O

Carbohydrate Reactions
 O

Important reactions
of carbohydrate molecules
 Carbonyl group  Oxidation to a
(alone) carboxylic acid
group
 Reduction to a
hydroxyl group
 Cyanohydrin
reaction (and
reaction with other
nucleophiles)
 O

Important reactions (cont.)

 Hydroxyl groups  Ester formation
 Ether formation
 Cyclic acetal
 Oxidation to
carbonyl
 Reduction to deoxy
 Replacement with
NH2, SH, or X
 O

Important reactions (cont.)

 Both carbonyl and  Cyclic hemiacetals
hydroxyl groups (pyranose/furanose)
 Formation of acetals
(glycosides)
 Aldose/ketose
isomerizations
 O

Important reactions
 Anomeric hydroxyl  Formation of
group glycosyl halides
 Oxidation to lactones
 O

Carbonyl group oxidation

 O

Tollens test

2Ag(NH3)2+ + RCHO + 3OH-

This is This is
reduced to oxidized to

2Ag(s) + RCOO- + 4NH3 + 2H2O

A silver
mirror
 O

Fehlings test

2Cu(OH)2 + RCHO
This is This is
reduced oxidized

RCOOH + Cu2O + H2O

A brick red
precipitate
 O

Tollen’s test Fehling’s test

 O

Variations on the
Fehlings and Tollens tests
 Nelson-Somogyi - uses an
arsenomolybdate reagent. You read the
optical density at 500 nm
 Benedict - uses copper citrate. This is a
less alkaline medium than the Fehlings
test
 O

Stoichiometric
sugar determination
RCHO + I2 + 3NaOH

RCOO-Na+ + 2NaI + 2H2O

This reaction utilizes oxidation of the aldehyde with
hypoiodite (IO-) at pH 9.5
 O

Glucose oxidase
O
OH
CH2OH glucose CH2OH
OH
O oxidase H
O OH-
H OH HO
H H
OH H O
OH H OH
HO H HO +
H
H OH H OH OH

OH

D-glucono- In alkaline
O2
1,5-lactone solution
H2O2 this exists
as
D-gluconate
 O

Oxidation of D-glucose
 This reaction is used to measure D-
glucose in foods, blood, and other
biological materials
 D-gluconic acid is found naturally (in
small amounts) in fruit juices and honey
 O

Lactones
CH2OH
CHOH
HO O
HH O
H
OH

D-glucono-1, 4,-lactone
O
OH

OH
-lactone
HO
OH

OH

OH
CH2OH
D-gluconic acid O
H
H O
OH H
HO
H OH

D-glucono-1, 5-lactone

-lactone
 O

Lactone stability
 The 1, 5-lactone can be isolated but the 1,
4-lactone is somewhat more stable and
often the only product isolated
 Gamma lactone is ~5.8% vs delta lactone
at 4.1% of the acid level
 O

Glucono-delta-lactone (GDL)
 A food acidulant
 Primary use is in baking industry where
it reacts slowly with sodium bicarbonate
to produce carbon dioxide
 Also GDL prevents discoloration of
refrigerated dough
 Produced by oxidation of β-D-
glucopyranose
 O

GDL as preservative
 Due to its general acidity, GDL will
decrease microbial action in such systems
as
– bread dough
– fish cakes
– surimi
 O

Other uses of GDL

 In meat
– Reduces the amount of nitrite used in curing
– Increases shelf life by inhibiting growth of
lactic acid bacteria
 In pasta and rice
– Improves color and texture
 O

Other uses of GDL

 In canned and frozen vegetables
– Improves stability and texture
 Can be used as a partial replacement for
vinegar in salad dressings
 Can be used as a protein coagulant in the
manufacture of cheese and tofu
 O

Examples of GDL in foods

Tofu Tofu angel hair substitute

 O

Carbonyl group reduction

 O

Glucose to glucitol (sorbitol)

H O
OH

OH OH

HO
reduction HO

OH NaBH4, OH
H2/Pt or Pd,
OH
H2/Raney Ni OH

OH OH

D-glucose D-glucitol
(sorbitol)
Yield  100%
 O

Sorbitol
 Note the suffix -itol
 Sorbitol is widely distributed in nature
but in relatively low concentration
 Was discovered in the berries of the
mountain ash (Sorbus aucuparia)
 Physical form: syrup or crystal
 O

Sorbitol food uses

 Functions as a
humectant (water
retainer)
 Non-cariogenic (S.
mutans cannot
metabolize)
 About 50% as sweet as
sucrose
 Largest use (non-food)
– Toothpaste (cooling
effect; endothermic
heat of solution)
 O

Sorbitol food uses (cont.)

 Sugarless gums
 Sugarless mints
 Sugarless hard candies
 Sugarless cough drops
 A cryoprotectant in surimi
 A starting material for sorbitan esters
(emulsifiers)
 O

Sorbitol and Vitamin C

OH

OH
Acetobacter H

HO
suboxydans OH
H
O
OH
H OH
OH H CH 2 OH
OH H
OH
L-sorbose
OH

D-glucitol

OH

H OH H
O O
OH OH
H H
HO O H OH
H OH
OH OH H O
L-ascorbic acid lactone
2-Keto-L-gulonic acid
 O

Vitamin C synthesis
 High yields at each step allow Vitamin C
as a low cost product
 L-ascorbic acid functions
– Collagen formation
– Fatty acid metabolism
– Brain function
– Drug detoxification
 O

Other Vitamin C functions

 Plants
– Respiration
– Growth
– Maintenance of CO2 balance
 Foods
– General antioxidant function, especially in
aqueous systems
 Natural occurrence
– Rose hips, persimmon, citrus fruits
 O

Mannitol
OH
OH OH

O
OH HO

HO reduction
HO HO
+
OH
OH OH

OH
OH OH

OH OH OH

D-fructose D-glucitol D-mannitol

 O

Mannitol preparation
 Laboratory methods
– Hydrogenation of D-mannose
– Fermentation
 Commercially
– Hydrogenolysis of sucrose
 O

Sucrose hydrogenolysis
Sucrose

D-glucose D-fructose

Isomerization Hydrogenation

D-glucose, Hydrogenation Mannitol,

D-fructose, Glucitol
D-mannose
 O

Mannitol
characteristics and uses
 Not a humectant
 Crystallizes easily, moderately water
soluble
 Candies
– Anti-caking agent
– Dusting agent
 Sweetness: 65% of that of sucrose
 O

Mannitol uses (cont.)

 Sugarfree
– Chocolates
– Mints
– Cough drops
– Hard and soft candies
 O

Xylitol
H O

OH

OH OH
hydrogenation
HO HO

OH OH

OH OH

D-xylose Xylitol

(from hemicellulose
xylans)
 O

Xylitol characteristics
 Has an endothermic heat of solution,
therefore it produces a cooling sensation
in the mouth
 Sweetness: 70-95% of that of sucrose
 Non-cariogenic
 O

Maltitol CH 2 OH
O H
H
H
OH H
HO O H
H OH
H OH
H CH 2 OH M a lto s e
HO H

H
O
OH
(from corn syrup)

h y d r o g e n a tio n

CH 2 OH
OH C -6
O
H H HO C -5
H
OH H
HO O H C -4 M a ltito l
H OH
OH C -3

HO C -2
OH C -1
 O

Maltitol uses
 Reduced calorie candies
 Miscellaneous other foods

 Also included here are isomaltitol and

hydrogenated starch hydrolyzates
 O

Erythritol

•Non-cariogenic
•0.2 kcal/gram No D or L designation
•Strong cooling effect in the mouth This is a meso compound
•RS ~ 0.5
•Not hygroscopic
 O

Erythritol in Truvia
 O

Cyclitols
 Only contain hydroxyl groups
 But unlike alditols, these are cyclic
 Most are 6 membered rings
(polyhydroxycyclohexanes and –hexenes)
 Myo-Inositol
 O

Myo-inositol
 Derivatives such as phosphate esters,
ethers, and deoxy forms also occur
 Myo-inositol occurs free in virtually all
plants
 Isomers also exist
 Phytic acid
 O

Phytic acid
 Mostly occurs as phytates, mixed
potassium-magnesium salts
 Generally an anti-nutritional factor as it
binds minerals, especially calcium and
doesn’t allow them to be absorbed
 Recent studies have shown some positive
effects related to cancer, heart disease,
antioxidant activity and contribution to
fecal bulk
 O

Oxidation of
non-anomeric hydroxyl groups
 O
H O HO O

H OH H OH

HO H 30% HNO3 HO H

HO H HO H

H OH H OH

OH HO O

D-galactose Galactaric acid

The product of this
75% yield
type of oxidation of
an aldose is an
H O HO O aldaric acid
H OH 30% HNO3 H OH
HO H HO H
H OH H OH
H OH H OH

OH HO O

D-glucose
D-glucaric acid
(Saccharic acid)

50-65% yield
 O

Periodate oxidation
CH2OH CH2OH
OR OR
H 2IO4- H
H H
OH H H
HO H O H
H OH O

+ HCOOH
 O

Periodate oxidation
 This reaction can be used to
quantitatively measure the number of
adjacent hydroxyls in a molecule
 It is used in the determination of
polysaccharide structure
 The reaction is most rapid at pH 3-5
 O

Dinitrogen tetroxide (N2O4)

 This reagent is specific for oxidation of
primary alcohol groups
 If you start with an aldose, you get an
alduronic acid
 O

Dinitrogen tetroxide
O
OH
CH2OH
H OH H OH
N2 O 4
H H
OH H OH H
HO O HO O
H OH CH3 H OH CH3

Methyl -D-
Methyl -D-
glucopyranoside
glucopyranosiduronic
acid
 O

Aldobiouronic acid
 When the substrate is a disaccharide, the
product is an aldobiouronic acid (a
disaccharide with a uronic acid at its
non-reducing end)
H
H O OH H
H H O OH
OH H H
CH2OH H O OH OH H
OO H OH H
H
OH
H
H
N204 H OO H OH
H
HO H OH H
H OH HO H
H OH

An aldobiouronic acid
 O

Uronic acid preparation

 Laboratory preparation of uronic acids
typically involves the use of oxygen in
aqueous solution with platinum or
palladium catalysts
 O

Galactose oxidase
OH
OH
H OO CH3
H H O O CH
OH H H
3
CH2OH H O H OH H
OO H OH galactose H
OH
H oxidase OH OO H OH
OH H H
H H OH H
H OH H H
H OH

IO-

HO

H O O CH
O OH H
3
OH H
H
OH OO H OH
H
OH H
H H
H OH
 O

Hydrogen peroxide
 A non-specific oxidant
 Depolymerizes oligo- or polysaccharides
 Involves a free radical mechanism
 Employs an Fe+2 catalyst
 Fe+2 + HO-OH  Fe+3 + HO. + OH-
 O

.OH

H CH 2 OH
O OH CH 2 OH
H . O OH
OH H H
H OH H
H
O H OH
O H OH
CH 2 OH
O CH 2 OH
H O
H H
OH H H
HO H OH H
HO H
H OH
H OH

CH 2 OH
O OH
O H
OH H
H
H OH

CH 2 OH
H O
H .
OH H
HO
H OH
 O

Esterification
 O

Esters
 Reaction of alcohol with acid anhydride
or acid chloride forms an ester
 Usually done in the presence of a base
such as triethylamine, pyridine, sodium
acetate, sodium hydroxide, etc. This is
known as the Schotten-Bauman
technique and is done to shift the
equilibrium toward the product ester
 O

Use of acid anhydrides

O O
O
R OH +
R R
alcohol
acid anhydride

O O

O +
HO R
R R
fatty acid
ester
 O

Use of acid chlorides

O

R OH + Cl
R
alcohol
acid chloride

O + HCl
R R

ester
 O

Uses of esters
 Esters may be used
– In determination of the monosaccharide
composition of polysaccharides
– In analysis of the sugar composition of foods
– As blocking groups in sugar synthesis
 O

Alditol acetates
 Sometimes aldoses are reduced to sugar
alcohols before acetylation
 The resulting alditol acetates are
– Volatile
– Thermostable
– Ideal for determination by gas
chromatography (GC), where they are
identified by retention time or mass
spectrometry
 O

Alditol acetates

OH O Ac
H O
Ac2O,
H OH H O Ac
H OH
NaBH4 HO H C5H5N Ac O H
HO H
H OH H O Ac
H OH
H OH H O Ac
H OH
OH O Ac
OH
Acetylated
D-glucose D-glucitol
D-glucitol
 O

Esters-General information
 Esters occur in nature
– Acetates, e.g., xanthan
– Succinate half esters
 Esters are extensively used as OH
protecting and blocking groups in
carbohydrate synthesis
– Easily prepared
– Usually crystalline
– Easily removed with base
 O

Sugar phosphates
PO3H2 PO3H2
O PO3H2
O
O OH O O
H H
H H OH
OH H
HO H HO OH
H OH H

D-Glucose D-Fructose
6-phosphate 1,6-bisphosphate
 O

Sugar phosphates
 Sugar phosphates are common
intermediates in sugar metabolism, e.g.,
the Embden-Meyerhof pathway
 Monophosphate esters are also found
naturally in certain starches, e.g., potato
 Modified food starch phosphates (cross-
linked starches) are highly stable to
viscosity breakdown
 O

Starch phosphates
Starch
Starch
O
O

+ O P O Na
O P O Na

OH O
Monostarch phosphate
Starch
(potato starch)
Distarch phosphate
(cross-linked starch)
 O

Ethers
 Williamson ether synthesis
 SN2 mechanism
– RO-Na+ + R’X  ROR’ + NaX
 O

Williamson ether synthesis

SN2 reaction video

 O

4-O-Methyl D-glucuronate
 This ether is a O OH
constituent of various
hemicelluloses and
gum arabic H O OH
H
OH H
H3C O H
H OH
 O

Etherification
 Etherification of some polysaccharides
– Modifies their properties
– Makes them more useful
 Examples (derivatives of cellulose)
– Methyl (-CH3)
– Carboxymethyl (-CH2COO-Na+)
– Hydroxypropyl (-CH2CH(OH)CH3)
 O

Methylation analysis
 Ethers are very useful in methylation
analysis of polysaccharide structure
determination
 With this method it is possible to
determine the nature of the linkages
between the component sugars, e.g., 12,
1  3, 1 4, or 1 6
 O

Methylation analysis
 Typically involves
– Permethylation
– Acid hydrolysis
– Acetylation
– GC-MS analysis
 The hydroxyl groups involved in the
glycosidic linkage are the only ones not
methylated
 O

Internal ethers
(anhydro sugars)
O
R
O

O
O
R A 1C4ring

O
R
A 3, 6-anhydro ring such as
appears in iota-carrageenan
 O

Anhydrosugars
 Typically this internal ether is a 3, 6
linkage
 Such structures are found in seaweed
polysaccharides such as
– agar
– furcellaran
– -carrageenan
– -carrageenan
 O

Non-ionic surfactants
(emulsifiers)
 Sorbitol
– Cyclic dehydration accompanying
esterification with fatty acids produces
sorbitan esters
– The products of these reactions are mixtures
 O

Sorbitan monostearate
 A mixture of
– C16 and C18 esters of sorbitol
– 1, 5-anhydro-D-glucitol
– 1, 4-anhydro-D-glucitol
– and 1,4:3,6-dianhydro-D-glucitol
(isosorbide)
 This mixture may be modified with
ethylene oxide to produce ethoxylated
sorbitan esters (non-ionic detergents)
 O

Sorbitan derivatives
 The mono-, di-, and CH2OH

triesters of these H
HOH2C
HOHC

structures are the HO

H
H
O

H
HO
H H
O
H

HO
Spans H
OH
H
H
OH
H

 Polyoxyethylenated HO

derivatives are the O

Polysorbates and
Tweens O

OH
 O

Cyclic acetals
 The hydroxyl groups on carbohydrates
react with aldehydes or ketones to form
cyclic acetals
 Common carbonyl compounds include
acetone and benzaldehyde
 Sometimes such acetals occur naturally,
as in xanthan gum
 O

Cyclic acetals
CH3
HO
O H
O O
OH
O HO H OH
H H H
The 4,6-O-(1-carboxyethylidene)--D-mannopyranosyl unit
of the polysaccharide xanthan. The acetal is derived from
pyruvic acid.
 O

Cyclic acetals
 Such acetals are good
blocking groups
CH2OH
because they react OH
O
with two hydroxyls H3C
O
H
H
at once and are later H O
easily removable H3C H O

with dilute acid CH3

H3C
 O

Browning reactions
 Alternative names
– Non-enzymatic
– Non-enzymic
– Maillard
 O

w/ sugar
w/o sugar
 O

Two major types of

non-enzymatic browning reactions
 Caramelization

 Maillard browning

Louis Camille Maillard (1878-1936)

 O

Maillard browning
 This is the common browning of foods on
heating or storage
 Requires a reducing sugar and a free
amino group
 The colors (and aromas) so produced
may be desirable or undesirable or only
desirable below a certain level
 Thus, we need top know how to control
this reaction
 O

Food sources of browning

 Soy sauce
 Non-fat dry milk
 Bread crust
 Used to be a problem in dried egg white
but no longer (use of glucose oxidase)
 O

Aromas
 Browning reactions also contribute
flavors in
– Milk chocolate
– Caramels
– Toffees
– Fudges
 O

36% 64%

0.022%

Furanoses are present are

very low levels
 O

Maillard reaction
 O

Decomposition of the
Amadori compound

Control
point
 O

Strecker degradation
H3C CH3 O H3C
O O
CH3
+ H3C OH CO2
+ O
NH2 H3C
O L-valine H
NH2
H3C H3C
methylpropanal
2, 3-butanedione
3-amino-2-butanone

H3C CH3
O H2N N
H3C CH3
O2
+
H3C CH3
N
NH2 O
H3C CH3 tetramethylpyrazine
 O

Maillard browning
reaction variables
 Temperature
– Generally, the higher the temperature, the
greater the browning

 pH
– As pH goes down, so does browning. Due to
protonation of the reactive amino group,
making it unreactive
 O

Under near neutral conditions

O  -
..
NH2 CH
+
 O

Under acidic conditions

H + O-
..
NH2 CH
+
No reaction is possible with the nitrogen
lone pair as it is already reacting with the
hydrogen ion
 O

Maillard browning
reaction variables
 Water

0%, min 30%, max 100%, min

%water
 O

Maillard browning
reaction variables
 Oxygen
– Seems to have little effect

 Metal ions
– Cu(I), Cu(II) and Fe(II), Fe(III) speed up the
reaction. Other metals seems to have little
effect. Can control the with metal chelators
(e.g., EDTA).
 O

Maillard browning
reaction variables
Sugar Open chain Melanoidin
Glucose 1 1
Fructose -- 1.2
Galactose 3.4 2.0
Xylose 7.1 5.5
Arabinose 7.1 6.7
Ribose 354 Very high
 O

A practical example of the

effect of Maillard browning in cookies
 O

Control of browning
 Create unfavorable conditions for the
reactions to take place
– Water--low or high
– pH--lower, decreases browning
– Temperature-- the lower the temperature,
the less the browning
 O

Control of browning
 Removal of substrates
– Glucose oxidase--removes the open chain
form by converting it to gluconic acid. This
technology is used in preserving dried egg
whites.
– Ribose oxidase--This enzyme activity occurs
in Lactobaccilus pentoaceticum and is used to
prevent fish from browning.
 O

Sulfite browning inhibitors

 Principally sulfur dioxide or bisulfite.
These sulfur compounds react with HMF
near the end of the decomposition of the
Amadori compound and divert it to a
non-reactive product, that is one that
cannot be converted into melanoidin
pigments
 O

Sulfite inhibitor reaction

O
R O
H

HMF
-
HSO 3

H
m e la n o id in R O
p ig m e n ts -
SO3
O
H
sta b le --
w ill n o t p ro ce e d
to fo rm p ig m e n ts
 O

Maillard overview
Control point
Pigment
Aromas
 O

Why worry about

Maillard browning?
 Aesthetics
– Overly browned foods are not aesthetically
attractive and may not be accepted
– Also, due to the pyrazines produced by the
Strecker degradation, these highly browned
foods may have odd, off flavors and also not
be accepted
 O

Why worry about

Maillard browning?
 Nutritional effects
– Amino acids that participate in the Maillard
reactions are lost from a nutritional point of
view. This may be especially important
where the amino acid is very reactive and in
foods where it is already in very low
concentration. This would be the case for L-
lysine in cereals.
 O

Reason for the

reactivity of L-lysine
O Reactive epsilon amino
O
H3C
OH H2N
OH
NH2
NH2
L-alanine Alpha amino
L-lysine
 O

Glucose and lysine

 O

Why worry about

Maillard browning?
 Mutagenicity
– This is not yet settled in the literature. Some
researchers find mutagenic products in
browned model systems, others do not. But
there is still the possibility that highly
browned food may contain potential
mutagens.
 O

Reactions during caramelization

 Anomeric equilibration (alpha, beta)
 Pyran, furan equilibration
 Sucrose inversion
– Hydrolysis
 Aldose, ketose interconversions
 Inter- and intramolecular condensation
 Dehydration
 Skeletal fragmentation
 Browning
 O

Thermolytic products
 Vigorous heating HO
H C
HOH2
H
O

produces HO H OH
H OH
intramolecular
H

condensation heat - water

products such as
levoglucosan
O

OH O 1, 6-anhydro-D-glucopyranose
(Levoglucosan)
OH OH
 O

Caramelization
 Flavoring caramelization
buffer
Sucrose syrup Inversion, fragmentation
heat (maltol, etc.)
 O

Flavor compounds
O O
O O
O
H

O O O O
H H
Maltol
Isomaltol 2H-4-Hydroxy-5-
methylfuran-3-one
 O

Flavor compounds
 Maltol and isomaltol contribute to fresh
baked bread flavor
 2H-4-Hydroxy-5-methylfuran-3-one is
characteristic of burnt meat flavors and
is also a flavor and sweetener enhancer
 O

Caramelization
 Caramel pigments

Glucose Partially heat Caramel

syrups neutralized Colloids
pH 4
 O

Caramel pigments
 During the reaction, unsaturated ring
compounds are produced which condense
and produce the polymer pigments
(caramel colloids)
 Specific catalysts will produce specific
caramel colors, solubilities and acidities
 O

Pigment uses
 Colas
 Baked goods
 Syrups
 Candies
 Pet foods
 Dry seasonings
 Beer and other alcoholic beverages
 O

Pigment structure
 Unknown
 Contains
– hydroxyl groups
– carboxylic groups
– ketone groups
– hydroxylated carbon-carbon double bonds
– phenolic groups
 O

Pigment production
 The formation rate of the pigments
increases with increasing temperature
and pH
 In the absence of buffering salts, large
amounts of humin (C125H188O8), a bitter
substance, is produced
 Minimal production of humin is desired
 O

Acrylamide
 A study in 2002 detected small amounts
of acrylamide in foods
 The concern is the acrylamide is a known
neurotoxin and carcinogen
 Initial reports targeted starchy foods but
it is now known that acrylamide is not
directly formed from starch nor is it
required for it’s formation
 O

Where found?
 O

Consumption?
 O

Common factor
 High heat processing
– Not formed in boiled foods (100oC), e.g.
boiled potatoes
– Neurotoxic “no observed adverse effect”
level was >>>> than what could be achieved
by food consumption
 Main concern was carcinogenic potential
– Human intake is 1000 times lower than
carcinogenic level in rats
– No known case of cancer from eating
acrylamide containing foods
Formation
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Details on next slide
 O

Formation (cont.)
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Formation
 L-asparagine is required
– The carbons in acrylamide come from
asparagine
 Promoting formation
– Free asparagine
– Reducing sugars
– High temps (>130oC, best at 200oC)
– Low moisture
– pH 5.5-7.0
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Reducing acrylamide formation

 Reduce asparagine
– Select cultivars low in asparagine or use
asparaginase to remove
 Reduce reducing sugars
– Select cultivars, avoid cold storage, blanch
 Lower temp
– But this can decrease brownness
 Lower pH
– Dip in citric acid, may cause off taste
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Reducing acrylamide
formation
 Add other amino acids
– Provides competition for asparagine, e.g.
glycine or glutamine
 Degradation of formed acrylamide
– Prolonged heating, e. g., coffee roasting