Carbohydrates

Topics
Sugars: Glucose & maltose

Carbohydrate digestion

Carbohydrate structure and function

Colorimetric determination of glucose

Carbohydrates
Carbohydrates are contained in stodgy foods in the diet e.g. bread,
pasta and rice.
Carbohydrate is comprised of Carbon(C), Hydrogen(H) and
Oxygen(O).
The basic building blocks of carbohydrates are monosaccharides.
Monosaccharides all have the same basic formula (CH2O)n.

One common monosaccharide, is

glucose with a formula of C6H12O6

Glucose structures
Glucose can exist as an open chain form, but generally exists
as a ring form.
CH2OH

C OH

OH

HO

OH

C
O

The 6-membered ring

contains an oxygen
atom

CH2OH
H

O
H

HO OH

H OH
OH

a-glucose

The chemical groups

shown in green, are
above the plane of the
ring

Alpha and beta-glucose

6CH OH
2
5
O
H
H
H
4
1
H OH
HO OH
2
3
OH
H
a-glucose

CH2OH
O
H
HO OH
H

OH

H H
OH

b-glucose

In position 1 the alpha prefix refers to the hydroxyl (OH) being below
the plane of the ring.
The carbon atoms are numbered, as shown, in green, this is to help
describe how the sugar units can combine.

Maltose and amylose

Maltose is a disaccharide of glucose.
Many monosaccharides linked
together result in a polysaccharide.
Amylose is a glucose polysaccharide.

maltose

The bond formed is called a

glycosidic bond i.e. a bond between
2 sugar units.
amylose

Formation of di- and ploysaccharides

The glucose units link together by a condensation reaction, i.e. water is formed
as a product.
CH2OH
O
H
H
H

CH2OH
O
H
H
H

HO OH
H

H OH

HO OH

OH

H OH

OH

glucose + glucose

Condensation Reaction
CH2OH
O
H
H
H
HO OH
H

H
OH

CH2OH
O
H
H
H
O

OH

H OH

OH

maltose + water

+ H2O

Reducing sugars
Glucose, fructose (found in fruit), maltose and galactose are
reducing sugars.
When heated with Benedicts solution an orange precipitate forms.
Cu(II) ions in Benedicts solution are reduced to Cu(I) ions.

Sucrose is a non-reducing sugar and does

not give an orange precipitate when
heated with Benedicts solution.

Heating sucrose with dilute acid breaks it

down into two monosaccharides (alphaglucose and fructose), which are reducing
sugars which will then give a positive
result with Benedicts solution.

Carbohydrate digestion
Glucose, used in respiration, comes from the carbohydrates we
ingest and digest.
Glucose can be absorbed rapidly in the small intestine but starch is
too large a molecule and must be broken down into smaller units
(digested) by hydrolytic enzymes.
starch

amylase

maltose

maltase

glucose

Structure of Starch

Starch consists of amylose and

amylopectin.
Amylose is a polysaccharide. It
consists of glucose linked together
linearly with a-1,4 linkages.

amylose

amylopectin

a-1,4 linkages means that the linkage is from carbon number 1 on the first
glucose to carbon number 4 on the next glucose unit. a reflects the position of
the hydroxyl on the free carbon 1 at the terminal.
CH2OH
O
HH
H
HO OH
H

H
OH

CH2OH
O
H H
H

CH2OH
O
H
H
H
O

OH

OH

OH

H O

OH

More
glucose
molecules

Alpha-amylase and Maltase

Alpha-amylase and
maltase are enzymes that
break down amylose and
maltose respectively.
They work on different
substrates, but both
hydrolyse the a-1,4
linkages that hold the
glucose units together.

CH2OH
O
H
H
H
HO OH
H

CH2OH

H
O

OH

CH2OH
O
H
H
H

Hydrolysis reflects the fact

that an OH and H group
HO OH
have been added.

H OH
OH

O
H

Either OH for
maltose or
more glucose
units for
amylose

OH

OH

CH2OH
H
+

O
H

HO OH

H OH
OH

Structure of Cellulose
Cellulose is a linear, unbranched polysaccharide.
It consists of glucose, joined by beta-1,4 linkages.
300-15,000 glucose molecules per cellulose
molecule and forms parallel hydrogen bonded
microfibrils.
CH2OH
O
H
OH
H
HO OH
H

The linkages are formed by a condensation

reaction

H H
OH

More glucose molecules

O
O

Starch v cellulose comparison

Starch

Cellulose

Monosaccharide unit

glucose

glucose

Linkage

alpha-1,4

beta,1-4

Function

Energy store in
plant tissue

Structural role in plant

tissue

Hydrolysis in humans?

Yes,by the
enzyme aamylase

No

Structure of Glycogen

Glycogen is a very large, branched polymer

of glucose.
There is an a-1,4 linkage between two
glucose units and an a-1,6 linkage at the
branch points. Branch points occur at
approximately 1 residue in every 10.

CH2OH
H

O
H

HO OH
H

H OH
OH

Carbohydrate structure: function

Category
Monosaccharide
(made of 1sugar
molecule)

Example
glucose
fructose
galactose

Site
fruit
fruit, nectar
milk

Structure
H

OH

OH

a-glucose
Disaccharide
(made of 2
monosaccharides
joined together)

maltose = a-glucose + a-glucose

sucrose = glucose
+ fructose
lactose = glucose
+ galactose

germinating
seeds
phloem tissue,
fruit and milk

b-glucose

fructose

O
maltose

Polysaccharide
(made of many
monosaccharides
joined together)

starch
glycogen
cellulose
chitin

=
=
=
=

polymer of glucose
polymer of a-glucose
polymer of b-glucose
polymer of
glucosamine
(glucose with an
amino acid attached)

chloroplast
stroma
muscle cells
plant cell wall
exoskeleton of
arthropods

O
O
cellulose

Carbohydrate structure: function

Carbohydrate

Structure

Structure: Function

Starch
Is the main storage
polysaccharide
in plants.

Made of two polymers of a-glucose; amylose

and amylopectin

Insoluble in water, therefore good storage compound

e.g. in stroma of chloroplasts
Amylose helix forms a compact shape allows tight
packing
Amylopectin has many protruding ends (glucose
molecules) which can be hydrolysed rapidly allows
rapid release of glucose to provide energy via
respiration.

amylose

amylopectin

Glycogen
Is the main storage
polysaccharide
of animal and
fungal cells.
Cellulose
Is the structural
polysaccharide
in plants.

Similar structure to amylopectin (in that it is a

polymer of a-glucose) of starch but has many
more branches and the branches are shorter.
Glycogen is even more compact than
amylopectin.

Compact structure allows faster hydrolysis than starch animals may need emergency glucose faster
than plants.

Hydrogen bonding prevents water entering the molecule,

therefore resistant to enzyme hydrolysis which makes it
an excellent structural polysaccharide.
The long, unbranched fibrous structure provides great
mechanical strength.

Carbohydrate summary
The structural formula for simple carbohydrates may be written
(CH2O)n.
Glucose is a monosaccharide that can be used as an energy
source.
Glucose is stored as a polymer - glycogen in animals, starch in
plants.
Maltose is a disaccharide of glucose linked by a glycosidic bond.
Glycosidic linkages are formed by condensation reactions, and
broken by hydrolysis reactions.

Colorimetric determination of glucose by

the neocuproine assay (1)
Safety
Gloves, glasses and lab.coats should be worn.
Care should be taken handling all chemicals. Neopuroine hydrochloride is classed as
an irritant, cupric sulphate pentahydrate is classed as harmful if swallowed and an
irritant, sodium carbonate is irritating to eyes.
Be aware of local disposal rules. Cupric sulphate pentahydrate is toxic to the aquatic
environment.

Sensitivity: ~20ng 2mg glucose in 200ml (approximately 0.5 50mM)

Reagents
(A)Dissolve 40g of anhydrous sodium carbonate, 16g glycine and 450mg cupric sulphate
pentahydrate in 600ml of water and make up to 1 litre with more water. This solution is
stable.
(B)Dissolve 0.15g neocuproine hydrochloride in 100ml water.
This solution is stable for months if stored in a dark bottle.
Standard solution 50mM glucose

Colorimetric determination of glucose by

the neocuproine assay (2)
Example Set-up
Test-tube

Glucose
standard (ml)

Water
(ml)

Control - 0mM

200

Standard- 10mM

40

160

Standard- 20mM

80

120

Standard- 30mM

120

80

Standard- 40mM

160

40

Standard- 50mM

200

unknown

200-X

Tubes should be set-up in triplicate

Colorimetric determination of glucose by

the neocuproine assay (3)
Experimental
1.
2.
3.
4.
5.
6.
7.
8.

9.

Set up test-tubes as shown above.

Add 400ml Reagent A and 400ml Reagent B to each tube, mix well.
Heat the solutions at 100C for exactly 12 minutes and cool rapidly to room
temperature ( by putting in an ice-bath for example).
Add 1.0ml of water to each tube, washing down any condensate formed, mix well.
Read the absorbance at 450nm on a spectrophotometer.
Average the results, and subtract the absorbance for the control sample from all other
samples ( both standards and unknown).
Plot the resulting absorbance of the standards (y-axis) against concentration of
glucose (x-axis).
Use the graph to calculate the concentration of glucose in the unknown sample.
i.e. Unknown has the same absorbance as 20mM glucose standard, however 100ml
unknown was added along with 100ml water in the set-up. This would give a glucose
concentration of 20 X (dilution factor of 2) =40mM.
If the unknown sample lies outwith the absorbance covered by the standard samples,
then the experiment must be repeated. The unknown sample can be diluted, or the
standard curve extended (in this example down to 1mM) so the unknown sample will
come within the absorbance range.

Colorimetric determination of glucose by

the neocuproine assay (4)
Notes:
1. If a precipitate forms during the final cooling it may be simply
re-dissolved by slight warming.
2. Non-carbohydrate reducing agents interfere in this assay.
Description of mechanism
The copper (2+)ion is reduced to the copper (+) ion state in the
presence of a reducing sugar. The neocuproine binds the copper (+) ion
and results in colour formation.

Quiz
Which of the following is a non-reducing sugar?
?

lactose

sucrose

galactose

fructose

glucose

The name of the bond joining two glucose molecules in maltose is:
glycosidic

Quiz
Starch is composed of?
?

amylose and glycogen

glucose and amylopectin

a polysaccharide and a disaccharide

amylose and amylopectin

True or false:
Amylase is a protein and the hydrolytic enzyme responsible for digesting starch? True
Glucose is soluble and is therefore stored in the human body as starch? False
The formation of the disaccharide maltose involves a condensation reaction? True