https://www.inosr.

net/inosr-experimental-sciences/ Ugwu et al

The Effects of Freezing on the Nutritional Composition of

ABSTRACT
The degradation of fish in the storage condition of the freezer is influenced by several factors which include the
fishing species, the rate at which the fish is frozen, the temperature at which it is stored, the duration of storage, the
type of freezing applied, and enzymatic processes. The freezing and frozen storage lead to macro- and microscale
changes of fish muscles, with major changes being lipid oxidation inducing rancidity, protein denaturation and
aggregation causing toughness, red color fading and freezer burn. The process of freezing involves transforming
water to ice which in turn results into an increase in the concentration of dissolved substances, adjusting the acid-
base balance. It can lead to pH changes of up to 1 unit, often towards acidity and in some cases up to 10 units due to
salt and other compounds precipitation. The described changes permanently alter the properties of the physical-
chemical nature of frozen fish including the convenience and storage conditions of the food, thereby directing
attention to the necessity of new approaches in technologies.
Keywords: Frozen storage, fish degradation, nutritional composition, enzymatic processes, freezing effects.

INTRODUCTION
Fish and fishery products are subject to different Microwave ovens transform regular electricity into
cooking method to increase their hygienic quality high-frequency microwaves that water, fat, and
through destroying pathogenic microorganism and sugar can absorb, thus water molecules vibrating
elevating their flavor and taste [1]. In the course of lead to food heating [5]. The martensite
cooking, some chemical and physical changes take transformation, advocated in the patents by [6, 7],
place which can either be favorable or unfavorable succeeded in increasing the toughness to 85 of the
for the nutritional value of the food. For example, hot rolled Zr steel produced by pure rolling. The
protein denaturation during cooking contributes to change is wrought by the triggering of partial
increased digestibility; it usually results in the recrystallization by plastic deformation, with partial
decreased content of thermolabile compounds, fat- rather than complete recrystallization for food
soluble vitamins or polyunsaturated fatty acids [1, 2, heating and food preservation several types of
3]. Fish frying through the traditional means of food electromagnetic waves have been reported by
preparation improves the sensory values of the food different researchers. Microwave heating has been
by giving off aroma compounds, tempting color, well studied but infrared heating is somewhat still
crust and taste [2]. On the other hand, the modern unexplored [8]. Infrared radiation has many
customers are paying more attention to health risks benefits over conventional heating methods such as
connected with high oil consumption for example, shorter heating time, small equipment size, faster
obesity and heart diseases. These considerations can processing, flavor retention, vitamin retention and
diminish the marketability of coated products, which minimal migration of solvent from the core to outer
can take on quite a large amount of cooking oil region [9]. Freezing is used more frequently for fish
during the flash-frying process, up to 30% of their and fish products as it as better preservation
weight according to [4]. Furthermore, the repeated properties than other storage temperatures, in
use and overheating of oil during frying can regard to taste and nutritional essence. What is
generate various lipid degradation compounds that more, the cryogenic treatment reduces the microbial
may then negatively impact human health [4]. or enzymatic activity, thus increasing the lifespan of
the preserved objects [10]. Commonly used frozen

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fish products like fish cutlets, fish fingers, and fish troponin are myofibrillar proteins responsible for
burgers provide constant, predictable quality, easy muscle contraction.
transportation, and are close to fresh counterparts Frozen storage leads to freezing denaturation of fish
[11]. Eating fish also increases meal consumption proteins, particularly myofibrillar proteins. This
aside from the nutritional quality it provides to the denaturation results in the formation of protein-
body. As such, fish products in forms such as ‘ready protein bonds and the development of high-
to cook’ and ‘ready to eat’ have great potential as molecular-weight polymers that become
condiments [12]. Fish fingers, fish cutlets, and fish unextractable in salt solutions. The increase in salt
burgers are the products which can bring different concentration in the unfrozen phase contributes to
varieties of healthy foods and to increase per capita protein denaturation alongside physicochemical
consumption [11]. Further studies are necessary for changes during frozen storage. Protein insolubility
producing novel aquatic products which by the mean poses a more significant challenge than lipid
time are accepted by local consumers and storing life oxidation in lean fish. The extent of protein
is prolonged. denaturation depends on various factors such as fish
Principles of Freezing species, nutritional status, rigor stage,
Freezing is a process that lowers the temperature pretreatments, freezing velocity, lipid oxidation, and
below the freezing point, allowing most of the water storage temperature. Protein denaturation is less
to turn into ice. The freezing point depends on the pronounced when freezing occurs in the pre-rigor
substances dissolved in the tissue fluid, with fish stage compared to the post-rigor stage of fish.
containing 75–80% water [13, 4]. During freezing, Denatured proteins also lose their enzymatic
latent heat is removed during the phase transition of activity.
water from liquid to solid, along with sensible heat Secondary reactions occur between proteins and
reduction as the temperature decreases. various reactants, leading to lower extractability and
The presence of dissolved and colloidal materials in reduced functionality of myofibrillar proteins during
the tissue fluid decreases the freezing point below frozen storage [14]. Marine animal muscles are
0°C. Each food must be cooled down to its specific reported to be more susceptible to protein
cryohydric point to freeze completely. Seafood denaturation by freezing compared to mammalian
typically starts freezing at temperatures between – muscle [15]. Changes in the functional properties of
1°C and –3°C. The freezing process involves the frozen fish muscle are attributed to conformational
concentration of organic and inorganic salts, which transitions of muscle proteins, resulting in protein
depress the freezing point. Most water (90–95%) is aggregation involving hydrophobic interactions,
frozen at –25°C, with the critical zone for ice hydrogen bonding, and the formation of covalent,
formation occurring between –1°C and –5°C to non-disulfide bonds. Additionally, alterations in the
avoid the formation of large ice crystals that can structure of water and/or protein-water
disrupt cell walls. interactions, along with the transfer of water to
The freezing process of fish occurs in three stages. larger spatial domains, contribute to these changes
First, the product's temperature decreases just below [16]. Protein denaturation during freezing or frozen
0°C, removing sensible heat. Second, the storage is considered to be influenced by salt
temperature remains constant at around –1°C concentration and denaturing due to the freezing out
during the 'thermal arrest period,' where latent heat of water molecules, which alter the conformation of
is removed. It's crucial for the fish to pass through proteins. Furthermore, free fatty acids formed
this period quickly to ensure a high-quality frozen during freezing affect the stability of myofibrillar
product. In the last stage, the temperature sharply proteins [17].
decreases, allowing for freezing of the remaining Functional properties of proteins, including
water, followed by further cooling to the desired solubility and extractability of the myofibrillar
subfreezing temperature to remove the sensible heat fraction, are related to many functional properties.
of the frozen food [4, 13]. Freezing, particularly at a slow rate, initially results
Chemical and nutritional changes from freezing in a noticeable decrease in myosin-actin affinity due
on Fish Quality to myosin head denaturation, followed by a
Fish muscle comprises dark and light muscles, with continued decrease in ATPase activity and
dark muscle predominantly used for sustained denaturation of the myosin tail [18]. Frozen storage
activity, particularly in swimmers like tuna and leads to conformational changes in protein
mackerel, which have higher amounts of dark muscle molecules, resulting in a loss of functionality such as
compared to other fish species. Light muscle is water holding capacity, viscosity, gel-forming
present in all fish species. Sarcoplasmic proteins, ability, and lipid emulsifying capacity. Undesirable
abundant in pelagic fish, include enzymes and characteristics such as odor, consistency, hardness,
oxygen carriers. Myosin, actin, tropomyosin, and

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and stickiness are observed in frozen products due to hydroperoxides in the presence of oxygen and pro-
protein denaturation. oxidant molecules such as heme pigments and trace
The enzyme responsible for trimethylamine oxide elements. The rate of oxidation increases with the
(TMAO) breakdown to dimethylamine (DMA) and amount of unsaturation and double bonds of fatty
formaldehyde (FA) remains active down to acids, as well as temperature and light exposure.
temperatures as low as –8°C. Formaldehyde levels in Further oxidation leads to the decomposition of
fish muscle serve as an index of frozen storage hydroperoxides into multiple compounds.
deterioration. In species where formaldehyde is Non-volatile and volatile compounds, varying in
produced, such as gadoids, muscle texture and molecular weight and polarity and bearing different
functionality deteriorate faster during frozen storage oxygenated functions such as hydroperoxy, hydroxy,
due to DMA and FA formation, accompanied by a aldehyde, epoxy, and ketone functions, are formed
decrease in myofibrillar protein extractability [19]. during frozen storage of fish [25]. Aldehydes can
FA reacts with different functional groups of protein cross-link with proteins, leading to muscle tissue
side chains to form intra- and intermolecular hardening. Malondialdehyde and gluteraldehyde,
methylene bridges, leading to protein denaturation end-products of lipid oxidation, interact with other
and aggregation [20]. FA accumulation is higher in compounds such as amines, nucleosides, nucleic
dark muscles of fish, resulting in a dry and cottony acids, proteins, amino acids, and phospholipids to
texture upon chewing. Additionally, FA participates form polymers, resulting in structural damage and
in the formation of interaction compounds with changes in functionality [26]. Maintaining the
fluorescent properties [21]. The proportion of storage temperature of frozen fish as low as possible
disulfide and non-disulfide covalent bonds with is essential to inhibit oxidation of highly unsaturated
proteins also increases with frozen storage time, lipids, directly related to the production of off-
contributing to protein denaturation and flavors. Lipid oxidation leads to undesirable changes
aggregation [20]. in taste, odor, texture, muscle functionality, and a
[20] reported a continuous decrease in sulphydryl reduction in nutritional quality. Fatty fish has a shelf
groups with a concomitant increase in disulfide bond life of 4–6 months at –18°C, while lean fish can be
formation in lizardfish, croaker, threadfin bream, and preserved for 7 to 12 months [27].
bigeye snapper during extended frozen storage. Endogenous fish enzymes may remain active during
Proteins are susceptible to oxidative reactions, frozen storage, even at –20°C, leading to lysosomal
where exposure to oxygen leads to the destruction of enzyme leakage as the lipid peroxidation enzyme
amino acids and the formation of protein-lipid bonds. system remains active at temperatures below the
This results in the formation of protein-centered freezing point of fish tissue. Phospholipase may be
radicals due to structured aggregates between activated by freezing, initiating phospholipid
carbonyl groups of oxidized lipids and protein hydrolysis in frozen fish muscle. Lipases and
molecules [22]. phospholipases play significant roles in lipid
Oxidation of proteins leads to modification of amino hydrolysis, leading to the formation of free fatty
acid residue side chains, formation of protein acids. Both fatty and lean fish species exhibit
carbonyls, cleavage of peptide bonds, formation of significant lipid hydrolysis during frozen storage,
covalent intermolecular cross-linked derivatives, and resulting in rancid flavor and deleterious effects on
loss of sulfhydryl groups, impacting muscle quality ATPase activity. Free fatty acids interact with
by altering solubility, protein functionality, essential proteins, contributing to texture deterioration and
amino acid content, and digestibility [23]. During lipid oxidation, forming higher-molecular-weight
frozen storage of fish, lipid hydrolysis and oxidation lipids (triglycerides and phospholipids) more rapidly
occur, influencing protein denaturation, texture [21, 18, 28].
changes, functionality loss, and fluorescence Frozen storage can disrupt muscle cells, releasing
development. Lipid oxidation compounds react with mitochondrial and lysosomal enzymes into the
proteins, peptides, free amino acids, and sarcoplasm, leading to the loss of water-soluble
phospholipids [21]. proteins, vitamins, and minerals during thawing,
The hydrophobic effect of free fatty acids on proteins with increased leakage of expressible fluid as the
and the interaction of oxidized lipids with cysteine- proportion of denatured proteins increases. Lipid
SH, the epsilon-NH3 group of lysine, and N- oxidation affects the nutritional value of frozen food
terminus groups of aspartic acid, tyrosine, by triggering oxidative interactions with sulfur-
methionine, and arginine in fish proteins influence containing proteins, resulting in nutritional losses.
the solubility of myofibrillar and sarcoplasmic Additionally, protein oxidation leads to the loss of
proteins [24]. Highly unsaturated fatty acids, essential amino acids and decreases product
abundant in fish lipids, are prone to lipid oxidation. digestibility [29].
These polyunsaturated fatty acids are oxidized to

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CONCLUSION
The freezing process effectively retards biochemical primarily due to oxidation reactions, and freezer
and physical changes in foods, but it cannot entirely burn. The conversion of water to ice during freezing
eliminate undesirable alterations. Deterioration of increases the concentration of dissolved materials,
fish during frozen storage is influenced by various leading to changes in acid-base equilibrium. This can
factors, including fish species, freezing rate, result in pH shifts of up to 1 unit, usually towards
temperature, duration of storage, freezing method, acidity, and in some cases, drastic pH changes up to
and enzymatic degradation. Chemical and structural 2 units due to the precipitation of salts and other
changes occur in fish muscle during freezing and compounds such as phosphate. These changes
frozen storage, including oxidation of lipids leading irreversibly affect the physicochemical properties of
to rancid odor and flavor, toughening due to protein food systems, emphasizing the need for enhanced
denaturation and aggregation, discoloration technologies to improve the quality of frozen fish.
REFERENCES
1. Alizadeh, E., Chapleau, N., de Lamballerie, M. 9. Ranjan, R., Irudayaraj, J., & Jun, S. (2002).
& Le‐Bail, A. (2007). Effect of different freezing Simulation of Infrared Drying Process: Drying
processes on the microstructure of Atlantic Technology: An International Journal. 20(2),
salmon (Salmo salar) fillets. Innovative Food 363 - 379.
Science and Emerging Technologies, 8: 493– 10. Mart´ın-Belloso, O., & Elez-Mart´ınez, P.
499. (2005). Enzymatic Inactivation by Pulsed
2. Esther, U. A., Okechukwu, P.C. U., Emmanuel, Electric Fields Emerging Technologies for
I. O., Patrick, M. A., Chinyere, N. U., Okon, M. Food Processing. ed. D-W Sun, pp. 155–81.
B. (2023). Nutritional Care in Diabetes London: Acad.
Mellitus: A Comprehensive Guide. 11. Vanitha, S. M., Chaurasia, S. N. S., Singh, P.
International Journal of Innovative and M., & Naik, P. S. (2013). Vegetable
Applied Research, 11(12) 16-25. statistics. Tech. Bull. IIVR 51, 250.
3. Esther, U. A., Emmanuel, I. O., Okechukwu, P. 12. Reddy, S. G., Reddy, R. R., Bronkhorst, E. M.,
C. U., Chinyere, N. U., Daniel, E. U., Prasad, R., Jagtman, A. M., & Bergé, S. (2012).
Awotunde, O. S., Dorcas, I. A. (2023). Health related quality of life of patients with
Nutritional Requirements During Pregnancy: nonsyndromic orofacial clefts. J Oral Maxillofac
A Comprehensive Overview. International Surg Med Pathol., 24: 6–10.
Journal of Innovative and Applied Research, 13. Gökoglu, N. (2002). Su Ürünleri İşleme
11(12) 26-34. Teknolojisi, pp. 157, Su Vakfı Yayınları,
4. Venugopal, V. (2006). Quick freezing and Istanbul.
individually quick-frozen products. Seafood 14. Benjakul, S., Visessanguan, B., Thongkaewa, C.
Processing, pp. 446, Taylor and Francis, CRC & Tanaka, M. (2003). Comparative study on
Press, New York physicochemical changes of muscle proteins
5. Garcia-Arias, M. T., Pontes, E. A., Garcia- from some tropical fish during frozen storage.
Linares, M. C., Garcia-Fernandez, M.C., & Food Research International, 36: 787–795.
Muniz, F.J. (2003). Cooking-freezing and 15. Diaz‐Tenorio, L., Garcia‐Carreno, F.L. &
reheating (CFR) of sardine (Sardina pilchardus) Pacheco‐Aguilar, R. (2007). Comparison of
fillets. Effect of different cooking and reheating freezing and thawing treatments on muscle
procedures on the proximate and fatty acid properties of whiteleg shrimp (Litopenaeus
compositions. Food Chem. 83(3): 349–356. vannamei). Journal of Food Biochemistry, 31:
6. Kawakita, Y., Abe, H., Hodate, K., Iguchi, A., 563–576.
Kobayashi, M., Mori, T., Kasai, K., Tanai, Y., 16. Sánchez‐Alonso, I., Martinez, I.,
Kanbe, Y., & Mashiyama, H. (2001). The Sánchez‐Valencia, J. & Careche, M. (2012).
relation between plasma leptin concentrations Estimation of freezing storage time and quality
and carcass lipid contents in Japanese Black changes in hake (Merluccius merluccius, L.) by
steers. In Livestock Production Science, vol. 73, low field NMR. Food Chemistry, 135: 1626–
2001, p. 25–34. 1634.
7. Hokari, M., Kuroda, S., Chiba, Y., Maruichi, K., 17. Kolbe, E. & Kramer, D. (2007). Planning for
& Iwasaki, Y. (2009). Synergistic effects of seafood freezing, pp. 112. Alaska Sea Grant
granulocyte-colony stimulating factor on bone College Program, Cooper Publishing, Alaska.
marrow stromal cell transplantation for mice 18. Sun, D. (2006). Handbook of Frozen Food
cerebral infarct. Cytokine. 46, 260-6. Processing and Packaging, pp. 732, CRC Press
8. Sepulveda, D.R., & Barbosa-Cánovas, G.V. Taylor & Francis Group, New York.
(2016). Heat transfer in food products.

64
https://www.inosr.net/inosr-experimental-sciences/ Ugwu et al
19. Tejada, M., Mohamed, G.F., Huidobro, A. & International, 34. 47-53. 10.1016/S0963-
Garcia, M.L. (2003). Effect of frozen storage of 9969(00)00127-7.
hake, sardine and mixed minces on natural 25. Serón-Arbeloa, C., Puzo-Foncillas, J., Garces-
actomyosin extracted in salt solutions. Journal Gimenez, T., et al. (2011). A Retrospective
of the Science of Food and Agriculture, 83: Study about the Influence of Early Nutritional
1380–1388. Support on Mortality and Nosocomial Infection
20. Benjakul, S., Visessanguan, W., Ishizaki, S. & in the Critical Care Setting. Clinical Nutrition,
Tanaka, M. (2000). Differences in gelation 30, 346-350.
characteristics of natural actomyosin from two http://dx.doi.org/10.1016/j.clnu.2010.11.004
species of bigeye snapper, Priacanthus tayenus 26. Pereira de Abreu, D.A., Losada, P.P., Maroto, J.
and Priacanthus macracanthus. Journal of Food & Cruz, J.M. (2010). Evaluation of the
Science, 66: 1311–1318. effectiveness of a new active packaging film
21. Aubourg, S.P. (1999). Lipid damage detection containing natural antioxidants (from barley
during frozen storage of an underutilized fish husks) that retard lipid damage in frozen
species. Food Research International, 32: 497– Atlantic salmon (Salmo salar L.). Food
502. Research International, 43: 1277–1282.
22. Saeed, S. & Howell, N. K. (2002). Effect of lipid 27. Hui, Y.H., Legaretta, I. G., Lim, M.H., Murrell,
oxidation and frozen storage on muscle K.D. & Nip, W. (2004). Handbook of Frozen
proteins of Atlantic mackerel (Scomber Foods, pp. 1293. CRC Press, Boca Raton.
scombrus). Journal of the Science of Food and 28. Barthet, V. J., Gordon, V., & Dany, J. (2008).
Agriculture, 82: 579–586 Evaluation of a colorimetric method for
23. Lund, M. N., Heinonen, M., Baron, C. P., & measuring the content of FFA in marine and
Estévez, M. (2011). Protein oxidation in muscle vegetable oils. Food Chemistry, 111, 1064−
foods: a review. Mol Nutr Food Res. 2011 1068.
Jan;55(1):83-95. doi: 10.1002/mnfr.201000453. 29. Özkan, M., Yemenicioglu, A., & Cemeroğlu, B.
Epub 2010 Dec 1. PMID: 21207515. (2005). Degradation of various fruit juice
24. Siddaiah, D., Reddy, G. V., Raju, C. V. anthocyanins by hydrogen peroxide. Food
Chandrasekhar, T. C. (2001). Changes in lipids, Research International, 38. 1015-1021.
proteins and kamaboko forming ability of silver 10.1016/j.foodres.2005.03.013.
carp (Hypophthalmichthys molitrix) mince
during frozen storage. Food Research

65