EXPERIMENT 5: ISOLATION AND 3.

Cellubiose- Glucose + Glucose (β-1,4)

CHARACTERIZATION OF CARBOHYDRATES 4. Lactose- Glucose + Galactose

Carbohydrates
 Are polyhydroxyaldehydes and Homopolysaccharides Heteropolysaccharides
polyhyroxyketones Starch α-1,4/α – Hyaluronic acid
 Cn(H2O)n 1,6
 Monosaccharides as their building blocks. Glycogen α – 1,4 and Dermatan (skin)
 Other types of carbohydrates: α-1,6
- Oligosaccharides Cellulose β -1,4 Keratan (nails)
- Polysaccharides
Monosaccharides Chitin NAG Agarose, peptidoglycan
 Also known as the simple sugars.
 Cannot be further hydrolyzed.
Oligosaccharide
 Simplest monosaccharide contains three
1. Dextrin: product of starch degradation.
carbon called trioses.
Polysaccharides/ Glycans
A. Hexoses
1. Glucose
Qualitative Test
 Aldohexose
 Dextrose, blood sugar, physiologic
1. Molisch test
sugar
 General Test for carbohydrates.
2. Fructose
 Reagent: Molisch Reagent (α-
 Ketohexose
Naphthol in 95% ethanol)
 Reducing sugar
 (+) Purple/ Red ring
 Fruit sugar, sweet fruit and honey
 Principle: Dehydration
 Aka Levulose, the sweetest
monosaccharide.
Carbohydrates react with conc. sulfuric
3. Galactose
acid get dehydrated to form furfural and
 Readily absorbed in the intestines,
its derivatives.
biosynthesized in mammary glands.

4. Mannose

Test for Hexoses

Hexose Test

Glucose Moore’s test

Fructose Seliwanoff’s Test Furfural

Galactose Mucic Test

Mannose Osazone Test

B. Pentose 5-HydrixymethylFurfural
1. Arabinose- found in Gum Arabic/ Acacia
2. Fehlings test:
2. Ribose- found in RNA
3. Deoxyribose- found in DNA
 Reagent: Fehling’s Reagent ( consist of
bistartratocuprate II).
Disaccharide
 Fehlings solutions is always freshly
- Composed of two monosaccharide
prepared.
units.
o Fehlings A: Copper (II) sulfate
1. Sucrose- Glucose + Fructose
pentahydrate
2. Maltose- Glucose + Glucose (α-1,4)

Prepared by: Shanny G. Estera RPh

 o Fehlings B: Potassium sodium tartrate.
 The bistartratocuprate(II) complex
oxidizes the aldehyde to a carboxylate
anion, and in the process the copper (II)
are reduced to copper (I) giving a Red ppt
as a positive result.
o Principle: Reduction Reaction
o (+) Brick red ppt
3. Benedicts Test
 Test for simple carbohydrates, it
identifies reducing sugars which
have free ketone or aldehyde functional
groups.
 Can be used to test presence of glucose
in urine.
 Sodium carbonate provides the
alkaline conditions which are required
for the redox reaction. Sodium citrate
complexes with the copper (II) ions so
that they do not deteriorate to copper(I)
ions during storage.
 Benedicts reagent: NaOH + CuSO4
 Principle: Reduction rxn
 (+) Brick red
4. Barfoed’s Test
 Used for distinguishing monosaccharides
from reducing disaccharides.
Monosaccharides usually react in about 1-2
minute while the reducing disaccharides take
much longer time between 7-12 minutes to
react with the reagent.
 Principle: Reduction rxn
 Reagent: Cupric acetate in acetic acid
 (+) Brick Red ppt.
Prepared by: Shanny G. Estera RPh
4. Tollen’s Test
 Used to test the presence of aldehyde.
 In this reaction the aldehyde group of
the sugar is oxidized to form a
carboxylic acid while the Ag 1+ is reduced
to silver metal, which deposits as a thin
film on the inner surface of the glass.
 Principle: Redox reaction
 Reagent: Silver + Ammonia
 (+) Silver mirror
5.
Picric Acid
Test
 Test for
the presence of reducing sugars.
 The reducing sugars react with Picric Acid
(toxic yellow crystalline solid)  also chemically
known as 2,4,6-trinitrophenol (TNP) to form a
red colored Picramic Acid.
 Reducing sugars reduce picric acid(yellow
solution) to picramic acid (mahogany red
solution).
 Reagent: Picric Acid
 (+) Mahogany Red
6. Osazone/Kowarsky Test
 Monosaccharide reacts with phenyl
hydrazine which is a crystalline
compound. The sugars which will
reduce as a result of this reaction will
result in the formation of osazones.
 Reagent: Phenylhydrazine
 (+) Yellow ppt
  (+) Insoluble ppt

10. Bial’s Test

- Test used to distinguish pentoses from
hexoses.
- Pentoses ( such as ribose sugar) form furfural
in acidic medium which condense with orcinol
in presence of ferric ion to give blue green
colored complex which is soluble in butyl
alcohol.
- Reagent: Resorcinol + Iron
- (+) Blue-green/ Blue soln
7. Seliwanoff’s Test
- Specific test for ketones.
- Ketoses undergo dehydration to yield
furfural derivatives more rapidly than
aldoses.
- The test reagent causes the dehydration of
ketohexoses to form 5-
hydroxymethylfurfural.
- Reagent: Resorcinol in Conc. HCl
- (+) Red/ Pink

8. Keller-Killiani Test
 Test used to identify presence of
deoxysugar, cardiac glycosides.
 Reagent: Acetic acid, Fe chloride,
Sulfuric acid
 (+) Reddish Brown

Glycogen Storage Disease

9. Mucic Acid Test
 Test for Galactose, Oxidation of most
monosaccharides by nitric acid yields
soluble dicarboxylic acids, however
galactose yields insoluble mucic acid.
 Reagent: Nitric Acid

Prepared by: Shanny G. Estera RPh

0 Glycogen synthase

I Von Glucose-6-phosphatase
Gierke

II Pompe Acid maltase/α-1,4-

glucosidase If ever may mga katanungan or di
naiintindihan just message me para
III Forbes- Debranching enzyme
Cori mabigyan natin ng kasagutan 😊
IV Andersen Branching enzyme
- SHANNY ESTERA RPh

V McArdle Muscle phosphorylase

VI Hers Glycogen
phosphorylase

VII Tarui Muscle

phosphofructokinase

VIII Liver phosphorylase

kinase

Prepared by: Shanny G. Estera RPh