changes during freezing-physical and chemical changes.

INTRODUCTION
One of the greatest challenges for food technologists is to
maintain the quality of food products for an extended period.
The principles of low-temperature preservation have been
employed for many years. It renders advantageous negative
effect of reduced temperature on various chemical and
biochemical reactions responsible for food spoilage, as well as
on microbial growth and spore germination. Freezing is a
well-known long-term preservation process widely used in the
food industry. This is because changes in the nutritional or
sensory characteristics of foods are small if appropriate
freezing and storage procedures are followed. The freezing of
foods normally consists of pre-freezing treatments, freezing,
frozen storage, and thawing, each of which must be properly
conducted to obtain optimum results. A decrease in
temperature generally decreases the rate of chemical reactions
that are responsible for the deterioration in food quality over
time; therefore freezing is frequently used to extend the shelf
life of food products. When a product is frozen, the formed
ice crystals may cause cell rupture and alterations in the
transport properties of cell membranes, which have practical
consequences in terms of leaching of cellular substances from
tissues as well as water loss, leading normally to
disappointing consequences in terms of texture. There is a
general acceptance that high freezing rates retain the quality
of a food product better than lower freezing rates since
evidence tends to show that relatively slow freezing causes
large ice crystals to form exclusively in extracellular areas,
while high freezing rates produce small uniformly distributed
ice crystals. The formation of ice may result in textural
changes and disruption of cell compartments that cause the
release of chemically reactive components. Furthermore, the
removal of water during ice formation concentrates the solutes
in an unfrozen matrix, which can affect reaction conditions,
such as pH and ionic strength. Therefore in order to extend the
shelf life of frozen food products, it is crucial to understand
the chemical reactions that can occur in food components that
can lead to quality deterioration.
28.2 EFFECT OF FREEZING ON PRINCIPAL
CONSTITUENTS OF FOODS
The effect of freezing on the food components is diverse, and
some components are affected more than others. For example,
protein can be irreversibly denatured by freezing, whereas
carbohydrates are generally more stable. Other common
chemical changes that can proceed during freezing and frozen
storage are lipid oxidation, enzymatic browning, flavour
deterioration, and the degradation of pigments and vitamins.
The main goal of the freezing process is to extend the shelf
life of a raw material or product beyond that achievable at
temperatures above the initial freezing point of the material.
Therefore, it is important to understand the modifications that
can occur during freezing in food components and that can
further lead to quality degradation. This chapter focuses on
chemical and biochemical reactions that affect the quality of
frozen food systems. These reactions and specific examples in
food are summarized in Table 28.1.
Changes that occur in foods during freezing, storage and
thawing can be both chemical and physical in nature. Various
chemical, enzymatic and physical changes are promoted as a
result of the concentration of components (concentration
effects) in the unfrozen water phase within the frozen foods.
For example:
  Chemical changes such as oxidative rancidity or
oxidation of flavour components, pigments and
vitamins.
 Enzymatic reactions such as enzymatic
browning or lipolytic rancidity.
 Meats become tougher due to protein
denaturation by chemical effects and cell
breakage by ice crystals
In freezing foods, the objective is to promote the formation of
tiny ice crystals rather than the formation of fewer but larger
ice crystals that cause cellular damage. Ice crystal damage can
lead to loss of water from the food product once it is thawed.
The drip that is found in thawed strawberries or beef is due in
part to ice crystal damage to the cells, leading to leakage of
cellular fluids into extracellular spaces, and to the loss of
water-holding capacity of food components as a result of
concentration effects.
Other undesirable changes include formation of package ice
and freeze dehydration which is popularly called freezer burn
and can produce unsightly food surfaces and loss of nutrients.
"Freezer burn" is a misnomer since the food does not "burn"
in the freezer but rather takes on an appearance of having been
burned because of the moisture loss that occurs during this
freeze dehydration.
28.2.1 WATER
Water is an essential constituent of most foods. It is present in
a very wide range, varying, for example, from 4% in milk
powder up to 95% in tomato and lettuce. Water may exist as
an intracellular or extracellular component in vegetable and
animal products, as a dispersing medium or solvent in a
variety of products, as the dispersed phase in some emulsified
products such as butter and margarine, or as a minor
constituent in other foods. The conversion of water into ice
during freezing has the advantage of fixing the tissue structure
and separating the water fraction in the form of ice crystals in
such a way that water is not available as a solvent or cannot
take part in deterioration reactions. On the other hand, ice
crystals formed during freezing can affect quality parameters
such as color, texture, and flavor. Meanwhile, in the
remaining unfrozen portion, the concentration of dissolved
substances increases, while the water activity of a product
decreases. Usually, this part of water is non-freezable and,
therefore, not available for chemical reactions or as
plasticizers. The water that does not freeze is normally
considered to be the critical water content above which
deteriorative changes may occur. Critical water is a rather
unusual substance having high boiling and low freezing
points, high specific heat, high latent heats of fusion and
vaporization, high surface tension, high polarity, and unusual
density changes. The considerable difference in the densities
of water and ice may result in structural damage to foods
when they are frozen, being more likely in plant tissue with its
rigid structure and poorly aligned cells than in muscle with its
pliable consistency and the parallel arrangement of cells.
Table-28.1: Chemical Reaction of Food Components
during Freezing that Affect Food Quality

Food Mechanism of Effect on Studies in

Components degradation quality food

Protein Denaturation Degradation Toughenin

of texture g and
 functional
changes,
particularly
loss of
protein
solubility
in fish,
Loss of
and
protein
functional
solubility,
properties
emulsifying
capacity
Loss of
water
holding
capacity of
meat for
processing

Lipid Hydrolysis Release of Short chain

FFA— FFA giving
known to rancid
odour in
contribute to dairy
unpleasant product
flavours
Medium
chain FFA
caused a
‘‘sweaty’’
flavour in
 mutton
Toughenin
g of
muscles in
frozen
Indian
sardine

Unstable
hydroperoxid
es break
down to
reactive Off-
compounds flavours
Oxidation shelled
and interact oysters
Autoxidation with other
Detection
Enzymatic components of rancid
oxidation to produce flavour in
off flavours, silver
discoloration pomfret
, and
toughening
of muscle
protein

Carbohydrate Hydrolysis Increases the Sucrose

s amount of hydrolysis
smaller
molecular
 weight
components
—leads to
lower
melting Firmness of
temperatures ice cream
decreased
as
Change of hydrolysis
texture progressed

Greenness
in Brussels
sprouts
decreased
Stability of
Colour Green green
pigments chlorophyll colour in
forms kiwifruit
olivebrown Frozen
(a) Pheophytinizati pheophytin blanched
Chlorophyll on spinach had
in the
a higher
presence of amount
acid or heat
of
pheophytin
than fresh

(b) Enzymatic Glucosidase Loss of

 hydrolyses
glycosidic
linkages and anthocyani
produces n in
reaction
sugars raspberry
in the late
and aglycone cultivar
compounds was more
severe than
the early
cultivars
Anthocyanin Depending
on the pH of
the food,
Red colour
different hue of sour
Structure of forms of cherry
anthocyanin anthocyanin weakened
exist, during
depends on pH
value usually from frozen
red to blue as storage
pH
increases

(c) Oxidation The loss of Loss of

Carotenoids pigments carotenoid
causes fading in salmon
of colour and
loss of
nutritive
 value

Flavour Enzymatic Change of Change in

compounds degradation flavour the
profile compositio
n of
aromatic
compounds
of
strawberrie
s
Change in
Lipid oxidation aroma
Produces off- profile of
flavours frozen
green peas
Green and
fatty off-
flavour
notes in
frozen trout
Leaching of Weakens and due to
components changes the breakdown
during the sensory products of
blanching unsaturated
Perception FFA
process
Decrease
of organic
acids in
green beans
and Padron
 peppers
Change in
volatile
compositio
n of guava
Effect of
Effect of heat heat
Changes in
concentrati
on of
odorants
during heat
treatment

Micronutrient Loss of
s vitamins C
Oxidation Loss of and B6 due
(a) Vitamins nutritional to
value blanching
because in french
of the loss of fries
vitamins Oxidation
of AA in
peas, lima
beans, corn
and green
Generally beans
(b) Minerals stable
Unchange
d mineral
compositio
 n in
artichokes,
green
beans, and
peas after
freezing
Mineral
content of
Loss mainly boiled fresh
through vegetables
leaching was not
different
from frozen
vegetables

When water freezes at atmospheric pressure, it expands nearly

9%. The degree of expansion varies considerably owing to the
following factors:
 Moisture content: Higher moisture contents
produce greater changes in volume.
 Cell arrangement: Intercellular air spaces,
which are common in plant tissue. These spaces
can probably accommodate growing crystals,
and thereby minimize changes in the specimen
exterior dimensions; for example, whole
strawberries increase in volume by 3%, whereas
coarsely ground strawberries increase by 8.2%,
when both are frozen to −20°C
 Concentration of solutes: High concentrations
reduce the freezing point and do not freeze or
expand at commercial freezing temperatures.
  Freezer temperature: It determines the amount
of unfrozen water and hence the degree of
expansion.
 Crystallized components: It includes ice, fats,
and solutes, which contract when they are
cooled; this reduces the volume of food.
28.2.2 PROTEINS
Proteins may undergo changes during freezing and frozen
storage, primarily because of denaturation. Denaturation can
be defined as a major change in the native structure that does
not involve alteration of the amino acid sequences and usually
involves the loss of biological activity and significant changes
in some physical or functional properties such as solubility.
Oxidative processes during storage can also contribute to
protein denaturation; oxidizing agents (e.g., enzymes,
transition metals) can react with proteins via lipid and
nonlipid radicals. For example, the addition of
malonaldehyde, a commonly occurring product of lipid
oxidation, to trout myosin solutions during storage at −4°C
was found to accelerate protein denaturation. Fish protein is
particularly sensitive to denaturation where the protein
develops cross links between adjacent protein molecules that
effectively stop the thawed fish protein to reabsorb water to
recreate the pre-frozen gel structure. This denatured protein
has a much tougher and rubbery texture than the native
protein. The textural changes that occur in fish proteins have
been attributed to changes in the myofibrils. The rate at which
fish or beef muscle is frozen also influences the degree of
protein denaturation. Although rapid freezing generally results
in less denaturation than slower freezing, intermediate
freezing rates can be more detrimental than slow freezing, as
judged by textural changes and the solubility of actomyosin.
For example, cod fillets frozen at intermediate rates developed
intracellular ice crystals large enough to damage the cellular
membranes.
Freezing and frozen storage do not significantly affect the
nutritional value of meat and fish proteins. However, on
thawing frozen meat and fish, substantial amounts of intra-
and extracellular fluids and their associated water-soluble
proteins and other nutrients may be lost (the so-called drip
loss). The volume of drip loss on thawing of meat and fish is
highly variable, usually of the order of 2%–10% of net
weight; however, in exceptional circumstances, up to 15% of
the weight of the product may be lost. Nevertheless, it was
observed for fish that if the product is stored for an
appropriate short time and at a sufficiently low temperature,
the subsequently thawed fish would rehydrate with the protein
returning to its original gel condition. The caseinate micelles
of milk, which are quite stable to heat, may also be
destabilized by freezing. On frozen storage of milk, the
stability of caseinate progressively decreases and this may
lead to complete coagulation. Enzymes have also been linked
to protein denaturation, as it is known that low temperature
decreases the activity of enzymes in tissue, but does not
inactivate them.
28.2.3 LIPIDS
Lipids in food exhibit unique physical and chemical
properties. Their compositions, crystalline structure, melting
properties, and ability to associate with water and other
nonlipid molecules are especially important to their functional
properties in many foods. During processing, storage, and
handling of foods, lipids undergo complex chemical changes
and react with other food constituents, producing numerous
compounds both desirable and deleterious to food quality. The
process of auto oxidation and the resulting deterioration in
flavor of fats and fatty foods are often described by the term
rancidity. In particular, the unsaturated bonds present in all
fats and oils represent active centers that, among other things,
may react with oxygen. This reaction leads to the formation of
primary, secondary, and tertiary oxidation products that may
make the fats or fat-containing foods unsuitable for
consumption.
Lipids can degrade in frozen systems by means of hydrolysis
and oxidation. Lipid oxidation is indeed one of the major
causes of food spoilage. It is of great economic concern to the
food industry because it leads to the development of various
off flavors and off-odors. In addition, oxidative reactions can
decrease the nutritional quality of foods. Lipids in foods can
be oxidized by both enzymatic and non enzymatic
mechanisms. One of the enzyme that is considered important
in lipid oxidation is lipoxygenase, which has recognition for
its off-flavor development in vegetable. Lipoxygenase is the
main enzyme responsible for pigment bleaching and off-odors
in frozen vegetables; if the enzyme is not inactivated before
freezing by blanching, it can generate offensive flavors and
loss of pigment color. At temperatures below −10°C, both
enzymatic and non enzymatic reactions associated with lipid
oxidation are decelerated. However, in the range from 0°C to
−10°C, decreased oxidative stabilities have been noted.
Unless the rate is very slow, the rate of freezing has been
found to have little influence on the oxidative stability of
frozen products. Instead, storage temperatures play a
dominant role in dictating the stability of food products,
including muscle foods. The order of time/temperature
holding treatments, on the other hand, markedly influences the
development of rancidity. The hydrolysis of lipids or lipolysis
results in the release of free fatty acids. Freezing can facilitate
lipid oxidation, partly because the competing reactions of
microbiological spoilage are avoided and partly because of the
concentration effects. Thus, lipid oxidation is relatively more
important in frozen muscle tissue than in fresh tissue. Lipid
degradation can be reduced in frozen foods by lowering the
storage temperature, excluding oxygen (e.g., use of vacuum
packaging), adding antioxidants (e.g., butylated
hydroxytoluene or BHT as well as natural vitamin E), and
supplementing the diet of animals with antioxidants.
28.2.4 VITAMINS
Freezing is considered as one of the best food preservation
methods when judged on the basis of nutrients retention.
However, it is well known that significant amounts of some
vitamins can be lost from processing prior to freezing (e.g.,
peeling and trimming, leaching especially during blanching),
chemical degradation, and thawing. The stability of vitamins
in foods is generally influenced by pH and the presence of
oxygen, light, metals, reducing agents, and heat. It has been
reported that for some frozen foods such as strawberries, the
total and biologically active ascorbic acid remain at
essentially the same level for a year or longer if the foods are
stored below −18°C, although vitamin C losses have also been
found to occur at temperatures as low as −23°C. The
conversion to the partially active dehydroascorbic acid and the
totally inactive 2,3-diketogulonic acid increases with
increasing storage temperature; complete conversion
practically occurs in 8 months at −10°C and in less than 2
months at −2°C. Such findings were instrumental in
establishing −18°C as the upper limit for frozen food storage
and for using biologically active ascorbic acid as a general
indicator of quality deterioration during frozen storage. For
peaches and boysenberries, a 10°C rise in the temperature
from −18°C to −7°C caused the rate of vitamin C degradation
to increase by a factor of 30–70. Vitamin C and thiamine
(vitamin B1) have been studied extensively since they are
water soluble, highly susceptible to chemical degradation, and
present in many foods; they are also required in the diet and
are sometimes deficient in the diet. Therefore, it is generally
assumed that if these vitamins are retained, all other nutrients
would also be well retained.
28.2.5 CARBOHYDRATES AND MINERALS
Carbohydrates occur in plant and animal tissues in many
different forms and levels. In animal organisms, the main
sugar is glucose and the storage carbohydrate is glycogen; in
milk, it is almost exclusively the disaccharide lactose. In plant
organisms, approximately 75% of the solid matter is
carbohydrate. The total carbohydrate content can be as low as
2% of the fresh weight in some fruits or nuts, more than 30%
in starchy vegetables, and over 60% in some pulses and
cereals. In plants, the storage carbohydrate is starch, while the
structural polysaccharide is cellulose. The nutritive value of
carbohydrates is not significantly affected during handling of
fresh foods and the subsequent processing and distribution of
frozen foods. In general, carbohydrates are susceptible to
hydrolysis during frozen storage, which can still occur at
temperatures as low as −22°C. Like B vitamins and proteins,
carbohydrates are less affected by process and more by loss
through drip following a freeze–thaw cycle. Sugar hydrolysis
increases the number of solutes in the food matrix, resulting in
a reduction in the amount of ice in the product, which may
alter certain physical properties; for example, the firmness of
ice cream was found to inversely relate to the degree of
hydrolysis. Blanching and freezing can cause changes in
texture and the pectic composition of certain foods. Both
treatments produce a gradual breakdown in the protoplasmic
structure organization, with a subsequent loss of turgor
pressure, release of pectic substances, and final softening
effect.
Minerals present in any form (e.g., chemical compounds,
molecular complexes, and free ions) can dramatically affect
the color, texture, flavor, and stability of foods. Minerals are
chemically stable under typical conditions of handling and
processing, and nutrient losses are negligible, provided that
losses by physical means (e.g., leaching) are avoided.
Nevertheless, no changes were observed in six mineral
elements (Ca, Cu, Mg, Mn, Ni, and Zn) between fresh and
frozen artichokes, green beans, and peas; boiled fresh
vegetables and boiled frozen vegetables also exhibited similar
mineral contents.
28.3 CONCLUSION
Freezing is complex process involving physical and chemical
changes that might greatly affect the food quality. Further to
minimize the changes in food components during freezing it is
imperative to to freeze a product quickly to –18°C and store it
at the same temperature throughout the cold chain.