 Enzymatic (polyphenoloxidase).

Fresh cut
vegetables, fruit eg. Apples, potatoes, non-
toxic, no flavor
(When the tissue is cut or peeled, it rapidly
darkens on exposure to air as a result of
conversion of phenolic compounds to brown
melanoidins)
 Caramelization. Sugars at very high
temperatures.
 THE MAILLARD REACTION
Nonenzymatic Browning
 Two major types of non-
enzymatic browning reactions

1. Caramelization
2. Maillard browning
Browning Reactions
Caramelization
 Caramelization is the browning of sugar, a
process used extensively in cooking for the
resulting nutty flavor and brown color.


Caramelization in
sucrose; all brown color
is a result of browning,
as sucrose is normally
colorless
Caramelization
 Caramel is produced commercially as a coloring
material and flavoring materials.
 Carbohydrate used : sucrose (most often), D-
fructose, D-glucose, invert sugars may also be used
 Reaction is facilitated by small amount of acids and
certain salts.
 Acids : food grade sulfuric, phosphoric, acetic,
citric acid
 Bases: ammonium, sodium, potassium and calcium
hydroxides
 Salts : ammonium, sodium, potassium carbonates
4 classes of caramel
 Class 1 caramel (plain caramel or caustic
caramel)
Prepared by heating a carbohydrate without
a source of either ammonium or sulfite ions
 Class 2 caramel (caustic sulfite caramel)
Prepared by heating a carbohydrate in the
presence of sulfite ions but in the absence of
ammonium ions
This caramel used to add color to beers &
other alcoholic beverages is reddish brown,
has a solution pH of 3-4
4 classes of caramel
 Class 3 caramel (ammonium caramel)
Prepared by heating a carbohydrate in the presence
of ammonium ions but in the absence of sulfite ions
This caramel used in bakery products, syrups and
puddings is reddish brown, has a solution pH of 4.2-
4.8
 Class 4 caramel (sulfite ammonium caramel)

Prepared by heating a carbohydrate in the presence

of ammonium ions and sulfite ions
This caramel used in cola soft drinks, acidic
baverages, candies, pet foods is brown, has a solution
pH of 2-4.5
Caramelization
 Some important molecules that are
produced during caramelization are
Furans (have a nutty aroma)
-Hydroxymethylfurfural (HMF) is found in
honey, juices, milk but also in cigarettes.
-Hydroxyacetylfuran (HAF) has a sweet
aroma and a low odor threshold.
Caramelization
Diacetyl (smells like butter)
-Diacetyl is mainly responsible for a buttery
or butterscotch flavour
- eg. 2,3-butanedione
Maltol (toasty)
-Maltol has a toasty flavor and used as a
flavor enhancer in breads and cakes.
Esters and lactones which have a sweet rum
like flavor
 “thesequence of events that begins with
reaction of the amino group of amino acids
with a glycosidic hydroxyl group of sugars;
the sequence terminates with the formation
of brown nitrogenous polymers or
melanoidins”
 John deMan
Maillard browning
 Common browning of foods on heating or on
storage that is usually due to a chemical
reaction between reducing sugars mainly D-
glucose and a primary amino group (a free
amino acid or amino group on a side chain of
a protein molecule
 Also called as nonenzymatic browning to
differentiate it from the more rapid, enzyme
catalyzed browning commonly observed in
freshly cut fruits and vegetables such as
apples and potatoes
 Desirable:
– Color - bread crust, syrup, meat
-Flavor - coffee, cocoa, meats

 Undesirable
– Color - changes in color during storage
-Flavor - changes during processing and storage
-Nutritional loss - essential amino acids
-Toxicity/mutagenicity
 Occurs between reducing sugars and amines
at high temperatures
 Produces flavor
 Produces color
 Produces toxic products
 Destroys nutrients (lysine)
Five steps are involved in the
process
1.The production of an N-substituted
glycosylamine from an aldose or ketose
reacting with a primary amino group of an
amino acid, peptide, or protein.
2.Rearrangement of the glycosylamine by an
Amadori rearrangement type of reaction to
yield an aldoseamine or ketoseamine.
3.A second rearrangement of the ketoseamine
with a second mole of aldose to form a
diketoseamine, or the reaction of an
aldoseamine with a second mole of amino acid
to yield a diamino sugar
Five steps are involved in the
process
4. Degradation of the amino sugars with loss of
one or more molecules of water to give amino
or nonamino compounds.
5. Condensation of the compounds formed in
Step 4 with each other or with amino
compounds to form brown pigments and
polymers.
Formation of Glycosylamines in the
Browning Reaction

D-glucose Glycosylamines

The mechanism as shown is thought to involve

addition of the amine to the carbonyl group of
the open-chain form of the sugar, elimination of a
molecule of water, and closure of the ring.
Amadori rearrangement
 The Amadori rearrangement of the
glycosylamines involves the presence of an
acid catalyst and leads to the formation of
ketoseamine or 1-amino-1-deoxyketose
 The ketoseamines are relatively stable
compounds, which are formed in maximum
yield in systems with 18 percent water
content
 Amadori rearrangement
The mechanism of
the Amadori
rearrangement
involves
protonation of the
nitrogen atom at
carbon 1
Glycosylamines

Amadori
compound-
ketoseamine
 HMF (hydroxymethylfurfural)

Conversion of amidori product to HMF

 HMF (hydroxymethylfurfural)

Amines •Polymerize to a dark colored,

insoluble material containing
nitrogen called melanoidin.
•Pyrolytic products
Strecker aldehydes
 O OH
OH
O
O
O CH3 CH3

maltol isomaltol
 Reactionsbetween - dicarbonyl compounds
formed in the Maillard reaction and amines.

 Thereaction yields aminoketones, aldehydes

and carbon dioxide.
 O O O
R -H2O -H2O O
COOH R
O H2N N -H2O H
OH
H
H R Stecker
dicarbonyl amine NH3 CO2
COOH

O O
-H2O -H2O O
COOH R
N -H2O H
OH
H R Stecker aldehyde
ine NH3 CO2
COOH
Rich nutty, meaty flavors
 Asparagine

O CH2
C O
H
H2N CH
O
C
Strecker product H2N H2

Acrylamide
(neurotoxicant and a weak
human carcinogen.
 Level of acrylamide (< 1.5 ppm) have been
reported in variety of food product that are
made by frying, baking and roasting.
 Acrylamide is not detected in unheated or
boiled foodstuffs
 Acrylamide is a known neurotoxicant and a
weak human carcinogen.
Food Ppb Acrylamide
Almonds 236-457
Breads 0-343
Cookies 36-432
French fries 20-1325
Potato chips 117-196
 Acrylamide is derived from the reaction
between reducing sugar and the a-amino
group of free L-asparagine
 The reaction occurs via Schiff base
intermediates, which then undergoes
decarboxylation, followed by carbon-carbon
bond cleavage to form acrylamide, whose
atoms are derived from L-asparagine
Acrylamide formation requires a
minimum T of 120oC.
Cannot occur in high moisture
foods
pH 4-8
2 strategies to minimize formation of
acylamide in foods:
1. Removal of either one or both the
substrates (eg. soaking in water, reduce
60% of acrylamide level within processed
potato product by removing reaction
substrates, reducing sugar & free
asparagine)
2. Alteration of processing condition
 Reagent modification (eg. Protonation of
asparagine by lowering the pH,protonation of
the a-amino group of asparagine, reducing its
nucleophilic potential ; conversion of
asparagine to aspartic acid with asparaginase
; incorporation of salts have been shown to
mitigate acrylamide formation)
 Optimization of thermal processing
condition (Temperature, Time)-minimize
acrlamide level
 Occurs in many fruits and vegetables
 When the tissue is cut or peeled, it rapidly
darkens on exposure to air as a result of
conversion of phenolic compounds to brown
melanoidins
 Cu cofactor
 Active between pH 5-7
 Phenolic compounds are widely distributed in
the plant kingdom and are considered to be
secondary metabolites.
 Structurally they contain an aromatic ring
bearing one or more hydroxyl groups.
 Polyphenol oxidase (PPO) catalyses the
oxidation of phenols to o-quinones, which
are highly reactive compounds.
 O-quinones thus formed undergo
spontaneous polymerization to produce
high-molecular-weight compounds or
brown pigments (melanins).
 These melanins may in turn react with
amino acids and proteins leading to
enhancement of the brown colour
produced.
Quinone formation is both
dependant of enzyme and oxygen
Once the reaction starts, the
subsequent reactions occur
spontaneously and no longer depend
of oxygen or enzyme
 Enzymatic browning can be inhibited by
targeting the enzyme, the substrates (oxygen
and polyphenols) or the products of the
reaction.
 i) Inhibition targeted toward the enzyme
 ii) Inhibition targeted toward the substrate
 iii) Inhibition targeted toward the products
 Inhibitors which act directly on polyphenol
oxidase
 Chelators
– azide, cyanide, carbon monoxide, halide ions
and tropolone, is well documented for the
inhibition of polyphenol oxidase from various
sources.
- phosphates, EDTA, organic acids
 Inactivationof the polyphenol oxidases by
heat treatments such as steam blanching is
effectively applied for the control of
browning in fruits and vegetables to be
canned or frozen. Heat treatments are not
however practically applicable in the storage
of fresh produce.
 Theelimination of oxygen from the cut
surface of fruits or vegetables greatly retards
the browning reaction. Exclusion of oxygen is
possible by immersion in water, syrup, or by
vacuum treatment.
 Vacuum packaging of pre-peeled potatoes
to exclude oxygen, was observed to
extend their shelf life.
 Vacuum packaged products however
rapidly undergo browning upon exposure
to air.
 Anaerobic conditions created by vacuum
packaging are a cause for safety concern in that
they are potentially capable of supporting the
growth of Clostridium botulinum and the
production of its toxin
 Sulphated polysaccharides also have an
inhibitory effect on browning
 Apart from possible complexation, sulphate
groups are believed to exert their inhibitory
effect through chelation of the copper group of
the polyphenol oxidase
 Specific adsorbents, which undergo
complexation with the phenolic substrate may
be applied in the physical elimination of
phenolic compounds from food systems.
 The use of cyclodextrins for the removal of
phenolic compounds from raw fruit and
vegetable juices has been patented in the United
States
 Cyclodextrins are thought to inhibit polyphenol
oxidase activity through the formation of
inclusion complexes with polyphenols
 Enzymatic modification of phenolic substrates
may serve to inhibit polyphenol oxidase activity.
 O-methyltransferase for example converts o-
dihydroxy phenolics to the corresponding
methoxy derivatives, which do not serve as
substrates for polyphenol oxidase
 Similarly, protocatechuate 3,4-dioxigenase
prevents the browning of apple juice due to
substrate modification
 These enzyme modification techniques are not
however feasible in commercial use, owing to
the high cost of these enzymes.
O-quinones, which are the products
of diphenol oxidation, are capable
of reacting with each other,
resulting in the formation of dimers
of the original phenol.
 These dimers, which possess an o-
diphenolic structure
 Ascorbic acid, thiol compounds, sulphites
are capable of inhibiting dimer formation
by reducing o-quinones to o-diphenols, or
through the formation of colourless
addition products.
 Mechanisms involved in the control of
enzymatic browning by sulphites are
shown in Figure
 The primary role of reducing agents
such as sulphiting agents in the
inhibition of enzymatic browning is to
reduce the pigment precursors
(quinones) to colourless, less-reactive
diphenols.
51
 Although sulphites are very effective in
controlling browning, they are subject to
regulatory restrictions owing to their potentially
adverse effects on health.
 Many reports have described allergic reactions
in humans, following the ingestion of sulphite-
treated foods by hypersensitive asthmatics.
 Food and Agriculture Organization (FAO)
recommend an acceptable sulphite daily intake
of 0-0.7 mg sulphur dioxide per kg of body
weight.
 The FDA has proposed maximum residual sulphur
dioxide levels for certain foods. In accordance
with these proposed limits,
 residual sulphur dioxide levels for fruit juices,
dehydrated potatoes, and dried fruit, are 300,
500, and 2000 ppm respectively (Federal
Register, 1988).
 Shrimp products having residual sulphite levels in
excess of 100 ppm are considered unsafe
(Federal Register, 1985).
 Ascorbic acid also acts as an oxygen scavenger
for the removal of molecular oxygen in
polyphenol oxidase reactions.
 Polyphenol oxidase inhibition by ascorbic acid
has been attributed to the reduction of
enzymatically formed o-quinones to diphenols
 Ascorbic acid is however irreversibly oxidized to
dehydroascorbic acid during the reduction
process, thus allowing browning to occur upon its
depletion
 More stable forms of ascorbic acid derivatives :
erythrobic acid, 2- and 3-phosphate derivatives
of ascorbic acid, phosphinate esters of ascorbic
acid, and ascorbyl-6-fatty acid esters of ascorbic
acid
 quinone

diphenol
 An important characteristic of sugars is their
ability to form crystals.
 In the commercial production of sugars,
crystallization is an important step in the
purification of sugar. The purer a solution of
a sugar, the easier it will crystallize .
 In certain foods, crystallization is
undesirable, such as the crystallization of
lactose in sweetened condensed milk or ice
cream
 Crystal growth of sucrose consists of two
steps:
(1) the mass transfer of sucrose molecules
to the surface of the crystal, which is a
first-order process; and
(2) the incorporation of the molecules in
the crystal surface, a second-order process.
 Lactose can occur in two crystalline forms,
the -hydrate and the -anhydrous forms.
 The most common form is the -hydrate
(C12H22O11-H2O), which can be obtained by
crystallization from a supersaturated solution
below 93.50C.
 When crystallization is carried out above
93.50C, the crystals formed are of -
anhydrous type.
The solubility of lactose is less than
most other sugars, which may
present problems in a number of
foods containing lactose
When milk is concentrated 3:1, the
concentration of lactose approaches
its final solubility.
 When this product is cooled or when sucrose
is added, crystals of a-hydrate may develop.
 Such lactose crystals are very hard and
sharp; when left undisturbed they may
develop to a large size, causing a sensation
of grittiness or sandiness in the mouth. This
same phenomenon limits the amount of milk
solids that can be incorporated into ice
cream.
 Thecrystals of a-hydrate lactose usually
occur in a prism or tomahawk shape.
The concentration of sugar in the
solution is increasing due to the
evaporation of water
 There is no change in the sugar
concentration
 But there is a decrease in temperature
 At some point (a unique combination of
concentration and temperature) the solution
will become saturated
 Asthe temperature drops below the
saturation point, the solution becomes
supersaturated (contain more dissolved
substances that can be dissolved)
 But when should we induce crystal
formation?
 At a relatively high temperature, or
 At a relatively low temperature?
 The answer depends on understanding about
sugar crystal growth
Sugar
Sugar
Sugar
Sugar
Induce
here

Induce
here

Large
crystals, Small crystals,
coarse smooth
 This is important because in candy (and
frozen desserts) crystal size = texture
 Large crystals = coarse, crunchy texture
 Small crystals = smooth, velvety texture
 Starch can be hydrolyzed by acid or enzymes
or by a combination of acid and enzyme
treatments.
 A large variety of products can be obtained
from starch hydrolysis using various starches
such as corn, wheat, potato, and cassava
(tapioca) starch.

Cassava starch
Corn starch Potato starch
 Glucosesyrups, are hydrolysis products of
starch with varying amounts of glucose
monomer, dimer, oligosaccharides , and
polysaccharides.

 The degree of hydrolysis is

expressed as dextrose equivalent (DE),
defined as the amount of reducing sugars
present as dextrose and calculated as a
percentage of the total dry matter.
 Glucose syrups have a DE greater than 20 and
less than 80.
DE< 20 -the products are referred as
maltodextrins
DE>80 -the products are referred as
hydrolyzates.
 The properties of maltodextrins are
influenced by the nature of the starch used;
those of hydrolyzates are not affected by the
type of starch.
 Theinitial step in starch hydrolysis involves
the use of a heat-stable a-amylase. This
enzyme randomly attacks a-1, 4 glycosidic
bonds resulting in rapid decrease in
viscosity.
 These enzymes can be used at temperatures
as high as 1050C.
reaction produces maltodextrins,
 This
which can be used as important functional
food ingredients ; fillers , stabilizers,
and thickeners.
 The next step is saccharification by
using a series of enzymes that hydrolyze
either the a-1,4 linkages of amylose or the
a-1, 6 linkages of the branched
amylopectin.
 The action of the various starch-degrading
enzymes is shown in Figure 1
 In addition to products containing high levels
of glucose (95 to 97 percent), sweeteners
with DE as follows can be produced.
40 to 45 (maltose)
50 to 55 (high maltose),
55 to 70 (high conversion syrup)

 High dextrose syrups can be obtained by

saccharification with amyloglucosidase.
 The next step is the isomerization of
glucose to fructose.
 This process is done with glucose
isomerase in enzyme reactors.
 The conversion is reversible and the
equilibrium is at 50 percent conversion.
 High-fructose syrups are produced with 42 or
55 percent fructose.
debranching
enzyme.
 Polyols or sugar alcohols occur in nature and
are produced industrially from the
corresponding saccharides by catalytic
hydrogenation.
 Sorbitol, the most widely distributed natural
polyol, is found in many fruits such as plums,
berries, cherries, apples, and pears. It is a
component of fruit juices, fruit wines, and
other fruit products.
 It is commercially produced by catalytic
hydrogenation of D-glucose.
 Starch

Enzymatic hydrolysis

Dextrose syrup Maltose syrup

Hydrogenation

Sorbitol syrup

Crystallization

sorbitol