Carbohydrates!

What are carbohydrates?

General formula: Cn(H2O)n
Examples: sugar, starch, fibers
Go foods primary source of
energy
In biochemistry, they are known as
saccharides

Carbohydrate composition
Monosaccharides
Simplest carbohydrates; single sugar
molecule
Number of stereoisomers: 2n
Where n = # of chiral carbons

Carbohydrate composition
Disaccharides
Linked by O-glycosidic bond
Interaction between the anomeric carbon
(electrophile) and the lone pair e (nucleophile) from the primary alcohol
Reducing disaccharide is produced if there is
one free anomeric carbon outside of the
glycosidic bond

Carbohydrate composition
Oligosaccharides
Carbohydrates that have 2-10 sugar
molecules
glycoproteins, glycolipids

Carbohydrate composition
Polysaccharides
Carbohydrates that have more than
10 sugar molecules
Storage polysaccharides: glycogen,
starch

Monosaccharides
Monosaccharide classification:
Aldoses: terminal aldehyde
Ketoses: keto-group (position-dependent)
Both are functional isomers
Both contain a primary alcohol attached
to the last carbon

Monosaccharides
Monosaccharides
Simplest aldose: D-glyceraldehyde
Simplest ketose: dihydroxyacetone
Fischer
projection:

Monosaccharides
Epimers:
D-glucose and D-mannose are 2-epimers
D-glucose and D-galactose are 4-epimers
Fischer
projection:

Hemiacetals and Hemiketals

Hemi half
Formed by addition of alcohol
(nucleophile) to the carbonyl
(electrophile)
Responsible for the formation of the
closed chain configuration of saccharides
Pyranose and furanose

Hemiacetals and Hemiketals

Monosaccharides
Pyranose (6-membered ring) or furanose (5-membered
ring)
glucose (dextrose), fructose, galactose
glucopyranose, fructofuranose,
galactopyranose
Haworth projection:

Monosaccharides
Mostly furanose
Ribose and xylose -> ribofuranose and xylofuranose

Haworth projection:

Anomers
Differentiation between
alpha- and beta- form of
anomeric carbon (based
on the OH group)
In their cyclic form,
anomers differ in the
orientation of the OH
group of the primary
carbon

Monosaccharides
Chair and Envelope conformation

Monosaccharides
Are reducing agents:
All aldoses have reducing property
Due to the free anomeric carbon

All ketoses have reducing property

Can be hydrolyzed to aldose

Disaccharides
Maltose ( 1-4 glycosidic bond between 2 glucose
molecules)
O-(-D-glucopyranosyl)1,4--D-glucopyranose

Disaccharides
Cellobiose (-1,4 glycosidic bond between 2 glucose
molecules)
O-(-D-glucopyranosyl)1,4--D-glucopyranose

Disaccharides
Sucrose (table sugar) (-1 of glucose and -2 of fructose)
non-reducing

O-(-D-glucopyranosyl)1,2--D-fructofuranose

Disaccharides
Lactose (milk sugar) (-1,4 between galactose and
glucose)
O-(-D-galactopyranosyl)1,4--D-glucopyranose

Polysaccharides
Glycogen and starch
Homopolysaccharides hydrolysis will yield only one kind of
monosaccharide
GLYCOGEN energy storage liver / muscle
STARCH energy storage in plants
CELLULOSE structural

Overall non-reducing (contains a very small

concentration of anomeric carbons in the entire
structure)

Polysaccharides
Amylose helical
6 glucose residues per turn
-1,4 glycosidic bonds

Amylopectin
-1,4 glycosidic bonds and -1,6 glycosidic branches every 2430 residues

Glycogen
-1,4 glycosidic bonds and -1,6 glycosidic branches every 812 residues

Isolation of Polysaccharides
starch from cassava
Addition of H2O

glycogen from chicken liver

Addition of HOAc
Addition of EtOH

Hydrolysis of the Isolates

Acid Hydrolysis complete hydrolysis
Monosaccharide product

Enzymatic Hydrolysis incomplete

hydrolysis
Breaks specific bonds
Salivary amylase will break down -1,4
glycosidic bonds

Iodine Test

Molischs Test
Reagents
H2SO4, EtOH and -naphtol

Principle involved
Strong acid to dehydrate monosaccharide to produce furfural
or hydroxymethylfurfural
Intermediate furfural derivatives will condense with -naphtol

Visual positive result

Purple ring at interphase for all carbohydrates

Bials-Orcinol Test
Reagents
HCl, FeCl3 and orcinol

Principle involved
Strong acid to dehydrate monosaccharide to produce furfural
or hydroxymethylfurfural
Will then condense with orcinol

Visual positive result

Blue-green solution for aldopentoses

Seliwanoffs Test
Reagents
HCl, resorcinol

Principle involved
Strong acid to dehydrate monosaccharide to produce furfural
or hydroxymethylfurfural
Will then condense with resorcinol

Visual positive result

Cherry-red solution for ketohexoses

Redox reactions of aldoses

Benedicts Test
Reagents
CuSO4, Na2CO3, Na3C6H5O7

Principle involved
Cu2+ oxidizing agent
Cupric ions reduced to cuprous, form Cu2O precipitate
Sodium citrate used to keep Cu ions in solution

Visual positive result

Brick-red precipitate for all reducing saccharides
All monosaccharides are reducing saccharides

Barfoeds Test
Reagents
Cu(CH3COO)2, acetic acid

Principle involved
Cu2+ oxidizing agent
Cupric ions reduced to cuprous, form Cu2O precipitate

Visual positive result

Brick-red precipitate for all reducing monosaccharides
Monosaccharide (< 3 mins)
Disaccharide (~10 mins) -> considered as a negative result

Mucic Acid Test

Reagents
Concentrated HNO3

Principle involved
HNO3 is a strong oxidizing agent
formyl functional group will get oxidized to carboxylic acid
Primary alcohol group will get oxidized to carboxylic acid

Visual positive result

Test for galactose (galactaric acid / mucic acid -> sugar acid
product)
Plane of symmetry not optically active
D-aldoronic acid

Phenylhydrazone Acid Test

Reagents
phenylhydrazine

Principle involved

phenylhydrazine is a strong oxidizing agent

formyl functional group will get oxidized to carboxylic acid
Secondary alcohol will get oxidized to ketone
Product is phenylhydrazone
Phenylhydrazone + phenylhydrazine = osazone

Visual positive result

Yellow orange crystals (osazones)
D-glucosazone (for glucose and fructose and mannose)
No sucrosazone

Mechanism for Phenylhydrazone

Test

Nelsons Test
Nelsons A and Nelsons B
Nelsons A: Rochelle salt sodium potassium tartrate, Na 2CO3,
NaHCO3, Na2SO4
Will also keep Cu ions in solution

Nelsons B: CuSO45H2O, H2SO4

Nelsons A and Nelsons B -> tartaric acid product

Colorimetry of Absorbance is based on the intensity of

the product of arsenomolybdate and the copper oxide
solution
blue / blue-green solution

Thin Layer Chromatography

Mobile phase:
Para-anisaldehyde: visualizing agent
Green colored spots based on complex with the sugar

The more OH groups, the more polar

glucose > maltose > dextrin in Rf value