Reactions of Monosaccharides

1. Mutarotation :-
When an aldohexose is first dissolved in water and
the polarised light is passed through the solution the initial
optical rotation shown by the sugar gradually changes until
a constant fixed rotation characteristic of the sugar is
reached. This phenomenon of change of rotation ic called as
mutarotation.
When D- Glucose is crystallised from water or dilute
alcohol at room temperature , α-D-Glucose separates &
when its fresh solution is made , its specific rotation of
polarised light will be +112degree initially ,but after 12-18
hrs it changes to a fixed rotation of +52.5 degree.
When D-Glucose is crystallised from water at 98 degree
Celsius β-D-Glucose separates & when its solution is made is
made, its initial rotation will be +19 degree which within
few hours changes to a fixed rotation of +52.5 degree.
This change in the optical rotation is due conversion of
α-form into a equilibrium mixture of α & β – form. This
means that glucose existing in isomeric form in solution
changes into the same equilibrium mixture regardless of
which form is dissolved. ie,in glucose solution at equilibrium
2/3rd of the sugar exist as the β-form & 1/3rd as α-form.
2. Reaction with acids :-
Polysaccharides and the compound carbohydrates
when boiled with dilute mineral acid such as HCL & H2SO4
gets hydrolysed into their constituent monosaccharides.
Monosaccharides when treated with conc. H2SO4
undergoes dehydration with removal of 3 H2O molecules &
form furfural derivatives. ie, pentoses with 12%HCL with
release of 3 H2O molecules form furfural and the hexose
with conc. H2SO4 or HCL with removal of 3 H2O molecules
form hydroxymethyl furfural.
The furfural derivatives with certain phenolic compounds
condense to form coloured products (compounds with
characteristic colours). Thus it forms the basis for certain
test used for detection of sugars.
Example :-
1. Molisch’s test : with α-naphthol in alchoholic
solution sugars gives purple or red violet ring.
This is a sensitive test not a specific test. All
sugars gives the + test. So it is used as a general
test for carbohydrates.

2. Seliwanoff’s test : with resorcinol gives a cherry

red colour. This is characteristic of D-fructose.
3 . Reaction with alkalies :-
With alkalies monosaccharides reacts in various ways.
a) In mild alkaline medium the sugar changes to cyclic α
& β forms with an equilibrium between the two
isomeric forms. On standing rearrangement occurs &
an equilibrated mixture of glucose , fructose &
mannose. This is because in weak alkaline solution
they are all interconvertible . It is because all three
sugars give the same enediol which tautomerises to all
three sugars. That is in mild alkaline solution the +H
atom attached to the second carbon atom shifts to the
first carbon atom & result in the formation of double
bond b/w 1 &2 carbon atom (ene group) and to these
double bonded carbon atoms two hydroxyl groups are
attached (diol). This compound is called enediol. The
rearrangement of +H atom of enediol form give the
isomers of the original sugar . The interconversion of
sugars through a common enediol form is called Lobry
de Bruyen-Van Ekenstein transformation .
The enediols are highly reactive so they are
easily oxidised by O2 & other oxidising agents to form
sugar acids by breaking at the double bond . That is
they readily reduce oxidising ions such as Cu++,
Hg+,Bi+++,Ag+ & Fe(CN)6---. This reducing action sugar
in alkaline medium is used for both qualitative &
quantitative determination of sugars. Reagents
 containing Cu++ ion is used. Benedict’s reaction used to
detect the presence of sugar in urine is based on this
action. ie, when sugar solution is boiled with Benedict’s
reagent containing sodium carbonate, sodium citrate
& copper sulphate, sugar gets converted in to enediols,
the Cu++ ions take electrons from enediol &oxidise
into sugar acid & in turn gets reduced to Cu+ ions, this
in turn combine with the OH- ions to form yellow
cuprous hydroxide which on heating gets converted to
red cuprous oxide. The appearance of yellow to red ppt
indicates reduction & the quantity of sugar present can
be roughly estimated from colour & amount of ppt .
Any sugar with free Aldehyde or Keto group reduce the
Benedict’s reagent. As Glucose is a reducing sugar it
gives + test.
b) In conc. Alkali the sugars caramelises & gives various
decomposition products ,yellow &brown pigments ,
salts may be formed, many double bond b/w c-atoms
are formed & C to C bonds may rupture.
4.OSAZONE FORMATION :-
When a mixture of phenylhydrazinehydrochloride &
sodium acetate is added to a sugar solution and heated in a
boiling waterbath for 30mt to 45mt and then cooled slowly
by itself, crystals are formed. They are called the osazone. It
is a useful means of preparing crystalline derivatives of
sugars. All reducing sugar form osazones with excess
phenylhydrazine at boiling temperature.
Osazones are

 Insoluble
 Have characteristic melting points
 Have characteristic crystal form or structure
 Have characteristic precipitation time.
Thus osazones are valuable in identification of sugars.
3 steps are involved in the osazone formation
a. Aldohexose or ketose reacts with phenylhydrazine to
form phenylhydrozone with release of a water
molecule.
b. Phenylhydrozone then react with a molecule of
phenylhydrazine to form intermediate Keto –
derivative with release of ammonia molecule &
aniline.
c. This intermediate Keto – derivative then reacts with
another molecule of phenylhydrazine and form
osazone with release of H2O molecule.
The reaction involves the carbonyl carbon (aldehyde
or ketone group) & the adjacent carbon.
Each sugar will have characteristic crystal form of
osazones.
 Glucosazone are fine , yellow needle shaped
crystals arranged like a broom, aggregates or
 sheaves or crosses . It is typically described as
bundle of hay. MP is 204 to 205degree Celsius.
The glucose , fructose & mannose form the same
osazone crystals.
 Lactosazone crystals are irregular clusters of
needle and look like a powder puff.ie, hedgehog
or pincushion with pins or flower of touch –me-
not- plant.
 Maltosazone are star shaped , sunflower shaped
or petal shaped crystals.
Osazones are used to differentiate sugars in
biological fluid like urine.

5. Oxidation of sugars:-
When oxidised under different conditions , the
Aldoses form
a) Monobasic Aldonic acids or
b) Dibasic saccharic acids or
c) Monobasic Uronic acids containing aldehyde groups
When an aldose is oxidised with the hypobromous
acid (HOBr,Br2/H2O) the aldehyde group gets
converted into a carboxyl group and the aldonic acid
is formed . Thus glucose is oxidised to gluconic acid,
mannose to mannonic acid & galactose to galactonic
acid.
When aldoses is heated with conc.HNO3 under
proper conditions the first & last carbon atoms
simultaneously oxidise to form dicarboxylic acids
known as saccharic or aldaric acids .ie, both aldehyde
& primary alcohol group is converted into carboxylic
groups (-COOH) . Glucose is oxidised to glucosaccharic
acid, mannose to mannaric acid & galactose to mucic
acid. The mucic acid forms insoluble crystals & is the
basis for a test for identification of galactose.
When aldehyde group of aldose is protected and
the molecule is oxidised the last carbon becomes
COOH group ie, the primary alcohol group is converted
to _COOH group, without oxidation of aldehyde group
and uronic acid is produced . Due to the presence of
free _CHO group they exert reducing action. The
glucose is oxidised to glucuronic acid , mannose to
mannuronic acid & galactose to galacturonic acid. The
glucuronic acid is formed in the liver by uronic acid
pathway, an alternative pathway for glucose
oxidation. The glucuronic acid is used by the body for
conjugation with insoluble molecules to make them
soluble for detoxification purpose ie,the glucuronic
acid conjugates with toxic substances, drugs,
hormones, even bilirubin & converts them to a soluble
nontoxic substance, a glucuronide, which is excreted in
urine & also for synthesis of heteropolysaccharides.
 6. Reduction of sugars:-
When treated with reducing agents such as
sodium amalgam , hydrogen reduce the sugars to alcohol.
Aldose forms corresponding alcohol & ketose forms two
alcohols because of appearance of a new asymmetric
carbon atom in the process.eg:-

 Glucose is reduced to sorbitol

 Mannose to mannitol
 Galactose to dulcitol
 Ribose to ribitol
 Fructose to sorbitol & mannitol
Sorbitol, mannitol & dulcitol are used to identify
bacterial colonies. Mannitol is also used to reduce
intracranial tension by forced diuresis . The sorbitol &
dulcitol when accumulate in abnormal amount produces
changes in the tissues by osmotic effect. Eg:- cataract of
lens.

7. Formation of esters or acetylation:-

Due to the presence of alcoholic group -OH groups ,
the sugars react with anhydrides & chlorides of many
organic & inorganic acids like acetic acid , phosphoric acids,
sulphuric acid & benzoic acids to form esters of
corresponding acids.ie,esterified to form acetates,
phosphates,sulphates,benzoates etc. Sugar phosphates are
of great biological Importance. Metabolism of sugars inside
the body starts with Phosphorylation . Glucose-6-phosphate
& Glucose -1-phosphate are important intermediaries of
glucose metabolism.

8. Forms iodo –compounds:-

An aldose when heated with conc.HI it loses all of its
oxygen and is converted into an iodo-compound.
Conc. HI
Glucose → Iodohexane (C6H13I)
As the resulting compound is a straight chain compound
relating to normal hexane , it means that there is’nt any
branched chains in structure of glucose.

9. Glycoside formation:-
Glycosides are compounds containing a
carbohydrate & a non carbohydrate group. It is formed
when the hemi- acetal group ( hydroxyl group of anomeric
carbon ) of a monasaccharide condenses with an alcohol or
phenol group . The non carbohydrate group is called
aglycone. The aglycone may be methyl alcohol, glycerol,
phenol, adenine, sterols, hydroquinones, anthraquinones.
The glycosides are named according to the carbohydrate
present in them. If contains glucose called glucoside ; if
galactose called galactoside and so on. Glycoside do not
reduce Benedict’s reagent because the sugar group is
masked. But if hydrolysed by boiling with dilute acid the
sugar group become free & can then reduce copper. Αlpha –
glycosides are hydrolysed by maltase from yeast and β –
glycosides by Emulsin from almonds. Thus α & β forms can
be distinguished by enzyme hydrolysis.
Examples of glycosides of biomedical importance :-
Glycosides are present in many drugs, spices and in
the constituents of animal tissues. They are widely
distributed in the plant kingdom.
a) Phlorhizin (glucose + phloretin) → obtained from rose
bark & root & bark of apple tree. It blocks the
transport of sugar across the mucosal cells of small
intestine & also renal tubular epithelium; it displaces
Na+ from the binding site of carrier protein and
prevents the binding of sugar molecule and produces
glycosuria. It produces renal damage in animal.
b) Digitonin (4 galactose + xylose + digitogenin, a steroid)
→ obtained from leaves of foxglove. It is a cardiac
stimulant & used in cardiac insufficiency.
c) Plant indicant (glucose + indoxyl ) → obtained from
leaves of indigofera is used as stain.
d) Ouabain → obtained from strophanthus species is a
sodium pump inhibitor. It inhibits active transport of
Na+ in cardiac muscle.
Other monosaccharides of biological
importance:-
1) Deoxy sugars:-
They represents the sugars in which the
oxygen of the hydroxyl group are removed. Thus
–CHOH or –CH2OH becomes –CH2 or –CH3.
Important are
 2-deoxy –D-ribose → found in the DNA
 6-deoxy-L-galactose or L-fucose found as
constituent of glycoproteins, blood group
antigens & bacterial polysaccharides.
The deoxy sugars will not reduce & will not form
osazone. Fuelgen staining is specific for 2-deoxy sugars in
tissues. So for the DNA. It is based on the reaction of 2-
deoxy sugar with Schiff reagent (Dye Fuchsine is decolorized
by sulphurous acid. Basic fuchsine- a triphenylmethane dye
whose dominant component is pararosanilin ). In case of
DNA cells are hydrolysed with hydrochloric acid to produce
apurinic acid & the stained with Schiff reagent to produce
magenta stained nuclei.
2) Pentoses :-
They are sugars containing 5 carbon atoms.
Important are
a) D-ribose : the constituent of RNA, coenzymes
FAD(flavin-adenine dinucleotide),
 NAD(nicotinamide-adenine dinucleotide),
CoA(coenzyme), ATP(adenosine tri phosphate).
b) D -2- deoxyribose : constituent of DNA.
c) D-ribulose : its phosphates esters is an
intermediate in HMP shunt pathway.
d) Arabinose is present in cherries & in gycoproteins.
e) D-xylulose : its phosphates esters is an
intermediate in HMP shunt pathway. L- xylulose is
a metabolite of D- glucuronic acid ( uronic acid)
pathway. In a hereditary disorder in metabolism it
is excreted in urine called pentosuria.
( lyxose –aldopentose,epimeric with arabinose &
xylose; lyxulose- keto derivative of lyxose;
lyxoflavin-similar to riboflavin except D-lyxitol
present in place of D-ribitol.)
f) Xylose is seen in proteoglycans.
g) D –lyxose : is constituent of lyxoflavin isolated
from human heart muscle.
3) Amino sugars or hexosamines :-
Sugars containing amino group (NH2) in their
structure is called amino sugars. The amino sugars
does not show any reducing property and wont
form osazones.
Two types of amino sugars are physiologically
important. They are
1. Glycosylamine
2. Glycosamine (glycamine)
 In case of glycosylamine the anomeric –OH group is
replaced by the NH2 group. Eg:- Ribosylamine , a derivative
of it is involved in the synthesis of purines.
In case of glycosamine the alcoholic - OH group of
sugar molecule is replaced by the amino group. The amino
group is added to the second carbon atom in case of
hexoses. Glucosamine & galactosamine are two naturally
occurring glycosamine. The amino group in the sugar can be
further acetylated to produce N - acetylated sugars such as
N -acetyl-glucosamine (GluNac) & N-acetyl-galactosamine
(GalNac), etc.
 Glucosamine is seen in hyaluronic acid , heparin ,
blood group substance.
 Glucosamine is the chief organic constituent of
cell wall of fungi & shells of crustacean (crabs,
lobsters etc. It occurs as chitin made up of
repeated units of GluNac. So glucosamine is
called chitosamine.
 Galactosamine occurs as GalNac in chondroitin
sulphates present in the cartilages, bones,
tendons, and heart valve. So Galactosamine is
called chondrosamine.
 Certain antibiotics such as erythromycin,
carbomycin, contain amino sugars. The amino
 sugars are related to the antibiotic activity of
these drugs.
 Mannosamine is a constituent of glycoproteins.
 D –mannosamine condense with the pyruvic acid
to form a amino sugar acid called Neuraminic
acid. It is found in the nature as N –acetyl
Neuraminic acid (NANA) known as Sialic acids.
Neuraminic acid & sialic acids occur in no. of
muucopolysaccharides & in glycolipids like
gangliosides. A no. of nitrogenous
oligosaccharides which contain neuraminic acid
are found in human milk. Certain bacterial cell
walls contain muramic acid. Neuraminidase is the
enzyme which hydrolyses to split NANA from the
compound.
 Muramic acid is structurally a condension product
of D-Glucosamine & lactic acid.