EXPERIMENTAL REPORT OF

BIOCHEMISTRY
CARBOHYDRATE TEST

By :

Name : Intan Dwi Puspitasari

Class : PBU 2015

NIM : 15030204092

Mathematic and Science Faculty

SCHOOL YEAR

2015/2016

1
 TABLE OF CONTENTS

CHAPTER I

PREFACE PAGE

1. Background of Experimental.....3
2. Stating Problem......3
3. Aims of Experimental4
4. Benefits......4

CHAPTER II

LITERATURE REVIEW

5. Carbohydrate and the test......5

CHAPTER III

DESIGN OF EXPERIMENTAL

6. Tools and Materials.11

7. Procedure12
8. Workflow...13

CHAPTER IV

RESULT OF EXPERIMENTAL

9. Data of Experimental...22
10. Data Analyzity.28
11. Discussion....29

CHAPTER V

ENCLOSURE

12. Summary..32
13. Suggestion.......32

REFFERENCE38

APPENDIX.39

CHAPTER I
2
 PREFACE
1.1 Background of Experimental

Experiments in Biochemistry is designed as a result of learning biochemistry theory.

Experiments of wide scope and uniform quality have been provided. Opportunities are
presented to become skilled in the use of common and special biochemical apparatus, to
become proficient in the quantitative determination of biological substances in a variety of
naturally occurring materials, and to gain experience in the analysis of typical biological
compounds. The students understanding of, and appreciation for, the 'development, use, and
application of biochemical procedures is furthered by detailed explanations and discussions
of the principles underlying chemical reactions and techniques and of background and related
current investi-gations. The more than 900 references to the original and the review literature
should aid in increasing the biochemical subject matter. Among the new experiments which
have been found analyses of carbohydrates, identification of carbohydrates in a mixture of
carbohydrates.

To determine existence of carbohydrate in the pure used molisch reagent. This reacton
based on forming of furfural or that derivats from carbohydrate which dehydration by sulfat
concentrated. In alkali condition, saccharide will form enidid which easy to oxidation. All of
monosaccharide and disaccharide except sucrose and fructose will react Precise directions for
the preparatpositive when its done benedict test. To determine any sugar which contain of
keton used seliwanoff test. And to indicate polysaccharide we use Iodine test. All of this test
is to know the kind of carbohydrate which is consist in the fruit, because in the same way to
the process of fruits ripe commonly carbohydrate in the fruit have change the composition
because of the enzyme activity.Ion of special reagents, complete lists of apparatus and
reagents, precautions in the conduct of experiments, discussions of biochemical principles
and techniques, and a summary of the results are given in the Appendix..

1.2 Stating Problem

1. How to identify carbohydrate (Monosaccharide,Disaccharide,Polysaccharide) in the

experimental?
2. How to indicate any substance which reduction in alkali condition ?
3. How to differentiate sacaride which can reduct and cant reduct?
4. How to indicate any ketose?
5. How to indicate any polysaccharide (especially amylum)?
6. How to indicate any carbohydrate in material which doesnt know in general?

1.3 Aims of Experimental

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 1. To identify carbohydrate (Monosaccharide,Disaccharide,Polysaccharide) in the
experimental.
2. To indicate any substance which reduction in alkali condition.
3. To differentiate sacaride which can reduct and cant reduct.
4. To indicate any ketose.
5. To indicate any polysaccharide (especially amylum).
6. To indicate any carbohydrate in material which doesnt know in general.

1.4 Benefits

1. We can identify carbohydrate (Monosaccharide,Disaccharide,Polysaccharide) in the

experimental.
2. We can indicate any substance which reduction in alkali condition.
3. We can differentiate sacaride which can reduct and cant reduct.
4. We can indicate any ketose.
5. We can indicate any polysaccharide (especially amylum).
6. We can indicate any carbohydrate in material which doesnt know in general.

CHAPTER II

LITERATURE REVIEW
A. THE CARBOHYDRATES

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 Carbohydrates are the most abundant biomolecules belonging to class of organic
compounds found in living organisms on earth. Each year, more than 100 billion metric tons
of CO2and H2O are converted into cellulose and other plant products due to photosynthesis.
Living matter is largely made of biomolecules consisting of water and complex polymers of
amino acids, lipids, nucleotides and carbohydrates. Carbohydrates are most special of themin
that they remain associated with the three other polymers mentioned. Carbohydrates are
linked with amino acid polymers (proteins) forming glycoproteins and with lipids as
glycolipids. Carbohydrates are present in DNA and RNA,which are essentially polymers of
D-ribose-phosphate and 2-deoxy-D-ribose phosphate to which purines and pyrimidines bases
are attached at the C-1 reducing position. Carbohydrates are a widely diverse group of
compounds that are ubiquitous in nature. More than 75% of the dry weight of the plant world
is carbohydrate in nature - particularly cellulose, hemicellulose and lignin.

Existence of sugars is confirmed before life itself appeared on earth. It is highly

probable that there was a relative abundance of various sugars and their phosphates in the
prebiotic world, where the basic building blocks must have polymerized and assembled,
ultimately to forma self-sustaining, self-reproducing, and adaptive entity. The reality or
possibility that carbohydrates will be found in fossils is almostzero because they are
relatively unstable, capable of being dehydrated and of combining with other molecules. At
higher temperatures they caramelize and char. It has been known for over a century that many
sugars formed fromformaldehyde in alkaline solution. Several alternative, synthetic pathways
and reaction conditions have been proposed but specific details probably never be
established. Condensation of formaldehyde, a prebiotic constituent, led to formation of
glycol-aldehyde, trioses, tetroses, pentoses and hexoses in the laboratory. UV light, electric
discharges and ionization radiation at the right pH and temperature may also promote
synthesis of sugars. The phosphorylation of sugars is an indispensable step in the assembly of
the building blocks of living matter. Cyanogen, a likely prebiotic constituent, is capable of
catalyzing the synthesis of glucose mono- and diphosphate fromglucose and orthophosphate.

Carbohydrates comprise a comprehensive group of naturally occurring substances,

which include innumerable sugars and sugar derivatives, as well as high-molecular weight
carbohydrates (polysaccharides) like starch and cellulose in plants and glycogen in animals. A
polysaccharide molecule is composed of a large number of sugar or sugar-like units.
Carbohydrates are of great importance in biology. The unique reaction, which makes life
possible on Earth, namely the assimilation of the green plants, produces sugar, fromwhich
originate, not only all carbohydrates but, indirectly, also all other components of living
organisms.

The important role of carbohydrates, generally, in the metabolismof living organisms,

is well known. The biological breakdown of carbohydrates (often spoken of as "combustion")
supplies the principal part of the energy that every organismneeds for various vital processes.
It is not surprising; therefore, that the carbohydrates and their metabolismhave been the
subject of comprehensive and in many respects successful biochemical and medical research
for a long time.

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 This chapter is created in view of giving an idea about sugars (monosaccharides to
polysaccharides; their derivatives, their important reactions, structures, function and
biological importance. Efforts have made toexplain the chemistry and organization of
biomolecules in terms of stereioisomerism, optical isomerism,anomeric forms, mutarotations
etc. Information about polysaccharides in plants and living organisms, blood sugars and rare
sugars is also included.

Importance of carbohydrates

Carbohydrates are of great importance in biology. The unique reaction, which makes
life possible on the Earth, namely the assimilation of the green plants, produces sugar,
fromwhich originate, not only all carbohydrates but, directly or indirectly, all other
components of living organisms. The carbohydrates are a major source of metabolic energy,
both for plants and for animals that depend on plants for food. Aside fromthe sugars and
starch that meet this vital nutritional role, carbohydrates also serve as a structural material
(cellulose), a component of the energy transport compound ATP, recognition sites on cell
surfaces, and one of three essential components of DNA and RNA. Importance can be
considered under following headings;

Metabolic/Nutritional

The important role of carbohydrates, generally, in the metabolismof living organisms,

is well known. The biological breakdown of carbohydrates (often spoken of as "combustion")
supplies the principal part of the energy that every organismneeds for various processes.
Carbohydrates and their metabolismhas been the subject of biochemical and medical research
for a long time. Carbohydrates play a major role in promoting health fitness, forma major part
of food and help a great deal in building body strength, by generating energy. They are one
among the three prominent macronutrients that serve as excellent energy providers, the other
two being fats and proteins. Carbohydrate intake can takeplace in different forms like sugar,
starch, fibers etc., which are a dietary staple in most parts of the world, and the oxidation of
carbohydrates is the central energy-yielding pathway in most nonphotosynthetic organisms.
The functions of carbohydrates are multiple and it is owing to this fact that it becomes all the
more necessary to incorporate carbohydrates in our meal. For instant for energy generation,
sugars and starch act as the perfect fuel that enables us to carry out our physical activities
efficiently and effectively. Fiber does wonders in keeping your bowel function going smooth.
Carbohydrates add on to the taste and appearance of food item,thus making the dish tempting
and mouthwatering. They are sometimes used as flavors and sweeteners. Carbohydrates aid
in regulating blood glucose and also do good to our body by breaking down fatty acids, thus
preventing ketosis. Talking about the importance of carbohydrates, apart fromits direct
benefits, there is alsoan added advantage of carbohydrate consumption and that is that
carbohydrates are found in different foods, which if eaten, also pave way for consuming other
essential nutrients. Therefore,it is preferable to go in for distinctive carbohydrate food
sources.

Structural

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 The structural diversity possible by linking the different, common sugar is immense:
theoretically far greater than that of proteins, which largely consist of 22 amino acids linked
by a single type of peptide bond. Linkages between sugars can occur through a glycoside
linkage between the anomeric, first carbon of a sugar in either or configuration with any of
a variety of hydroxyl groups on the adjacent sugar. In fact, many possible combinations of
sugars do not seemto exist. Insoluble carbohydrate polymers serve as structuraland protective
elements in the cell walls of bacteria and plants and in the connective tissues of animals.
Plant cell walls are complex arrangements of cellulose, hemicellulose and lignin. This
contributes significantly to the overall digestibility of the fiber. The proportion of each
component depends on species and age of the plant. Cellulose is the primary structural
component of plants. It is found primarily in the cells walls and is a primary fiber component
of animal feeds.

Classification

Carbohydrates are called saccharides or, if they are relatively small, sugars.
Classifications of carbohydrates are outlined in the following table.

The compounds carbohydrates have common samefunctional groups, glyceraldehydes and

gulose are classifed as aldoses and ribulose and dihydroxyacetone as ketoses (Fig. 2). All of
these compounds are alcohols with many hydroxyl groups. They are polyhydroxylated and
either aldehydes or ketones.

The important groups of carbohydrates that will be studied are the simple sugars
(monosaccharides), typified by glucose, the complex sugars (disaccharides), typified by cane
sugar and lactose, and the polysaccharides, typified by starch and cellu1ose. In each of
these divisions it is necessary to study also other compounds closely related to the types-

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fructose, galactose, and arabinose, related to glucose, maltose, related to lactose, and
glycogen, the dextrins, and inulin, related to starch.

MONOSACCHARIDE

The simplest and smallest unit of the carbohydrates is the monosaccharide, (mono = one,
saccharide = sugar) fromwhich disaccharides, oligosaccharides, and polysaccharides are
constructed. Monosaccharides are either aldehydes or ketones, with one or more hydroxyl
groups; the six-carbon monosaccharides glucose (an aldohexose) and fructose (a keto hexose)
have five hydroxyl groups. (Fig. 3) The carbon atoms, to which hydroxyl groups are attached,
are often chiral centers, and stereoisomerismis common among monosaccharides

1.GLUCOSE

The tests for glucose depend upon both 'chemical properties and physical properties. On
account of its easy oxidisability, it brings about certain" reductions" and is hence a reducing
sugar. It reacts with phenyl-hydrazine to form a characteristic yellow crystalline " osazone,"
which can be identified under the micro-scope. It is decomposed by certain ferments in yeast
with production of ethyl alcohol and carbon dioxide gas, so that the evolution of gas serves as
a test for it. It rotates the plane of polarisation of polarised light, and can therefore be tested
for in the polarimeter. It is readily soluble in water. The following tests should be carried out
with a per cent. solution of' glucose :

-Experiment 1. Add to 5 C.c. of glucose solution in a test-tube 2 drops of the specIal

Molisch's reagent (made by dissolving 5 gm. of a-naphthol in 100 C.c. of ethyl alcohol).
Shake. Pour carefully down the side of the inclined test-tube 5 C.c. of concentrated
sulphuric acid. With care scarcely any mixing takes place. A reddish-violet colour is
produced at the surface between the two solutions. (A greenish colour is to be neglected.)
This test is given by all soluble carbohydrates, and a similar colour is developed on the
surface of insoluble carbohydrates under appropriate conditions. Reduction Tests Copper
solutions are reduced with precipitation of cuprous oxide (red or yellow according to
variations in the experimental conditions), certain silver solutions are reduced with
precipitation of metallic silver, and certain bismuth solutions with precipitation of metallic
bis-muth. Such results are due to the ease with which glucose and related sugars can be

8
oxidised. This is usually attributed to the presence (or potential presence) of an aldehyde or
ketone radical in the carbohydrate molecule.

-Experiment 2.Reduction of Benedict's Solution This has the following composi-tion: 17'3
gm. copper sulpha~e, 173 gm. sodmmcitrate, 100 gm. anhydrous sodium carbonate,
dissolved in water, and water added to 1 litre. The substitution of citrate for tartrate gIves a
much more permanent solution. Experiment 4. Transfer 5 C.c. of Benedict's solution to a
test-tube and add exactly 8 drops (using a small piece of glass tubing or a pipette as a
dropper) of the glucose solution. Boil for two minutes. Allow to cool slowly. A precipitate
gradually forms throughout the solution, red, yellow or green, depending upon the
concentration of the glucose solu-tion used. In absence of a reducing substance the solution
remains clear.

2. FRUCTOSE

Carry out Experiments 1 to 8 with a 1 per cent. solution of fructose. Positive results will be
obtained in every test. Specific Tests for Fructose:

-Experiment 1. SELIWANOFF'S REACTION. To 5 C.c. of the' special (S~liwanoff's)

reagent (0'5 gm. of resorcinol dissolved in a litre of 10 per cent. hydrochlorIC acid) add a
few drops of the fructose solution. Boil for not more than thirty seconds. A red colour
develops, and then a brown-red precipitate separates. (DIlute solutions of fructose do not
give the precipitate.) Decant off the solution and add alcohol. The precipitate dissolves to a
red solution. Repeat thjs test with 1 per cent. glucose solutIOn, and note that no red colour
develops during thirty seconds' boiling, but that longer boiling does produce it.

THE DlSACCHARIDES

A disaccharide consists of two monosaccharides joined by an O-glycosidic bond.

Disaccharides can be homo- and heterodisaccharide (Fig. 5). Three most abundant
disaccharides are sucrose, lactose, and maltose. In sucrose the anomeric carbon atoms of a
glucose unit and a fructose unit are joined. Lactose, the disaccharide of milk, consists of
galactose joined to glucose by a (14) glycosidic linkage. In maltose, (14) glycosidic
linkage joins two glucose units. Sucrose and lactose are heterosaccharides and maltose is
homosaccharide.

3. SUCROSE

Experiment 1. Make up 100 c.c. of a 5 per cent. solution of sucrose. Measure the rotation.
Take 50 c.c. of this solu-tion (accurately measured) in a shallow porcelain dish, add 2 c.c. of
concentrated hydrochloric acid and heat on tile water-bath for forty-five minutes. Cool,
exactly neutralise with 10 per cent. sodium hydroxide, transfer to a 50 c.c. graduated flask,
with rinsings, and make up to the original 50 c.c. volume. Again measure the rotation.
Calculate from the results whether or not the change represents complete transformation of
the sucrose to glucose and fructose. Then. with the hydrolysed solution, carry out Benedict's,

9
Fehling's, Barfoed's and Seliwanoff's tests, and show that these are all positive. The student
himself should hydrolyse a little sucrose on the small scale with acid, cool, neutralise and
carry out the tests just enumerated.

4. LACTOSE

Experiment 1. Make up a 1 per cent. solution of lactose. Note the hard, gritty feel of the
crystals and their relative insolubility as compared with glucose and fructose. Repeat
Experimcnts 1 to 8 with this solution. Not that positive results are obtained with all these
tests except Bar-foed's and the fermentation test. Barfoed's test is not given by disaccharides.
A good baker's yeast (free from foreign strains) does not ferment lactose. Note the
microscopic appearance of the lactosazone and compare it with the photograph in Plate I.
Repeat the mucic acid test (Experiment 13). A positive result is obtained. Lactose is first
hydrolysed to glucose and galactose, and these are oxidised respectively to saccharic and
mucic acids. The saccharic acid remains in solution.

5. MALTOSE

Experiment 1. Repeat experiments 1 to 8 with a 1 per cent. solution of maltose. Note that all
the tests are posi-tive except Barfoed's. Note the microscopic appearance of the osazone and
compare it with the photograph in Plate I. Note. Glucosazone crystals commence to separate
from the hot solution, but ]aetosazone and maltosazonc crystals only commence to separate
when the solution is allowed to cool. 'fhis fact help;; to differentiate hctween them.

POLYSACCHARIDES

Polysaccharides are relatively complex carbohydrates. They are polymers made up of many
monosaccharides joined together by glycosidic bonds. They are, therefore, very large, often
branched, macromolecules. They tend to be amorphous, insoluble in water, and have no
sweet taste. When all the monosaccharides ina polysaccharide are of the sametype, the
polysaccharide is called a homopolysaccharide and when more than one type of
monosaccharide is present, they are called heteropolysaccharides. Examples include storage
polysaccharides such as starch and glycogen and structural polysaccharides such as cellulose,
and chitin. Xylan a hemicellulose is a heteropolysaccharide. Polysaccharides have a general
formula of Cn(H2O)n-1where n can be any number between 200 and 2500. Considering that
the repeating units in the polymer backbone are often six-carbon monosaccharides, the
general formula can also be represented as (C6H10O5)nwhere

n={40...3000}.

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 6.AMYLUM

Experiment 1. Scrape a raw potato, stir up the scrapings vigorously in a littlc water and strain
through cheese cloth into a small beaker. The starch granules rapidly settle. Decant off the
liquid, wash with cold water, and again decant off the liquid. Shake up the residue with
water, transfer a little to a slide and examine under the low power of the micro-o little rice
and wheat starch suspended in water and trans-ferred to slides. Note that each has a
characteristically different appearance.

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 CHAPTER III

DESIGN OF EXPERIMENT
3.1 Tools and Material

Molischs Test

A. Tolls : Reaction tube, drop pipette, reaction tube rack, reaction tube clamp,
measuring glass.

B. Materials : Concentrated H2SO4 , molisch reagent (-naphtol 10% solutions in

ethanol/methanol), carbohydrate solutions.

Benedicts Test

A. Tolls : Reaction tube, drop pipette, reaction tube rack, reaction tube clamp,
measuring glass, waterbath.

B. Materials : Benedict reagent, carbohydrate solutions( Glucose 1%, Fructose 1%,

Amylum 1%, Sucrose 1%, Maltose 1%, Lactose 1%, Cellulose 1%)

Seliwanoffs Test

A. Tolls : Reaction tube, drop pipette, reaction tube rack, reaction tube clamp,
measuring glass, waterbath, timer.

B. Materials : Seliwanoff reagent (0,05% resorsinol in HCL 3 N), carbohydrate

solutions( Glucose 1%, Fructose 1%, Amylum 1%, Sucrose 1%, Maltose 1%, Lactose
1%, Cellulose 1%).

Iodines Test

A. Tolls : Reaction tube, drop pipette, reaction tube rack, reaction tube clamp,
measuring glass.

B. Materials : Iodine solutions 1 M (10 grams KI in 1 L water + 2,5 gram iodine ),

Amylum 1%, Cellulose 1%.

Carbohydrates Test in to Fruits

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 A. Tolls : Reaction tube, drop pipette, reaction tube rack, reaction tube clamp,
measuring glass, waterbath,

B. Materials : Seliwanoff reagent, Benedict reagent,Molish reagent, Iodine,

Concentrated H2SO4, the petri cup.

3.2 Procedure

Molischs Test

1. Prepare all of the carbohydrate to be solutions which have concentration 1%.

2. Added 2 ml solutions of carbohydrate 1% in to different reaction tube.
3. Added 2-3 drops of Molisch reagent, shake gently for 5 seconds.
4. Aslant that reaction tube, droped 1 ml ( 20 drops of H2SO4 through the
tube reactions wall. Upright reaction tube and observe any purple red ring in
the border of both solutions.

Benedicts Test

1. Prepare all of the carbohydrate to be solutions which have concentration 1%.

2. Added 2 ml benedict reagent in to reaction tube.
3. Added 5 drops of glucose 1% then heat it in to waterbath in 5 minutes, let it
cooler and compare the changes of the color that happened.
4. Do this test with the same method to another carbohydrate 1%.

Seliwanoffs Test

1. Prepare all of the carbohydrate to be solutions which have concentration 1%.

2. Added 1 ml seliwanoff reagent in to reaction tube.
3. Added 2 drops of amylum 1%. In the same time, reaction tube placed in to
waterbath until the color changed (note the speed of form color from each
reaction tube).
4. Do this test with the same method to another carbohydrate solutions.

Iodines Test

1. Prepare three reaction tube, added 3 ml amylum solutions 1% to each other.

2. Added 2 drops water in to first tube, 2 drops HCL in to second tube, 2 drops
NaOH in to third tube. Mix all of tube.Then added 5 drops of Iodine
Solutions. .Observe the changes that happened.
3. Heats the color tube and makes it cold. Observe the changes that happened.

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 4. Do the test to the solutions of Cellulose 1% and glycogen 1%.

Carbohydrates Test in to Fruits

1. In Molisch test, benedict test , seliwanoff test, the method is the same with the
experiment before.
2. For Iodine test
Each extract of unripe, ripe and well ripe fruits placed in the petri cup.
Added 1 drop of iodine to each extract, then note the change of color.

3.3 Work Flow

Molischs Test

Amylum 1%

-Put 2 ml in to test tube

- Added 2-3 drops of Molisch

reagent
Amylum 1% + Molisch
reagent

-shake gently for 5 minutes

-Incline the test tube

- droped 1 ml ( 20 drops of H2SO4 through the tube reactions

Amylum 1% + Molich reagent +

H2SO4
-Upright the test tube

- observe thechanges of the color that happened

Discoloration

Benedicts Test

1. Glucose Solution 5. Fructose Solution

2 ml of in
-Put benedict reagent
to reaction tube. 2 ml of benedict
--Put reagent
in to reaction tube.

-Added 5 drops of Glucose -Added 5 drops of Fructose

1%. 1%.

-Heats in to waterbath for -Heats in to waterbath for

about 5 minutes. about 5 minutes.
14
-Let it cooler -Let it cooler

-Comparing the color -Comparing the color

 2 color layers, they are in
the top is colorness,
bottom is blue and there is
maroon color and thin (+)
sediment.
2. Lactose Solution
2 ml of benedict reagent
-Put in to reaction tube.
-Added 5 drops of Lactose 1%.
-Heats in to waterbath for
about 5 minutes.
-Let it cooler
-Comparing the color
alteration and the occur of
Thesediment.
color is blue and there
is maroon color sediment.
3. Maltose Solution
2 ml of benedict reagent
-Put in to reaction tube.
-Added 5 drops of Sucrose
1%.
-Heats in to waterbath for
about 5 minutes.
-Let it cooler
-Comparing the color
alteration and the occur of
2 color layers, the tops is
greenish blue and the
bottom is blu. The sediment
color is maroon and thin (+
+)
The color is light blue and
the sediment is maroon
color and thick (+++)
6. Sucrose Solution
2 ml of benedict reagent
-Put in to reaction tube.
-Added 5 drops of Sucrose
1%.
-Heats in to waterbath for
about 5 minutes.
-Let it cooler
-Comparing the color
The color is blueand
alteration andthe
nooccur of
sediment
7. Amylum Solution
2 ml of benedict reagent
-Put in to reaction tube.
-Added 5 drops of Sucrose
1%.
-Heats in to waterbath for
about 5 minutes.
-Let it cooler
-Comparing the color
alteration and the occur of
The color is blu and have no
sediment.
15
 4. Cellulose Solution

2 ml of benedict reagent

-Put in to reaction tube.

-Added 5 drops of Sucrose

1%.

-Heats in to waterbath for

about 5 minutes.

-Let it cooler

-Comparing the color

alteration and the occur of
The color is blu and have no
sediment.

Seliwanoffs Test

1. Glucose Solution 5. Fructose Solution

1 ml of seliwanoff reagent 1 ml of seliwanoff reagent

-Put in to reaction tube. -Put in to reaction tube.

-Added 2 drops of Glucose -Added 2 drops of Glucose

1%. 1%.

-In the same time, reaction -In the same time, reaction
tube placed in waterbath until tube placed in waterbath until
the case of change color. the case of change color.

-Note the speed of the -Note the speed of the

formation of color formation of color
The color is yellowish (++) The color is orange (+++) , 16
, and the speed is 21 and the speed is 6 minutes
minutes 33 seconds 22 seconds
2. Lactose Solution 6. Sucrose Solution

1 ml of seliwanoff reagent 1 ml of seliwanoff reagent

-Put in to reaction tube. -Put in to reaction tube.

-Added 2 drops of Glucose -Added 2 drops of Glucose

1%. 1%.

-In the same time, reaction -In the same time, reaction
tube placed in waterbath until tube placed in waterbath until
the case of change color. the case of change color.

-Note the speed of the -Note the speed of the

formation of color formation of color
The color is yellowish (++ The color is orange (++) ,
+) , and the speed is 22 and the speed is 9 minutes
minutes 52 seconds

3. Maltose Solution 7. Amylum Solution

1 ml of seliwanoff reagent 1 ml of seliwanoff reagent

-Put in to reaction tube. -Put in to reaction tube.

-Added 2 drops of Glucose -Added 2 drops of Glucose

1%. 1%.

-In the same time, reaction -In the same time, reaction
tube placed in waterbath until tube placed in waterbath until
the case of change color. the case of change color.

-Note the speed of the -Note the speed of the

formation of color. formation of color.

The color is yellowish (+++ The color is yellowish (+) ,

+) , and the speed is 15 and the speed is 24 minutes
minutes 30 seconds 45 seconds.

17
 4. Cellulose Solution

1 ml of seliwanoff reagent

-Put in to reaction tube.

-Added 2 drops of Glucose

1%.

-In the same time, reaction

tube placed in waterbath until
the case of change color.

-Note the speed of the

formation of color.

The color is yellowish (+++

++) , and the speed is 22
minutes 25 seconds.

Iodines Test

1. Amylum Solution

3 ml of Amylum
Solution 1 %

Put it in reaction tube using a

-Added 2 drops of
water using a pipette.
1st Reaction Tube 2nd Reaction Tube
-Mix this solution and
added 5 drops of
Iodine Solutions.
-Observed the
changes.
-when the color
change then heated,
after that makes it
cold.
-Added 2measuring
drops of HCLglass -Added 2 drops of
using a pipette. NaOH using a
3pipette.
rd
Reaction Tube
-Mix this solution and
added 5 drops of -Mix this solution and
Iodine Solutions. added 5 drops of
Iodine Solutions.
-Observed the
changes. -Observed the
changes.
-when the color
change then heated, -when the color 18
after that makes it change then heated,
cold. after that makes it
cold.
The color is The color is The color is clear
blackish blue blackish blue

2. Cellulose Solution

3 ml of Cellulose
Solution 1%

Put it in reaction tube using a

measuring glass

1st Reaction Tube 2nd Reaction Tube 3rd Reaction Tube

-Added 2 drops of -Added 2 drops of HCL -Added 2 drops of
water using a pipette. using a pipette. NaOH using a
pipette.
-Mix this solution and -Mix this solution and
added 5 drops of added 5 drops of -Mix this solution and
Iodine Solutions. Iodine Solutions. added 5 drops of
Iodine Solutions.
-Observed the -Observed the
changes. changes. -Observed the
changes.
-when the color -when the color
change then heated, change then heated, -when the color 19
after that makes it after that makes it change then heated,
cold. cold. after that makes it
cold.
 The color is clear The color is The color is clear
blackish blue

Carbohydrates Test in to Fruits

1. Molisch Test

2 ml of 1% unripe 2 ml of 1% ripe 2 ml of 1% well ripe

avocado solution Avocado Solution avocado solution
-Put in to raction - Put in to raction - Put in to raction
tube. tube. tube.

-Added 2-3 drops of -Added 2-3 drops of -Added 2-3 drops of

Molisch reagent. Molisch reagent. Molisch reagent.

-Mix rapidly in 5 -Mix rapidly in 5 -Mix rapidly in 5

minutes minutes minutes

-Aslant that reaction -Aslant that reaction -Aslant that reaction

tube tube tube

-Added 1 ml (20 -Added 1 ml (20 -Added 1 ml (20

drops) of H2SO4 drops) of H2SO4 drops) of H2SO4
through reaction through reaction tube through reaction tube
20
Theres purple ring Theres purple ring Theres purple ring
(+) between both (++) between both (+++) between both
solution. solution. solution.

2. Benedict Test

2 ml of Benedict 2 ml of Benedict 2 ml of Benedict

reagent reagent reagent

-Put in to reaction -Put in to reaction -Put in to reaction

tube. tube. tube.

-Added 5 drops of -Added 5 drops of -Added 5 drops of

1% unripe avocado 1% ripe avocado 1% well ripe
solution. solution. avocado solution.

-Heats in to -Heats in to -Heats in to

waterbath about 5 waterbath about 5 waterbath about 5
minutes. minutes. minutes.

-Let it cool. -Let it cool. -Let it cool.

-Compared the -Compared the -Compared the

Theres maroon Theres maroon Theres maroon
sediment(+++) sediment(++) sediment(+)
3. Seliwanoff Test

1 ml Seliwanoff 1 ml Seliwanoff 1 ml Seliwanoff

reagent reagent reagent

-Put in to reaction -Put in to reaction -Put in to reaction

tube. tube. tube.

-Added 2 drops of -Added 2 drops of -Added 2 drops of

1% unripe avocado 1% ripe avocado 1% well ripe
solution, In the same solution, In the same avocado solution, In
time placed reaction time placed reaction the same time
tube in to waterbath tube in to waterbath placed reaction tube
until formed color. until formed color. in to waterbath until
formed color.
-Note the speed of -Note the speed of
formed color. formed color. -Note the speed of
formed color.

Color change Orange Color change Color change

in 5 minutes 40 Orange in 8 minutes Orange in 7 minutes
seconds. 01 seconds. 44 seconds.
21
 4. Iodine Test

1 drop of 1% unripe 1 drop of 1% ripe 1 drop of 1% well

Avocado solution Avocado solution ripe Avocado
solution
-Put in to petri cup -Put in to petri cup -Put in to petri cup

-Added 1 drop of -Added 1 drop of -Added 1 drop of

Iodine Iodine Iodine

-write the change of -write the change of -write the change of

Theres green Theres green Theres yellow

solution (+) solution (+++) green solution

CHAPTER IV

RESULT OF EXPERIMENT
4.1 Data of Experimental

Table 1. Test of 1% Carbohydrate Solutions with Molisch Reagent

N Carbohydrate Activity Result

O Solution
Before After
Glucose 1% 2 ml glucose 1%+ -Glucose color= clear Ring = +
1 Molisch reagent+ -Molisch reagent= dark
concentrated H2SO4 brown. Clouded = ++
-Glucose-molisch
reagent= clear but there
are dark brown
suspended.

22
2 Fructose 1% 2 ml fructose 1%+ -Fructose color= clear Ring = +++
Molisch reagent+ -Molisch reagent= dark
concentrated H2SO4 brown. Clouded = +
-Fructose + molisch
regent= clear but there
are dark brown
suspended.
3 Sucrose 1% 2 ml sucrose 1%+ -Sucrose color= clear Ring = +++++
Molisch reagent+ -Molisch reagent= dark
concentrated H2SO4 brown. Clouded = ++
-Sucrose + molisch ++
regent= clear but there
are dark brown
suspended.
4 Maltose 1% 2 ml maltose 1%+ -Maltose color= clear Ring = +++
Molisch reagent+ -Molisch reagent= dark
concentrated H2SO4 brown. Clouded = ++
-Maltose + molisch +
regent= clouded but
there are dark brown
suspended.
5 Lactose 1% 2 ml lactose 1%+ -Lactose color= clear Ring = ++
Molisch reagent+ -Molisch reagent= dark
concentrated H2SO4 brown. Clouded =+ +
-Lactose + molisch
regent= clouded but
there are dark brown
suspended.
6 Amylum 1% 2 ml amylum 1%+ -Amylum color= white Ring = ++
Molisch reagent+ cloudy
concentrated H2SO4 -Molisch reagent= dark Clouded = ++
brown. +
-Amylum + molisch
regent= clouded but
there are dark brown
suspended.
7 Cellulose 1% 2 ml cellulose 1%+ -Cellulose color= white Ring = ++++
Molisch reagent+ cloudy
concentrated H2SO4 -Molisch reagent= dark Clouded = ++
brown. +++
-Cellulose+ molisch
regent= clouded but
there are dark brown
suspended.

23
 Table 2. Test of 1% Carbohydrate Solutions with Benedict Reagent

N Carbohydrate Activity Result

O Solution
Before After
Glucose 1% Benedict reagent + -Before heated: -2 color layers:
1 5 drops of 1% color is blue. Top:
glucose solution and -after heated: colorness
then heat in to two layers Bottom:
waterbath for about (colorness on blue
5 minutes. the top and -sediment:
blue on the Maroon
bottom). color and
Sediment : thin(+)
black red
2 Fructose 1% Benedict reagent + Before heated: color -color : Light blue
5 drops of 1% is blue. -sediment:
fructose solution -after heated: Maroon color and
and then heat in to Color: thick(+++)
waterbath for about maroon (++)
5 minutes. Sediment :
maroon (+)
3 Sucrose 1% Benedict reagent + Before heated: color -color: blue
5 drops of 1% is blue. -no sediment
sucrose solution and -after heated:
then heat in to Constant
waterbath for about color and
5 minutes. there is no
sediment.
4 Maltose 1% Benedict reagent + Before heated: color -2 color layers:
5 drops of 1% is blue. Top:
maltose solution and -after heated: colorness
then heat in to three layers Bottom:
waterbath for about (maroon on blue
5 minutes. the top -sediment:
,greenish Maroon color and
blue on the thin(+)
middle and
darkblue on
the bottom).
There is no
sediment.

24
 5 Lactose 1% Benedict reagent + Before heated: color -2 color layers:
5 drops of 1% is blue. Top:
lactose solution and -after heated: greenish
then heat in to color: blue
waterbath for about greenish Bottom:
5 minutes. blue. blue
there is no -sediment:
sediment. Maroon color and
thin(++)
6 Amylum 1% Benedict reagent + Before heated: color -color : blue
5 drops of 1% is blue. - no sediment:
amylum solution -after heated:
and then heat in to color: blue
waterbath for about there is no
5 minutes. sediment
7 Cellulose 1% Benedict reagent + Before heated: color -color : blue
5 drops of 1% is blue. - no sediment:
cellulose solution -after heated:
and then heat in to color : blue
waterbath for about there is no
5 minutes. sediment.

Table 3. Test of 1% Carbohydrate Solutions with Seliwanoff Reagent

NO Carbohydrate Activity Result

Solution
Before After Time

25
 Glucose 1% 1 ml seliwanoff + 2 -Seliwanoff Yellowish 21
1 drops of 1% glucose reagent: not (++) minutes
solution. Then colored 33
heated. -Amylum seconds
solution: not
colored
2 Fructose 1% 1 ml seliwanoff + 2 -Seliwanoff Orange (++ 6 minutes
drops of 1% reagent: not +) 22
fructose solution. colored seconds
Then heated. -Amylum
solution: not
colored
3 Sucrose 1% 1 ml seliwanoff + 2 -Seliwanoff Orange (++) 29
drops of 1% sucrose reagent: not minutes
solution. Then colored 59
heated. -Amylum seconds
solution: not
colored
4 Maltose 1% 1 ml seliwanoff + 2 -Seliwanoff Yellowish 15
drops of 1% maltose reagent: not (++++) minutes
solution. Then colored 30
heated. -Amylum seconds
solution: not
colored
5 Lactose 1% 1 ml seliwanoff + 2 -Seliwanoff Yellowish 22
drops of 1% lactose reagent: not (+++) minutes
solution. Then colored 52
heated. -Amylum seconds
solution: not
colored
6 Amylum 1% 1 ml seliwanoff + 2 -Seliwanoff Yellowish 24
drops of 1% reagent: not (+) minutes
amylum solution. colored 45
Then heated. -Amylum seconds
solution:
clouded
7 Cellulose 1% 1 ml seliwanoff + 2 -Seliwanoff Yellowish 22
drops of 1% reagent: not (+++++) minutes
cellulose solution. colored 25
Then heated. -Amylum seconds
solution:
clouded

Table 4. Test of 1% Carbohydrate Solutions with Iodine

26
 N Carbohydrate Activity Result
O Solution
Before After
Amylum 1% 3 ml amylum 1 % + 2 -amylum:
1 drops of water + 5 drops of clouded
Iodine , then heated. -aquades:
clouded
-iodine: blackish
green (+++)
3 ml amylum 1 % + 2 -amylum:
drops of HCL + 5 drops of clouded
Iodine , then heated. -aquades:
clouded
-iodine: blackish
green (++)
3 ml amylum 1 % + 2 -amylum:
drops of NaOH + 5 drops clouded
of Iodine , then heated. -aquades:
clouded
-iodine: clouded
2 Cellulose 1% 3 ml cellulose 1 % + 2 -amylum: clear
drops of water + 5 drops of -aquades: clear
Iodine , then heated. -iodine: blackish
purple (+++)

3 ml cellulose 1 % + 2 -amylum: clear

drops of HCL + 5 drops of -aquades:
Iodine , then heated. clouded
-iodine: blackish
purple (++)
3 ml cellulose 1 % + 2 -amylum: clear
drops of NaOH + 5 drops -aquades: clear
of Iodine , then heated. -iodine: clear

Table 5. Test of 1% Carbohydrate Solutions in to Fruits

27
N Activity Result
O
Before After
Molisch Test - Brownish green - There is purple
1 solution (+) ring between
both of solution
a. Unripe Avocado (+)
b. Ripe Avocado - yellowish green - There is purple
solution (++) ring between
both of solution
(++)
c. Well ripe Avocado - Dark green - There is purple
solution (+++) ring between
both of solution
(+++)
2 Benedict Test - Brownish green - There is maroon
solution (+) sediment (+++)

a. Unripe Avocado
b. Ripe Avocado - yellowish green - There is maroon
solution (++) sediment (++)

c. Well ripe Avocado - Dark green - There is maroon

solution (+++) sediment (+)

3 Seliwanoff Test - Brownish green - Color change

solution (+) become orange at
a. Unripe Avocado 5 minutes 40
seconds
b. Ripe Avocado - yellowish green - Color change
solution (++) become orange at
8 minutes 01
seconds
c. Well ripe Avocado - Dark green - Color change
solution (+++) become orange at
7 minutes 44
seconds
4 Iodine Test - Brownish green - There is green
solution (+) solution (+)
a. Unripe Avocado

28
 b. Ripe Avocado - yellowish green There is green solution
solution (++) (+++)

c. Well ripe Avocado - Dark green There is yellowish green

solution (+++) solution.

4.2 Data Analyzity

Table 1. Test of 1% Carbohydrate Solutions with Molisch Reagent

2 ml Amylum 1% + Molich reagent + H2SO4 will reaction and change the color
become clouded and there are dark brown suspended. It happens not only on amylum but
also lactose,cellulose, maltose,fructose,sucrose, and glucose solutions. It is because to
determine existence of carbohydrate in the pure used Molisch Test. This reaction based on
forming of furfural or that derivats from carbohydrate which dehydration by the sulfat
concentration. The results will react with -naphtol form compounds purple redness.
Example saccharide with added by sulfat concentrated will dehydration to be furfural
compounds or that derivats like hidroximetil furfural. So the test was showing that the
solution is pure carbohydrate.

Table 2. Test of 1% Carbohydrate Solutions with Benedict Reagent

Benedict reagent + drops of 1% glucose solution and then heat in to water bath in 5
minutes. The result is there are two layers of that solution and there is a maroon sediment.
Beside glucose, the solution that can changes and make sediment are fructose, lactose and
maltose solution. In alkali condition, saccharide will form enidid which easy to oxidation.
All of the monosaccharide and disaccharide (except sucrose,trekalosa and polysaccharide
such as amylum and cellulose) will react positive when its done benedict test. Copper
solutions alkali when reducted by carbohydrate which have free aldehid or keton will
form Cupro Oxide (Cu20) which have green color,red orange, maroon and any maroon
sediment in the bottom of reaction tube. Amylum, cellulose and sucrose didnt have
sediment in the end of reaction because they are negative reaction with benedict reagent.

Table 3. Test of 1% Carbohydrate Solutions with Seliwanoff Reagent

1 ml Seliwanoff reagent + 2 drops of amylum 1 % then heated in to waterbath. The

change of color is yellowish and the time to reaction is 24 minutes 45 seconds. Wheter
fructose and sucrose when added by seliwanoff the color change to be orange, it means
that the solution conist of ketose. And both of solution are quickly in time when heated in
to waterbath the other solution. Because the determine any sugar which contain of keton
used seliwanoff test. This reaction principle based on formed 4-Hidroxil Metil Furfural
which will form a purple substance which any resorsinol (1,3-dihidroxi benzene). This
reaction specifically to ketose with characteristic by the result of reaction to change in to
29
 red color. So beside fructose and sucrose, other solutions are have ketose, but the
composition is less than both of them so the color become yellowish and get long time to
reaction.

Table 4. Test of 1% Carbohydrate Solutions with Iodine

2ml amylum 1%+2 drops of water+5 drops of Iodine the heated. In other tube we
change the water to be HCl or NaOH. Than the changes of color after heated is blacklish
blue and clear. This reaction also happened in cellulose. This is happened because the
amylum-iodide complex is formed as charge-recall electrons are charged particles-is
transferred between amylum and iodide ions-tri-iodide or pentaiodide.

The transfer of charge between the amylum and the iodide ion changes the spacing
between the energy level/orbitals. This change results in the amylum-iodide complex
absorbing light at a different wavelength-than any other species aforementioned-resulting
in an intense purple color; biologist call this color blue-back. So , the reaction is positive
between amylum and iodine, and also the cellulose, it marked by the change of color to be
blackish blue.

Table 5. Test of 1% Carbohydrate Solutions in to Fruits

This test consist of molisch test, benedict, seliwanoff and iodine. But the solution is
fruits. The fruits that we choose is avocado in vary conditions, they are ripe, unripe and
well ripe. In molish test the third condition fruit have the same method and the result
there is purple ring,which have the level concentration in color. In our test the high
concentration in color is well ripe. Actually the color shows about polysaccharide that
should be in the unripe avocado. This result can be different with the teory because of
false in added, or maybe the time is too quickly so the reaction not finished. Because in
this test to know the kind of carbohydrate which consist in the fruits, in the same way to
the process of fruits ripe commonly carbohydrate in the fruit have change the composition
because of the enzyme activity. In the wellripe fruit and sweet will find much glucose and
fructose, in the unripe fruit will find carbohydrate in the shape of amylum which possible
finally of other carbohydrate. And when the test of iodine, the color that should be
indicated after reaction is blackish-purple but the result of the test is still green color. The
reason maybe the solutions of iodine that less and the reaction not finished perfectly.

4.3 Discussion

Table 1. Test of 1% Carbohydrate Solutions with Molisch Reagent

1. Why formed the ring red purple on the materials that contains the carbohydrate?
Answer:
It is because to determine existence of carbohydrate in the pure used Molisch Test.
This reaction based on forming of furfural or that derivats from carbohydrate which
dehydration by the sulfat concentration. The results will react with -naphtol form
compounds purple redness. Example saccharide with added by sulfat concentrated will
30
 dehydration to be furfural compounds or that derivats like hidroximetil furfural. So the
test was showing that the solution is pure carbohydrate.

2. Is the same the intensity of the red purple ring in test you use in the experiment?
Explain!
Anwer:
Rather the same,and change the color become clouded and there are dark brown
suspended. It happens not only on amylum but also lactose,cellulose,
maltose,fructose,sucrose, and glucose solutions. It is because to determine existence
of carbohydrate in the pure used Molisch Test

Table 2. Test of 1% Carbohydrate Solutions with Benedict Reagent

1. What is the color of sediment formed? Why is it happened?
Answer :
In alkali condition, saccharide will form enidid which easy to oxidation. All of the
monosaccharide and disaccharide (except sucrose,trekalosa and polysaccharide such
as amylum and cellulose) will react positive when its done benedict test. Copper
solutions alkali when reducted by carbohydrate which have free aldehid or keton will
form Cupro Oxide (Cu20) which have green color,red orange, maroon and any
maroon sediment in the bottom of reaction tube

2. In benedict test, why is the sucrose not including in to the sugar reduction?
Answer :
. because sucrose didnt have sediment in the end of reaction because they are
negative reaction with benedict reagent

Table 3. Test of 1% Carbohydrate Solutions with Seliwanoff Reagent

1. Which group from carbohydrate that have positive reaction to the seliwanoff test?
Why?
Answer:
Wheter fructose and sucrose when added by seliwanoff the color change to be orange,
it means that the solution conist of ketose

2. Can seliwanoff test used to differentiate sucrose from fructose?

Answer:

So beside fructose and sucrose, other solutions are have ketose, but the composition is
less than both of them so the color become yellowish and get long time to reaction.

Table 4. Test of 1% Carbohydrate Solutions with Iodine

1. Why is the color changed often it heated?
Answer :

31
 .This is happened because the amylum-iodide complex is formed as charge-recall
electrons are charged particles-is transferred between amylum and iodide ions-tri-
iodide or pentaiodide

2. Which substance beside amylum that give the color to the iodine?
Answer :
There is Cellulose solution

Table 5. Test of 1% Carbohydrate Solutions in to Fruits

1. Why is the well ripe still found of carbohydrate in form polisacaride?
Answer :
In the well ripe fruit and sweet will find much glucose and fructose, in the unripe fruit
will find carbohydrate in the shape of amylum which possible finally of other
carbohydrate. And because of the polysaccharide chain doesnt hang loosely all.

2. Explain the process of amylum hydrolysis in enzymatic?

Answer :
the kind of carbohydrate which consist in the fruits, in the same way to the process of
fruits ripe commonly carbohydrate in the fruit have change the composition because
of the enzyme activity.

CHAPTER V

ENCLOSURE
5.1 Summary

32
 1. We can identify carbohydrate (Monosaccharide,Disaccharide,Polysaccharide) in the
experimental.
2. Indicate any substance which reduction in alkali condition.
3. Differentiate sacaride which can reduct and cant reduct.
4. Indicate any ketose.
5. Indicate any polysaccharide (especially amylum).
6. Indicate any carbohydrate in material which doesnt know in general such as in fruits.

5.2 Suggestion

After we know how to indicated the unknown fruit or maybe unknown solution with
the several test,it can be help us when we feel hesitant to eat or to taste something. We can
use this method to enactive and keep our body from the snakes or drinks that can be destroy
our digestion.

REFFERENCE
Arbianto,Purwo. 1994. Biokimia Konsep Konsep Dasar . Jakarta:Tanpa
Penerbit

Isnawati . 2009.Biokimia.Surabaya:Unesa University Press

33
 Leningher,albert L . 1975. Biochemistry Second Edition : The Molecular
Basis Of Cell Structure And Function.New york :worth publisher

Leningher,albert L . 1982. Dasar Dasar Biokimia Jilid 1 .Jakarta

:Penerbit Erlangga

DUNN ,MAX S.1951.Experiment in Biochemistry.hal 77

Khowala ,Suman, dkk.2008. BIOMOLECULES: (INTRODUCTION, STRUCTURE &

FUNCTION) .Carbohydrates, pages 1-33.

MORROW ,CLARENCE AUSTIN .1935. BIOCHEMICAL LABORATORY

METHODS FOR STUDENTS OF THE BIOLOGICAL SCIENCES

http://www.brilliantbiologystudent.weebly.com/iodine-test--for-starch.html (acces on
29/03/2016 at 20:00)

APPENDIX
1. MOLISCH TEST

34
 Figure Figure Figure
6.1.1.Molisch 6.1.2.Molisch test 6.1.3.Molisch test
test in 1% in 1% fructose in 1% sucrose
glucose solution solution

Figure Figure Figure

6.1.4.Molisch test 6.1.5.Molisch test 6.1.6.Molisch test
in 1% maltose in 1% lactose in 1% amylum
solution solution solution

Figure
6.1.7.Molisch test
in 1% cellulose
solution
2. BENEDICT TEST

a. glucose solution

35
1. 2ml benedict reagent + glukose 1% ( before heated into waterbath )

2. Glukose 1% (after heated) 3. Glukose 1% (let until cool)

two layers (colourless on the top 2 layers colour

colourless in the top
and blue on the bottom) & brick in the bottom) & brick
red sediment
red sediment

b. Fructose solution
1. 2ml benedict reagent + fructose 1% (before heated into water bath )

36
 2. fruktose 1% (after heated ) 3. Fruktose 1% (let until cool)

Brick red colour and brick red sediment brick red colour and brick
red sediment

c. Sucrose solution
1. 2ml benedict reagent + sucrose 1% (before heated into water bath )

37
 2. Sucrose 1% (after heated ) 3. Sucrose 1 % (let until cool )

Constant colour and there is no sediment blue and no sediment

d. Maltose solution
1. 2ml benedict reagent + maltose 1% (before heated into waterbath )

38
2. maltose 1% (after heated ) 3. Maltose 1% (let until cool )

3 layers colour ( brick in top , 2 layers colour greenish blue in the

top blue
Greenish blue in the middle , and in the bottom and there is brick red
sediment
Dark blue in the bottom )

e. Lactose solution

39
 1. 2ml i benedict reagent + lactose 1% (before heated into waterbath )

2. lactose 1% (after heated ) 3. lactose 1 % ( let until cool )

Greenish blue colour and there is no sediment blue colour and there is brick
red sediment

f. Amylum solution
1. 2ml benedict reagent + amylum 1% ( before heated into waterbath )

40
2. amylum 1% (after heated ) 3. Amylum 1% ( let until
cool )

41
 Blue colour blue colour there is no
sediment

g. Cellulose experiment
1. 2ml benedict reagent + cellulose 1% (before heated into waterbath )

2. cellulose 1% (after heated ) 3. cellulose 1 % (let until cool

)

Blue colour blue colour there is no

sediment

42
3. SELIWANOFF TEST

43
Amylum 1% + Cellulose 1% + Fructose 1% +
Seliwanoff reagent Seliwanoff reagent Seliwanoff reagent
after heated after heated after heated

Glucose 1% + Lactose 1% + Maltose 1% +

Seliwanoff reagent Seliwanoff reagent Seliwanoff reagent
after heated after heated after heated

4. IODINE TEST

44
Amylum+aquades Amylum+HCl+io Amylum+NaOH+i
+iodine and after dine and after odine and after
heated the color heated the color heated the color
change be blackish change be is clear.
blue blackish blue

Selulose+aquades Selulose+HCl+io Selulose+NaOH+i

+iodine and after dine and after odine and after
heated the color heated the color heated the color
change be blackish change be is clear.
blue blackish blue

45
5. CARBOHYDRATE TEST ON FRUITS

pour avocado solution into

Meterials of xperiment :
reaction tube
unripe, ripe and well ripe
avocado

Solution of molish reagent

added avocado solution Benedict reagent pour into
rection tube

Added benedict reagent Heated tube reaction

with avocado solution contain benedict and
avocado solution into
waterbath

Heated reaction tube that

Seliwanoff reagent pour contain seliwanoff and
into reaction tube avocado solution into
waterbath

46
Pour avocado solution into
Avocado solution added
petri cup
iodine reagent.

47