CARBOHYDRATES

Carbohydrate - polyhydroxy aldehyde, ketones and their derivatives. Functions - structure material storage material for carbon and energy.

CONCEPTS OF ISOMERS

Two or more different compounds which contain the same number and types of atoms and the same molecular weights.

Projection formula (spatial arrangement to represent 3dimensional structure):

HO H OH H2 OH
D-glyceraldehyde (hydroxy group at the highest numbered asymmetric carbon atom is written to the right):
HO HO H H 2 OH

L-glyceraldehyde (hydroxy group at the highest numbered asymmetric carbon atom is written to the left): D-Erythrose is the mirror image of L-Erythrose.

Glyceraldehydes: Model compounds of monosaccharides

D-Glyceraldehyde L-Glyceraldehyde

D-Erythrose

L-Threose

POLARIMETER
ixed

Light

Nicol rism

lane olarized Light

Analyzer ell (Sample) Nicol rism ye

Dextrorotatory -plane polarized light rotated to clockwise (or to the right) Levoratatory - plane polarized light rotated to counterclockwise.

FISHER PROJECTION FORM

H C H C OH HO C H H C OH H C OH CH2 OH O H C H C OH HO C H H C OH H C CH2 OH O or OH HO C H C OH HO C H H C OH H C CH 2 OH O H

Anomeric Carbon --- The carbon atom which is involved in hemiacetal or acetal formation.

Anomers: tereoisomers formed when ring is formed (E, F).

H H HO H HO

O OH H OH H H 2 OH

H H O HO H

OH OH H OH H H 2 OH or O

HO H HO H

H OH H OH H H 2 OH

E is same side with ring

HAWORTH PROJECTION FORMULAS FOR SUGARS

C C C C C C 4

C 5 1 3

E - D - Glucopyranose

HAWORTH PROJECTION FORMULAS FOR SUGARS

H C H HO H H C C C C CH
2

OH OH O H HO

H C H C C C C CH
2

OH OH H OH O

H OH OH H

OH

E -D-Glucofuranose (0.5%)
HO

H C H C C C C CH
2

O OH H OH OH OH

E -D-Glucopyranose (35%)

H H

Aldehydo-D-glucose (0.03%)

HO C H HO H H C C C C CH
2

H OH O H

HO C H HO H C C C C CH

H OH H OH O

OH OH

OH

F -D-Glucofuranose (0.5%)

F -D-Glucopyranose (0.5%)

MONOSACCHARIDE
Hexoses 1. Glucose (dextrose) --- rotate the polarized light to the right.
H C H C HO C H C H C CH 2 OH OH H OH O 4 HO 3 OH 6 CH2 OH O 5 OH 1 2 OH OH

Glucose (dextrose)
Most common monosaccharide. Commercially from starch. Cereal roots, legumes, animal tissue, and liver. Mutarotation --The optical changes of glucose in water solution to constant value E20D = +520 E - D - glucose " E20D = 113 D - glucose E20D = 52  F - D - glucose E20D = = 19

At equilibrium = 35% of E - form and 65% of F - form.

2.

Galactose --- Component of lactose (milk sugar)

C
2

E-D-Galactopyranose

3.

Fructose (levulsoe) --- Rotation in polarimeter is left

C C C C C C

C C C C C

C C or C C
2

D-Fructose

F-D-Fructose

E-D-Fructose

Fructose (levulsoe)

F - D - Fructofuranose
CH2 OH C O HO C H H C OH H C OH CH2 OH CH2 OH OH C HO C H H C OH H C H C H OH O

E - D - Fructofuranose
H O OH OH OH OH CH2OH

Naturally-occurring free form

Disaccharides (anydrides of 2 monosaccharides):

Maltose: 4-0-E-D-Glucopyranosyl (1"4) E-DGlucopyranose

Cellobiose 4-0-F-D-Glucopyranosyl (1"4)-F-D-Glucopyranose

CH2 OH O OH HO OH O

CH2 OH O OH OH OH

Sucrose
2-0-E-D-Glucopyranosyl F-D-Fructofuranoside
C
2

1 C 2

5 3 4 C 6
2

Invert Sugar --- when sucrose in solution, the rotation changes from detrorotatory (+66.5) to levorotatory (-19.8). So, sucrose is called Invert Sugar. Sucrose has been hydrolyzed into glucose and fructose.

Lactose
Principal sugar in milk
CH2 OH OH OH OH O O OH OH OH CH2 OH O

4-0-F-D-Galactopyranosyl (1"4)-E-D- Glucopyranose 4-0-F-D-Galactopyranosyl (1->4)-F-D-Glucopyranose

RELATIVE SWEETNESS OF DIFFERENT SUGARS

Sucrose Glucose Fructose Lactose Invert Sugar Maltose Galactose

100 74 174 16 126 32 32

Oligosaccharide
Raffinose (Galactose + Glucose + Fructose) 6-0-E-D-Galactopyranosyl (1->6)-2-0-E-D-Glucopyranosyl (1>2)-F-D-Fructofuranoside

2O

O O

O O O O O

2O

O O
2O

O O O

Mel iose

ucrose Moiety

Stachyose (Galactose + Galactose + Glucose + Fructose)

6-0-E-D-Galactopyranosyl (1"6)-6-0-E-D-Galactopyranosyl (1-> 6) -2-0-E-D-Glucopyranosyl-F-D-Fructofuranoside Flatulence Factor

POLYSACCHARIDE
Anhydrides of Monosaccharides Through ether F linkages. Cellulose --- polymer of F-D-Glucose (1, 4) linkage. Repeating cellobiose moiety.

CH2 OH O OH OH OH O

CH2 OH O OH O

CH2 OH O OH

CH2 OH O

OH

OH

OH

OH

STARCH
The reserve carbohydrate of plants. Occurs as granules in the cell. Made of amylose and amylopectin. Amylose --- ploymer of E-D- Glucose (1"4) linkagestraight-chain.
CH2 OH O OH O OH OH OH OH OH O CH2 OH O OH CH2 OH O OH CH2 OH O

O
OH

Amylopectin

Polymer of E-D-Glucose (1->4) linkage in addition to E-D-Glucose (1"6) linkage.

The length of linear units in amylopectin is only 25. E-(1"4) linkage (25) to E-(1"6) linkage.

GLYCOGEN
Animal starch. E - (1 " 4) linkage and E - (1 -> 6) linkage 12 : 1

PECTIN
Polymer of 4-0-E-D-Galacturonic acid (1 -> 4) & Methylgalacturonate

DEGREE OF METHYLATION (DM)

The higher the degree of methylation, the higher the temperature at which the gel forms.

For gel formation at least 50% of the carboxyl group are methylated. Pectin (about 74 DM) is used in jams. For jellies DM is about 60.

FUNCTIONAL PROPERTIES OF POLYSACCHARIDES (GUMS)

1. 2. 3. 4. 5.

Viscosity control Texture control Emulsifying agent Water-binding capacity Stabilizer

CORN SYRUP
Enzyme Corn Starch H (
+

Corn Syrup High Glucose Maltose Dextrose

Dextrose quivalent (D ) =

Gra of Reducing Sugar Gra of Total Sugar

x1

The higher the D , generally the greater the glucose content in corn syrup.

HIGH FRUCTOSE SYRUPS

Glucose Isomerase Glucose in Syru Fructose

Generally: Glucose Fructose Maltose Isomaltose Higher Saccharides

50% 42% 1.5% 1.5% 5.0%

CORN SYRUP
% Dry Weight High Maltose Syrup Glucose Maltose Triose Tetrose Pentose 9 52 13 2 24 Low DE 14 12 10 9 55 High DE 43 32 3 5 12

MODIFIED STARCHES
1. Pregelatinized Starch --- simple precooked and roll dried to give product that readily disperses in cold water.

2. Thin-boiling or Acid-modified Starch --- suspending granular starch in a very dilute acid under somewhat below its gelatinization temperature (somewhat mild).

3. Oxidized Starches --- oxidation of starch with alkaline hypochloride to get -coo- at C6.

CARBOHYDRATE DETERMINATION
1. Monosaccharides and Oligosaccharides A. Enzymatic Method 1. 2. B. 1. 2. 3. 2. Glucose oxidase Hexokinase Paper or thin layer chromatography Gas chromatography Liquid column chromatography

Chromatography Method

Polysaccharides

Glucose Oxidase System

Glucose Oxidase D-Glucose + O2 Peroxidase H2O2+ 0 - Dianisidine (Colorless) 2 H2O + Oxidized 0-Dianisidine (Brown) Gluconic Acid + H2O2

H3 CO H2 N

OCH3 NH 2

H3 CO HN

OCH3 NH

Hexokinase System
Hexokinase Glucose + ATP G - 6 - P DH G - 6 - P + NADP+ Gluconate - 6 - phosphate + NADPH + H+ G - 6 - P + ADP

NADP+ : (Nicotinamide Adenine Dinucleotide Phosphate) The amount of NADPH formed in this reaction is stoichiometric with the amount of glucose. NADPH is measured by its absorption at 334 nm.

QUALITATIVE ANALYSES OF CARBOHYDRATES

Thin Layer chromatography Silica gel as stationary phase (250 Qm) Quantitative by densitomer (time - varies with solvent, generally overnight):

Solvent For Spot A Rf = A S C B A A B S

Gas Liquid Chromatography Requirement: compounds must be volatile. Sugars form Trimethylsilyl Ether:
CH 3 O Si CH 3 CH 3 CH3 Cl Si CH3 CH 3

Ether linkage

Trimethyl chlorosilane

Liquid Chromatography
Carbohydrate Analysis (Waters Assocs. 840380 Stationary Phase) Solvent: H2O/CH3CN

Glucose

Fructose Maltose Maltotriose

Retention time

POLYSACCHARIDES
Usually, quantitation depends on chemical or enzymatic hydrolyses of them, followed by analysis of the monomers. Starch 1. Amylose + I- -> Blue complex

Use spectrometer for determining the amylose - iodine blue color - according to spectrophotometric reading. 2. Amylopectin + I- -> Reddish color.