Experiment 4 : Test for

carbohydrates
A) Physical tests

1) Solubility of / arabinose / fructose / glucose / sucrose / starch / dextrin /

|ArabinoseFructoseGlucoseSucroseStarchDextr#

o%Fclfsdubieefsoubeef.name/Insoube
H2O soluble soluble soluble soluble Insoluble Insoluble

Alcohol Insoluble Insoluble Insoluble Insoluble Insoluble Insoluble

l
WATER 10% NaCl

Arabinos Fructose Glucose Sucrose Starch Dextrin

Arabinos Fructose Glucose Sucrose Starch Dextrin

Ethanol
Saccharides are e mostly
polar molecules .

Water is polar and therefore

dissolves the saccharides

90% Water which

10% Nacl has

dissolves saccharides

Alcohol is mostly non

-

polar
Arabinos Fructose Glucose Sucrose Starch Dextrin because of long carbon

chains and therefore cannot

dissolve the saccharides

Polysaccharides are hydrophobic and

cannot easily dissolve .

2) Dialysis using 0.1M Glucose ,
o.IM sucrose ,
o >
.
% starch .

Set -

up
"

semipermeable theaopethiengoemitienmeabfeetimuemebranaen
.q-
. are wins
rubber band .

*
Dip the test tube into a beaker 1 flask with water .

The of test tube must be

dipped in first
opening the

Maisch test for the dialysate ( the solution inside the beaker / flask )

AddzdropsmolischreagentCAlpha-NapthollMBIMLConc.AZ
Violet rings are produced in o.IM Glucose and o -
l H sucrose
but not in o . > % starch ( lighter intensity than normal molisch .

Glucose sucrose the

and
pass through semi -

permeable membrane
the
and into water but starch is a
big molecule and does

not pass through the semi -

permeable membrane .

0.7% Starch 0.1M Sucrose 0.1M Glucose

Dialysate Dialysate Dialysate
Molisch Molisch Molisch
Conc. H2SO4 Conc. H2SO4 Conc. H2SO4
B Furfural
.
reactions of carbohydrates

1) Motion Test ( Alpha -

Napthol reaction ) -

General test for Carbohydrates

AddzdropsmolischreagentCAlpha-NapthollBBIMLConc.AZ

0.02 M 0.02M 0.05%

Water Glucose Sucrose Starch
Molisch Molisch Molisch Molisch
Conc. H2SO4 Conc. H2SO4 Conc. H2SO4 Conc. H2S04

* Yiaglbothygirnaqseareproducedinallsolutionscontaini.no
Lp
. Due to the dehydration of the carbohydrate by H2S 04 to form

↳ furfural which

Reaction occurs
condenses

for
with molisch

carbohydrates
reagent to form purple condensation

larger than tetroses .

(4 C atoms )

0
pH

K¥ #o
Ho
1-1
it
②② Cio Hiott
Furfural 5
hydroxy methyl furfural
a Nap thot
-

2- Fur aldehyde
at

1
Molisch reagent .

"
from
Furfural is dehydration pentose :
from hexose
a

'

Hydroxy methyl
furfural is dehydration

and forms
hapthol furfural a
molecules of react with either
z
double bond Oxygen of furfural
bond by breaking
.
2) Thymol Test

3-6drops3%thymolinalcoholB5-IomLconc.HolM2minBc

0.02M 0.02M 0.05%

Glucose Sucrose Starch
3% Thymol 3% Thymol 3% Thymol
Conc. HCl Conc. HCl Conc. HCl
Boil Boil Boil

¥%%I:E%miii:rEauan
The solution will turn to a Carmine red color

Starch * Carmine red ( lighter )

-

4am Due to formation of furfural with the addition of Hd and

heat .
Fural will condense with thymol and form a more stable
structure than in Molisch 's .

40 Reaction occurs for carbohydrates larger than tetrose .

OH CAS

O / GHz

Potato
11
At
Furfural
HOT it,
His OH
g
hydroxy methyl furfural
-

5- Methyl
Furaldehyde ( pro pan y 1) phenol
2- -2 -2 -

thymol
forms
molecules of thymol react with either furfural and a
z
double bond Oxygen of furfural
bond by breaking
.

Thymol is more stable than d -

napthol due to its

of not volatile f?)

nature being
3) Seliwanoff 's Reaction ( Resorcin -

HCl tests )

intuitionally
5mLSeliwanoff'sreagentBwaterbath

Water 0.01M 0.01M 0.01M

Glucose Fructose Arabinose
Seliwanoff’s Seliwanoff’s Seliwanoff’s Seliwanoff’s
Boil Boil Boil Boil

:÷:÷÷÷÷:→.
The seliwanoff reagent is a mix of resorcinol and HCl

Resorcinol will react with ketose ( a carbohydrate containing

a ketone )

¥÷÷÷÷:÷÷÷÷÷÷÷i¥i÷÷÷
"

Utz

④
R -

c -

H K -

C -
Ho OH
" Il I

O O
OH Cee Hak
Aldose ketose Resorcinol

Fructose is a ketose and when dehydrated form

5 hydroxy methyl furfural which react with Resorcinol in 2 minutes
-

Ketoses undergo dehydration at a faster rate than aldoses .

④ '
CHO

Furfural
2-
Furaldehydeo
11

HO
TH
-11
5-
hydroxy methyl furfural
4) Moore's Test ( Action of concentrated alkali )

la 1mL
#
5% glucose and 1mL conc . NaOH and boil .

4• Yellow 1
orange
/ dark brown color
Also caramel color

This is due to reducing sugars except sucrose

Reducing sugar is a sugar with a free aldehyde or ketone

All monosaccharides are reducing sugars .

Reducing sugar is a reducing agent ,

therefore it loses Itt or electron

during redox reaction .

On
Based on the liberation of aldehydes which polymerize to form a

resinous substance ,
caramel .

Positive :
Glucose , galactose ,
maltose ,
fructose ,
lactose
Negative : sucrose ,
glycogen ,
starch

to aldoses with a base so ketoses

ketoses transform ,

caramel odor and color

will also give
.

ALDO -
KETO ISOMERISM

ftp.
5% Glucose
Conc. NaOH
Direct Boiling
5) Token 's Phloroglucin reaction

5mLPhlorogluoinSolutionBwaterbathinboilingHz

Water 0.01M 0.01M 0.01M

Phloroglucin Glucose Fructose Arabinose
Boil Phloroglucin Phloroglucin Phloroglucin
Boil Boil Boil

¥rYa%÷oiieiwisn9iown9oiEr%neex%e
Positive test if the color is betadine -
like .

Water -

light Yellow color

Arabinose reddish -

brown color

th Phlorogluoin behaves as a ketone ; differentiates pentose

from hexose .

OH

ca Ha Oz I
Phlorogluoinol
*
✓
Benzene
-
I
, 3,5
-

trio I
1315 tri
hydroxy benzene
-

OH
µ
O

④ '
CHO Ho
-11
it

Furfural
5
hydroxy methyl furfural
-

2- Furaldehyde

to Furfural which red

Pentoses denigrate gives a

color or ppt #
with Phloroglucinol
4. Phloroglucide
 SACCHARIDES USED IN 4 35

Reducing Monosaccharide Fructose ketohexose

-
-

Glucose -

Aldohexose
-

Galactose -

Aldo hexose -

isomer of Glucose
-

Arabinose -

Aldo pentose

Reducing Di saccharide Maltose ( 2 Glucose )

"

-
-

Lactose ( Galactose be Glucose )

Non -

Reducing Di -

saccharide -

Sucrose ( Glucose be Fructose )

4. can
undergo hydrolysis to produce component
sugars

Polysaccharide :
Starch
Dextrin