JOANNA ERLINDA V.

TAYLAN DATE PERFORMED: 11- 13 and 15-18

BSN 1A DATE SUBMITTED: 11 – 20 - 18

EXPERIMENT NO. 5

CARBOHYDRATES

I. INTRODUCTION

Carbohydrates are naturally occurring gases in plants (where they are produced

photosynthetically), animals, and microorganisms that has different roles involving structures

and metabolism. Carbohydrates are aldehydes or ketones of higher polyhydric alcohols or

components that yield these derivatives on hydrolysis (Prasad, 2010). Carbohydrates is also

known as “hydrate carbon” since it contain carbon, oxygen and hydrogen with the ratio

(CH2O)n. One feature of carbohydrate is the presence of large number of functional groups.

Carbohydrates are classified according to the number of simple sugar units they comprise.

Monosaccharides are the simplest carbohydrates for they contain a single polyhydroxy

aldehyde or ketone. Because of its size, monosaccharides cannot be broken down into simpler

units by hydrolysis. They are usually building blocks of disaccharides or polysaccharides for

instance glucose, fructose, xylose and galactose. Whereas, disaccharides are water-soluble

carbohydrates having two monosaccharide units bonded covalently to each other. Sucrose,

table sugar, lactose, milk sugar, and maltose are examples of disaccharides. On the other hand,

oligosaccharides are carbohydrates containing two to ten monosaccharide unit attached

together by covalent bonds. As for polysaccharides, a polymeric carbohydrate, they contain

many monosaccharide units bonded covalently. In the experiment done, the carbohydrates

involved were glucose, xylose, fructose, galactose, lactose, maltose and sucrose. As
aforementioned, glucose, xylose, fructose and galactose are monosaccharide sugars. Glucose

is the most abundant monosaccharide in nature and most important component of human diet.

Xylose, is a natural sugar usually found in berries, spinach and pears. It is known to be a great

alternative to white sugar. . Fructose, the sweetest-tasting of all sugars, is a biochemically the

most important ketohexose (Stoker, 2010). Galactose, a sugar commonly found in lactose or

milk sugar, is classified as a monosaccharide, an aldose, a hexose, and a reducing sugar.

Meanwhile the disaccharides used in the experiment were lactose, maltose and glucose.

Lactose, the main carbohydrate found in milk, is made up of the monosaccharides galactose

and lactose joined by a glycosidic linkage. Maltose, commonly known as malt sugar, is formed

by two glucose units. Sucrose, common table sugar, is the most abundant of all disaccharides

and is produced commercially from the juice of sugar cane and sugar beets (Stoker, 2010).

II. FLOWCHART OF PROCEDURE

A. MACROSCOPIC APPEARANCE
Observe powdered samples of:
 Glucose
 Galactose
 Fructose
 Lactose
 Sucrose
 Xylose

USING MICROSCOPE

GLUCOSE

Crystallization is the process of formation of solid sugar crystals from a sugar solution and it
offers a particularly fascinating and rewarding experience for the beginner microscope
enthusiast, and there is basically no limit to the scope for exploration, even with relatively
simple apparatus.
GALACTOSE

It is not normally found in nature, but is mostly hydrolyzed from the disaccharide lactose,
which is found in milk, as part of a disaccharide made by glycosidic linkage to a glucose
molecule. The lactose disaccharide from milk is a major energy source for almost all animals,
including human

FRUCTOSE

Fructose is a water-based mounting medium, which is suitable for a wide variety of specimens.
It is safe to use and it is easy and cheap to make. Fructose syrup is not suitable for making
slides that last for many years, but is should be sufficient for classroom usage where students
would like to re-examine their specimens over and over again over a period of time.

LACTOSE

The physical stability and soli-state recrystallization of spray-dried ‘amorphous’ lactose

particles were visualized using environmentally controlled atomic force microscopy and
conventional optical microscopy.

SUCROSE

It is a natural osmolyte accumulated in the cells of organisms as they adapt to environmental

stress.

XYLOSE

It is a sugar first isolated from wood, and named for it. Xylose is classified as a monosaccharide
of the aldopentose type which means it contain five carbon atoms and includes an aldehyde
functional group.
 GLUCOSE GALACTOSE

FRUCTOSE LACTOSE

SUCROSE XYLOSE
B. SOLUBILITY

1. 1 mL H2O

2. 1 mL 10% NaCl Solution

1 2 3 4 5 6
3. 1 mL 0.2 % HCl Solution

4. 1 mL 95% ethyl alcohol

Glu Gal Fru Lac Suc Lyx

Add a pinch each of solid sugar

Shake and observe solubility

C. CONFORMITY

Glu Gal Fru Lac Suc Lyx 1 mL of 5% sugar solution +

1 mL 6 N NaOH
1 2 3 4 5 6
 Heat in H2O Bath

Note color changed and odor produced

2. MOLICH”S TEST

1 mL of 5% sugar solution
+ 1 drop Molich’s reagent
+1 mL cone H2SO4
(inclined position)

Glu Gal Fru Lac Suc Lyx

Note color produced at the interface of two liquids

3. SELIWANOFF’S TEST

1 mL of Seliwanoff’s
reagent + 5 drops 5 %
Glu Gal Fru Lac Suc Lyx
sugar solution
 Heat in boiling water

Note change in appearance

D. REDUCING TESTS

Glu Gal Fru Lac Suc Lyx

5 mL sugar solution
samples
1.) TROMMER’S TEST
+ 05 mL 6 N NaOH
+ 1 drop by drop 10% CuSO4 solution

Heat in boiling water bath for 3 minutes

2.) FEHLING’S TEST

6 t.t 0.5 mL sugar samples

+0.5 mL Fehling’s A
+0.5 mL Fehling’s A

Heat in boiling water bath 6-8 minutes

Note color change

3.) BENEDICT’S TEST

6 t.t 0.5 mL sugar samples

1 mL Benedict’s reagent

Mix

Heat is boiling water bath 6-8 minutes

Note result
 Observe

Heat again for 5 minutes

4.) BARFOED’S TEST

6 t.t 0.5 mL sugar samples

1 mL Barfoed’s reagent

Heat in water bath for 1 minute

Allow to cool for 15 minutes

Note changes
III. DATA AND RESULTS

REDUCTIONS TESTS FOR SUGARS

A. GLUCOSE
OSERVATIONS
Trommer’s test Initial color: light violet to orange
After heating: Matcha green
Fehling’s test Initial color: light blue
After heating: light brown
Benedict’s test Initial color: light blue
After heating: dark brown
Barfoed’s test Initial color: light blue
After heating: lighter blue

B. GALACTOSE
OSERVATIONS
Trommer’s test Initial color: indigo to peach
After heating: mint green
Fehling’s test Initial color: light blue
After heating: dark brown
Benedict’s test Initial color: light blue
After heating: light brown
Barfoed’s test Initial color: light blue
After heating: lighter blue green

C. FRUCTOSE
OSERVATIONS
Trommer’s test Color after heating: Matcha green

Fehling’s test Color after heating: dark brown

Benedict’s test Color after heating: dark brown
Barfoed’s test Color after heating: faded light blue green
D. LACTOSE
OSERVATIONS
Trommer’s test Initial color: indigo to peach
After heating: brown
Fehling’s test Initial color: light blue
After heating: brown
Benedict’s test Initial color: light blue
After heating: light brown
Barfoed’s test Initial color: light blue
 After heating: light blue

E. SUCROSE
OSERVATIONS
Trommer’s test Initial color: royal blue to blue
After heating: sky blue
Fehling’s test Initial color: light blue
After heating: light blue
Benedict’s test Initial color: light blue
After heating: light blue
Barfoed’s test Initial color: light blue
After heating: light blue

F. XYLOSE
OSERVATIONS
Trommer’s test Initial color: brown to orange
After heating: light blue green
Fehling’s test Initial color: yellow with blue
After heating: light brown
Benedict’s test Initial color: light blue
After heating: dark brown
Barfoed’s test Initial color: light blue
After heating: light blue

CONFIRMATORY TESTS FOR ALDOSES AND KETOSES

A. GLUCOSE
OBSERVATIONS
Moore’s test Color: umber brown
Odor: pleasant
Molich’s test Color: dark violet
Odor: pleasant
Seliwanoff’s test Color: slightly beige
Odor: pleasant

B. GALACTOSE
OBSERVATIONS
Moore’s test Color: Syrup
Odor: pleasant
Molich’s test Color: dark brown
Odor: pleasant
Seliwanoff’s test Color: slightly cream
Odor: pleasant
C. FRUCTOSE
OBSERVATIONS
Moore’s test Color: Gold (light yellow) (before heating)
Yellow orange (after heating)
Odor: pleasant
Molich’s test Color: brown
Odor: pleasant
Seliwanoff’s test Color: bright red
Odor: unpleasant

D. LACTOSE
OBSERVATIONS
Moore’s test Color: Tawny
Odor: pleasant
Molich’s test Color: violet
Odor: pleasant
Seliwanoff’s test Color: no change
Odor: pleasant

E. SUCROSE
OBSERVATIONS
Moore’s test Color: no change
Odor: pleasant
Molich’s test Color: violet
Odor: pleasant
Seliwanoff’s test Color: red
Odor: pleasant

F. XYLOSE
OBSERVATIONS
Moore’s test Color: cinnamon
Odor: pleasant
Molich’s test Color: dark violet
Odor: pleasant
Seliwanoff’s test Color: no change
Odor: pleasant
 SOLUBILITY OF SOME SUGARS IN DIFFERENT SOLVENTS

OBSERVATIONS
Water 10% NaCl 0.2 % HCl 95% EtOH
GLUCOSE Fully dissolved Fully dissolved Fully dissolved partially
(clear) (clear) (hazy) dissolved
(hazy)
GALACTOSE Fully dissolved Fully dissolved not dissolved partially
(clear) (clear) (sturdy) dissolved
(hazy)
FRUCTOSE Fully dissolved Fully dissolved partially not dissolved
(clear) (clear) dissolved (sturdy)
(hazy)
LACTOSE Fully dissolved partially partially partially
(clear) dissolved dissolved dissolved
(hazy) (hazy) (hazy)
SUCROSE Fully dissolved Fully dissolved partially not dissolved
(clear) dissolved (sturdy)
(clear)
(hazy)
XYLOSE Fully dissolved Fully dissolved Fully dissolved not dissolved
(clear) (clear) (clear) (sturdy)

IV. DISCUSSION AND CONCLUSION:

Moore’s Test
In table 1, We tested the presence of reducing sugar using Moore’s test. We observed
that Glucose produced an Umber brown (dark) color , Galactose produced Syrup in color which
is also a dark one. While fructose, Lactose and Xylose produced dark to light brown color. On
the other hand, Sucrose didn’t produce a color, it remained clear hence it is a non-reducing
sugar. A Positive result produces a brown solution when put in a water bath. Thus, Glucose,
Galactose, Fructose, Lactose and Xylose are positive on this test.

Molisch’s Test
It is a test for the presence of Carbohydrates. A positive reaction is indicated by
appearance of a purple ring at the interface between the acid and test layers. We obtained a
purple ring in all sugar, this is due to the formation of an unstable condensation product of beta-
naphthol with furfural (produced by the dehydration of the carbohydrate). Hence, all sugars are
all positive.

Seliwanoff’s Test
.Seliwanoff’s test 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 and if it contains an aldehyde group, it is an aldose. This test is
based on the fact that, when heated, ketoses are more rapidly dehydrated than aldoses. We
observed that after boiling the mixture, Glucose obtained a slightly beige color, Galactose
obtained a slightly cream color and Fructose obtained a bright red color, while Lactose and
Xylose remained clear, and Sucrose obtained light red. A positive reaction produces a red color,
so only Fructose and sucrose are positive for the test.

Barfoed’s Test
Barfoed’s test is similar to Fehling’s test, except that in Barfoed’s test, different types
of sugars react at different rates. Barfoed’s reagent is much milder than Fehling’s reagent.
Reducing monosaccharides react quickly with Barfoed’s reagent, but reducing disaccharides
react very slowly or not at all. Therefore, it is possible to distinguish between a reducing
monosaccharide and a reducing disaccharide using Barfoed’s reagent.
Glucose and fructose are both monosaccharides and reducing sugars. Therefore they formed
deep blue colouration in this reaction. Sucrose is a disaccharides however it is not a reducing
agent therefore there is no observed color change and the solutions remains blue.

Benedict’s Test
Benedict’s test is a semi quantitative test. This is also a test for the presence
of reducing sugars. The color formed depends upon the amount of reducing sugar
present in the mixture. So after boiling the mixture for 8 minutes, the Glucose,
Fructose, and Xylose produced a dark brown color, while Galactose and Lactose
produced light brown in color, but Sucrose remains its color. Thus, Sucrose is a
non reducing sugar which does not react with Benedicts reagent. The rest
obtained a positive results.

V. REFERENCES

https://en.wikipedia.org/wiki/Carbohydrate

http://people.umass.edu/~mcclemen/581Carbohydrates.html