH CH OH O OH OH
H
C C C C
+ CH2 O C
HOCH2 C O C
O
OH

2. The general reaction of Lipid


In this experiment, a sapling reaction was carried out. The sapling reaction
was a reaction involving fat or oil with an alkali which produced soap and
glycerol. This experiment was carried out by mixing NaOH, coconut oil, ethanol
and then heating it to become solid. The function is as a hydrolysis reagent to
form soap, coconut oil serves as a basic ingredient in making soap, and ethanol
functions for fat emulsifiers. The solid formed then clarified with saturated NaCl
aims to separate and glycerol. Glycerol does not experience precipitation due to its
high solution. Then dissolved and filtered and added with water to test the pH
using a universal indicator and produce Ph 12 which means the solution is
alkaline. then CaCl2 is added which serves to determine the effect of water on
soap. In the water the soap will produce a little foam. The reaction :
𝐶17 𝐻35 𝐶𝑂𝑂𝑁𝑎 + 𝐻2 𝑂 → 𝐶17 𝐻35 𝐶𝑂𝑂− + 𝑁𝑎+
Anions with fatty sulfuric acid react water and are hydrolyzed
𝐶17 𝐻35 𝐶𝑂𝑂− + 𝐻2 𝑂 → 𝐶17 𝐻35 𝐶𝑂𝑂𝐻 + 𝑂𝐻
The reaction that occur :
2𝐶17 𝐻35 𝐶𝑂𝑂𝑁𝑎 + 𝐶𝑎𝐶𝑙2 → 𝐶𝑎 (𝐶17 𝐻35 𝐶𝑂𝑂) + 2𝑁𝑎𝑐𝐶𝑙
3. The General Reaction Of Protein
In this egg white experiment diluted with distilled water, if the solution
obtained is not clear then add salt and then put into 4 different tubes.
a. First tube, deposition reaction: added 2 drops of ZnSO4 which produces a
cloudy solution and there is a white precipitate below. This indicates the
presence of negatively charged proteins and binds positively charged metals.
The reaction that occur :
CH3 O CH3

H2N C C OH + ZnSO4 H2N C C SO4 + Zn

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

H2N CH C HN CH C OH + H2OH N2 C C OH + H2N C C OH

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

S 2+ + H2 O + Pb2 O2_ → PbS + 4OH −

H. CONCLUSION AND SUGGESTION


1. CONCLUSION
a. Carbohydrate divided in some types, that is monosaccharide which consists of
glucose and fructose, disaccharide also known as sucrose and polysaccharide
also known as starch. There are some terst to know the reaction of
carbohydrates compounds that are luff test, molisch test, and seliwanof test. In
molisch test, all the results we obtained are not in accordance with the positive
test or theory. In luff test, only the glucose that is in accordance with theory,
and in seliwanof test, only the fructose and sucrose that are in accordance with
the theory.
b. In general reaction of lipid, there is saponification reaction, from this reaction it
can produce the soap from lipid with pH 12 and the solution is base.
c. In general reaction of protein there are precipitation reaction which in this
experiment obtained the precipitated or sediment, then biuret test, the positive
test is the solution is become purple but in this experiment become brown, the
next experiment is a solution that produce bed smell after adding NaOH and
heated. That’s the solution is acid. And then the solution that product the black
of color solution.
2. SUGGESTION
At the suggestion of this experiment, is for the next practitioner it is
needed to be focused to see every changes that occur in this experiment, and when
adding and mixing the chemicals should be done the right way. So, the results
obtained can be in accordance with the theory.
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