AIM : To study the different qualitative chemical test for carbohydrates .

Theory :
CARBOHYDRATE

Introduction : Carbohydrates are defined as poly-hydroxy aldehydes or poly-hydroxy

ketones or compounds that produce any of the above on hydrolysis, although they are
widely distributed both n animal and plant kingdom yet they are obtained mainly from
plants, they are formed in plants by a process known as photosynthesis and make up
about 70% of the solid plant material.
xCO2 + yH2O sunlight &chlorophylCx (H2O) + xO2

Carbohydrates are grouped into two major classes,ie. Simple sugar (saccharides) and
polysaccharides. Low molecular weight carbohydrates are crystalline soluble in water and sweet
in taste. Eg sucrose, fructose, the high molecular weight carbohydrates are
amorphous,tasteless and relatively less soluble in water. Eg starch, cellulose, gum, pectin,
inulin, etc.
Classification of carbohydrate :
1- Monosaccharides: These are sugar which can be further hydrolyzed to simple sugar.
However they are classified according to the number of c-atom in sugar molecules they are
further divided into :a) Bioses :They contain 2 c-atom, they dont occur free in nature.
b) Trioses :They contain 3 c-atom but in the forms of phosphoric esters. Eg.
Glyceraldehyde
c) Tetroses :They contain 4 c-atom. Eg. Erythrose
d) Pentoses : They are common in plants and are products of hydrolysis of
polysaccharides like hemicellulose, mucilage, gums. Eg. Xylose, Ribose
e) Hexoses :They are the monosaccharides containing 6 c-atom and are
abundentaly available carbohydrates of plants kingdom. Eg. Glucose, Galactose,
Fructose.
f) Heptoses :They contain 7 c-atoms vitally important in the photosynthesis of plant
and glucose metabolism of animals. Eg. Glucoheptose
2- Disaccharides: Carbohydrates which on hydrolysis yield 2 molecules of mono-saccharides
are called saccharides. Eg. Lactose, Maltose, Sucrose.
Sucrose hydrolysis Glucose + Fructose
Lactose hydrolysis Glucose + Galactose
Maltose hydrolysis Glucose + Glucose
Lactose and maltose are reducing sugar because of free aldehyde group of one of the
monosaccharide residue.
In sucrose, glycoside linkage is between C-1 of glucose and C-2 of fructose. So free aldehyde
or keto group is not available, thats why sucrose is non-reducing sugar.

3- Polysaccharides :They are made up of more than 10 monosaccharides units. Eg. Cellulose,
starch, glycogen.

Quantitative Test
Rapid test are available to establish the presence or absence of a carbohydrates in a sample.
These tests are based on specific color typically for these groups.
1. Molisch Test- It is a general test for all carbohydrates. A positive test indicates the
presence of carbohydrates in a test solution.
Glucose concH2SO4 ,3H2OHydroxymethyl furfuralNapthol Violet colored complex
Carbohydrates undergo dehydration when treated with conc. H2SO4bonds to yield
monosaccharides which in the presence of an acid get dehydrated to form Furfural and
its derivatives on condensation with -Napthol form a violet colored complex. All
carbohydrates except amino sugar give this test.
Procedure- 2ml of sugar solution add 2 drops of -napthol solution and mix. Add 2ml of
conc. H2SO4 along the side of the test tube very gently by keeping the tube slightly in a
inclined position so that two distinct layers are formed. Appearance of purple color at the
junction of two layers indicates the presence of carbohydrates in the sample.
2. Anthrone Test- Added conc. H2SO4 to glucose, it forms Hydroxy methyl furfural by
adding Anthrone reagent to form bluish green complex.
Glucose concH2SO4 ,3H2OHydroxymethyl furfural Anthrone reagent Bluish green complex
In this, furfural produced reacts with Anthrone reagent to give bluish green complex.
3. Iodine Test- It is a test of polysaccharides which absorbs I2 and form colored complex
starch gives blue color, on heating starch give blue color because starch 1 complex
dissociates.
Procedure- Take 1ml of sugar solution in a test tube. Add a drop of dilute HCl to acidify
the solution or 5 drops of Glacial acetic acid. Add 4-5 drops of I2 solution to it and mix the
contents gently. Note the color of the product. Gently warm the solution and cool it. Note
the change.
4. Benedicts Test-Reducing sugars have a free aldehyde or keto group. It undergo
tautomerisation inti emediol form under hot alkaline condition. It converts the cupric ion
of the benedict solution inti cuprous ion.
5. Fehlings Test- This test is used for reducing sugar solution, formation of red precipitate
or yellow precipitate of cuprous oxide indicates the presence of red sugar.
6. Picric Acid Test- Picric acid gets converted into picranic acid which gives red color.
7. Barfoeds Test- This test is used for distinguishing between monosaccharides and red
disaccharides, monosaccharides react fast within 1-2 minutes but disaccharides take
longer time i.e 7-12 minutes. This test is also based on the red property of sugar which
produces the red precipitate of Cu2O.
8. Bials Test- Useful for pentose sugar reaction is due to the formation of furfural in acid
medium which condenses with osazone in presence of ferric ion to give blue green color
complex.
9. Tollens Test- Galactose react with phloroglucinol in hot acidic medium to give cherry
red color.

10. Osazone Test- Several red sugar form characterstic shaped crystals on reaction with
phenyl hydrazine and hence they can be identified like maltose give sunflower like
crystal. Lactose give wooly cotton balls crystals.
11.KOH Reduction Test- Lactose form emediol and these reacts with KOH to give yellow
color which deepens on cooloing.