Objective

To perform fructose, glucose, lactose, galactose, ribose, ribulose, sucrose,

starch and two unknown saccharides solution tests with series of reagents
To determine the 2 unknown saccharide solution

Introduction

A carbohydrate is a biological molecule consisting of carbon (C), hydrogen (H)

and oxygen (O) atoms, usually with a hydrogenoxygen atom ratio of 2:1 (as in
water); in other words, with the empirical formula Cm(H2O)n (where m could be
different from n). Carbohydrate also known as 'saccharide', a group that includes
sugars, starch, and cellulose. The saccharides are divided into four chemical groups:
monosaccharides, disaccharides, oligosaccharides, and polysaccharides.
Monosaccharides and disaccharides, the smallest carbohydrates, are commonly
referred to as simple sugars. Disaccharides formed from 2 monosaccharides bonded
together via glycosidic linkage. Oligosaccharides is the polymerization of several
monosaccharides. While polysaccharides is a polymer of saccharides.
Carbohydrates have numerous roles in living organisms. Polysaccharides serve as
storage of energy in form of starch and glycogen. Carbohydrate also serve as
structural components in form of cellulose in plants and chitin in arthropods. The 5-
carbon monosaccharide ribose is an important component of coenzymes which
helps in TCA cycle, example of them are ATP, FAD and NAD. Furthermore, it is the
backbone of the genetic molecule known as RNA. The related deoxyribose is a
component of DNA. Saccharides and their derivatives include many other important
biomolecules that play key roles in the immune system, fertilization, preventing
pathogenesis, blood clotting, and development.
In this experiment, solution containing fructose, glucose, lactose, galactose,
ribose, ribulose, sucrose, starch and two unknown saccharides was given. This
solution will be tested by using series of reagents that will produce color changes
with specific functional groups of saccharides being tested.
Procedure
A. Molischs test for carbohydrate
Molisch's test is a sensitive chemical test for the presence of carbohydrates, based
on the dehydration of the carbohydrate by sulfuric acid or hydrochloric acid to
produce an aldehyde, which condenses with two molecules of phenol (usually -
naphthol, though other phenols (e.g. resorcinol, thymol) also give colored products),
resulting in a red- or purple-colored compound. Negative results show absence of
carbohydrate.
Methods :
1. 2 ml of each solutions was inserted into a clean test tubes.
2. 2 drops of Molisch reagent was added into test tube and was mixed
thoroughly
3. 5 ml a concentrated Sulphuric Acid was poured into each test tubes
4. Purple color was observed indicating positive results

B. Benedicts test for reducing sugar

Benedict's reagent is a chemical reagent commonly used to detect the presence of
reducing sugars, however other reducing substances also give a positive reaction. It
is a complex mixture of sodium carbonate, sodium citrate and copper(II) sulfate
pentahydrate. It is often used in place of Fehling's solution. A positive test with
Benedict's reagent is shown by a colour change from clear blue to a brick-red
precipitate.
Methods :
1. 1 ml of each solutions was added into each clean test tubes.
2. 5 ml of benedict reagent was added into each test tubes and mixed
thoroughly.
3. Test tubes was placed in boiling water bath for 3 minutes
4. Test tubes was removed to allow it cool.
5. Color changes was observed

C. Barfoeds test for monosaccharides

Barfoed's test is a chemical test used for detecting the presence of
monosaccharides. It is based on the reduction of copper(II) acetate to copper(I)
oxide (Cu2O), which forms a brick-red precipitate. The aldehyde group of the
monosaccharide which normally forms a cyclic hemiacetal is oxidized to the
carboxylate.
Methods :
1. 1 ml of each solutions was added into each clean test tubes.
 2. 3 ml of barfoed reagent was added into each test tubes and mixed
thoroughly.
3. Test tubes was placed in boiling water bath for 3 minutes
4. Test tubes was removed to allow it cool.
5. Color changes was observed
D. Lasker and Enkelwitz test for ketoses
Lasker and Enkelwitz test uses the same concept as benedicts test only it is done at
lower temperature. Color changes is the same with Benedicts test.
Methods :
1. 1 ml of each solutions was added into each clean test tubes.
2. 5 ml of benedict reagent was added into each test tubes and mixed
thoroughly.
3. Test tubes was placed in 55C water bath for 10 - 20 minutes
4. Test tubes was removed to allow it cool.
5. Color changes was observed. Positive result will have green, red or yellow
precipitate.

E. Seliwanoffs test for ketoses

It is a chemical test which distinguishes between aldose and ketose sugars. Ketoses
are distinguished from aldoses via their ketone/aldehyde functionality. If the sugar
contains a ketone group, it is a ketose. If a sugar contains an aldehyde group, it is
an aldose. This test relies on the principle that, when heated, ketoses are more
rapidly dehydrated than aldoses.
Methods :
1. 1 ml of each solutions was added into each clean test tubes.
2. 5 ml of Seliwanoff reagent was added into each test tubes and mixed
thoroughly.
3. Test tubes was placed in boiling water bath for 4 minutes
4. Test tubes was removed to allow it cool.
5. Color changes was observed. Positive results show orange or red color.

F. Bials test for pentoses

Bial's test is a chemical test for the presence of pentoses. It is named after Manfred
Bial, German Physician. The components include orcinol, hydrochloric acid, and
ferric chloride. A pentose, if present, will be dehydrated to form furfural which then
reacts with the orcinol to generate a colored substance. The solution will turn bluish
and a precipitate may form.
Methods :
1. 2 ml of each solutions was added into each clean test tubes.
2. 5 ml of Bials reagent was added into each test tubes and mixed thoroughly.
 3. Test tubes was placed in boiling water bath for less than 5 minutes
4. Test tubes was removed to allow it cool.
5. Color changes was observed. Positive results show changes from green to
blue precipitate.

G. Iodine test for starch and glycogen

The iodine test is used to test for the presence of starch. When treated with iodine
solutioniodine dissolved in an aqueous solution of potassium iodidethe triiodide
anion (I3) complexes with starch, producing an intense blue/purple colour.
However, the intensity of the color decreases with increasing temperature and with
the presence of water-miscible organic solvents such as ethanol. The test cannot be
performed at very low pH due to the hydrolysis of the starch under these conditions.
When the iodine solution comes into contact with starch, it turns dark blue/purple.
Methods :
1. 5 ml of each sample was added into each clean test tubes.
2. 2-3 drops of Lugols iodine reagent was added into each test tubes and was
mixed thoroughly
3. Brown-blue color for glycogen and blue-black color for starch was observed
indicating positive results.

References
Flitsch, Sabine L.; Ulijn, Rein V (2003). "Sugars tied to the spot". Nature. 421 (6920):
21920. doi:10.1038/421219a. PMID 12529622.
Jump up ^ Maton, Anthea; Jean Hopkins; Charles William McLaughlin; Susan
Johnson; Maryanna Quon Warner; David LaHart; Jill D. Wright (1993). Human Biology
and Health. Englewood Cliffs, New Jersey, USA: Prentice Hall. pp. 5259. ISBN 0-13-
981176-1.