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.

polymerize CO2 + H2O photosynthesis glucose chlorophyll light polymerize

cellulose giving structure to plants

clothing fiber wood

starch, plant seeds eaten by animals

CO2 + H2O + energy

glucose

glycogen (liver)

25-3

Monosaccharides
Monosaccharides

are classified by their number

Formula C3 H6 O3 C4 H8 O4 C5 H1 0 O5 C6 H1 2 O6 C7 H1 4 O7 C8 H1 6 O8

of carbon atoms:
N ame Trios e Tetros e Pen tos e Hexose Heptose Octose

25-4

Monosaccharides
There

are only two trioses:

CHO CHOH CH2 OH Glyceralde hyde (an aldotrios e) CH2 OH C= O CH2 OH D ihydroxyacetone (a k etotriose )

Chiral Center R and S stereoisomers

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 H C OH

convert to a Fisch er p rojection

CHO H
.

OH

CH2 OH (R )-Glycerald ehyde (three-dimensional represen tation )

CH2 OH (R)-Glyceraldeh yd e (Fis cher projection)

25-6

D,L Monosaccharides
In

1891, Emil Fischer made the arbitrary assignments of D- and L- to the enantiomers of glyceraldehyde.
CHO H OH CH2 OH CHO HO H CH2 OH

D -Glyce raldehyde L-Glyce raldehyde (R)-Glyceraldehyde (S )-Glyce raldehyde [ ]25 = +13.5 [ ]25 = -13.5
D D

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 Lglyceraldehyde; 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 Daldopentoses in the biological world:

CHO CHO H H OH OH CH2 OH D-Erythrose HO H CHO H OH CH2 OH D-Threos e H H H OH OH OH CH2 OH D-Ribose H H H CHO H OH OH CH2 OH 2-De oxy-Dribos e

D-aldotetroses Expect four stereoisomer 22 for aldotetroses. Two D and two L.

D-aldopentoses Expect total of 8 (=23) steroisomers. Four D, 25-9 four L.

D,L Monosaccharides
The

most abundant hexoses:

CHO H OH HO H H OH H OH CH2 OH D-Glu co s e CHO H OH HO H HO H H OH CH2 OH D -Gala cto s e CH2 OH C O HO H H OH H OH CH2 OH D -Fru cto s e

aldohexoses Expect 16 stereoisomers 24 for aldohexoses. Eight D and eight L.

ketohexose Expect 8 stereoisomers 24 for ketohexoses. Four D and four L.

25-10

D Aldohexoses binary display

All altruists gladly make gum in gallon tanks.

0= 000

1=001

2=010

L.Fieser

There is 3=011 also the L series, the mirror image 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 Dglucosamine.
CHO O CHO H H NHCCH3 H NH2 HO HO H HO H H H OH HO 4 H H H OH H OH CH 2 OH CH 2 OH D -Glu cosamine D -Mannosamine D -Galactosamine N-Acetyl-D glucos amin e CHO NH2 H OH OH CH2 OH CHO H2 N 2 H HO H H OH H OH CH2 OH

25-12

Physical Properties
Monosaccharides

are colorless crystalline solids, very soluble in water, but only slightly soluble in ethanol.
sweetness relative to sucrose:
Carbohydrate S w eetness Relative to S ucrose 1.74 1.25 1.00 0.97 0.74 0.33 0.32 0.16 Artificial Sw eetener Saccharin Aces ulfame-K Asp artame S w eetness Relative to S ucrose 450 200 160

Fructos e In vert su gar Sucrose (tab le sugar) Hon ey Glucose Maltos e Galactose Lactose (milk s ugar)

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
CHO H OH HO H H OH
1

red raw to sh ow th e -OH on carbon-5 close to the aldeh yd e on carbon-1

H 5 OH CH2 OH D -Glucose

Lay molecule on side. cis, b

top

CH2 OH OH H5 O H OH H C 1 H HO H anomeric carbon OH

trans,
anomeric carb on

CH2 OH O OH( b ) H H OH H HO H H OH b -D -Glucopyranose (b -D -Glucose)

CH2 OH OH H H + OH H HO OH( ) H OH -D -Glucopyranose ( -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 H H O H H H HOCH2 H O H H OH ( b)

OH ( ) OH OH -D -Rib ofuranose ( -D -Ribose)

OH H b -2-D eoxy-D -ribofu ran os e (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 Dfructose, a ketohexose.
1 2

CH 2 OH HOCH2
5

HOCH2
5

O H HO H

CH 2 OH
2

H HO

OH ( )

-D -Fructofuranose ( - D -Fructos e)

C=O HO H 4 H OH 5 H OH 6 CH 2 OH D -Fru ctose

3

O H HO H

OH ( b)
2

H HO

CH 2 OH
1

b - D -Fru ctofu ran os 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.
H OH HO HO H H O H OH H OH HO HO H H H OH H OH O OH H rotate about C-1 to C-2 bond H OH HO HO H H OH H OH H O H OH HO HO H O H H OH OH

Open chain form

b-D -Glu copyran os e (b -D -Glucos e)

H -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

H
4

O H

H OH
3

OH( b)
1

CH2 OH O
5 3 2

HO HO

HO H

H
2

OH( b) OH 1

OH

b-D -Glucopyranose (Haw orth projection)

b -D -Glu copyran os e (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 36 +52.7 +112.0 b-D -glucos e +18.7 +52.7 64 -D -galactos e b-D -galactose
CH2 OH O OH ( b )

+150.7 +52.8

+80.2 +80.2
HO HO

28 72
CH2 OH O

HO HO

OH b-D -Glu cop yranose [ ] D2 5 + 18.7

OH ( ) -D -Glucopyranose 25 25-24 [ ] D + 112

HO

Hemiacetals and Acetals, carbonyls and alcohols

Addition reaction .

(Unstable in Acid; Unstable in base)

Substitution reaction

(Unstable in Acid; Stable in base)

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 OH O OCH3 H H + H H H H H + CH OH + OH H 3 OH H OH H -H O 2 HO H HO OCH3 H HO H OH H OH H OH b-D -Glu copyran os e Methyl b -D -glu coMeth 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
The

anomeric 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 HN O N H U racil O N N H Cytosine NH2 HN O O CH3 N N NH2 N HN H2 N N O N N H

N H Thymine

N H Ad enin e

Guan ine

25-28

N-Glycosides
The b-N-glycoside formed between D-ribofuranose and cytosine.
NH2 N HOCH2 H H HO O O H H OH anomeric carbon N a b -N -glycosid ic bond

25-29

Reactions

25-30

Reduction to Alditols, aldehyde alcohol

The

carbonyl group of a monosaccharide can be reduced to an hydroxyl group by a variety of reducing agents, including NaBH4 and H2/M.
CH2 OH O OH CHO H OH HO H H OH H OH CH2 OH D -Glu cose CH2 OH H OH HO H H OH H OH CH2 OH D -Glucitol (D -Sorbitol)

HO HO

NaBH4

OH b-D -Glucop yranose

An alditol
25-31

Other alditols
Other alditols common in the biological world are:
CH2 OH HO H HO H H OH H OH CH2 OH D -Mann itol CH2 OH H OH HO H H OH CH2 OH Xylitol

CH2 OH H OH H OH CH2 OH Eryth ritol

25-32

Oxidations

Oxidation can be done in several ways. Tollens reagent (Ag+(NH3)2 or Benedicts 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 (1) C= O ( CHOH ) n CH2 OH A 2-k etose CHOH C-OH (2) ( CHOH ) n CH2 OH An enediol CHO CHOH (3) ( CHOH ) n CH2 OH An aldose COOH CHOH ( CHOH ) n CH2 OH An aldonic acid

Ketose to aldose conversion via keto enol tautomerism

Oxidation

Reducing sugar

25-35

Oxidation to carboxylic acids

25-37

Oxidation to Uronic Acids

Enzyme-catalyzed

oxidation of the terminal -OH group gives a -COOH group.

CHO H OH enzyme -catalyzed oxidation HO H H OH H OH CH2 OH D -Glucos e H HO H H CHO OH COOH O H HO OH HO OH OH COOH D -Glucuronic acid (a uronic acid)

OH

25-38

Oxidation by periodic acid, HIO4 or H5IO6

Periodic

acid cleaves the C-C bond of a glycol.

OH HO C OH O - 2 H2 O + I C OH HO OHOH
A 1,2-diol Periodic aci d

OH O I C O OHOH C O
A cycli c pe ri odi c es ter

C O C O + H 3 IO 4
Iodic acid

25-40

Oxidation by HIO4
It also cleaves -hydroxyketones
H H C OH C O R H H H5 IO 6 H C HO C R O O

+ H2 O

H C OH HO C OH R

-Hydroxyketone

Hydrated in termediate

and -hydroxyaldehydes.
O H C H C OH R -Hydroxyald ehyde + H2 O OH H C OH H C OH R Hydrated in termediate H5 IO6 OH H C O H C O R

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 OH

Analysis of A: 4 moles of periodic acid used. 4 bonds broken. Products: Formic acid from CHOH- or CHO-. Formaldehyde from CH2OHOHC-CO2H from CHOH-CO2H

HO

OH

OH

CH2OH

25-42

Another example
Oxidation

of 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 HO HO CH2 OH O OH
2 HIO 4 2 HIO
5

O O

C CH2 OH O C H OCH3

OCH3

OH O C H

Meth yl b -D -glucopyranoside

25-43

Osazones, Epimers

O CHO

3 PhNHNH2

CH=NNHPh

PhCHO
C=NNHPh O

CHOH

aldose

osazone

osone

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 Dmannose are epimers. NNHPH
CHO HC HO CHO H OH PhHNN H

HO

HO

HO

OH

OH

OH

OH

OH

OH

CH2OH

CH2OH

CH2OH

D glucose

D mannose

25-45

Glucose Assay: diabetes (background)

The

analytical 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
The

glucose oxidase method is completely specific for D-glucose.

25-47

Glucose Assay
The enzyme glucose oxidase is specific for b-Dglucose. 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 otoluidine 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 HO CHO

HO

OH

K. F.
HO

HO

OH OH OH

CH2OH

partial structure of arabinose

CH2OH

CH2OH

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??

CHO HO

CHO

OH

HO

+
OH

HO

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

OH

OH

CH2OH

CH2OH

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)

Substitution reaction

(Unstable in Acid; Stable in base)

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

Table

sugar, 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 glucose unit.

Used in hemiacetal link. Not the reducing aldohexose unit (not the carboxylic acid).

This glucose derivative was the free CHO unit, the Structure on next reducing slide. sugar.
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 -1,6glycosidic 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 wellnourished 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) (Daltropyranose). 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 Now thethe 2,6 b ano 25-80 mer First the D glucose Now the b 1,4 Now link D-Altrose