Carbohydrates

Chapter 25

25-1
Carbohydrates
 Carbohydrate: A polyhydroxyaldehyde, a
polyhydroxyketone, or a compound that gives
either of these compounds after hydrolysis.
 Monosaccharide: A carbohydrate that cannot be
hydrolyzed to a simpler carbohydrate.
• They have the general formula CnH2nOn, where n varies
from 3 to 8.
• Aldose: a monosaccharide containing an aldehyde
group.
• Ketose: a monosaccharide containing a ketone group.

25-2
Importance of Carbohydrates to us….

cellulose clothing fiber

polymerize giving structure wood
photosynthesis to plants
CO2
+ glucose
H2O polymerize
chlorophyll
light
starch, plant seeds

eaten by animals

CO2 glucose glycogen (liver)

+
H2O
+
energy
25-3
Monosaccharides
 Monosaccharides are classified by their number
of carbon atoms:
N ame Formula
Trios e C3 H6 O3
Tetros e C4 H8 O4
Pen tos e C5 H1 0 O5
Hexose C6 H1 2 O6
Heptose C7 H1 4 O7
Octose C8 H1 6 O8

25-4
Monosaccharides
 There are only two trioses:
CHO CH2 OH
Chiral Center
CHOH C= O
R and S
stereoisomers CH2 OH CH2 OH
Glyceraldehyde D ihydroxyacetone
(an aldotriose) (a ketotriose)

 These compounds are referred to simply as

trioses, tetroses, and so forth tellling the number
of carbon atoms present.

25-5
Review Fischer Projections
 Fischer
projection: A two dimensional
representation for showing the configuration of
carbohydrates.
• Horizontal lines represent bonds projecting forward.
• Vertical lines represent bonds projecting to the rear.
• The more highly oxidized carbon is shown at the top.

CHO convert to a CHO

Fisch er p rojection
H C OH H OH
CH2 OH CH2 OH
.

(R)-Glycerald ehyde (R)-Glyceraldeh yd e

(three-dimensional (Fis cher projection)
represen tation ) 25-6
D,L Monosaccharides
 In 1891, Emil Fischer made the arbitrary
assignments of D- and L- to the enantiomers of
glyceraldehyde.
CHO CHO
H OH HO H
CH2 OH CH2 OH
D -Glyceraldehyde L-Glyceraldehyde
(R)-Glyceraldehyde (S )-Glyceraldehyde
[]D25 = +13.5 []D25 = -13.5

25-7
D,L Monosaccharides
 According to the conventions proposed by
Fischer:
• D-monosaccharide: A monosaccharide that has the
same configuration at its penultimate (next to bottom)
carbon as D-glyceraldehyde; that is, its -OH is on the
right when written as a Fischer projection.
• L-monosaccharide: A monosaccharide that has the
same configuration at its penultimate carbon as L-
glyceraldehyde; that is, its -OH is on the left when
written as a Fischer projection.

25-8
D,L Monosaccharides
 D-aldotetroses
and the two most abundant D-
aldopentoses in the biological world:

CHO CHO
CHO CHO H OH H H
H OH HO H H OH H OH
H OH H OH H OH H OH
CH2 OH CH2 OH CH2 OH CH2 OH
D-Erythrose D-Threose D-Ribose 2-Deoxy-D-
ribose
D-aldotetroses
D-aldopentoses
Expect four stereoisomer Expect total of 8 (=23)
22 for aldotetroses. Two D steroisomers. Four D,
and two L. four L. 25-9
D,L Monosaccharides
 The most abundant hexoses:
CHO CHO CH2 OH
H OH H OH C O
HO H HO H HO H
H OH HO H H OH
H OH H OH H OH
CH2 OH CH2 OH CH2 OH
D-Glu co s e D -Gala cto s e D -Fru cto s e

aldohexoses ketohexose
Expect 16 stereoisomers 24 for Expect 8 stereoisomers 24
aldohexoses. Eight D and for ketohexoses. Four D
eight L. and four L.

25-10
 D Aldohexoses binary display

All
altruists
gladly
make gum
There is
in gallon
0= 000 1=001 2=010 3=011 also the L
tanks.
series,
the mirror
image
L.Fieser
structures

4=100 5=101 6=110 7=111 25-11

Amino Sugars
 Amino sugar: A sugar that contains an -NH2
group in place of an -OH group.
• Only three amino sugars are common in nature
• N-Acetyl-D-glucosamine is a derivative of D-
glucosamine.
CHO CHO CHO CHO O
H NH2 H2 N 2 H H NH2 H NHCCH3
HO H HO H HO H HO H
H OH H OH HO 4 H H OH
H OH H OH H OH H OH
CH2 OH CH2 OH CH2 OH CH2 OH
D -Glu cosamine D -Mannosamine D -Galactosamine N-Acetyl-D -
glucos amin e

25-12
Physical Properties
 Monosaccharides are colorless crystalline solids,
very soluble in water, but only slightly soluble in
ethanol.
• sweetness relative to sucrose:
S w eetness S w eetness
Relative to Artificial Relative to
Carbohydrate S ucrose Sw eetener S ucrose
Fructos e 1.74 Saccharin 450
In vert su gar 1.25 Aces ulfame-K 200
Sucrose (tab le sugar) 1.00 Asp artame 160
Hon ey 0.97
Glucose 0.74
Maltos e 0.33
Galactose 0.32
Lactose (milk s ugar) 0.16
25-13
Cyclic Structure
 Monosaccharides have hydroxyl and carbonyl
groups in the same molecule and those with five
or more carbons exist almost entirely as five- and
six-membered cyclic hemiacetals.
• Anomeric carbon: The new stereocenter created as a
result of cyclic hemiacetal formation.
• Anomers: Carbohydrates that differ in configuration at
their anomeric carbons named  and b.

25-14
Haworth Projections
 Haworth projections
• Five- and six-membered hemiacetals are represented
as planar pentagons or hexagons viewed through the
edge.
• They are commonly written with the anomeric carbon
on the right and the hemiacetal oxygen to the back
right.
• The designation b- means that the -OH on the
anomeric carbon is cis to the terminal -CH2OH;
- means that it is trans to the terminal -CH2OH.

25-15
 Haworth Projections
1
CHO red raw to sh ow th e -OH CH2 OH
H OH on carbon-5 close to the
aldeh yd e on carbon-1 H5 OH
HO H H O
OH H C
H OH Lay molecule HO 1 H
top

5
H OH on side. H OH
cis, b
CH2 OH trans, 
D -Glucose anomeric anomeric
CH2 OH carbon CH2 OH carb on
H O OH(b) H OH
H + H
OH H OH H
HO H HO OH()
H OH H OH
b-D -Glucopyranose -D -Glucopyranose
(b-D -Glucose) ( -D -Glucos e )
25-16
Haworth Projections
• Six-membered hemiacetal rings are shown by the infix
-pyran-.
• Five-membered hemiacetal rings are shown by the
infix -furan-.

O O
Furan Pyran

25-17
Conformational Formulas
• Five-membered rings are so close to being planar that
Haworth projections are adequate to represent
furanoses.
HOCH2 O H HOCH2 O OH ( b)
H H H H
H OH ( ) H H
OH OH OH H
-D -Rib ofuranose b-2-D eoxy-D -ribofu ran os e
(-D -Ribose) (b-2-D eoxy-D -ribose)

25-18
Conformational Formulas
• Other monosaccharides also form five-membered
cyclic hemiacetals.
• Here are the five-membered cyclic hemiacetals of D-
fructose, a ketohexose.
1
CH2 OH
1 2
HOCH2 O CH2 OH 3
C=O HOCH2 O OH ( b)
5 H HO 2 HO H 5 H HO 2
4
H OH( ) H OH H CH2 OH
5
HO H H OH HO H
1
6
CH2 OH
 -D -Fructofuranose D -Fru ctose b- D -Fru ctofu ran os e
( - D -Fructos e) (b- D -Fructose)

25-19
 Conformational Formulas; b to  conversion
• For pyranoses, the six-membered ring is more
accurately represented as a chair conformation.
OH
H OH H
H O H OH
HO HO Open chain
HO OH HO O
H OH H OH
form
H H H H
b-D -Glu copyran os e rotate about
(b-D -Glucos e) C-1 to C-2 bond

H OH OH
H
H OH H O
HO HO
HO H HO H
H OH H OH
H O H OH
-D -Glucop yranose
(-D -Glu cose)
25-22
Conformational Formulas
• The orientations of groups on carbons 1-5 in the
Haworth and chair projections of b-D-glucopyranose
are up-down-up-down-up.

6 CH OH
2 6
5 O CH2 OH
H OH(b) 4
H HO O
4 OH 1 HO 5
H
HO H 2 OH( b)
3 3
2 OH 1
H OH
b-D -Glucopyranose b-D -Glu copyran os e
(Haw orth projection) (chair conformation )

25-23
Mutarotation
 Mutarotation:The change in specific rotation that
occurs when an  or b form of a carbohydrate is
converted to an equilibrium mixture of the two.
[] after % Present at
Mon os accharide [] Mutarotation Equ ilib riu m
-D -glu cose +112.0 +52.7 36
b-D -glucos e +18.7 +52.7 64

-D -galactos e +150.7 +80.2 28

b-D -galactose +52.8 +80.2 72

CH2 OH CH2 OH
HO O HO O
HO HO
OH ( b)
OH HO
b-D -Glu cop yranose OH ( )
-D -Glucopyranose
[] D2 5 +18.7
[] D 25 +112 25-24
Hemiacetals and Acetals, carbonyls and alcohols

Addition
reaction
. (Unstable
in Acid;
Unstable in
base)

(Unstable
in Acid;
Stable in
base)
Substitution reaction 25-25
Glycosides, anomeric OH becomes OR, acetal formation.

 Glycoside: A carbohydrate in which the -OH of

the anomeric carbon is replaced by -OR.
• methyl b-D-glucopyranoside (methyl b-D-glucoside)
glycos idic
bond
CH2 OH CH2 OH CH2 OH
O OH O OCH3 OH
H + H H
H H H H
OH H + CH3 OH OH H + OH H
HO -H2 O H HO OCH3
H HO
H OH H OH H OH
b-D -Glu copyran os e Methyl b-D -glu co- Meth yl -D -glu co-
(b-D -Glu cose) pyran os ide p yranosid e
( Methyl b-D -glucos ide) (Methyl -D -glucos ide)

25-26
Glycosides, acetals
 Glycosidic bond: The bond from the anomeric
carbon of the glycoside to an -OR group.
 Glycosides are named by listing the name of the
alkyl or aryl group bonded to oxygen followed by
the name of the carbohydrate with the ending -e
replaced by -ide.
• methyl b-D-glucopyranoside
• methyl -D-ribofuranoside

25-27
N-Glycosides
 Theanomeric carbon of a cyclic hemiacetal also
undergoes reaction with the N-H group of an
amine to form an N-glycoside.
• N-glycosides of the following purine and pyrimidine
bases are structural units of nucleic acids.
O NH2 O NH2 O
CH3 N N
HN N HN N HN
O N O N O N N N H2 N N N
H H H H H
U racil Cytosine Thymine Ad enin e Guan ine

25-28
N-Glycosides
• The b-N-glycoside formed between D-ribofuranose and
cytosine.

NH2

N
a b-N -glycosid ic
bond
O N
HOCH2 O
H H
H H anomeric
HO OH carbon

25-29
Reactions

25-30
Reduction to Alditols, aldehyde  alcohol
 Thecarbonyl group of a monosaccharide can be
reduced to an hydroxyl group by a variety of
reducing agents, including NaBH4 and H2/M.
CHO CH2 OH
CH2 OH H OH H OH
HO O HO H HO H
HO NaBH4
OH H OH H OH
OH H OH H OH
b-D -Glucop yranose CH2 OH CH2 OH
D -Glu cose D -Glucitol
(D -Sorbitol)

An
alditol
25-31
Other alditols
• Other alditols common in the biological world are:
CH2 OH
HO H CH2 OH
CH2 OH HO H H OH
H OH H OH HO H
H OH H OH H OH
CH2 OH CH2 OH CH2 OH
Eryth ritol D -Mann itol Xylitol

25-32
Oxidations

Oxidation can be done in several ways.

Tollens reagent (Ag+(NH3)2 or Benedict’s solution
(Cu2+ tartrate complex). Not synthetically useful
due to side reactions.
Bromine water oxidizes aldoses (not ketoses) to
monocarboxylic acids (Aldonic Acids).
Nitric Acid oxidizes aldoses to dicarboxylic acids
(Aldaric acids).
Enzyme catalyzed oxidation of terminal OH to
carboxylic acid (Uronic Acid)
Periodic Acid oxidizes and breaks C C bonds.
Later for that. 25-33
Reducing Sugars
 Sugars with aldehyde (or ketone group) in
solution. The group can be oxidized and is
detected with Tollens or Benedicts solution.
Ketone groups converted to aldehyde via
tautomeric shifts (later).

25-34
Problem with Tollens
 2-Ketoses are also oxidized to aldonic acids in
basic solution (Tollens).
CH2 OH CHOH CHO COOH
C= O (1) C-OH (2) CHOH CHOH
(3)
( CHOH ) n ( CHOH ) n ( CHOH ) n ( CHOH ) n
CH2 OH CH2 OH CH2 OH CH2 OH
A 2-ketose An enediol An aldose An aldonic acid

Ketose to aldose conversion Oxidation

via keto enol tautomerism
Reducing sugar

25-35
Oxidation to carboxylic acids

25-37
Oxidation to Uronic Acids
 Enzyme-catalyzedoxidation of the terminal -OH
group gives a -COOH group.
CHO CHO
H OH enzyme-catalyzed H OH
COOH
HO H oxidation HO H O
HO
H OH H OH HO OH
H OH H OH OH
CH2 OH COOH
D -Glucose D -Glucuronic acid
(a uronic acid)

25-38
Oxidation by periodic acid, HIO4 or H5IO6
 Periodic acid cleaves the C-C bond of a glycol.
OH
C OH HO
O - 2 H2 O
+ I
C OH HO OHOH

A 1,2-diol Periodic acid

C O OHO C O
I + H3 IO 4
C O OHOH C O Iodic acid
A cyclic periodic
ester

25-40
Oxidation by HIO4
• It also cleaves -hydroxyketones
H H H
H C OH H C OH H5 IO 6 H C O
+ H2 O
C O HO C OH
HO C O
R R
R
-Hydroxyketone Hydrated
in termediate

• and -hydroxyaldehydes.
O OH OH
H C H C OH H C O
+ H2 O H5 IO6
H C OH H C OH
R R H C O
-Hydroxyald ehyde R
Hydrated
in termediate 25-41
Examples. Identify each of the glucose derivatives.

A + 4 HIO4 yielded 3 HCO2H + HCHO + OHC-CO2H

B + 5 HIO4 yielded 4 HCO2H + 2 HCHO

C + 3 HIO4 yielded 2 HCO2H + 2 OHC-CO2H

CO2H

Analysis of A:
H OH
4 moles of periodic acid used. 4 bonds
broken. HO H

Products:
H OH
Formic acid from –CHOH- or CHO-.
Formaldehyde from CH2OH- H OH

OHC-CO2H from –CHOH-CO2H CH2OH 25-42

Another example
 Oxidationof methyl b-D-glucoside consumes two
moles of HIO4 and produces one mole of formic
acid, which indicates three adjacent C-OH
groups.
periodic acid cleavage
at thes e tw o b on ds
H
CH2 OH CH2 OH
2 HIO C O
HO O 2 HIO
4 5 O
HO O C
OCH3 OH O OCH3
OH C
H
Meth yl b-D -glucopyranoside H

25-43
Osazones, Epimers

O
CHO CH=NNHPh
3 PhNHNH2 PhCHO
CHOH C=NNHPh
O

osazone
osone
aldose

25-44
 Use of osazone in structure determination
Fischer found that (+) glucose and (+) mannose yielded the
same osazone indicating that they differed only at the C2
configuration. Hence, if we know the configuration of (+)
glucose we immediately have that of (+) mannose.
Stereoisomers that differ in configuration at only one
stereogenic center are called epimers. D-glucose and D-
mannose are epimers. NNHPH
CHO CHO
HC

H OH HO H
PhHNN

HO H HO H
HO H

H OH H OH
H OH

H OH H OH
H OH

CH2OH CH2OH
CH2OH

D mannose
D glucose 25-45
Glucose Assay: diabetes (background)
 Theanalytical procedure most often performed in
the clinical chemistry laboratory is the
determination of glucose in blood, urine, or other
biological fluid.

25-46
Glucose Assay
 Theglucose oxidase method is completely
specific for D-glucose.

25-47
Glucose Assay
• The enzyme glucose oxidase is specific for b-D-
glucose.
• Molecular oxygen, O2, used in this reaction is reduced
to hydrogen peroxide H2O2.
• The concentration of H2O2 is determined
experimentally, and is proportional to the
concentration of glucose in the sample.
• In one procedure, the hydrogen peroxide oxidizes o-
toluidine to a colored product, whose concentration is
determined spectrophotometrically.

25-48
Killani-Fischer lengthening of chain

As lactones

Get both epimers.

25-49
Ruff Degradation shortening of chain

25-50
Fischer proof of structure of glucose

Emil Fischer received the 1902 Nobel prize for

determining the structure of glucose.

What was available to him in 1888?

•Theory of stereoisomerism
•Ruff degradation
•Oxidation to aldonic and aldaric acids
•Killani-Fischer synthesis
•Various aldohexoses and aldopentoses

25-51
Fischer proof of structure of glucose

25-52
Fischer started with the aldopentoses

25-53
Experiments on (-) arabinose

Must be an
OH here
25-54
Use KF to get aldohexoses
Fact: Killani Fischer synthesis on (-) arabinose yielded (+) glucose and (+) mannose

CHO CHO
CHO

OH HO
HO
K. F.

HO HO
+

OH

OH OH
CH2OH

CH2OH CH2OH
partial
structure of
arabinose
mixture of + glucose and + mannose,
both of unknown structure. Which is
glucose is unknown.

25-55
 Aldaric acids from glucose and mannose

Fact: nitric acid oxidation of either glucose or mannose

yields optically active aldaric acids. This locates the OH on
C4 since both aldaric acids have to be active.

C4 OH on right C4 OH on left 25-56

Which is glucose??

Which one, A or B, is
CHO CHO glucose is determined
by preparing the
OH HO
aldaric acids
HO HO
(dicarboxylic acids).
+

OH OH

OH OH

CH2OH CH2OH

A B

25-57
Final piece of data…
Fact: the aldaric acid from (+)glucose is also produced by
nitric acid oxidation of a different aldohexose, (+)gulose.

25-58
Where are we?
 We have determined the straight chain structure
of (+) glucose. But certain data indicates the
problem is not yet solved.
 Glucose does not give some reactions
characteristic of aldehydes
• A qualitative test, Schiff test, for aldehydes is negative.
• Bisulfite addition products cannot be made
 Mutarotation changes specific rotation.
 Glucose is converted into two acetals (the methyl
D-glucosides), not hemiacetals, by reaction with
one mole of methanol (acid).
Conclude: glucose is a cyclic hemiacetal.
25-59
Now to determine ring size.
We can methylate the various OH groups,
converting them into OMe. Two kinds of OH
•OH at anomeric carbon
•OH on backbone

One of the backbone OH groups may be

bonded to the anomeric carbon to form a
ring. We seek to detect which one.

First review characteristics of hemiacetals

and acetals.

25-60
Hemiacetals and Acetals, carbonyls and alcohols

Addition
reaction
. (Unstable
in Acid;
Unstable in
base)

(Unstable
in Acid;
Stable in
base)
Substitution reaction 25-61
Methylation to find ring size.

The observed 4 carbon and

5 carbon dicarboxylic acids
indicate free OH was on C5.
25-62
Conformation of the pyranose ring.

Ring flips can occur. Generally the

conformation with large groups equatorial
dominate.

Generally the CH2OH should be made equatorial

25-63
Extreme case: a-D-Idopyranose

25-64
disaccharides
 Sucrose, table sugar
 Maltose, from barley
 Lactose, milk sugar

25-65
Sucrose, table sugar
 Tablesugar, obtained from the juice of sugar
cane and sugar beet.

-1, b-2 bond

25-66
Lactose
 The principle sugar present in milk, 5 – 10%.

25-67
Maltose
 From malt, the juice of sprouted barley and other
cereal grains.

25-68
Structure Determinatin of (+) Maltose
 Experimental Facts
 C12H22O11
 Positive for Tollens and Fehlings solution,
reducing sugar
 Reacts with phenylhydrazine to yield osazone,
C12H20O9(NNHC6H5)2
 Oxidizes by bromine water to monocaboxylic
acid.
 Exists in two forms which undergo mutarotation.
Consistent with two aldoses linked together with
one hemiacetal group.
25-69
More data…..
 Maltose undergoes hydrolysis with aq. acid or
maltase to yield two D (+) glucose units. Two
glucose units joined together: glucose – acetal
linkage (glucoside) – glucose – hemiacetal.
 Maltase hydrolysis is characteristic of 
glucosides. Conclude something like

25-70
How to proceed…..
Label the rings and label the free OH groups.

Next
Slide

25-71
Hydrolysis products

Point of
attachment to
the other
Used in
glucose unit.
hemiacetal
link.
This glucose
Not the reducing
derivative was
aldohexose unit
the “free” CHO
(not the carboxylic
unit, the
acid).
reducing Structure on next
sugar. slide.
25-72
Maltose

25-73
Starch
 Starch is used for energy storage in plants
• It can be separated into two fractions; amylose and
amylopectin; each on complete hydrolysis gives only
D-glucose.
• Amylose: A polysaccharide composed of continuous,
unbranched chains of up to 4000 D-glucose units
joined by -1,4-glycosidic bonds.
• Amylopectin: A highly branched polymer of D-glucose;
chains consist of 24-30 units of D-glucose joined by -
1,4-glycosidic bonds and branches created by -1,6-
glycosidic bonds.

25-74
Glycogen
 Glycogen is the reserve carbohydrate for
animals.
• Like amylopectin, glycogen is a nonlinear polymer of
D-glucose units joined by -1,4- and -1,6-glycosidic
bonds bonds.
• The total amount of glycogen in the body of a well-
nourished adult is about 350 g (about 3/4 of a pound)
divided almost equally between liver and muscle.

25-75
Cellulose
 Cellulose: A linear polymer of D-glucose units
joined by b-1,4-glycosidic bonds.
• It has an average molecular weight of 400,000 g/mol,
corresponding to approximately 2800 D-glucose units
per molecule.
• Both rayon and acetate rayon are made from
chemically modified cellulose.

25-76
Acidic Polysaccharides
 Hyaluronic acid: An acidic polysaccharide
present in connective tissue, such as synovial
fluid and vitreous humor.

25-77
Acidic Polysaccharides
 Heparin
• Its best understood function is as an anticoagulant.
• Following is a pentasaccharide unit of heparin.

25-78
 An example…..
A synthetic polysaccharide is composed of three monosaccharides units
in the following order: (D-glucpyranose) – (L-fructofuranose) – (D-
altropyranose).
The linkages between the units and the structures of the individual D units are
given below where the wavy lines are used to avoid specifying configuration.

Add all substituents to the rings and show the linkages between the units.
You must show stereochemistry clearly.
25-79
 Solution

Now L fructose Now the b ano mer

Now D-Altrose
rst the D glucose…
Now the b 1,4 link Now the  2,6 25-80