CHEMLS103 | BIOCHEMISTRY MIDTERM

7 CONTENTS • Fiber is in vegetables, fruits, & whole

grain foods.
I. CARBOHYDRATES

II. Starch
• There are two different types of fiber --
CARBOHYDRATES
soluble and insoluble. Both are
WHAT ARE CARBOHYDRATES
Definition: important for health, digestion, and
preventing diseases.
a biological compound containing carbon,
hydrogen, and oxygen that is an important
source of food and energy Simple Carbohydrates:

Your body uses carbohydrates to make glucose

• Can be found naturally or as added
which is fuel that gives your body energy.
sugars

Glucose can be used immediately or stored.

• Added sugars have fewer nutrients
than foods with naturally-occurring
sugars
Healthier foods high in carbohydrates are ones
higher in dietary fiber without added sugar.

Carbohydrates can be found in the • Examples of ingredients as added

following: sugar: brown sugar, corn sweetener,
Fruits corn syrup, dextrose, fructose, fruit
juice concentrates, glucose, high
Vegetables fructose corn syrup, honey.
Breads, cereals, and other grains
Carbohydrates in foods occur in various
Milk & milk products
forms, including the following:
Foods containing added sugar Dietary fiber - a type of carbohydrate
that the body cannot
Two Types of Carbohydrates easily digest. It occurs naturally in
Complex Carbohydrates: fruits, vegetables, nuts,
seeds, beans, and whole grains.
• Starch & dietary fiber
• Starch is in certain vegetables like Total sugars - which include sugars that
potatoes, dry beans, cereals, and corn. occur naturally in
foods, such as dairy products, as well
as added sugars, which

are common in baked goods, sweets,
and desserts. The body
very easily digests and absorbs sugars.
Sugar alcohols - a type of carbohydrate
that the body does not
fully absorb. They have a sweet taste
and fewer calories than
sugar. Sugar alcohols are added to
foods as reduced-calorie
sweeteners, such as in chewing gum,
baked goods, and sweets.
Functions of Carbohydrates
The main function of carbohydrates is
to provide energy and food to the
body and to the nervous system.
Carbohydrates are known as one of the
basic components of food, including
sugars, starch, and fiber which are
abundantly found in grains, fruits and
milk products.
Carbohydrates are also known as
starch, simple sugars, complex
carbohydrates and so on.
It is also involved in fat metabolism
and prevents ketosis.
Inhibits the breakdown of proteins for
energy as they are the primary source
of energy.
An enzyme by name amylase assists in
the breakdown of starch into glucose,
finally to produce energy for
metabolism.
Sources of Carbohydrates
CHEMLS103 | BIOCHEMISTRY MIDTERM
1. Simple sugars are found in the
form of fructose in many fruits.
2. Galactose is present in all dairy
products.
3. Lactose is abundantly found in
milk and other dairy products.
4. Maltose is present in cereal,
beer, potatoes, processed
cheese, pasta, etc.
5. Sucrose is naturally obtained
from sugar and honey
containing small
amounts of vitamins and minerals.
Classification of Carbohydrates
The carbohydrates are further
classified into simple and
complex which is mainly based on their
chemical structure and
degree of polymerization.
1. Monosaccharides
Glucose is an example of a
carbohydrate monomer or
monosaccharide. Other examples of
monosaccharides include
mannose, galactose, fructose,
2. Disaccharides
Two monosaccharides combine to
form a disaccharide.
Examples of carbohydrates having two
monomers include-
Sucrose, Lactose, Maltose.
 CHEMLS103 | BIOCHEMISTRY MIDTERM

3. Oligosaccharides the molecule (mono, di, and

polysaccharides).
Carbohydrates formed by the
condensation of 2-9 monomers CHEMICAL TESTS FOR
are called oligosaccharides. By this CARBOHYDRATES
Numerous tests have been devised for
convention, trioses,
the determination of the properties
pentoses, hexoses are all
and for the
oligosaccharides.
differentiation of carbohydrates.

Carbohydrates Molisch Test

Carbohydrates are one of the three
principal classes of foods, and is a Seliwanoff Test
major source of
energy in our diets. Benedict Test

Carbohydrates are the major Fehling’s Test

constituents of plants, and are also
found in some animal
tissues. MOLISCH TEST
This is a general test for carbohydrates.
This test is based on the reaction of a
Carbohydrates are compounds
carbohydrate with concentrated
composed of carbon, hydrogen and
sulfuric acid to form a furfural or
oxygen containing an
hydroxyfurfural which
aldehyde or ketone functional group
then reacts with the Molisch reagent
and many alcohol groups, and can also
(α-naphthol) to yield a colored
be called polyhydroxy aldehydes or
product, generally a
polyhydroxy ketones. Molecules that
purple ring at the interface of the two
when hydrolyzed yield
layers.
polyhydroxy aldehydes or ketones are
also considered carbohydrate
PRINCIPLE OF TEST
In Molisch’s test, the carbohydrate (if
Carbohydrates can be characterized in
present) undergoes dehydration upon
a number of different ways based on
the
the number
introduction of concentrated
of carbons (pentose, hexose), the
hydrochloric or sulphuric acid, resulting
functional group (aldose, ketose),
in the
formation of rings
formation of an aldehyde. This
(furanose, pyranose), and isomers (D
aldehyde undergoes condensation
or L) and the number of
along with two
monosaccharide units in

phenol-type molecules (such as ∝-
naphthol, resorcinol, and thymol),
resulting
RESULTS
Positive reaction for Molisch’s test is
given by almost all carbohydrates
(exceptions include tetroses & trioses).
It can be noted that even some
glycoproteins and nucleic acid give
positive results for this test (since they
tend
to undergo hydrolysis when exposed to
strong mineral acids and form
monosaccharides).
SELIWANOFF TEST
This test distinguishes a ketohexose
from an aldohexose and disaccharides.
The test is
based on the fact that, when heated,
ketoses are more rapidly dehydrated
than aldoses.
The reaction between a ketohexose
and the Seliwanoff reagent (resorcinol
and dilute
HCl) occurs rapidly (within 1 minute) in
boiling water and results in the
formation of a
reddish colored precipitate. The color
intensifies with further heating. Other
carbohydrates and aldohexoses will
give a faint reddish color if the heating
is prolonged
due to the conversion of glucose to
fructose by the HCl or hydrolysis of a
disaccharide
or polysaccharide to yield fructose
PRINCIPLE OF TEST
CHEMLS103 | BIOCHEMISTRY MIDTERM
Seliwanoff’s test is used to distinguish
aldoses from ketoses. On treatment
with
concentrated Acid, ketoses are
dehydrated more rapidly to give
furfural derivatives and on
condensation with resorcinol give
cherry red complex. The test will be
answered by
fructose, sucrose and other keto
containing carbohydrates.
RESULT INTERPRETATION
Positive seliwanoff’s test: Fructose and
sucrose
Negative seliwanoff’s test: glucose,
distilled water
BENEDICTS TEST
This tests for reducing sugars. Free
aldehyde groups, α-hydroxy ketones
and hemiacetals
are capable of reducing metal ions in
solution. All common monosaccharides
are
reducing sugars. Some easily
hydrolyzed disaccharides and
polysaccharides will give a
positive test, while harder to hydrolyze
di- and poly sacchrides may initially
In the Benedicts test Cu +2 ions (blue in
solution) are reduced to Cu +1 ions
that form
Cu2O which is a brick-red precipitate.
Depending on the amount of
precipitate
produced, a positive Benedicts test can
appear green, yellow, orange, or red.
For more

information see Hein.
PRINCIPLE OF BENEDICT’S TEST
Benedict’s test is performed by heating
the reducing sugar solution with
Benedict‘s
reagent. The presence of the alkaline
sodium carbonate converts the sugar
into a
strong reducing agent called enediols.
During the reaction, enediols decrease
the
cupric particles (Cu2+) present in
Benedict’s reagent to cuprous particles
(Cu+)
which appear as red copper oxide
(Cu2O) which is insoluble in water.
When
Benedict’s reagent solution and
reducing sugars are heated together,
the solution
changes its color to orange-red/ brick
red precipitate. The red-colored
cuprous
oxide is insoluble in water and hence,
separate out from the solution. When
the
concentration of the reducing sugar is
high in the solution, then the color
becomes
more intense (reddish) and the volume
of the precipitate increases in the test
tube
making it clearly visible.
Benedict’s solution is a deep-blue
alkaline chemical reagent used to
test for the presence of the aldehyde
functional group -CHO which
consists of copper sulfate pentahydrate
(CuSO4. 5H2O), sodium
CHEMLS103 | BIOCHEMISTRY MIDTERM
carbonate (Na2CO3), sodium citrate
(Na3C6H5O7) and distilled water.
Sodium carbonate renders alkaline
conditions which are required for
the redox reaction, while sodium
citrate is a complexing agent which
complexes with the copper (II) ions to
avoid degradation into copper
(I) ions during storage.
FEHLING’S TEST
Fehling’s Solution (deep blue colored) is used
to determine the presence of reducing sugars
and aldehydes. Perform this test with fructose,
glucose, maltose and sucrose.
PRINCIPLE OF FEHLING’S TEST
Fehling’s test is one of the sensitive test for
detection of reducing sugars. Fehling’s
reagents comprises of two solution Fehling’s
solution A and solution B. Fehling’s
solution A is aqueous copper sulphate and
Fehling’s solution B is alkaline sodium
potassium tartarate ( Rochelle salt). Rochelle
salts (sodium potassium tartarate) present in
the reagent acts as the chelating agent in this
reaction.These two solution are mixed in
equal amount before test. On heating an
aldehyde or reducing sugar with Fehling’s
solution give reddish brown prepitate.
Formation of red precipitate of cuprous oxide
denotes the presence of reducing sugar.
RESULTS
Positive Fehling’s test: reddish brown ppt (
glucose, fructose, lactose)

Negative Fehling’s test: No red ppt (sucrose,
starch)
OTHER TEST
IODINE TEST
Iodine reacts with polysaccharides to give
colored complexes. Specifically amylose gives a
deep blue-black color, amylopectin, cellulose,
form red to brown colors, and glycogen
produces a reddish colored precipitate.
Monosaccharides and Disaccharides are two
small to trap the iodine molecules and do not
form dark colored complexes. The iodine test
can be used to distinguish between
mono/disaccharides and polysaccharides. It
may also be used to follow the course of a
hydrolysis reaction.
Iodine test is an indicator for the presence of
starch. Iodine solution (iodine dissolved in an
aqueous solution of potassium iodide) reacts
with starch producing a blue-black color. Apply
this test to all the polysaccharides provided.
BIAL TEST
This test is used to distinguish between
pentoses and hexoses. The Bial reagent
contains orcinol (5-methyl rescorcinol)
dissolved in concentrated HCl with a small
amount of iron (III) chloride present. When
mixed with a monosaccharide, pentoses are
converted to a furfural to yield a blue-green
product. Hexoses, with prolonged heating,
yield a muddy red precipitate.
The components of this reagent are resorcinol,
HCl, and
CHEMLS103 | BIOCHEMISTRY MIDTERM
ferric chloride. In this test, the pentose is
dehydrated to form furfural and the solution
turns bluish and a precipitate may form.
Perform this test with ribose and glucose.
BARFOEDS TEST
The Barfoeds test is similar to the Benedicts
test, but differs in the specific reagents
used (copper (II) acetate and acetic acid) which
is less reactive than the Benedicts
reagent. It is used to differentiate between
monosaccharides and disaccharides.
When boiled in water, monosaccharides will
react in under five minutes, while
disaccharides will take longer. If heated long
enough (10-15 minutes) disaccharides
will give a positive result due to hydrolysis to
form monosaccharides. As in the
Benedicts test above, the primary reaction is
the reduction of Cu +2 ions to Cu2O
which forms a brick red precipitate.
Barfoed’s reagent, cupric acetate in acetic acid,
is slightly acidic and is balanced so that is can
only be reduced by monosaccharides but not
less powerful reducing sugars. Disaccharides
may also react with this reagent, but the
reaction is much slower when compared to
monosaccharides. Perform this test with
glucose, maltose and sucrose.
SYNTHESIS AND DEGRADATION OF STARCH
STARCH
• is a polysaccharide made up of many
glucose residues bound by a-glycosidic
bonds. In the plant world, it serves as a
storage energy substrate.
 CHEMLS103 | BIOCHEMISTRY MIDTERM

• It consists of two fractions: amylose and Amylose Amylopectin

amylopectin. content in content in
• Starch is long chains of sugar molecules starch is starch is
linked together like a chain. about 20% about 80%
Straight Branched
chain structure
AMYLOSE structure
Has a 1-4 Has a 1-4
glycosidic glycosidic
• It is a component of starch forming linear Linkages linkages and
chain polymers of D-glucose units. a 1-6
glycosidic
Amylose accounts for 20% of starch and is
linkages
insoluble in cold water.
Less soluble More soluble
• It has many uses in cooking and is also in water in water
used in some other industries Gives a dark Gives a
• Water Soluble blue/black reddish
color when brown color
iodine when iodine
AMYLOPECTIN solution is solution is
added added
• It is a component of starch Can be Cannot be
forming branched-chain hydrolyzed hydrolyzed
with a with a
polymers of D-glucose units.
amylase and amylase and
ß amylase ß amylase
• It has both a 1-4 and a 1-6 enzymes enzymes
glycosidic linkages and completely completely
accounts for 80% of starch Forms a gel Forms a gel
when hot when hot
• Pure amylopectin is readily water is water is
soluble in cold water. added Does added
not form a
• It can be readily digested by gel when
the human body which hot water is
added
contributes to its positive as
well as negative aspects.

AMYLOS CONCLUSION
VERSUS
AMYLOPECTIN • Amylose and amylopectin are two types
Amylose is a Amylopectin of polysaccharides that can be found in
straight is a branched starch granules. They have both
chain chain structural and chemical differences as
polymer of polymer of D- well as similarities. The main difference
D- glucose glucose units
between amylose and amylopectin is
units
 CHEMLS103 | BIOCHEMISTRY MIDTERM

that amylose is a straight chain polymer

whereas amylopectin is a branched
chain polymer.
TWO MAIN PROCESS IN
DEGRADATION OF STARCH
• Amylopectin is less soluble in water.
But amylopectin is soluble in hot water
• The initial step in random degradation
with swelling. It can from starch gel or
is the splitting of large chains into
paste when it cools down.
various smaller sized segments.

• This drastically reduces the viscosity of

SIMILARITIES BETWEEN AMYLOSE AND gelatinized starch solution, resulting in
AMYLOPECTIN Liquefaction - thinning of the solution.

• Both are polvsaccharide molecules.

• The final stages of degradation are
• Both are composed of D-glucose units. mainly the formation of mono-, di-, and
tri-saccharides - called as
• Both molecules have a 1-4 glycosidic Saccharification, due to the formation
linkages. of saccharides.
❖ Eg: Bacterial alpha-amylase - randomly
• Both types are found in starch attacks only the alpha-1,4 bonds-
granules. belongs to the liquefying category. ~

❖ The fungal alpha-amylase belongs to

CHEMICAL TESTS
the saccharifying category.

• lodine solution is used to test for

FACTORS AFFECTNG BIODEGRADATION
starch; a dark blue color indicates the
* Light, Water and 02(if aerobic)
presence of starch
* Environmental factor(physical and chemical)

* Temperature (Thermophiles, Mesophils)

AMYLASE
* Nutritional factor
• Starch molecules are broken down by
enzyme known as amylase.

Starch is a major glucose storage compound

• Bacteria and fungi secrete amylases for
extracellular digestion. When they have present in plants. It belongs to the category
broken down the insoluble starch, the
soluble end products such as (glucose of biopolymers and is polysaccharide in
or maltose) are absorbed into their
cells. nature. It is made up of thousands of

repeating units of glucose.

 CHEMLS103 | BIOCHEMISTRY MIDTERM

It cannot be synthesized in animals Amylose starch is a linear chain of alpha D-

glucose subunits that usually contains around
STRUCTURE 300 to 3000 glucosyl residues or even more.

AMYLOPECTIN
It is a polymer of glucose molecules. It is

present in plants in the form of starch

granules.
It is also called as amylopectin starch.
Starch is made up two components both of Amylopectin makes around 70-80% of starch. It
is a branched
which have different structures:
polymer of alpha D-glucose subunits that are
linked via the same 1-4 glycosidic bonds as in
Amylose amylose.

However, amylopectin also has alpha 1-6

glycosidic
Amylopectin
bonds among the glucose molecules at the
branch
AMYLOSE points.

PROPERTIES

It is also sometimes called amylose starch. It is

a
The amylose starch have following properties
polysaccharide made up of alpha-D-glucose
monomers.

Amylose makes around 20-30% of starch. It is Amylose is more crystalline in nature

an

unbranched compound having linear chains of

It is insoluble in cold water
glucose

molecules. these glucose molecules are linked

via alpha 1-4 Amylose is more resistant to digestion due to
lack of
glycosidic bonds.
 CHEMLS103 | BIOCHEMISTRY MIDTERM

branching

It gives blue color with iodine solution

Amylose decreases the gel strength of starch

IMPORTANCE

The properties of amylopectin are as follows

Plants
It dissolves in cold as well as hot water

Amylopectin is readily digestible due to the In plants, starch acts as the main energy
presence storage
of extensive branching compound. They store excess glucose during

daytime in the form of starch and use it as an

It increases the gel strength and solubility of energy source during the night.
starch as

mole water molecules can infiltrate due to

branching It provides energy to the embryo.

THE COLLECTIVE PROPERTIES OF STARCH ANIMALS

It undergoes hydrolysis to yield constituent Starch is the primary source of carbohydrates

sugars for

animals.

It becomes insoluble in water and forms gel

It provides energy to the animals.

It undergoes decomposition upon heating and

forms
It provides carbon skeleton to the animals that
dextrin can be
 CHEMLS103 | BIOCHEMISTRY MIDTERM

used in the synthesis of various other

compounds in
Paper Industry
their bodies.

HUMANS Starch is used to increase the strength of paper

It is used as a bounding powder for boards

Like other animals, it is the major
carbohydrate

source for humans. It is used in glues and adhesives to increase

their

binding strength
It is present in the human diet in the form of

grains, cereals, rice, potatoes, etc.

It is used in bioplastic production

FOOD INDUSTRY
The biosynthesis of starch is coupled with
the Krebs cycle and involves the following
steps

Starch is used as a sweetening agent in

beverages

Synthesis of ADP-glucose

It is used as a bulking agent for fairy products

Elongation of chain by starch synthase

It is used in ice creams as it lowers their

freezing point
Introduction of branches by starch branching
enzymes

It gives brown effect to caramels Starch is degraded in plants during the

night to obtain glucose for energy

It is added as a preservative in jams Phosphorylation

 CHEMLS103 | BIOCHEMISTRY MIDTERM

Reserve starch, an energy storage for

germination, a major component
Release of maltose by the action of Amylase
enzyme of food and feed, and an industrial starting
material, is formed in

amyloplasts.
Joining of maltotriose and degradation of the
resulting

compound

Release of phosphorylated residues

FACTS ABOUT STARCH

Starch, a polysaccharide , is synthesized in the
chloroplast.

Starch formation occurs in plastids.

Starch granules are classified as transitory or

reserve.

Transitory starch granules accumulate for only

a short period of time

before they are degraded.

Starch forms in leaf chloroplasts during the

day.

hydrolyzed and transported to other plant

parts at night as simple sugar.