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Carbohydrate Reactions
Important reactions
of carbohydrate molecules
Carbonyl group
(alone)
Oxidation to a
carboxylic acid
group
Reduction to a
hydroxyl group
Cyanohydrin
reaction (and
reaction with other
nucleophiles)
Important reactions (cont.)
Hydroxyl groups Ester formation
Ether formation
Cyclic acetal
Oxidation to
carbonyl
Reduction to deoxy
Replacement with
NH
2
, SH, or X
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Important reactions (cont.)
Both carbonyl and
hydroxyl groups
Cyclic hemiacetals
(pyranose/furanose)
Formation of acetals
(glycosides)
Aldose/ketose
isomerizations
Important reactions
Anomeric hydroxyl
group
Formation of
glycosyl halides
Oxidation to lactones
Carbonyl group oxidation
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Tollens test
2Ag(NH
3
)
2
+
+ RCHO + 3OH
-
2Ag(s) + RCOO
-
+ 4NH
3
+ 2H
2
O
A silver
mirror
This is
reduced to
This is
oxidized to
Fehlings test
2Cu(OH)
2
+ RCHO
RCOOH + Cu
2
O + H
2
O
A brick red
precipitate
This is
reduced
This is
oxidized
Tollens test Fehlings test
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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
Stoichiometric
sugar determination
RCHO + I
2
+ 3NaOH
RCOO
-
Na
+
+ 2NaI + 2H
2
O
This reaction utilizes oxidation of the aldehyde with
hypoiodite (IO
-
) at pH 9.5
Glucose oxidase
O
2
H
2
O
2
In alkaline
solution
this exists
as
D-gluconate
D-glucono-
1,5-lactone
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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
Lactones
-lactone
o-lactone
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
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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 -D-
glucopyranose
GDL as preservative
Due to its general acidity, GDL will
decrease microbial action in such systems
as
bread dough
fish cakes
surimi
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
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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
Carbonyl group reduction
Glucose to glucitol (sorbitol)
D-glucose D-glucitol
(sorbitol)
reduction
NaBH
4
,
H
2
/Pt or Pd,
H
2
/Raney Ni
Yield ~ 100%
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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
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)
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)
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Sorbitol and Vitamin C
Acetobacter
suboxydans
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
Other Vitamin C functions
Plants
Respiration
Growth
Maintenance of CO
2
balance
Foods
General antioxidant function, especially in
aqueous systems
Natural occurrence
Rose hips, persimmon, citrus fruits
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Mannitol
Mannitol preparation
Laboratory methods
Hydrogenation of D-mannose
Fermentation
Commercially
Hydrogenolysis of sucrose
Sucrose hydrogenolysis
Sucrose
D-glucose
D-fructose
D-glucose,
D-fructose,
D-mannose
Mannitol,
Glucitol
Isomerization
Hydrogenation
Hydrogenation
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Mannitol
characteristics and uses
Not a humectant
Crystallizes easily, moderately water
soluble
Candies
Anti-caking agent
Dusting agent
Sweetness: 65% of that of sucrose
Mannitol uses (cont.)
Sugarfree
Chocolates
Mints
Cough drops
Hard and soft candies
Xylitol
hydrogenation
(from hemicellulose
xylans)
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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
Maltitol
(from corn syrup)
Maltitol uses
Reduced calorie candies
Miscellaneous other foods
Also included here are isomaltitol and
hydrogenated starch hydrolyzates
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Erythritol
Non-cariogenic
0.2 kcal/gram
Strong cooling effect in the mouth
RS ~0.5
Not hygroscopic
No D or L designation
This is a meso compound
Cyclitols
Only contain hydroxyl groups
But unlike alditols, these are cyclic
Most are 6 membered rings
(polyhydroxycyclohexanes and hexenes)
Myo-Inositol
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
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Phytic acid
Mostly occurs as phytates, mixed
potassium-magnesium salts
Generally an anti-nutritional factor as it
binds minerals, especially calcium and
doesnt allow them to be absorbed
Recent studies have shown some positive
effects related to cancer, heart disease,
antioxidant activity and contribution to
fecal bulk
Oxidation of
non-anomeric hydroxyl groups
The product of this
type of oxidation of
an aldose is an
aldaric acid
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Periodate oxidation
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
Dinitrogen tetroxide (N
2
O
4
)
This reagent is specific for oxidation of
primary alcohol groups
If you start with an aldose, you get an
alduronic acid
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Dinitrogen tetroxide
Methyl o-D-
glucopyranoside
Methyl o-D-
glucopyranosiduronic
acid
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)
An aldobiouronic acid
N
2
0
4
Uronic acid preparation
Laboratory preparation of uronic acids
typically involves the use of oxygen in
aqueous solution with platinum or
palladium catalysts
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Galactose oxidase
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
-
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Esterification
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
Use of acid anhydrides
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Use of acid chlorides
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
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
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Alditol acetates
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
Sugar phosphates
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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 starces) are highly stable to
viscosity breakdown
Starch phosphates
Ethers
Williamson ether synthesis
S
N
2 mechanism
RO
-
Na
+
+ RX ROR + NaX
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Williamson ether synthesis
S
N
2 reaction video
4-O-Methyl D-glucuronate
This ether is a
constituent of various
hemicelluloses and
gum arabic
Etherification
Etherification of some polysaccharides
Modifies their properties
Makes them more useful
Examples (derivatives of cellulose)
Methyl (-CH
3
)
Carboxymethyl (-CH
2
COO
-
Na
+
)
Hydroxypropyl (-CH
2
CH(OH)CH
3
)
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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., 12,
1 3, 1 4, or 1 6
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
Internal ethers
(anhydro sugars)
A
1
C
4
ring
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Anhydrosugars
Typically this internal ether is a 3, 6
linkage
Such structures are found in seaweed
polysaccharides such as
agar
furcellaran
k-carrageenan
i-carrageenan
Non-ionic surfactants
(emulsifiers)
Sorbitol
Cyclic dehydration accompanying
esterification with fatty acids produces
sorbitan esters
The products of these reactions are mixtures
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)
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Sorbitan derivatives
The mono-, di-, and
triesters of these
structures are the
Spans
Polyoxyethylenated
derivatives are the
Polysorbates and
Tweens
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
Cyclic acetals
The 4,6-O-(1-carboxyethylidene)-|-D-mannopyranosyl unit
of the polysaccharide xanthan. The acetal is derived from
pyruvic acid.
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Cyclic acetals
Such acetals are good
blocking groups
because they react
with two hydroxyls
at once and are later
easily removable
with dilute acid
Browning reactions
Alternative names
Non-enzymatic
Non-enzymic
Maillard
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
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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)
Aromas
Browning reactions also contribute
flavors in
Milk chocolate
Caramels
Toffees
Fudges
~36% ~64%
0.022%
Furanoses are present are
very low levels
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Maillard reaction
Decomposition of the
Amadori compound
Control
point
Strecker degradation
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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
Under near neutral conditions
..
NH
2
CH
O
Under acidic conditions
..
NH
2
CH
O
H
+
No reaction is possible with the nitrogen
lone pair as it is already reacting with the
hydrogen ion
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Maillard browning
reaction variables
Water
%water
0%, min 100%, min 30%, max
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).
Maillard browning
reaction variables
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A practical example of the
effect of Maillard browning in cookies
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
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.
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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
Sulfite inhibitor reaction
Maillard overview
Control point
Pigment
Aromas
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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
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.
Reason for the
reactivity of L-lysine
Alpha amino
Reactive epsilon amino
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Glucose and lysine
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.
Reactions during caramelization
Anomeric equilibration (alpha, beta)
Pyran, furan equilibration
Sucrose inversion
Hydrolysis
Aldose, ketose interconversions
Inter- and intramolecular condensation
Dehydration
Skeletal fragmentation
Browning
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Thermolytic products
Vigorous heating
produces
intramolecular
condensation
products such as
levoglucosan
Caramelization
Flavoring caramelization
Sucrose syrup
Inversion, fragmentation
buffer
heat
(maltol, etc.)
Flavor compounds
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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
Caramelization
Caramel pigments
Glucose Partially Caramel
syrups neutralized Colloids
pH 4
heat
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
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Pigment uses
Colas
Baked goods
Syrups
Candies
Pet foods
Dry seasonings
Beer and other alcoholic beverages
Pigment structure
Unknown
Contains
hydroxyl groups
carboxylic groups
ketone groups
hydroxylated carbon-carbon double bonds
phenolic groups
Pigment production
The formation rate of the pigments
increases with increasing temperature
and pH
In the absence of buffering salts, large
amounts of humin (C
125
H
188
O
8
), a bitter
substance, is produced
Minimal production of humin is desired
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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 its formation
Where found?
Consumption?
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Common factor
High heat processing
Not formed in boiled foods (100
o
C), 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
Formation
L-asparagine is required
The carbons in acrylamide come from
asparagine
Promoting formation
Free asparagine
Reducing sugars
High temps (>130
o
C, best at 200
o
C)
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
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