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Effect of Tilapia Protein Isolate on Water-Holding Capacity and Quality of Tilapia Fish Muscle

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Permanent Link: http://ufdc.ufl.edu/UFE0018281/00001

Material Information

Title: Effect of Tilapia Protein Isolate on Water-Holding Capacity and Quality of Tilapia Fish Muscle
Physical Description: Mixed Material
Copyright Date: 2008

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Material Information

Title: Effect of Tilapia Protein Isolate on Water-Holding Capacity and Quality of Tilapia Fish Muscle
Physical Description: Mixed Material
Copyright Date: 2008

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EFFECT OF PROTEINT ISOLATE ON WATER-HOLDING CAPACITY AND QUALITY OF
TILAPIA FISH MUSCLE




















By

SAQIB HUSSAIN


A THESIS PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
MASTER OF SCIENCE

UNIVERSITY OF FLORIDA

2007


























Copyright 2007

by

Saqib Hussain

































To my family and friends









ACKNOWLEDGMENTS

I thank Dr. Hordur Kristinsson for his continuous support, advice, and help on this proj ect.

It has been a honor and privilege to work with him. I thank my committee members Dr. Maurice

Marshall and Dr Sally Williams for their guidance and suggestions.

Finally I thank my lab mates especially Sivakumar Raghavan and Holly Petty for their

support and help during my studies and research work.













TABLE OF CONTENTS


page

ACKNOWLEDGMENTS .............. ...............4.....


LIST OF TABLES ................. ...............7.__. .....


LIST OF FIGURES .............. ...............8.....


AB S TRAC T ............._. .......... ..............._ 12...


CHAPTERS


1 INTRODUCTION ................. ...............14.......... ......


2 LITERATURE REVIEW ................. ...............16................


W ater Control in M uscle .................. .......... ............1

Physico-Chemical Principles of Water-Holding .............. ...............16....
Isoelectric Point ............. ...... ._ ...............16....
Role of pH and NaCl .................... .____ ........._ ....__ .......__ .............17
Other Additives used to Improve Water-Holding Capacity .............. ....................1
Phosphates ................. ...............19.._._._......
Polysaccarides ..................... ...............22.
Soy Proteins and Whey Proteins .............. ...............23....
Use of Fish Proteins as Water Binders ................ ...............24..............

Obj ectives ................. ...............26.......... .....

3 MATERIALS AND METHODS .............. ...............28....


Raw Material .................... ...............28.
Alkali-aided Protein Isolation............... ...............2

Preparation of Protein Solutions ..........._...__........ ...............28....
Viscosity Measurement .............. ...............29....
pH Measurements .............. ...............29....
Protein Solubility ................. .......... ...............29.......
Protein Composition Analysi s ................ ...............30................
Cook Losses................... ......................3
Water-Holding Capacity (WHC) ................. ...............30........... ....
Drip Losses ................. ...............3.. 1..............
Color Analysi s ................ ...............31........... ....
Texture Analysis............... ...............32
Rheology Studies ............._. ...._... ...............32....
Statistical Analysis............... ...............33


4 RE SULT S AND DI SCU SSION ............... ...............3











Study I......................... ... .. .. ... ... ..........................3
Changes in Solubility and Viscosity of Tilapia Protein Isolate Solutions.......................38
Effect of PI Solutions on Cook and Drip Losses at Different Protein Concentrations ...41
Study II ...................... ..... ......... ..... .......4
Effect of PI Solutions on Cook and Drip Loss at Different Inj section Levels. ................45
PI Solution affecting Color of Uncooked and Cooked Tilapia Fillets ........._._...............5 1
PI Solutions Affecting Texture of Uncooked and Cooked Tilapia Fillets ........._._.........59
PI Solution Affecting Gel Forming (G') Ability of Tilapia Muscle ............... .... ..........._64
Study III ......... ........... .__ ...........__ ..............._ .. ...............6
Effect of Solution without Protein Isolate (PI) on Cook and Drip Losses ......................68
Effect of Solution without Protein Isolate (PI) on Color of Cooked and Uncooked
Sam ples ................... ......._. .. ........... ...... ..... .. ... .........72
Effect of Solution without Protein Isolate (PI) On Texture of Cooked and
U ncooked Sam ples.............. ... ... .. ..............................8
Effect of Solution without Protein Isolate On Gel Forming (G') Ability of Tilapia
M uscle............... ... .. ....... .... .... .........................8
Comparison between Fillets Inj ected With Solutions Containing Protein Isolate (PI)
and W without PI ........._._. ._......_.. ...............87....

CONCLU SION ........._._.._......_.. ...............92.....

LIST OF REFERENCE S ........._._.._......_.. ...............93....

BIOGRAPHICAL SKETCH .............. ...............99....











LIST OF TABLES


Table page

4-1 Uooked fish fillets inj ected with different PI solutions (pH study) ................. ................89

4-2 Cooked fish fillets inj ected with different PI solutions (pH study) ................ ................89

4-3 Uncooked fish fillets inj ected with different inj section solutions without PI (pH study)...89

4-4 Cooked fish fillets inj ected with different inj section solutions without PI (pH study).......89

4-5 Green weight drip losses of fish fillets inj ected with different PI solutions ................... ...90

4-6 Injected weight drip losses of fish fillets inj ected with different PI solutions .................90

4-7 Green weight cook losses of fi sh fillets inj ected with different PI solutions ................... ..90

4-8 Injected weight cook losses of fish fillets injected with different PI solutions. ...............90

4-9 Green weight drip losses of fish fillets inj ected with different inj section solutions
without PI. ........... ..... .._ ...............90....

4-10 Injected weight drip losses of fish fillets inj ected with different inj section solutions
without PI. ........... ..... .._ ...............90....

4-11 Green weight cook losses of fish fillets inj ected with different inj section solutions
without PI. ....._._................. ........__ ..........9

4-12 Inj ected weight cook losses of fish fillets inj ected with different inj section solutions
without PI. ........... ..... .._ ...............91....










LIST OF FIGURES


Figure page

3-1 Outline of study I. Protein solutions with protein isolate (PI) concentration. (1 to 5%)
inj ected in fish fillets to obtain 10% inj section levels ................. ............... 33..........

3-2 Outline of study II. 5% PI solution injected in fish fillets to obtain 5, 10 and 15%
inj section level s ................. ...............3.. 4.............

3-3 Outline of study III. Fish fillets injected with solution containing no PI to obtain 5,
10 and 15% inj section level s ................. ...............3.. 5.........

4-1 Solubility of tilapia protein isolates solution at different protein isolate (PI)
concentration. The pH values were: PI = 5.5, PI + NaCl = 5.5, PI + TPP = 8.2, PI +
TPP + NaCl = 8.2, PI (pH adjusted) = 8.2, PI + NaCl (pH adjusted) = 8.2.......................38

4-2 Viscosity of PI solutions at different protein concentrations .......__........... ........ .......41

4-3 Cook losses (%) based on green weight for inj ected tilapia samples at different
protein concentrations ................. ...............42.................

4-4 Cook losses (%) based on inj ected weight for inj ected tilapia samples at different
protein concentrations ................. ...............43.................

4-5 Drip losses (%) based on green weight for inj ected tilapia samples at different
protein concentrations ................. ...............44.................

4-6 Cook losses (%) based on inj ected weight for inj ected tilapia samples at different
protein concentrations ................. ...............45.................

4-7 Cook losses (%) based on green weight for tilapia samples inj ected to gain 5, 10 and
15% weight increase with a 5% PI solution ................. ...............47...........

4-8 Cook losses (%) based on inj ected weight for tilapia samples inj ected to gain 5, 10
and 15% weight increase with a 5% PI solution ................. ...............48...........

4-9 Drip losses (%) based on green weight for tilapia samples inj ected to gain 5, 10 and
15% weight increase with a 5% PI solution ................. ...............48...........

4-10 Cook losses (%) based on inj ected weight for tilapia samples inj ected to gain 5, 10
and 15% weight increase with a 5% PI solution ................. ...............49...........

4-11 Water-holding capacity of tilapia fillets inj ected to give 5, 10 and 15% weight
increase with a 5% PI solution............... ...............51

4-12 Lightness (L* value) of white cooked muscle for tilapia samples injected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 53..........










4-13 Redness (a* value) of white cooked muscle for tilapia samples injected to give 5, 10
and 15% weight increase with a 5% PI solution ................. ...............54...........

4-14 Yellowness (b* value) of white cooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 54..........

4-15 Lightness (L* value) of white uncooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 55..........

4-16 Redness (a* value) of white uncooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 55..........

4-17 Yellowness (b*" value) white uncooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 56..........

4-18 Lightness (L* value) of dark cooked muscle for tilapia samples inj ected to give 5, 10
and 15% weight increase with a 5% PI solution ................. ...............56...........

4-19 Redness (a* value) of dark cooked muscle for tilapia samples inj ected to give 5, 10
and 15% weight increase with a 5% PI solution ................. ...............57...........

4-20 Yellowness (b* value) of dark cooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 57..........

4-21 Lightness (L* value) of dark uncooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 58..........

4-22 Redness (a* value) of dark uncooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution ................. ............... 58..........

4-23 Yellowness (b* value) of dark uncooked muscle for tilapia samples inj ected to give
5, 10 and 15% weight increase with a 5% PI solution ................. ......... ................59

4-24 Firmness of cooked muscle for tilapia samples inj ected to give 5, 10 and 15% weight
increase with a 5% PI solution............... ...............61

4-25 Springiness of cooked muscle for tilapia samples inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution ................. ...............62........... ..

4-26 Gumminess of cooked muscle for tilapia samples inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution ................. ...............62........... ..

4-27 Firmness of uncooked muscle for tilapia samples inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution. .............. ...............63....

4-28 Springiness of uncooked for inj ected tilapia samples inj ected to give 5, 10 and 1 5%
weight increase with a 5% PI solution ................. ...............63........... ..










4-29 Gumminess of uncooked muscle for tilapia samples inj ected to give 5, 10 and 1 5%
weight increase with a 5% PI solution ................. ...............64......_... ..

4-30 Gel formation of minced tilapia fillets inj ected with PI solution with tripolyphophate
(TPP). Heated from 5-80oC and then cooled from 80-5oC .............. ....................6

4-3 1 Gel formation of uncooked tilapia muscle (at 9.2oC) inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution ................. ...............67........... ..

4-32 Gel formation of uncooked tilapia muscle (at 80. 1oC) inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution ................. ...............67........... ..

4-33 Gel formation of uncooked tilapia muscle (at 5.0oC) inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution ................. ...............68........... ..

4-34 Cook loss (%) based on inj ected weight of tilapia fillets inj ected with Sodium
Chloride (NaC1), TPP and NaCl + TPP solutions to obtain 5, 10 and 15% injection
level s .............. ...............70....

4-3 5 Cook loss (%) based on green weight of tilapia fillets injected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..70

4-36 Drip loss (%) based on inj ected weight of tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..71

4-37 Drip loss (%) based on green weight of tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% Injection levels............... .................7

4-3 8 Water-holding capacity of tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels............... ...............72.

4-39 Lightness (L* value) of white uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..74

4-40 Redness (a* value) of white uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..74

4-41 Yellowness (b* value) of white uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..75

4-42 Lightness (L* value) of dark uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..75

4-43 Redness (a* value) of dark uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..76

4-44 Yellowness (b* value) of dark uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..76










4-45 Lightness (L* value) of white cooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ........._..... ........_........77

4-46 Redness (a* value) of white cooked tilapia fillets inj ected with NaC1, TPP and NaCl
+ TPP solutions to obtain 5, 10 and 15% injection levels .............. .....................7

4-47 Yellowness (b* value) of white cooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ........._...... ........_........78

4-48 Lightness (L* value) of dark cooked tilapia fillets inj ected with NaC1, TPP and NaCl
+ TPP solutions to obtain 5, 10 and 15% injection levels .............. .....................7

4-49 Redness (a* value) of dark cooked tilapia fillets inj ected with NaC1, TPP and NaCl +
TPP solutions to obtain 5, 10 and 15% injection levels............... ...............79.

4-50 Yellowness (b* value) of dark cooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ........._...... ........_........79

4-5 1 Hardness of uncooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels............... ...............81.

4-52 Springiness of uncooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels............... ...............81.

4-53 Gumminess of uncooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels............... ...............82.

4-54 Hardness of cooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP solutions
to obtain 5, 10 and 15% inj section levels ................. ...............82...........

4-55 Springiness of cooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels............... ...............83.

4-56 Gumminess of cooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels............... ...............83.

4-57 Gel formation of uncooked tilapia fillets at 9.2oC inj ected with NaC1, TPP and NaCl
+ TPP solutions to obtain 5, 10 and 15% injection levels .............. .....................8

4-58 Gel formation of uncooked tilapia fillets (at 80. 1oC) inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..86

4-59 Gel formation of uncooked tilapia fillets (at 5.0oC) inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% inj section levels ................. ................ ..87

4-60 Protein composition of exudates of tilapia fillets injected with PI solutions after
cooking performed my SDS-PAGE electrophoresis............... ............9









Abstract of Thesis Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Mater in Science

EFFECT OF TILAPIA PROTEIN ISOLATE ON WATER-HOLDING CAPACITY AND
QUALITY OF TRLAPIA FISH MUSCLE

By

Saqib Hussain

May 2007

Chair: Hordur G. Kristinsson
Major Department: Food Science and Human Nutrition

Several million tons of Esh and fish byproducts are being wasted every year. These

byproducts could be converted into useful functional protein isolates using an alkali-aided

extraction method. In brief, alkali processing involves adjusting the pH of Eish proteins to 1 1.0 to

solubilize myofibrillar proteins, followed by filtration to eliminate bones and connective tissue.

The solubilized proteins are then precipitated at pH 5.5 to obtain a functional protein isolate.

Loss in water-holding capacity affects the quality and texture of meat products and costs millions

of dollars to the fish and meat industry annually. Currently, chemicals like phosphates, and

proteins from sources such as whey and soy, are being used to improve the water-holding

capacity of fish muscle. The obj ective was to study the effect of protein isolates prepared from

tilapia (Oreochromis niloticus) on the water-holding capacity, drip loss, texture, color and

theology of tilapia muscle. Improving the quality attributes of tilapia would increase their

economic value and would benefit the fish industry.

Four combinations of protein isolate solutions, namely, tilapia protein isolates (PI), PI +

NaC1, PI + sodium tripolyphosphate (TPP), and PI + NaCl + TPP were inj ected into tilapia

fillets. The concentration of PI in the inj section solutions were between 1 to 5%. Among the

different PI concentrations, a level of 5 % was chosen for all treatments, since minimum drip loss









and maximum cook yield was obtained at this level. The amount of protein isolate solution

inj ected into the fillets was 5, 10 and 15% based on tilapia muscle weight. Both the cook and

drip loss increased in the order, 5 < 10 < 15% inj section level. The cook yield obtained using PI +

TPP was significantly higher (P < 0.05) than those obtained using protein isolate without TPP at

10 and 15% injection levels. Texture (firmness, gumminess and springiness) and color (L*, a*

and b* values) analyses performed on cooked and uncooked tilapia fillets showed no significant

difference (P > 0.05) among the different protein isolate treatments. Storage modulus (G') of

tilapia fillets inj ected with protein isolates were significantly different (P < 0.05) from those

fillets inj ected with protein isolates containing TPP. Water-holding capacity was not significantly

different (P > 0.05) at 5 and 10% inj section levels but was significantly lower (P < 0.05) at 15%

inj section level for fillets inj ected with PI without TPP. These results were compared with tilapia

fillets inj ected with TPP, NaC1, and TPP + NaCl solutions without PI. PI solutions with TPP

showed higher cook yield and reduced drip loss compared to inj section solution with TPP.

However, PI solutions without TPP showed lower cook yield and higher drip loss compared to

TPP solution. It could be concluded that PI from tilapia is a good alternate for TPP as PI could

reduce cook and drip loss but had no effect on color and texture of tilapia fillets.









CHAPTER 1
INTTRODUCTION

Currently the demand for fish exceeds what our natural resources can safely supply,

leading to great pressure on traditional methods of Esh supply. Another problem facing the

seafood industry is large amounts of Esh and fish byproducts, which are being discarded without

any attempt of recovery. Many common fish stocks are declining and the demand of quality fish

and fish protein will exceed supply in the near feature (1. Global production of farmed fish

increased more than double in the past 15 years. It is believed that such growth relieves pressure

on ocean fisheries, but the opposite is true for some types of aquaculture. Farming carnivorous

species requires large inputs of wild fish for feed. Some aquaculture systems also reduce wild

fish supplies through habitat modification, wild seedstock collection and other ecological

impacts (2). Undesirable raw materials and byproducts can be a source of high quality fish

proteins, providing proper protein extraction methods are available. For the industry, it is

necessary to utilize byproducts in a better way so that profitability can be improved (3). New

successful fish protein extraction method has been recently developed. This method is based on

using either acid or alkali to solubilize fish proteins and then recovering them with isoelectric

precipitation. It has been shown that the acid-aided process recovers more protein types than the

alkali-aided process during isoelectric precipitation, which indicates that it leads to more protein

denaturation and thus more aggregation at the isoelectric point. It has further been shown that the

alkali-aided process more effectively removes heme proteins than the acid-aided process and

produces a more functional protein isolate (1.

One of the maj or problems with any muscle food product, including sea foods, is the loss

in the ability of muscle and muscle proteins to hold water. Loss in water-holding capacity costs

millions of dollars to the fish and meat industry annually. The ability of fresh meat to retain









moisture is one of the most important quality characteristic of raw products (4). Traditionally

phosphates, salts, starches, gums and plant source proteins have been used to improve water-

holding capacity in fish muscle. Addition of phosphates leads to very effective water uptake in

fish products (5). However, there are significant water losses after cooking with phosphate-

treated products. Phosphates also have strict labeling issues and have a history of abuse in the

seafood industry. Increasing salt content in muscle is also effective in increasing water-holding

of both cooked and raw products as well impacting the firmness of muscle products (6). Both

additions of salts and phosphates elevate the sodium content of the muscle products, which is an

increasingly less desirable practice in the food industry. Use of modified starches and gums as

well as non-meat protein sources has increased substantially over the years, but carries an

unfavorable label and may lead to an unnatural texture of the product. Using proteins isolated

from fish and fish byproducts to improve water-holding capacity of fish muscle would also

provide a natural alternative to the above ingredients.

One of many ways to describe quality of fish is to measure the ability of the muscle to

hold water. This is an important attribute for several reasons: (1) weight decrease due to loss of

water is of economic importance, (2) accumulated exudate is unattractive to consumers, and, (3)

water is important for muscle texture (3).









CHAPTER 2
LITERATURE REVIEW

Water Control in Muscle

Losses in water can occur by evaporation, drip loss, or during cooking. Such losses

inevitably produce shrinkage of meat muscle. The content of water and its distribution have a

profound influence on properties of meat, especially its toughness, juiciness, firmness, and

appearance (7). Much of the water in the muscle is entrapped in the structures of the cell,

including the intra- and extra myofibrillar spaces. As rigor progresses, the space for water to be

held in the myofibrils is reduced and fluid can be forced into the extra myofibrillar spaces where

it is more easily lost as drip (3).

The bulk of water inside a muscle fiber is free and the less mobile water is bound to

muscle proteins. Very little water is held by the myofibrillar proteins themselves; it is the

structure developed by the myofibrils, which holds much of the water. During cooking, the thick

fi1aments degrade and lateral shrinkage of the muscle fiber takes place followed by longitudinal

shrinkage. Water losses in cooked muscle increases concentration of structural components of

the muscle and connective tissue in the shrunken meat, which increases the toughness of meat

(7).

Physico-Chemical Principles of Water-Holding

Isoelectric Point

The maj ority of water affected by the process of converting muscle to meat is the

entrapped water. Influencing the retention of entrapped water includes manipulation of the net

charge of myofibrillar proteins, the structure of muscle cell and its components. During the

conversion of muscle to meat, lactic acid builds up in the tissue leading to reduction in pH of the

meat. Once the pH reaches the isoelectric point of maj or proteins, especially myosin (pl=5.4), the









net charge of proteins becomes zero and the repulsion of structures within the myofibril is

reduced, allowing structures to pack more closely and results in water losses from meat muscle

(4). Preventing this pH drop and thus maintaining the pH above the isoelectric point will

therefore lead to more water-holding. This is because above the pl the muscle proteins will carry

a negative charge, which leads to swelling and effective water binding to the proteins as well as

entrapment of water in the charged swollen protein matrix.

Role of pH and NaCI

The water-holding capacity has been reported to be influenced by a number of factors,

including ultimate pH, protein denaturation, intra- and interfascicular spacing and sarcomere

lengths (3). Final muscle pH before cooking has a maj or effect on protein functionality and

other quality attributes. Higher final muscle pH improves the solubility of sarcoplasmic proteins,

water-holding capacity, cooked color, and emulsion stability of cooked muscle compared to

initial muscle pH (8). High concentration of NaCl (2% to 3%) is required to solubilize

myofibrillar proteins. Presence of a low concentration of NaCl and low pH results in decreased

elasticity and increased water loss due to low ionic strength (9). An increase in muscle gel pH

above neutrality without addition of NaCl was shown to lead to a dramatic increase in water-

holding capacity and water uptake (10).

It has been shown that NaCl concentration, pH, heating temperature and interaction among

these factors influenced the loss of water in salmon muscle (3). The interaction effect among low

pH, low salt concentration and high temperature was strongest; addition of salt extracted the

myofibrillar proteins and resulted in a homogenous protein matrix with few intact fibers. When

heated to about 30oC, the muscle fiber structure results in enlarged pores and gaps. Low salt

concentration and low pH resulted in more pores and gaps in the muscle fiber structure and

enhanced liquid loss (3).









Both pH and ionic strength played a significant role in retention of water when washed

minced chicken muscle was cooked to form a gel. In gels with no added salt, the water loss upon

gel formation dropped from 30.2% to 7.7% as the pH of the gel was raised from pH 6.4 to 6.6.

From pH 7 to 7.4, water loss was at a minimum, only about 3 to 5%. Even at the point of

minimal water loss, gels with salt still loss significantly more water than gels without salts.

Enormous uptake of water as function of pH occurred when gels with no added salt were

immersed in deionized distilled water and was a strong indication of electrostatic repulsive

forces at work (10).

Texture of meat products is dependent on the gelation characteristics of myofibrillar

protein. A study was performed to investigate the impact of pH (5.6, 6.0, 6.5, and 7.0) on heat-

induced gelation properties of myofibrillar proteins from porcine semi membranous muscle.

Results indicated protein denaturation and gel formation are pH dependent. Furthermore, rate of

gelation appeared to influence water-holding capacity (11). Therefore, pH can be altered in meat

products to reach the desired extent of gel strength at a given temperature, or pH must be

monitored to ensure consistent quality from product to product.

Fillets of Atlantic cod (Gadus morhua) were wet-salted in brines of pH 6.5 and 8.5

containing different combinations of NaC1, KC1, MgCl2 and CaCl2, and then dry-salted in NaC1.

The composition and pH of the brines slightly affected the protein composition of the maj or

extract constituents and the functional properties of the muscle after dry-salting. Brining at

alkaline pH produced a larger variety of water-soluble proteins, particularly actin, than at pH 6.5.

Furthermore, the compositions of the protein fractions extracted with 0.86 M NaCl were very

similar for both pHs, irrespective of the composition of the brine; in this case, myosin heavy









chains were absent in both extracts due to aggregation caused by a massive uptake of salt by the

muscle (12).



The amount of water uptake depends upon the concentration of NaC1. At concentration of

0.2 M, muscle volume is at minimum. If the salt concentration is reduced below 40 mM, the

volume increases up to a maximum of two-fold. As the salt concentration raises above

physiological ionic strength, there is a progressive increase in amount of swelling, achieving a

maximum value of about IM in meat muscle (7). As mentioned above, the effect of salts is

highly influenced by the pH of the muscle.

Cod fillets were salted in brines with different pH values (6.5 and 8.5) and saline

compositions. Partial replacement of 50% NaCl with 50% KCl reduced penetration ofNa+ into

muscle, as did the addition of small amounts of CaCl2 (0.8%) and/or MgCl2 (0.4%) to pH 6.5

brines. The use of 0.4% MgCl2 at pH 6.5 negatively affected functional properties and further

hindered salt penetration into the muscle. The use of KCl in pH 8.5 brines increased hardness,

negatively affecting protein water-extractability. Moreover, the addition of divalent salts, at basic

pHs, slightly decreased water-holding capacity (13).

Other Additives used to Improve Water-Holding Capacity

Phosphates

Since the control and retention of water in muscle foods is of maj or economic and quality

importance, much effort has been put into finding ways to maximize the water-holding capacity

of muscle. Phosphates are used to improve water-holding capacity of fish muscle. Several

commercial phosphates are used in the meat and fish industry and they differ from one another in

their properties. Pyrophosphate is the smallest member of the series and the most active form. Its

use in industry is limited due to low solubility. Triphosphate is widely used due to its higher









solubility. Its effect on water-holding in meat muscle may be the result of conversion via

hydrolysis to pyrophosphate (14). Long chain polyphosphates are also used in the industry (7).

Several studies have demonstrated the effectiveness of using phosphates. During iced

storage, protein solubility and water uptake ability of cod mince was higher with sodium

hexametaphosphate than with sodium tripolyphosphate. Water uptake ability increased

significantly with sodium hexametaphosphate concentration up to 0.7% but decreased above

that, although protein solubility continued to increase (15).

The effects of NaCl and pyrophosphate were examined on beef muscle tissue and isolated

beef myofibrils with various concentration of NaC1, with and without 10mM pyrophosphate.

Higher concentration of NaCl increased the extraction of titin, myosin, and other myofibrillar

proteins from beef tissue and the inclusion of 10 mM pyrophosphate enhanced the extraction of

those proteins (16).

Combinations of sodium chloride and different phosphates were used to study the effect of

phosphate type, ionic strength, and pH on the binding ability of restructured beef rolls. The

results showed that binding ability increased linearly with increasing ionic strength and pH until

a maximum value was reached. Approximately 80% of the increase in binding ability was due to

the increase in ionic strength and pH. Above an ionic strength of 0. 15, polyphosphates (chain

length > 1.0) produced a synergistic increase in binding ability with increasing ionic strength.

The extent of the synergistic effect decreased linearly as the chain length of the phosphate

increased (17).

Polyphosphate at three concentrations (0, 3 and 5%) was inj ected in pork steaks cooked by

grilling to an internal core temperature of 72.5 or 80oC. Polyphosphate improved water-holding,

and generally produced tender and juicy meat than control streaks, although pork flavor intensity









was reduced and abnormal flavor intensity increased. Raising the core temperature from 72.5 to

80oC increased cooking loss from 35 to 42%, reduced tenderness, juiciness, abnormal flavors and

increased pork flavor intensity. Steaks containing 5% polyphosphate and cooked to 80oC were

more tender and juicy than steaks without polyphosphates cooked to a lower core temperature

(18).

The addition of polyphosphate allows swelling of myofibrils and myosin extraction to

occur at lower NaCl concentration than in its absence. The maximum amount of swelling of

isolated myofibrils is, however, much reduced, probably due to extraction of myosin.

Polyphosphates probably act by assisting NaCl in depolymerization of thick fi1aments. However,

they also promote the dissociation of actomyosin and thereby, enhance the redistribution of

myosin in meat and the extraction of myosin onto the surface of meat pieces (7). Permeation of

myosin throughout the meat in the presence of polyphosphate may lead to the formation of heat-

set myosin gel on cooking, which stabilize myofibrils against shrinkage and itself entraps water,

thereby reducing cooking losses. Polyphosphate-treated fish fillets have been found to lose less

weight than non-phosphated fillets after dry-salting and storage up to 3 weeks. However, the

quality of polyphosphate-treated fillets was poor compared to untreated fillets (5).

Sodium tripolyphosphate (STPP) and tetra sodium pyrophosphate (TSPP) performed better

than sodium hexametaphosphate (SHMP) in terms of water retention, water binding, and cook

yield in beef biceps femoris muscles. Although there were no differences between phosphate

concentrations for water-holding capacity or cook yield, increased phosphate concentrations did

allow for an increased ability to adsorb additional water, as well as increased sensory-evaluated

overall tenderness. Phosphate inj section at 18% levels allowed for improved sensory tenderness

compared to injection at 12% level. Additionally, the 18% level compared to 12% did increase









moisture percentage without exhibiting decreased water retention or decreased cook yields,

suggesting potentially increased overall yields with an increased inj section level (19).

Phosphates have been abused in the industry for years, and excessive use leads to water-

losses on preparation and adverse flavor effects. In addition to this, there are strict regulations on

the use of phosphates in many countries and many consumers have highly negative feelings with

respect to their use in sea foods.

Polysaccarides

A few alternatives to replace phosphate in sea foods and meats are commercially available,

including polysaccharides (e.g., modified starches and gums) with limited success in improving

water-holding capacity (3). These ingredients have multiple functionalities in muscle food

products. They can serve as thickening agents where they increase the viscosity of products.

They can function as gelling agents to change the structure of products form liquid to solid.

They can serve as stabilizers to maintain stable conditions for a longer period of time, and as

suspending agents to keep particles in suspension. Gums also react with proteins in foods and

alter their theological properties, solubility and gelling characteristics (20). In one study, it was

shown that incorporation of 0.5% carrageenan into cured turkey thigh meat products

significantly increased cook yield, visual appearance, and sliceability, and decreased freeze and

thaw purge compared with control samples. High bind values were obtained with a blend of

0.5% carrageenan and 2% starch compared with 0.5% carrageenan only (21).

Low fat meatballs (10% fat, formulated with 10% water, 3.2% spice mixture and 0.5-1%

carrageenan or guar gum) were evaluated for cooking characteristics and compared with controls

of higher fat content. A reduction in the fat level from 25% to 10% improved all cooking

parameters with respect to better cooking yield and fat retention. Addition of increasing levels of









carrageenan to low fat meatballs was more effective than guar gum for the textural properties

after cooking (22).

Soy Proteins and Whey Proteins

Much work has been done with processing of different soy protein products. Initial

viscosity, hardness and water-holding capacity has a positive correlation with water absorption

capacity of commercial soy protein isolates (23). It has been shown that soy protein isolates may

be used in emulsified fish products to improve water- and fat-binding properties of products (5).

Soy proteins were less effective than salt and phosphate in water-holding on thawing in frozen

cod fillets. When soy protein were combined with salt and phosphates, the performance was

better than salt and phosphate alone, however the appearance was adversely affected in frozen

cod fillets (9).

Karmas and Turk (24) investigated the water binding of cooked fish treated with a

combination of (a) sodium, calcium, potassium and isoelectric soy protein isolate; (b) sodium

and calcium whey protein concentrate; and (c) sodium caseinate. All proteins increased the water

binding of cooked fish. The ions associated with the whey and soy proteins did not affect the

water binding significantly. Sodium soy proteinate and sodium caseinate were both better water

binders than the sodium whey proteinate.

The effects of setting conditions and soy protein isolate (SPI) on textural properties of

surimi produced from grass carp were investigated. Protein concentration was the maj or factor

affecting the gel strength of grass carp surimi. Breaking force and distance of grass carp surimi

gels decreased with increase of protein ratio from SPI at 30oC and 40oC for 60 min setting and

heating at 85oC for 30 min, but the breaking force obtained for addition of 100 g/kg SPI protein

to grass carp surimi was higher than that for surimi alone at 60oC for 60 min incubation and

heating at 85oC for 30 min (25).










The pale, soft, and exudative (PSE) condition in pork meat has been associated with

protein denaturation that leads to loss of functional properties. The effect of soy glycinin addition

to model systems of actomyosin from PSE or normal meat on biophysical properties was

evaluated. Although the functional properties depended on actomyosin concentration, a 75%

actomyosin and 25% glycinin mixture presented acceptable properties with both types of

actomyosin. This combination was used to investigate protein-protein interactions. Gel strength

correlated with intermediate alpha-helical content and protein aggregation. Changes in protein

secondary structure probably added to protein denaturation and appear to be necessary to

improve gelation and water retention in PSE actomyosin-glycinin combinations (26).

Changes in textural and microstructural properties of washed and unwashed frozen fish

mince were studied as a function of the addition of non-Hish proteins (soy protein isolate, milk

protein isolate, egg white, and wheat gluten at 2, 4 or 6%) and 6% crystalline sorbitol (27). Soy

and milk proteins and sorbitol reduced the hardness of frozen fish mince, while egg white and

wheat gluten made the texture firmer without rubberiness developing after frozen storage. All

non-fish proteins and sorbitol stabilized the myofibrillar organization by reducing freeze-induced

contraction of myofibrils. The mechanisms of reducing texture hardening appear to be different

between sorbitol and non-fish proteins. Water binding properties and dispersibility made the

difference among non-fish proteins in reducing freeze-contraction of myofibrils. Non-fish

proteins not only reduced texture hardening during frozen storage, but also modified texture

during cooking as they underwent thermal gelation specific to each protein used (27).

Use of Fish Proteins as Water Binders

Use of proteins from fish in fish products would be an ideal alternative to the above

compounds to improve water-holding. Fish proteins possess properties that make them good

agents for water-holding, gelation, fat binding, emulsification and foaming (28). Little work has









been published on the functional characteristics of fish proteins added to food systems. However,

it has been shown that salmon protein hydrolysates reduced drip in salmon mince patties after

freezing (28). An earlier study reported the use of capelin protein hydrolysates to improve water-

holding capacity in minced pork on cooking (29). Hydrolyzed cod proteins gave slightly lower

gained weight with combination of salt and phosphates in salted cod fillets. However, the

combination of salt, phosphates and soy protein resulted in higher yield (5).

Protein hydrolysates from mechanically separated seal meat were used as a phosphate

alternative in meat products in order to improve water-binding capacity. The cook loss of

mechanically separated seal meat was lowest at 3% seal protein hydrolysate, similar to that of

polyphosphates at the same level. Losses were much higher at 0.5%, the maximum allowable

limit of polyphosphate. When compared with different phosphates, drip volume resulting from

use of seal protein hydrolysate was lower than that for samples treated with sodium

pyrophosphate, sodium tripolyphosphate and sodium hexametaphosphate (30).

Protein hydrolysates were prepared from male and spent capelin by the enzyme

preparations Alcalase, Neutrase and papain. Protein recovery varied from 51.6 to 70.6% by

enzymes. Alcalase served best for preparation of capelin protein hydrolysates, which had similar

amino acid composition of the starting capelin, except for lower levels of methionine and

tryptophan. The product had excellent solubility over pH range of 2-1 1 and when incorporated

up to 3% in meat model systems, resulted in 4% increase of cook yield (29). Hydrolysates

however have bitterness problems and may result in undesirable colors after oxidation (28).

Furthermore, they may be uneconomical with respect to production and may carry a negative

label. The use of unhydrolyzed fish protein isolate in water control has not been reported in the

literature.









A recent process has been developed where intact proteins can be economically and

efficiently extracted from low value muscle sources. This process involves extracting fish

proteins from diluted slurry of the fish raw material using either low pH (pH 2.5-2.8) or high pH

(10.8-11). Solubilized proteins are then separated from other constituents of the muscle via high

speed centrifugation or filtration. The proteins are then recovered and partially dewatered with

isoelectric precipitation and centrifugation or filtration. Proteins isolated by traditional methods

not only yield low amount of proteins but the final product has significant levels of impurities,

including fat and heme proteins, and also has often low functionality. Proteins isolated with acid-

aided and alkali-aided isolation method yield higher amount of protein, remove almost all the fat,

including membrane lipids, largely remove catalysts for oxidation (namely the alkaline-aided

process), and produce proteins with exceptional functional properties. A number of studies on

several species have shown that these proteins have excellent functionality and quality (1).

Preliminary studies have also demonstrated that these proteins have an excellent ability to bind

water when incorporated into fish muscle products. Therefore, there is great interest in

investigating this specific application of these proteins as it could become an alternative method

to improve fish quality.

Obj ectives

The main objective of the study was to investigate how fish proteins, extracted with the

alkali-aided process, influence water-holding capacity and quality of raw and cooked fish

muscle. The hypothesis being tested was that fish protein isolate has a better effect on water-

holding compared to traditional ingredients used for this purpose.

The above hypothesis was tested by incorporating the protein isolate (PI) into fish fillets

under various solution conditions (, pH, NaC1, Tripoly phosphate (TPP)) and testing against

solutions without any proteins added. This study was divided into three parts. In study I, protein









isolate solutions (1 to 5% PI concentration) were inj ected in fish fillets. Four protein inj section

solutions (PI, PI + NaC1, PI + TPP and PI + NaCl + TPP) were used to obtain 10% inj section

level. Solubility and viscosity analysis were performed on protein injection solutions. Cook loss

and drip loss analyses were performed on protein inj ected fillets to determine optimum PI

concentration. In study II, 5% PI concentration was selected based on previous studies and 5, 10

and 15% inj section levels. The influence of the isolated proteins on the fish muscle was

investigated by measuring muscle pH, drip loss, cook loss, press loss, texture and gel formation

ability of isolated treated fish muscle. Furthermore, a protein compositional analysis was

performed on exudates from protein inj ected cooked muscle. In study III, fish fillets were

inj ected with solutions (NaC1, TPP and NaCl + TPP) without protein isolates to compare with

those inj ected with solution containing PI. Muscle pH, drip loss, cook loss, press loss, texture

and gel formation ability of isolated treated fish muscle and protein compositional analysis on

exudates from cooked muscle. The experimental plans as outlined in Figure 3-1, 2, 3.









CHAPTER 3
MATERIALS AND METHODS

Raw Material

For protein isolation and different treatments, tilapia fillets were obtained from a local

supplier (Rain Forest Aquaculture, Sunrise, FL) within 48-72 hours of harvest. Each fillet ranged

130-170 g in weight. The protein isolation and injection studies were performed at 40C or on ice.

Alkali-aided Protein Isolation

Fish fillets were ground at 40C in a Scoville grinder with 6 mm holes. Ground muscle was

diluted in deionized water (1:6) and homogenized with an Ultra-turrax 19 homogenizer at setting

10 for 1 min. The system was then adjusted to pH 11.0 by using 2M NaOH. The solution was

then passed through three filtering screens (first two 1 mm, and the last 0.5 mm) to remove

insoluble contents. The pH of the solution was then adjusted pH 5.5 (isoelectric point) by using

2M HC1. The protein isolate was then dewatered by using a cloth screen (1). All isolates was

held on ice and used within 48 h of production. The process flow is outlined in Figure 3-4.

Preparation of Protein Solutions

The moisture contents of tilapia protein isolate was analyzed by using moisture balance

(CSC Scientific company, Inc, Fairfax, VA USA). Four different protein solutions with different

concentrations (1%, 2%, 3%, 4% and 5%) were prepared on the basis of % solid contents of

tilapia protein isolate. Different amounts of the tri polyphosphate (TPP) were added to the

protein solution to obtain 5%, 10% and 15% inj section levels so that of final TTP concentration in

tilapia fish fillet was no more than 0.5 % (according to FDA recommendations). NaCl (1%) was

also added to the protein solutions. The pH of NaCl free protein solution and protein solution

with NaCl was adjusted to pH 8.00 and pH 8.15 respectively. These pH values were attained to

simulate the pH of protein solution with TPP and the combination of TPP and salt.









The solutions were inj ected by a hand operated inj ector which has 20 needles (gauge 18)

on a 2x6 inch platform. The fish fillets were inj ected in triplicate to obtain 5%, 10% and 15%

increase in weight.

Viscosity Measurement

A Brookfield digital viscometer DV-II (Brookfield engineering laboratories Inc,

Stoughton, MA USA) was used to determine the viscosity of protein solutions. All the

experiments were done at 100 rpm spindle speed and the measurements were taken in cP

(centipoises) or mPa.s (millipascal second). A sample of 500 mL protein solution was measured

into a 600 mL glass beaker immediately after the preparation, and the viscosity measurement

was taken immediately. All the measurements were done in triplicate.

pH Measurements

A pH meter (Model 220 Denver Instrument, Fort Collins, CO) was used for pH adjustment

of solutions and measuring muscle pH before and after inj ecting the sample as well before and

after cooking. For the pH measurement, 10 g of ground muscle was mixed with 90 mL of

deionized water at 4oC and homogenized with an Ultra-Turrax T19 homogenizer and pH of the

homogenate recorded. The pH meter was calibrated using pH 2, 4, 7, and 10 buffers at 4oC.

Protein Solubility

A Sorvall RC-5B Refrigerated Superspeed Centrifuge (DuPont Instruments, Wilmington,

DE) with a SS-34 rotor type was used to perform protein solubility analysis on different protein

solutions. Protein solutions were prepared and centrifuged at 10,000 x G for 15 min at 40C. The

concentration of protein in supernatants and in protein solution was measured by the Biuret

method (31). This method involves reaction between cupric chloride and proteins at alkaline pH,

which leads to the formation of color complex with absorption maxima at 540 nm. A standard

curve was constructed using bovine serum albumin (Sigma Chemical Co., St. Louis, MO).









Protein solubility was expressed as the difference between protein concentrations in supernatant

versus total protein concentration in protein solution. Triplicate samples were analyzed for

protein solubility from each protein solution used.

Protein Composition Analysis

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to

analyze protein and peptide composition of the proteins remaining in the supernatant after

centrifugation (i.e. soluble proteins) and proteins in the exudate from isolate treated fish muscle

after cooking (10). Protein concentration was measured using the modified Biuret method (31).

A pre-cast 4-20% Tris-HCI gradient gel (Bio-Rad Laboratories, Hercules, CA) was used to

analyze the composition of proteins and peptides from ~6.5-200 KDa. Protein samples (3

mg/mL) were prepared in LaemmLi buffer with mercaptoethanol and 10 CIL applied to each well.

The gels were run at 200V for 1 h, then fixed for 1 h in 12% TCA solution, stained in a EZ-Blue

stain solution (Sigma Chemicals, St. Louis, MO) for 1 h and destined in water until bands were

clearly visible. Gels were scanned and protein bands identified and quantified.

Cook Losses

To determine cook losses, the inj ected fillets were placed in Ziploc bags (Johnson & Son

Inc., Racine, WI). Then the samples were placed in a Precision@ water-bath (Jouan Inc.,

Winchester, VA) and heated at 80oC for 20 min (reaching an internal temperature of 1650F). The

water lost on heating (cook loss) was measured and analyzed for protein content and

composition, as described above. All the samples were analyzed in triplicate for each treatment.

Water-Holding Capacity (WHC)

A Sorvall RC-5B Refrigerated Superspeed Centrifuge (DuPont Instruments, Wilmington,

DE) with SM-24 rotor type was used to study water-holding capacity of inj ected tilapia fillets. A

single sheet of Whatman@ 3 filter paper was placed inside the centrifuge tubes to absorb









excessive moisture released during centrifugation. Filter paper was folded in a way to give the

shape of centrifuge tube; minced meat was placed inside the filter paper before centrifugation.

Triplicate fish fillets for each treatment, including control, were minced at 40C in a Scoville

grinder (Hamilton Beach, Washington, NC) with 6 mm holes. Approximately 10 g of minced

muscle was weighed accurately and immediately centrifuged at 269 x g (1500 rpm) for 15 min at

40C. After centrifugation, the samples were taken out and weighed again, excluding the moisture

absorbed by filter paper. The weight loss after centrifugation was divided by the initial weight

and expressed as %WHC (5).

Drip Losses

Drip loss of fish muscle was determined after injection. Three fish fillets for each treatment

were used. Fillets were weighed before being placed in Ziploc bags (Johnson & Son Inc., Racine,

WI). Then the fish fillets were injected with protein solution and stored for 1 day at 4oC. Drip

loss will be calculated as the percentage of original weight lost.

Color Analysis

A digital Color Machine Vision System (CMVS) was used to measure L* (lightness), a*

(redness) and b* yellownesss) values. This system consisted of a light box with illumination

from D50 fluorescent lamps, a CCD camera, a computer to acquire the images and processing

images with ColorExpert color image software (E&CS, Gainesville, FL). The settings of the

CCD camera were: brightness = 141, hue = 163, saturation = 67, exposure = 2043, gamma = 128

and sharpness = 135. The L* value measures lightness and darkness ranging from 0 (pure black)

to 100 (pure white). The a* value represents redness, where a negative (-) a* value corresponds

to greenness and a positive (+) a* value redness. On the other hand, a positive (+) b* value

represents yellowness and a negative (-) b* value blueness (32). The color analyses were

performed on cooked as well as uncooked fish fillets. Tilapia fillets were placed in light box and










digital camera captured picture of fish fillets. The interior side of a fish fillet was used to analyze

white muscle. Where as, the skin side of fish fillet was used to analyze dark muscle. The average

of three rectangular areas of interest was used to analyze white muscle. On the other hand, a

single rectangular area of interest was used to analyze dark muscle. All the analyses were

performed on triple samples and for proper comparison the same fish fillets were used to analyze

cooked and uncooked muscle.

Texture Analysis

Texture profile analysis (TPA) was performed on cooked and uncooked tilapia fillets by

Instron Universal Material Tester Model 4411 (Instron Corp., Canton, MA). Three samples (2

cm x 2 cm x 1.5 cm) from each tilapia fillets were analyzed using a #15 probe. The test consisted

of two successive compression ramps to a value of 70% of the unloaded specimen height.

Firmness (N), springiness (mm) and chewiness (N) was analyzed for each sample. All the

analyses were performed in triplicates.

Rheology Studies

The minced tilapia fillets inj ected with different protein isolate solutions were subj ected to

oscillatory theological testing. This test was used to investigate the gel forming ability of the

fish fillets. The theological studies were performed by using AR 2000-Advanced Rheometer

(TA Instruments, New Castle, DE). The tilapia fillets were processed to a paste in a Scoville

grinder (Hamilton Beach, Washington, NC) with 6 mm holes before being subj ected to the test.

Samples were placed between a temperature controlled plate and a oscillating plate (40mm

acrylic plate). The temperature ramps were set from 5-800C (heating ramp) and then 80-50C

(cooling ramp) at 30C/min. Variables such as, strain was set at 0.001, where as frequency (Hz)

was set at 0. 10. The gel forming ability of the system was studied by following the storage

modulus (G') as a function of temperature.









Statistical Analysis

Each experiment was performed in triplicate. Significant differences (P < 0.05) between

means within the treatment and among treatments were analyzed using Tukey's LSD test.

Analysis of variance (ANOVA) was used to test significant differences between treatments by

employing the statistical analysis system (SAS) computer program (10). Similar small in the

graphs represents no significant difference (P > 0.05) within and among treatments.


Protein Isolate Solution
(1,2,3,4 & 5%)



PI & PI + NaCl
PI PI + NaCl PI + TPP PI + NaCl + TPP (Hajse .0





Fish Fillets I a Fillet Injection
Solubility
Viscosity
Control Inj ected




Uncooked Cooked



*Drip loss Cook loss



Figure 3-1.Outline of study I. Protein solutions with PI concentration. (1 to 5%) inj ected in fish
fillets to obtain 10% inj section levels.










Protein Isolate Solution
(5%)


* Muscle pH
* Drip loss
* Texture analysis
* Color Analysis
* Rheology


* Cook loss
* Muscle pH
* Texture analysis
* Color Analysis
* Protein Composition


Fillet Inj section


1 1


Figure 3-2.Outline of study II. 5% PI solution inj ected in fish fillets to obtain 5, 10 and 15%
inj section levels.


PI PI + NaCl PI + TPP PI + NaCl + TPP
(pH adjusted 8.20) (pH adjusted 8.20)


Fish Fillets













































Figure 3-3. Outline of study III. Fish fillets inj ected with solution containing no PI to obtain 5, 10
and 15% injection levels.











Fish Muscle




Grind




Dilute (1:6) with
deionized water



Homogenize




Adjust pH to 11.0




Screen




Adjust pH to 5.5
(isoelectic point)



Dewater


Figure 3-4. Outline of the alkali-aided process to extract functional fish proteins.









CHAPTER 4
RESULTS AND DISCUSSION

The study was performed in three steps. In study I, tilapia fillets were inj ected with tilapia

PI solutions. The concentration of protein in the inj ected solutions ranged from 1 to 5% based on

total volume of the inj ected solution. Four different PI solutions were prepared for each protein

concentration. The different PI solutions were PI + 1% NaC1, PI + 0.5% TPP, PI + 1% NaCl +

0.5% TPP and PI without any added salt or TPP. NaCl was added based on the volume of PI

solution, while TPP was added based on the fillet weight. The pH of the inj ected solutions

containing TPP was 8.2 + 0. 1 and without TPP was 5.5 + 0. 1. The inj ected tilapia fillets were

analyzed for drip and cook loss. Solubility and viscosity analysis were also performed on the PI

solutions. The amount of PI inj ected into the fillets was 10% based on the weight of the fillet.

In study II, the pH of PI solutions were adjusted to 8.2 and were inj ected into tilapia

fillets at 5, 10 and 15% based on the weight of the fillet. The fillets were analyzed for cook and

drip loss. Cooked and uncooked fillets were analyzed for texture and color. Rheology studies

were done on uncooked minced tilapia fillets. Exudates obtained from cook loss were analyzed

using SDS-PAGE electrophoresis.

In study III, three solutions, a) TPP (0.5%) b) TPP (0.5%) + NaCl (1%) and c) NaCl

(1%), were injected into tilapia fillets at 5, 10 and 15% based on the weight of fillets. Texture

and color analyses were performed on tilapia fillets before and after cooking the fillets in water-

bath. Rheology studies were done on uncooked minced tilapia fillets. SDS-PAGE electrophoresis

was used to analyze the exudate obtained from cook loss.










Study I

Changes in Solubility and Viscosity of Tilapia Protein Isolate Solutions

Protein isolate (PI) solutions containing different protein concentrations were inj ected into

tilapia fillets. Various physical attributes of fillets such as cook loss and drip loss, and textural

characteristics of fillets were affected by protein concentration in PI, pH and the presence or

absence of NaC1, tri-poly phosphate (TPP) or a combination of NaCl and TPP (Figure 4-1).




-A- PI + TPP
18.0
+- PI + TPP + NaCI
16.0 + PI + NaCI (pH adjusted)

14.0~ _PI (pH adjusted)
-5 PI + NaCI
12.0 _g -IPI

S10.0

to8.0

6.0

4.0

2.0

0.0
1% PI 2% PI 3% PI 4% PI 5% PI




Figure 4-1. Solubility of tilapia protein isolates solution at different protein isolate (PI)
concentration. The pH values were: PI = 5.5, PI + NaCl = 5.5, PI + TPP = 8.2, PI + TPP + NaCl
= 8.2, PI (pH adjusted) = 8.2, PI + NaCl (pH adjusted) = 8.2.

The solubility of proteins was lowest for protein isolate (PI) solutions with no chemical

ingredients. In the absence of chemical additives, the pH of PI solution was around 5.5, which is

isoelectric pH of myofibrillar proteins. At isoelectric pH, the net charge of protein molecules

would be zero which could result in low solubility of PI (1). There was no significant difference









(P > 0.05) in solubility among different protein concentrations. However, when NaCl was added

to the protein solution, the solubility of PI increased significantly (P < 0.05). The increase in

solubility could be due to solubilization of myofibrillar proteins by the added salt. Among the

different protein concentrations, protein solubility was highest at 2% PI concentration and lowest

at 4% PI concentration. The addition of sodium tripolyphosphate (TPP) to PI solution in the

presence or absence of NaCl significantly increased (P < 0.05) the solubility of PI. At 1% and

2% protein concentrations, the solubility of PI solution with TPP was significantly higher (P <

0.05) than PI solution with NaCl + TPP. At 3 and 4% protein concentrations, there was no

significant difference (P > 0.05) among PI with TPP and PI with combinations of NaCl and TPP.

However, at 5% protein concentrations the solubility of PI solution with combinations of NaCl

and TPP was significantly higher (P < 0.05) than PI solution with only TPP. Also, the solubility

for PI solutions containing TPP or a combination of NaCl and TPP decreased with increased

protein concentration. Addition of TPP increased the pH to 8.2 (away from the iso-electric

point), which could result in an increased protein solubility (10). At 1% protein concentration,

there was no significant difference in the solubility of PI solutions in the presence or absence of

NaCl (pH adjusted to 8.20 f 0. 10). However, the solubility of PI solution with NaCl was

significantly lower at 2% protein concentration while no significant difference (P > 0.05) was

observed at 5% protein concentration. The solubility of PI without NaCl decreased significantly

with increase in protein concentration. At 5% protein concentration the solubility of PI with

NaCl (pH adjusted to 8.20 f 0.10) was significantly higher (P < 0.05) than other PI solutions.

Overall, the solubility of PI was lower at 5% protein concentration and higher at 1% protein

concentration. The amount of water available for interacting with protein is lower at 5% protein

concentration than at 1%, which cause protein to aggregate and results in lower solubility at 5%









concentration (1). Myofibrillar proteins are insoluble in solutions of ionic strength that are

similar to physiological conditions (33). Addition of NaCl could shift the isoelectric pH of

myosin leading to exposure of hydrophobic and sulfhydryl groups at the protein surface resulting

in loss of helical structure. Such conformational changes could create more surface area for

protein solubilization, thus increasing its solubility (34). Earlier, researchers have reported that

the pH of myofibrillar protein solution has to be kept above 7.0 in order to keep the protein in

solution. As pH 7.0 is above the isoelectric point of myofibrillar proteins (pH 5.5), the proteins

would carry a net negative charge and an electrostatic repulsive force would exist between

myofilaments (35). Swelling could cause by spaces within and between myofilaments and could

improve water-holding capacity (7, 14). Yongsawatdigul and Park (36) reported that rockfish

actomyosin was insoluble at pH 5 to 6 and was slightly soluble at pH 7 to 9.

Viscosity analyses (Figure 4-2) was performed on six different PI solutions (PI, PI + NaC1,

PI + TPP, PI + NaCl + TPP) and PI, PI + NaCl (pH adjusted to 8.20 f 0.10). At 1% and 2%

protein concentration, there was no significant difference (P > 0.05) among the six PI solutions.

At 3% and 4% protein concentrations the viscosity of PI (pH adjusted to 8.20 f 0. 10) was

significantly higher (P < 0.05) than remaining PI solutions. At 5% protein concentration the

viscosity of the following PI solutions: PI + TPP and PI + TPP + NaC1, were significantly higher

(P < 0.05) than PI and PI + NaCl solutions. An increase in viscosity with pH is expected as

increased electrostatic repulsion between similar charges could lead to an increase in

hydrodynamic volume of the muscle proteins (10). The viscosity was highest for the PI solution

adjusted to pH 8.20 f 0.10, while it was significantly reduced (P < 0.05) for PI + NaCl solution

(pH adjusted to 8.20 f 0.10). There was no significant difference (P > 0.05) among the viscosity

of PI, PI + NaC1, and PI pH adjusted to 8.20 f 0. 10 solutions at 5% protein concentration. PI and










PI + NaCl solutions showed low viscosity at all protein concentrations. This could be due to low

solubility of PI at pH 5.5, which is the isoelectric pH of PI solutions (1). At slightly higher pH

(pH 8.2), the viscosity of the protein solution increases due to increased positive and negative

charges of muscle proteins, leading to electrostatic repulsion between proteins (37). The

viscosity of various PI solutions containing NaCl is lower due to presence of Na+ ions in PI

solutions (Figure 4). Addition of salt reduces the repulsion between the negatively charged

protein molecules at higher pH (1). This results in reduced viscosity of PI solutions containing

salt (Figure 4).





3000 PI
9 PI + NaCI e
2500 -1 OPI + TPP
FI PI + TPP + NaCI
d 2000 -1 9 PI (pH adjusted)d
a 0i PI + NaCI (pH adjusted)
S1500

$ 1000

500 _b
aaaaaaaaaa aaaa a aaa a aa
0 ~F~...ii
1% PI 2% PI 3% PI 4% PI 5% PI



Figure 4-2. Viscosity of PI solutions at different protein concentrations.

Effect of PI Solutions on Cook and Drip Losses at Different Protein Concentrations

In the first study, four different PI solutions (PI, PI + NaC1, PI + TPP and PI + NaCl +

TPP) were inj ected into tilapia fillets at a 10% inj section level on the basis of initial weight of

tilapia fillets. Figure 4-3, shows the cook loss yield on the basis of green weight (weight of

tilapia fillets before inj section of PI solutions). When PI +TPP and PI + NaCl + TPP solutions

from 1% to 5% protein concentration were inj ected into tilapia fillets, they significantly reduced









(P < 0.05) the cook loss compared to control sample. At 1% and 2% protein concentration, PI

and PI + NaCl solutions significantly increased (P < 0.05) cook losses. Whereas at 3, 4 and 5%

protein concentration, there was no significant difference (P > 0.05) among tilapia fillets inj ected

with PI and PI + NaCl solutions and control samples. The PI and PI + NaCl solutions showed no

improvement in cook loss because the solubility of protein in the solution is minimal due to the

pH of solution. Lower cook loss was seen with tilapia fillets injected with PI solutions

containing TPP because TPP increased the pH of PI solutions and increased the solubility of

proteins. Overall, PI solutions at 5% level with combination of TPP and NaCl showed better

results when compared to 1- 4% PI solutions.


SProtein
9 Protein + Salt
O Protein + TPP
10.0 FI Protein + Salt + TPP

5.0 a a a

0.0a




S-10.0 --
- -
bcbc~bc bc
-15.0 b dfbe bcbcd

-20.0 def e

Control 1% ~2% 3% 4% 5%
-25.0


Figure 4-3. Cook losses (%) based on green weight for inj ected tilapia samples at different
protein concentrations.

Cook losses based on inj ected weight of tilapia fillets (figure 4-4) showed similar results

compared to cook losses based on green weight. Inj section of PI solutions containing TPP + NaCl

reduced cook loss at 1 to 5% protein concentration. Tilapia fillets inj ected with PI solutions (PI +










TPP and PI + NaCl + TPP) showed significantly reduced (P < 0.05) cook losses on the basis of

green weight compared to tilapia fillets inj ected with PI solutions (PI and PI + NaC1). Cook loss

for tilapia fillets inj ected with PI solutions (PI and PI + NaC1) were significantly higher (P <

0.05) at 1 and 2% protein concentration compared to 3, 4 and 5% protein concentration.




SProtein
lii Protein + Salt
0 Protein + TPP
FI Protein + Salt + TPP
0.0

-5.0

-1 0.0
a a --a aa
c a a a ab a
cn -15.0

~-20.0 -bc

-25.0 -1 Edf cdC cccd c

-30.0 -dfe

-35.0
1% 2% 3% 4% 5%




Figure 4-4. Cook losses (%) based on inj ected weight for inj ected tilapia samples at different
protein concentrations.

Drip losses based on green weight (Figure 4-5) significantly improved for tilapia fillets

inj ected with all four PI solutions (PI, PI + NaC1, PI + TPP and PI + NaCl + TPP) at 1 to 5%

protein concentration compared to control samples. Among inj ected tilapia fillets, those fillets

inj ected with PI + TPP and PI + NaCl + TPP) solutions showed significant reduction (P < 0.05)

in drip loss compared to fillets inj ected with PI and PI + NaCl solutions from 1 to 4% protein

concentration. At 5% protein concentration there was no significant difference (P > 0.05) among

fish fillets inj ected with PI, PI + NaCl and PI + NaCl + TPP solutions. Fish fillets inj ected with

PI solution containing PI + TPP showed significant reduction (P < 0.05) in drip loss compared to










fillets inj ected with PI and PI + NaCl solutions, but showed no significant difference (P > 0.05)

with those fillets inj ected with PI solution containing PI + NaCl + TPP.


SProtein
lii Protein + Salt
O Protein + TPP
14.0
El Protein + Salt + TPP
12.0
ab
ab ab a ab a abc b
10.0
a 8. c bcdabcdl bcded

p; 6 .0 TII e l e Fr e e
E 4.0
2.0



ffff
-4.0 Control 1% 2% 3% 4% 5%





Figure 4-5. Drip losses (%) based on green weight for inj ected tilapia samples at different protein
concentrations.

Drip losses based on inj ected weight of tilapia fillets (Figure 4-6) showed similar results

compared to drip losses based on green weight. PI solutions containing TPP and NaCl reduced

drip losses at 1 to 5% protein concentration but there was no significant difference (P > 0.05)

among the different protein concentrations. Tilapia fillets inj ected with PI + TPP and PI + NaCl

+ TPP solutions showed significantly reduced (P < 0.05) drip losses on the basis of green weight

compared to tilapia fillets inj ected with PI and PI + NaCl solutions.

Similar results were found when whole lobsters were inj ected with 0.3% sodium

tripolyphosphate and significantly decreased cook loss by 5% (38). Another study showed that

when turkey breast muscle was inj ected with salt and various types the phosphates, the presence










of salt and phosphates significantly reduced expressible moisture and cook loss (39). Wynveen et

al (40) showed decrease in drip loss when pork muscle with inj ected with phosphates.


SProtein
lii Protein + Salt
0 Protein + TPP
il Protein + Salt + TPP
0.0

-1.0

-2.0 -1 a -a bc a a-~
ab --.~ b
S-3.0 ab
acb
j3 -4.0
.$ bcd
E -5.0 -ebd
IC I de
-6.0 e cde

-7.0 e

-8.0
1% 2% 3% e 4% 5%
-9.0



Figure 4-6. Cook losses (%) based on inj ected weight for inj ected tilapia samples at different
protein concentrations.




Study II

Effect of PI Solutions on Cook and Drip Loss at Different Injection Levels

PI with 5% protein concentration was selected for further investigation since the overall

performance of this concentration was better than 1-4% protein concentration solutions. The PI

and PI + NaCl solutions was adjusted to pH 8.20 + 0. 10 to match the pH of the solutions

containing TPP. Figure 4-7 shows that at 5% injection level, all treatments significantly reduced

(P < 0.05) cook loss based on green weight but there was no significant difference (P > 0.05)

among all four treatments (PI + TPP, PI + TPP + NaC1, PI and PI + NaC1). At 10% inj section

level, there was significant reduction (P < 0.05) in cook loss based on green weight among all









treatments, where tilapia fillets inj ected with PI solutions containing TPP showed significantly (P

< 0.05) less cook loss compared to fish fillets inj ected with PI containing no TPP. At 15%

inj section level, tilapia fillets injected with PI solutions containing TPP showed significantly

lower (P < 0.05) cook loss compared to fish fillets inj ected with PI containing no TPP, where as

there was no significant difference (P > 0.05) among fish fillets inj ected with PI and PI + NaCl

solutions and the control sample. There was no significant difference (P > 0.05) among tilapia

fillets inj ected with PI + TPP and PI + NaCl + TPP solutions at 10% and 15% inj section levels. It

can be concluded from Figure 7 that overall performance at 10% inj section level was better than 5

and 15% inj section levels. PI solutions containing TPP performed better than those without TPP.

Since both type of PI solutions had similar pHs, TPP also acts as a buffer by resisting a change in

the pH of solution after inj section (1 7). Moreover, when soluble myofibrillar proteins extracted by

ionic strength, when mixed with tilapia protein isolate, will form a viscoelastic gel matrix, thus

contributing to entrapments of water in cooked muscle (41).

Figure 4-8, shows cook loss yield based on inj ected weight of tilapia fillets. Fish fillets

inj ected with PI + TPP and PI + NaCl + TPP solutions showed significantly lower (P < 0.05)

cook losses compared to fish fillets injected with PI and PI + NaCl solutions at 5, 10 and 15%

inj section levels. Tilapia fillets inj ected with PI + TPP and PI + NaCl + TPP solutions showed no

significant difference (P > 0.05) at 5, 10 and 15% inj section levels except for fish fillets inj ected

with PI solution containing PI + TPP which showed significantly higher (P < 0.05) cook loss at

15% inj section level compared to fish fillets inj ected with the same solution at 5% inj section level.

Tilapia fillets inj ected with PI and PI + NaCl solutions showed no significant difference (P >

0.05) at 5, 10 and 15% injection levels.










mPI + TPP
9 PI + NaCI + TPP
o PI
10.0 PI + NaCI

5..

5.0 i



-5.0 -I fg h
edg o
bcdde cde
-10.0

ab atabcd
-15.0
Control 5% injection 10% iniection 15% Iniection



Figure 4-7. Cook losses (%) based on green weight for tilapia samples inj ected to gain 5, 10 and
15% weight increase with a 5% PI solution.

Drip loss based on green weight (Figure 4-9) was significantly improved at all inj section

levels compared to control samples. At 5% inj section level, there was no significant difference (P

> 0.05) among tilapia fillets inj ected with four PI solutions (PI + TPP, PI + TPP + NaC1, PI and

PI + NaC1). At 10% and 15% inj section levels, tilapia fillets inj ected with PI + TPP and PI + NaCl

+ TPP solutions showed significantly lower (P < 0.05) drip losses compared to fish fillets

inj ected with PI and PI + NaCl solutions. Shahidi et al. (30) had earlier obtained similar results

with seal protein isolates. They found that meat muscle treated with seal protein hydrolysates

(3% concentration) show much higher drip loss when compared to meat muscle treated with

0.5% tripolyphosphate











mPI + TPP
9 PI + NaCI + TPP
o PI
5 PI + NaCI


0.0

-5.0

En-10.0

-15.0

-20.0


-25.0


15% Iniection


5% injection


10% injection


Figure 4-8. Cook losses (%) based on inj ected weight for tilapia samples injected to gain 5, 10
and 15% weight increase with a 5% PI solution.


a Pl + TPP
lii Pl + NaCI + TPP
o Pl
a* Pl + NaCI


18.0
16.0
14.0
12.0
10.0
8.0
6.0
4.0
2.0
0.0
-2.0


efg


bcd
bc
r bb


Control 5% Injection 10% Injection 15% Injection


Figure 4-9. Drip losses (%) based on green weight for tilapia samples inj ected to gain 5, 10 and
15% weight increase with a 5% PI solution.

Figure 4-10, shows cook loss yield based on injected weight of tilapia fillets. The fish

fillets inj ected with PI + TPP and PI + NaCl + TPP solutions shows significantly lower (P <










0.05) drip losses compared to fish fillets injected with PI and PI + NaCl solutions at 10 and 15%

inj section levels, except there was no significant difference (P > 0.05) at 10% inj section level for

tilapia fillets inj ected with PI + NaCl + TPP and PI + NaCl solutions. Tilapia fillets inj ected with

PI solutions (PI + TPP and PI + NaCl + TPP) showed no significant difference (P > 0.05) at 5%

inj section level.




m PI + TPP
9 PI + NaCI + TPP
o PI
1.0 a PI + NaCI



S-2.0 a a a ab aa
3.0 -abcd
ab
-4.0 cbcd
-5.0 cee
-6.0~ -

5% iniection 10% iniection 15% Iniection


Figure 4-10. Cook losses (%) based on inj ected weight for tilapia samples inj ected to gain 5, 10
and 15% weight increase with a 5% PI solution.

Water-holding capacity (Figure 4-1 1) was performed on minced tilapia muscle inj ected

with the four PI solutions at 5, 10 and 15% inj section levels. There was no significant difference

(P > 0.05) among tilapia fillets inj ected with PI + TPP and PI + NaCl + TPP and control samples.

Also, there was no significant difference (P > 0.05) among the control samples and those inj ected

with PI and PI + NaCl at 5% injection level. However, the moisture loss at 10 and 15% inj section

level for PI and PI + NaCl was significantly higher (P < 0.05) than the control. It can be

concluded that PI containing TPP had no significant effect on water-holding capacity of tilapia

fish muscle at 5, 10 and 15% injection levels, whereas PI without TPP had no effect at 5%









inj section level, but it significantly reduced WHC at 10 and 15% inj section levels. However, it has

been shown that beef tissue water-holding capacity (WHC) was increased by higher NaCl

concentrations and the presence of 10 mM pyrophosphate (16). Studies showed improved water-

holding capacity, reduced drip loss, thaw loss and cook loss when pork muscle was inj ected with

phosphates (40). Sherman (42) indicated that water retention of lean pork was increased by

phosphates. Another study showed the increased water-holding capacity of cod mince by

tripolyphosphate, which was attributed to increased pH of cod mince by tripolyphosphate (15).

Pink salmon fillets coated with soy protein concentrate and pink salmon protein powder showed

lower drip and cook loss than control fillets (43). Chemically modified soy proteins increased the

water-holding capacity and texture of meat sausage by improving the solubility (44). Freeze

dried meat binder make from frozen mackerel with reducing agents exhibited high binding

ability for pork chunks (45). Incorporation of 1.5% carrageenan in ham increased yield and

resulted in reduced juiciness. There was no synergistic effect on moisture retention due to a

combination of starch and carrageenan (46). Blue whiting fish was minced and treated with

hydrocolloids to measure the water-holding capacity. Results showed no improvement in WHC

when treated with carrageenan and xanthan (47). Modified starches were shown to be effective

water binders to improve cook yields in beef patties (48). In another study, fish protein

hydrolysate (FPH) was used to improve WHC of lizard fish surimi during storage. Fly fish FPH

improved WHC but mackerel FPH and chub mackerel FPH had no effect on WHC of lizard fish

surimi (49). The study showed that phosphates alone had better ability to increase water-holding

capacity then the mixture of phosphate and soy protein isolate, or the mixture of phosphate and

carrageenan at the same concentrations (50).










STPP
lii TPP + NaCI
O PI
35.00 li PI + NaCI
30.00 -aca
v, bcde abcd abcde
2 25.00 -cde decdeded
2 20.00




0.00
Control 5% Injection 10% Injection 15% Injection




Figure 4-1 1. Water-holding capacity of tilapia fillets inj ected to give 5, 10 and 15% weight
increase with a 5% PI solution.

PI Solution affecting Color of Uncooked and Cooked Tilapia Fillets

Color analyses were performed on dark muscle and white muscle for both cooked and

uncooked samples. The samples were statistically compared within treatment and among

different inj section levels. L*, a* and b* values were recorded to determine any difference in

color for the different PI solutions. L* and a* values for white cooked muscle (Figure 4-12, 13)

were not significantly different (P > 0.05) among tilapia fillets inj ected with all four solutions.

Also there was no significant difference (P > 0.05) among 5, 10 and 15% injection levels. Figure

4-14 shows that b* values for tilapia fillets injected with PI + TPP solutions at 5, 10 and 15%

inj section levels were significantly different (P < 0.05) compared to control sample. Tilapia fillets

inj ected with PI + TPP + NaC1, PI and PI + NaCl solutions showed no significant difference (P >

0.05) compared to control samples with respect to b* values.

Figures 4-15, 16, 17 showed the L*, a* and b* values of white uncooked muscle

respectively. L* values for white uncooked muscle (Figure 4-15) showed no significant

difference (P > 0.05) among tilapia fillets inj ected with all four solutions and also showed no









significant difference (P > 0.05) among 5 and 15% inj section levels. It can be concluded that PI

inj section has no effect on a~value (redness) and b~value yellownesss) of Eish fillets. However,

soy proteins caused yellow discoloration on the surface of the Cod fillets (9). At 10% inj section

level, L* values for fish fillets inj ected with PI + NaCl + TPP solutions were significantly higher

than those inj ected with PI + NaC1. It has been shown that addition of phosphates in cod fillets

resulted in white precipitate forming on the surface of muscle (5).

For dark cooked muscle (Figure 4-18), L* values were not significantly different (P >

0.05) at 5 and 10% injection levels for all four different treatments. However, at 15% injection

level, fish fillets inj ected with PI + TPP and PI + NaCl + TPP had significantly higher (P < 0.05)

L* values compared to tilapia fillets inj ected with PI + NaCl solutions. The a* value(Figure 4-

19) and b* values (Figure 4-20) for dark cooked muscle were not significantly different (P >

0.05) among tilapia fillets inj ected with PI + TPP, PI + TPP + NaC1, PI and PI + NaCl solutions

and also were not significantly different (P > 0.05) among 5, 10 and 15% injection levels.

Dark uncooked muscle (Figure 4-21) showed no significant differences (P > 0.05) in L*

values of tilapia fillets inj ected with PI + TPP, PI + TPP + NaCl and PI solutions when compared

with their respective control samples. However, tilapia fillets inj ected with PI + NaCl solutions

had significantly higher b* values at 5 and 15% inj section levels compared to control and 10%

inj section levels. The a* (Figure 4-22) and b* values (Figure 4-23) for dark uncooked muscle

were not significantly different (P > 0.05) among fillets inj ected with PI + TPP, PI + TPP +

NaC1, PI and PI + NaCl solutions, and also showed no significant difference (P > 0.05) among

5%, 10% and 15% inj section levels. Color analysis of cooked and uncooked tilapia muscle shows

that there was no overall significant change in color for fish fillets after inj section with PI

solutions.










Studies have showed that lobster tail treated with 0.3% phosphate had significantly lower

b* value and a* value than control samples (38). Also, studies have showed that 0.3, 0.4 and

0.5% phosphate decreased L* values in all pork muscles compared to control samples (40).

Young et al (51) showed that the inj section of phosphate solutions darken the color of both beef

and turkey muscle. While Yoon et al. (27) showed that frankfurters incorporated with soy protein

and NaCl showed L* values compared to the control sample. Alkali aided protein isolation could

produce functional protein isolate (PI) with minimal contamination by eliminating the membrane

lipid, which helped to stabilize b* values result in maintaining higher whiteness values (52). A

study showed no effect on L* values (lightness) of pink salmon fillets coated with soy protein

concentrate and pink salmon protein powder. Coatings did not affect b* value yellownesss) and

a* value (redness) of pink salmon fillets. However all stored samples had higher L* values than

fresh raw salmon, which was attributed to carotenoids degradation during storage (43).

SPI + TPP
lii PI + TPP + NaCI
O PI
FI PI + NaCI
80-
7-aaaa aa a a a aa

70



60

55

50
Control 5% Injection 10% Injection 15% Injection



Figure 4-12. Lightness (L* value) of white cooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution.



























--'''


SPI + TPP
9 PI + TPP + NaCI
O PI
5 PI + NaCI


3.0

2.5

2.0

1.5


a
aTaaTT


I '


Control


5% Injection


10% Injection


15% Injection


Figure 4-13. Redness (a* value) of white cooked muscle for tilapia samples injected to give 5,
10 and 15% weight increase with a 5% PI solution.


SPI + TPP
9 PI + TPP + NaCI
O PI
5 PI + NaCI


9.0
8.0




3.0



2.0
1.0
0.0


Control


5% Injection 10% Injection 15% Injection


Figure 4-14. Yellowness (b* value) of white cooked muscle for tilapia samples inj ected to give
5, 10 and 15% weight increase with a 5% PI solution.


aa a


aab ab
cb










SPI + TPP
9 PI + TPP + NaCI
O PI
B PI + NaCI


64

62

. 60


52


Control 5% Injection 10% Injection 15% Injection


Figure 4-15. Lightness (L* value) of white uncooked muscle for tilapia samples inj ected to give
5, 10 and 15% weight increase with a 5% PI solution.


SPI + TPP
9 PI + TPP + NaCI
O PI

14 a 5 PI + NaCI


O t


Control 5% Injection 10% Injection 15% Injection


Figure 4-16. Redness (a* value) of white uncooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution.










HPI + TPP
9 PI + TPP + NaCI
O PI
8 PI + NaCI
ab


" 3.0
n


0.0


5% Injection 10% Injection 15% Injection


Figure 4-17. Yellowness (b* value) white uncooked muscle for tilapia samples injected to give 5,
10 and 15% weight increase with a 5% PI solution.


SPI + TPP
9 PI + TPP + NaCI
O PI
8 PI + NaCI
ab


abccd


50 -1 abcd
bcd a

40-





20


10-


15% Injection


Control 5% Injection 10% Injection


Figure 4-18. Lightness (L* value) of dark cooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution.


a
abab ab











Control











m PI + TPP
9 PI + TPP + NaCI
o PI
a PI + NaCI


S0.8-

0.5

0.3

0.0-


Control 5% Injection 10% Injection 15% Injection


Figure 4-19. Redness (a* value) of dark cooked muscle for tilapia samples inj ected to give 5, 10
and 15% weight increase with a 5% PI solution.


SPI
9 PI
O PI
li PI


+ TPP
+ TPP + NaCI

+ NaCI


3.0

2.5

S2.0

'a 1 .5






0.0


Control 5% Injection 10% Injection 15% Injection


Figure 4-20. Yellowness (b* value) of dark cooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution.














SPI + TPP
9 PI + TPP + NaCI
O PI
8 PI + NaCI


40-

30-

20

10

0


Control 5% Injection 10% Injection 15% Injection


Figure 4-21. Lightness (L* value) of dark uncooked muscle for tilapia samples inj ected to give
5, 10 and 15% weight increase with a 5% PI solution.




SPI + TPP
9 PI + TPP + NaCI
O PI
16 H PI + NaCI

14 -1 a
12 -1 aa aaa
a aa










Control 5% Injection 10% Injection 15% Injection



Figure 4-22. Redness (a* value) of dark uncooked muscle for tilapia samples inj ected to give 5,
10 and 15% weight increase with a 5% PI solution.













HPI + TPP
lii PI + TPP + NaCI
O PI
E' PI + NaCI
3.5
3.0 a
3. -a a a
a a, a a
2.5- a

2; 2.0 -1 a a aa

:,1.5 _a
1.0

0.5

0.0
Control 5% Injection 10% Injection 15% Injection



Figure 4-23. Yellowness (b* value) of dark uncooked muscle for tilapia samples inj ected to give
5, 10 and 15% weight increase with a 5% PI solution.

Chemically modified soy proteins increased the water-holding capacity and texture of meat

sausage but had no effect on color (44). Blue whiting Eish was minced and treated with

hydrocolloids. The L* values, b* values and a* values increased with addition of guar, alginate

and CMC (47). In other study, fish protein hydrolysate (FPH) when incorporated in lizard fish

surimi showed yellow color and decrease lightness during storage (49).

PI Solutions Affecting Texture of Uncooked and Cooked Tilapia Fillets

Texture analyses were performed on cooked and uncooked samples inj ected with PI

solutions to determine firmness (N), springiness (mm) and gumminess (N). Figure 4-24 showed

no significant change (P > 0.05) in firmness of cooked tilapia fillets after injecting with different

PI solutions compared to their respective control samples. However, tilapia fillets inj ected with

PI + TPP at 5 % inj section level showed significantly higher (P < 0.05) firmness (N) compared to

tilapia fillets inj ected with PI + TPP at 15% inj section level. The springiness of injected cooked









tilapia fillets (Figure 4-25) was not significantly different (P > 0.05) when compared to

respective control samples. However, springiness of tilapia fillets inj ected with PI + TPP at 10%

inj section level had significantly higher (P < 0.05) values compared to respective control samples.

At 5% and 10% inj section levels, tilapia fillets inj ected with PI + NaCl had significantly less (P <

0.05) springiness when compared to tilapia fillets inj ected with PI + TPP solutions at the same

inj section levels. At 15% inj section level tilapia fillets inj ected with PI + NaCl solutions had

significantly less (P < 0.05) springiness when compared to tilapia fillets inj ected with PI + NaCl

+ TPP. Cooked tilapia fillets inj ected with PI solutions showed no significant changes (P > 0.05)

in gumminess (Figure 4-26) when compared to their respective control samples. Also, there was

no significant change (P > 0.05) in gumminess when samples were compared within 5%, 10%

and 15% injection levels.

Texture analyses were also performed on uncooked tilapia fillets. There was no significant

difference (P > 0.05) in firmness (Figure 4-27) when tilapia fillets at 5, 10 and 15% inj section

levels were inj ected with PI and PI + NaCl solutions.. Tilapia fillets inj ected with PI + TPP

solutions at 15% inj section level had significantly less (P > 0.05) firmness when compared with

control samples. However, tilapia fillets inj ected with PI + NaCl + TPP solutions at 5% inj section

level were significantly (P < 0.05) firmer when compared with tilapia fillets inj ected with PI +

NaCl + TPP solutions at 15% inj section level. There was no significant difference (P > 0.05) in

springiness of uncooked tilapia fillets (Figure 4-28) injected with PI, PI + NaC1, PI + TPP at all

inj section levels. However, tilapia fillets injected with PI + NaCl + TPP solutions showed

significantly lower (P < 0.05) springiness at all inj section levels, compared to their respective

control samples. There was also no significant difference (P > 0.05) in gumminess of uncooked

tilapia fillets (Figure 4-29) inj ected with PI + TPP, PI and PI + NaCl solutions at all inj section










levels. However, tilapia fillets inj ected with PI + NaCl + TPP solutions had significantly lower

(P < 0.05) gumminess at all inj section levels, compared to respective control samples. It can be

concluded that PI + NaCl + TPP solutions significantly reduced (P < 0.05) springiness and

gumminess at all inj section levels and also significantly reduced firmness at the 15% inj section

level compared to respective control samples. Ramezani et al. (44) showed that chemically

modified soy proteins increased the water-holding capacity and texture of meat sausage.

However, incorporation of carrageenan at 1.5% increased yield but resulted in reduced juiciness

(46). When blue whiting fish treated with hydrocolloids, the hardness values increased with

addition of locust bean gum and carboxymethylcellulose (CMC) (47). TPA values for the pork

bolognas treated with carrageenan and soy protein concentrate were generally similar to the

control, indicating that addition of carrageenan and soy protein concentrate had no effect on

texture of pork bolognas (53). Carballo et al. (54) reported that addition of modified waxy corn

starch increased TPA hardness and chewiness in bologna sausage.

SPI +TPP
9 PI + NaCI + TPP
O PI
18 -ab ab a ab a ab 5 PI + Na1CI
16 ab ab ab .a
14 l ab b 1 b
S12 1 b



W 0



Control 5% Injection 10% Injection 15% Injection



Figure 4-24. Firmness of cooked muscle for tilapia samples inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution.













SPI +TPP
9 PI + NaCI + TPP
O PI
B PI +NaCI


1-
0-


Control 5% Injection 10% Injection 15% Injection


Figure 4-25. Springiness of cooked muscle for tilapia samples inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution.


Control 5% Injection 10% Injection 15% Injection


Figure 4-26. Gumminess of cooked muscle for tilapia samples inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution.


5PI +TPP

9 PI + NaCI +
TPP














OPI +TPP
OPI + NaCI + TPP
9PI
HPI + NaCI
abcde

T bede


abcde


25





15

0o


Control 5% Injection 10% Injection 15% Injection


Figure 4-27. Firmness of uncooked muscle for tilapia samples inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution.


API+TPP
9 PI+ NaCl + TIPP
0 PI
5 PI+ NaCl


12.00


10.00


0.00


Control 5% Injection 10 %Inje action 15% Injection


Figure 4-28. Springiness of uncooked for injected tilapia samples injected to give 5, 10 and 15%
weight increase with a 5% PI solution.














SPI+TPP
9 PI+ NaCI + TPP
6.00-
O PI

5.00 a PI+ NaCl
4.00-a b


abc c

1.00-


0~ .00 b
Control 5%nection 10Injct 15Ineto










demnsraed ha rplcemnt of 10% orj mtore muscl protein by soy prtein ioaemrel








reduced muscle protein gel strength. TPA hardness of minced chum salmon treated with mixture

of phosphate and carrageenan was higher than salmon treated with only phosphate and control

samples (50).

PI Solution Affecting Gel Forming (G') Ability of Tilapia Muscle

The G' (storage modulus) of minced tilapia fillets was monitored during a two step


temperature sweep experiment. In step I, minced fillet was heated from 5-80oC, and in step II,

minced sample was cooled from 80-5oC. Figure 4-30 showed notable differences in rheograms of

different inj section levels obtained by PI solution with TPP. The G' of all samples decreased









initially but increased after 50-55 oC. On cooling, all samples showed an increase in G'. This

could be due to interactions between the proteins which are strengthened with a decrease in

temperature.

Gel forming (G') ability of minced tilapia fish muscle inj ected with all PI solutions was

determined by heating fish sample from SoC to 80oC and then cooling fish sample from 80oC to

SoC at a rate of 3oC/min. The storage modulus, or G' (refers to elasticity) values of samples were

taken at initial (9.2oC), mid (80. 1oC) and final (5.0oC) stages during the heating and cooling

regime. G' values at the initial (9.2oC) stage of heating for tilapia fillets inj ected with PI + TPP

and PI + NaCl + TPP solutions were significantly lower (P < 0.05) (Figure 4-31) compared to

their control samples. At 5 and 10% inj section level, G' values for tilapia fillets inj ected with PI +

TPP and PI + NaCl + TPP solutions were significantly lower (P < 0.05) than tilapia fillets

inj ected with PI and PI + NaCl solutions. However, at 15% inj section level, G' values for tilapia

fillets inj ected with PI + TPP and PI + NaCl + TPP solutions were significantly lower (P < 0.05)

than tilapia fillets inj ected with PI + NaCl solutions. Fish fillets inj ected with PI solution with no

TPP had higher drip losses during storage compared to those inj ected with PI and TPP. Fish

fillets containing TPP held more water, thus resulting in lower G' value at initial temperature

(5oC).

When heated to 80. 1oC, the G' values of minced fillets inj ected with PI + TPP and PI +

NaCl + TPP solutions at 5 and 10% inj section levels (Figure 4-32) were significantly lower (P <

0.05) than their control samples, suggesting a lower ability of those samples to form firm gels on

heating. However, tilapia fillets inj ected with PI and PI + NaCl solutions showed no significant

difference (P > 0.05) at 5, 10 and 15% inj section level with respect to G' compared to the control

samples. This might be due to more drip losses from inj ected tilapia fillets. After the cooling step










(to ~5.0oC) G' values for fish fillets inj ected with PI + TPP solutions at 5 and 10% inj section level

were significantly lower (P < 0.05) than control sample and fish fillets inj ected with PI + TPP

solutions at 15% inj section level. Fish fillets injected with PI and PI + NaCl + TPP solutions

showed significantly (P < 0.05) lower G' values at 10 and 15% inj section level compared to their

control samples. However, there was no significant difference (P > 0.05) at 5% inj section level for

fish fillets inj ected with PI and PI + NaCl + TPP solutions compared to control samples (Figure

4-33). It can be concluded that addition of TPP to the inj section solution results in weaker gels

compared to PI solution containing no TPP.


160000

140000

120000

100000

80000

60000

40000

20000

0


+ Control
+- 5% In jection
- 10% In jection
+ 15% In jection


0 10 20 30 40 50
Temperature (C)


60 70 80


Figure 4-30. Gel formation of minced tilapia fillets inj ected with PI solution with TPP. Heated
from 5-80oC and then cooled from 80-5oC.











SPI + TPP
1.1PI + TPP + NaCI
O PI
60000 l i PI + NaCI


a
a
a
a










Control


50000 -

40000

30000

20000

10000

0-


5% Injection


10% Injection 15% Injection


Figure 4-3 1. Gel formation of uncooked tilapia muscle (at 9.2oC) inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution.


SPI + TPP
M PI + TPP + NaCI
O PI
60000~ -.PI + NacI

50000 a
abb
abc abcd
40000 -1 abc abc
bcd ac
bcd
a30000- I de ~ del C cde
i3 I e
20000

10000



Control 5% Injection 10% Injection 15% Injection



Figure 4-32. Gel formation of uncooked tilapia muscle (at 80. 1oC) inj ected to give 5, 10 and 15%
weight increase with a 5% PI solution.










STPP
lii TPP + NaCI
O PI
200000
ab l PI + NaCI
180000 -1 a
160000 _acbacdabc
abcd abcd
140000 I lT~ bcd bcd
;;120000 L de L c-d e
100000 -de
80000 -

60000
40000
20000

Control 5% Injection 10% Injection 15% Injection



Figure 4-33. Gel formation of uncooked tilapia muscle (at 5.0oC) injected to give 5, 10 and 15%
weight increase with a 5% PI solution.

McCord et al. (56) demonstrated that replacement of 10% or more muscle protein by soy

protein isolate markedly reduced muscle protein gel strength. The addition of tilapia protein

isolate in the tilapia muscle caused the dilution of muscle proteins. The interference of myosin to

myosin interaction may be the inferiority to the gel strength. In similar study, the addition of soy

protein isolate in carp surimi resulted in weaker gel formation (57). Lanier (58) reported that

substitution of fish surimi with whey protein concentrate or turkey breast mince with soy protein

concentrate decreased gel rigidity.

Study III

Effect of Solution without Protein Isolate (PI) on Cook and Drip Losses

Tilapia fillets were inj ected with PI free NaC1, TPP and NaCl + TPP solutions (adjusted to

the same concentration and pH as the solutions containing PI) to compare with those inj ected

with tilapia PI. Figure 4-34 shows that at 5%, 10% and 15% inj section levels, cook loss based on

inj ected weight was significantly higher for tilapia fillets inj ected with NaCl solution compared









to fish fillets inj ected with TPP and NaCl + TPP solutions. At 15% inj section level, the cook

losses for tilapia fillets inj ected with NaCl solution were significantly higher (P < 0.05) than 10%

inj section level, whereas cook losses at 10% inj section level were significantly higher (P < 0.05)

than 5% injection level. However, tilapia fillets injected with TPP and NaCl + TPP solutions

showed no significant difference at 5%, 10% and 15% injection levels. Figure 4-35 shows cook

losses based on green weight. At 5%, 10% and 15% inj section levels, fish fillets inj ected with

TPP and NaCl + TPP solutions lost significantly (P < 0.05) less water on cooking compared to

control samples. However, at 5% and 10% injection levels, fillets injected with NaCl solution

showed no significant difference (P > 0.05) in cook loss compared to control, while at 15%

inj section level, fillets lost significantly more water than control. NaCl solution inj ected into fish

fillets had no effect on pH of fish muscle compared to fillets inj ected with TPP and NaCl + TPP

solution. Phosphates are good buffers, which may assist in the depolymerization of thick

filaments and increase water uptake and retention (7). In a study, pork lions were inj ected with

polyphosphates at 5% and 10% inj section levels without salts. Polyphosphates improved water-

holding capacity and yield, lions were more tender and juicy than untreated controls (18).

Another study showed the use of NaCl and tripolyphosphate in needle inj ected pork lions

improved quality characteristic and increased yield (59).

Figure 4-36, shows drip loss yield based on inj ected weight of tilapia fillets. Fillets inj ected

with TPP and NaCl + TPP solutions had significantly lower (P < 0.05) drip losses compared to

fish fillets inj ected with NaCl solution at 10% and 15% inj section levels, except there was no

significant difference (P > 0.05) at 5% inj section level.

Drip loss based on green weight (Figure 4-37) was significantly less at 5%, 10% and 15%

inj section levels compared to control samples. At 5% inj section level, there was no significant












difference among tilapia fillets inj ected with NaC1, TPP and TPP + NaCl solutions. At 10% and


15% inj section levels, tilapia fillets inj ected TPP and NaCl + TPP solutions showed significantly


lower (P < 0.05) drip losses compared to fish fillets inj ected with the NaCl solution.


HNaCl M TPP

ONaCl + TPP


0.00-

-5.00-

-10.00-

-15.00-

-20.00-

-25.00-

-30.00 -


5%/ inicetion 10%/ inicetion e 15%~ Tninction


Figure 4-34. Cook loss (%) based on inj ected weight of tilapia fillets inj ected with NaC1, TPP
and NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.


SNaCI
GITPP
O NaCI + TPP


Control


cd
d
5% iniection 10% 15%~ Iniection


Figure 4-3 5. Cook loss (%) based on green weight of tilapia fillets injected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.


Water-holding capacity (Figure 4-3 8) analyses were performed on minced tilapia muscle


inj ected with NaC1, TPP and TPP + NaCl solutions. At 15% inj section level, there was no


10.0-

5.0 -

0.0 -



-10.0-

-15.0 -

-20.0 -










significant difference (P > 0.05) among tilapia fillets inj ected with TPP and NaCl + TPP

solutions and the control samples. However, at 5% and 10% injection levels, fillets injected with

TPP and NaCl + TPP solutions, respectively, improved water-holding capacity. There was no

significant difference (P > 0.05) among control samples and those inj ected with NaCl solution at

5% inj section level, but there was significantly higher (P < 0.05) weight loss at 10% and 15%

injection level. Phosphates bind to myofibrillar proteins, these phosphates can effectively

increase repulsions between myoHilaments allowing water-binding responsible for increased

water-holding capacity of brine-treated meat (41). The poultry industry and red meat industry

have taken advantage of the increases moisture retention through use of phosphates (51, 60). The

moisture binding improvements with the addition of phosphates can be attributed to increase in

pH and ionic strength, the binding of phosphates to meat proteins, and the dissociation of

actomyosin into action and myosin (37). Similar study showed soy protein concentrate mixed

with 3% carrageenan favorably affected the water-holding capacity of processed sausages

regardless of the fat contents (61).

SNaCI
a TPP
o NaCI + TPP
0.00

-2.00-
ab a
S-4.00- ab ab aa

-6.00
-8.00-

-10.00-c

-12.00
5% iniection 10% iniection 15% Iniection



Figure 4-36. Drip loss (%) based on inj ected weight of tilapia fillets injected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.











5NaCI
P TPP
O NaCI + TPP
14.0-
12.0
10.0_b
S8.0 -b
6.0 -deded
S4.0-def
2.0-
0.0 -id ~
-2.0 f f f
Control 5% Injection 10% Injection 15% Injection



Figure 4-37. Drip loss (%) based on green weight of tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% Injection levels.




5NaCl
9 TPP
ONaCl + TPP
35.0-
30.0-
c beab
25.0 c d dc






0.0
Control 5% Injection 10% Injection 15% Injection



Figure 4-3 8. Water-holding capacity of tilapia fillets injected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels.




Effect of Solution without Protein Isolate (PI) on Color of Cooked and Uncooked Samples

Color analyses were performed on dark muscle and white muscle of both cooked and

uncooked samples. The samples were statistically compared within treatment and among

different inj section levels. L*, a* and b* values were taken to determine any difference in color









by injecting different solutions. Figure 4-39, 40, 41 showed L*, a* and b* values of white

uncooked muscle, respectively. For white uncooked muscle, the L* values (Figure 20-1) were

not significantly different (P > 0.05) among tilapia fillets inj ected with NaC1, TPP and TPP +

NaCl solutions at all inj section levels. While at 10% inj section level, the L* values for fish fillets

inj ected with NaCl solution were significantly higher (P < 0.05) than control. However, NaCl

inj ected in chicken breast meat show decreased L*(lightness) before and after cooking (6). The

a* and b* values (Figures 4-42, 43, 44) showed no significant difference (P > 0.05) within

different treatments and among 5, 10 and 15 injection levels.

Uncooked dark muscle (Figures 4-42, 43, 44) inj ected with all solutions showed no

significant difference (P > 0.05) in L*,a* and b* values and also showed no significant

difference (P > 0.05) among 5, 10 and 15% inj section levels. It can be concluded that the inj section

solutions do not have a significant effect of the color of dark uncooked fish muscle.

In white cooked muscle, L* and b* values (Figures 4-45, 46, 47) showed no significant

difference (P > 0.05) among tilapia fillets inj ected with NaC1, TPP and TPP + NaCl solutions

with their respective control samples at 5, 10 and 15% injection levels. Whereas a~values for

fillets inj ected with TPP were significantly lower than those inj ected with NaCl solution at 5, 10

and 15% injection levels.

Cooked dark muscle (Figures 4-48, 49, 50) did not show any significant differences (P >

0.05) among tilapia fillets inj ected with NaC1, TPP and TPP + NaCl solutions with their

respective control samples at all inj section levels. The results were similar to that of Pietrasik et

al. (61), who had shown earlier that soy protein concentrate mixed with 3% carrageenan had no

influence on color of processed sausages.












5NaCI

9 TPP

O NaCI + TPP
abc a abc


so.oo -

70.00-

60.00-

50.00-

40.00-

30.00-

20.00-

10.00-

0.00 -


abc


a abc


bcd ab


Control 5% Injection 10% Injection 15% Injection


Figure 4-39. Lightness (L* value) of white uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.





5NaCI

9 TPP

16.00 O NaCI + TPP

14.00 _1 a abc
abc abc abab
abc bc bcab




12.00-

10.00






Control 5% Injection 10% Injection 15% Injection





Figure 4-40. Redness (a* value) of white uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.













SNaCI

9 TPP

7.00 a a NaCI + TPP


6.00-aa

5.00-a

4.00-


S3.00-

2.00-

1.00-

0.00
Control 5% Injection 10% Injection 15% Injection



Figure 4-41. Yellowness (b*" value) of white uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.


SNaCl

9 TPP

0 NaCI + TPP


45.00


~a T


40.00 -

35.00-

30.00-

~i25.00-

20.00-

15.00-

10.00-

5.00-

0.00 -


Control 5 %Inje cti on 10% Injection 15% Injection


Figure 4-42. Lightness (L* value) of dark uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.













SNaCl

9 TPP

0 NaCl + TPP


25.00


20.00-



15.00-



S10.00-



5.00-



0.00 -


C control 5 %Inje cti on 10% Injection 15% Injection


Figure 4-43. Redness (a* value) of dark uncooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.




SNaCl

9 TPP


O NaCI + TPP


4.00 -

3.50-

3.00-

2.50-




1.50-

1.00-

0.50-

0.00 -


Control 5% Injection 10% Injection 15% Injection


Figure 4-44. Yellowness (b* value) of dark uncooked tilapia fillets injected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.












5NaCI
9 TPP
O NaCIl + TPP


82.00 -

81.00

80.00 -

S79.00-

78.00-

77.00

76.00 -


Control 5% Injection 10% Injection


Figure 4-45. Lightness (L* value) of white cooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.





5NaCI
9 TPP

1.501 0NaCI +TPP


1.00


m -0.50 -

-1.00 -

-1.50 -

-2.00


cd
10% injection


bcd


5% injection


d
15% Injection


Control


Figure 4-46. Redness (a* value) of white cooked tilapia fillets inj ected with NaC1, TPP and NaCl
+ TPP solutions to obtain 5, 10 and 15% injection levels.


15% Injection


abc abc


abcd












5NaCI

9 TPP

O NaCI + TPP

a a













15% Injection


10.00 -
9.00 -
8.00 -
7.00
6.00
5.00-
4.00-
3.00-
2.00-
1.00-
0.00 -


a.


aa


Control 5% Injection 10% Injection


Figure 4-47. Yellowness (b* value) of white cooked tilapia fillets injected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.


SNaCI

9 TPP

0 NaCI + TPP


60.00


ahed


abcd
bcd


50.00

40.00

S30.00

20.00

10.00-

0.00 -


15% Injection


Control 5% Iniection 10% Injection


Figure 4-48. Lightness (L* value) of dark cooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.















SNaCI

9 TPP

O NaCI+ TPP


2.50

2.00-

S1.50 -

1.00

0.50

0.00 -


Control


Figure 4-49. Redness (a* value) of dark cooked tilapia fillets inj ected with NaC1, TPP and NaCl
+ TPP solutions to obtain 5, 10 and 15% injection levels.


SNaCI

9 TPP

0 NaCI+ TPP
















15% Injection


3.50

3.00

2.50

2.00

1.50


1.00

0.50

0.00 -


Control 5% Injection 10% Injection


Figure 4-50. Yellowness (b* value) of dark cooked tilapia fillets inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.


5% Injection


ab ab

ab










10% Injection 15% Injection









Effect of Solution without Protein Isolate (PI) On Texture of Cooked and Uncooked
Samples

Texture analyses were performed on uncooked tilapia fillets. There was no significant

difference (P > 0.05) in firmness (Figure 4-5 1) of fillets inj ected with NaC1, TPP and TPP +

NaCl solutions and among all inj section levels. Tilapia fillets inj ected with TTP solution at 15%

inj section level were significantly less (P < 0.05) firm when compared with its control samples.

There was no significant difference (P > 0.05) in springiness of uncooked tilapia fillets (Higure 4-

52) among all treatments and inj section levels with the exception of tilapia fillets inj ected with

NaCl + TPP solution at 5%. There was also no significant difference (P > 0.05) in gumminess of

uncooked tilapia fillets (figure 4-53) for all treatments and inj section levels.

Texture analyses were also performed on cooked tilapia fillets. There was no significant

difference (P > 0.05) in firmness for all treatments and inj section levels with the exception of

fillets injected with NaCl solution at 10% injection level (Figure 4-54). At 10 and 15% injection

levels there was a significant increase (P < 0.05) in springiness (Figure 4-55) for fillets inj ected

with NaCl + TPP solution compared to control. Also at 10 and 15% inj section levels the

springiness of cooked tilapia fillets injected with TPP and NaCl + TPP solutions was

significantly higher (P < 0.05) than those inj ected with NaCl solution. There was no significant

difference (P > 0.05) in gumminess of cooked tilapia fillets (Figure 4-56) inj ected with NaC1,

TPP and TPP + NaCl solutions at 5, 10 and 15% injection levels. At 10 and 15% injection levels,

gumminess of cooked tilapia fillets inj ected with TPP and NaCl + TPP solutions was

significantly higher (P < 0.05) than those inj ected with NaCl solution. Similar study showed soy

protein concentrate mixed with 3% carrageenan improved the texture of processed sausages (61).











5NaCI
P TPP

0 NaCI + TPP






abcd










15% Injection



TPP and NaCl + TPP


60.00


50.00


40.00


S30.00 -




10.00


0.00 -


Control 5% Injection 10% Injection


Figure 4-51i. Hardness of uncooked tilapia fillets inj ected with NaC1,
solutions to obtain 5, 10 and 15% injection levels.


5NaCI
9 TPP


O NaCI + TPP


O


Control 5% Injection 10% Injection 15% Injection


Figure 4-52. Springiness of uncooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels.











SNaCI
9 TPP
O NaCl + TPP


20








5


0


bb


b "


bb b


Control


5% Injection


10% Injection


15% Injection


Figure 4-53. Gumminess of uncooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels.







SNaCI
9 TPP


0 NaCl + TPP


30.00


25.00-


20z.00

S15.00




5.00-

0.00 -


Control 5% Injection 10% Injection 15% Injection


Figure 4-54. Hardness of cooked tilapia fillets inj ected with NaC1, TPP
solutions to obtain 5, 10 and 15% injection levels.


and NaCl + TPP














5NaCI
9 TPP
O NaCI + TPP


5-









1-

0


Control 5% Injection 10% Injection 15% Injection


Figure 4-55. Springiness of cooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels.




SNaCI
P TPP

O NaCI + TPP


2





0.5



0.


Control 5% Injection 10% Injection 15% Injection


Figure 4-56. Gumminess of cooked tilapia fillets inj ected with NaC1, TPP and NaCl + TPP
solutions to obtain 5, 10 and 15% injection levels.









Effect of Solution without Protein Isolate On Gel Forming (G') Ability of Tilapia Muscle

The gel forming (G') ability of minced tilapia fish muscle inj ected with NaC1, TPP and

TPP + NaCl solutions was determined as described before. G' values before heating (9.2oC)

showed that tilapia fillets inj ected with TPP and NaCl + TPP solutions were significantly lower

(P < 0.05) at 5, 10 and 15% injection levels compared to their control samples (Figure 4-57).

This might be due to higher water contents in the inj ected tilapia fish muscle. However, fillets

inj ected with NaCl solutions showed no significant difference (P > 0.05) at all inj section levels

when compared to control samples. Fish fillets inj ected with NaCl solution had higher drip loss,

and thus less moisture than the other inj ected samples, which might be the reason for higher G'

values at the start of the heating regime (9.2oC).

When heated to 80oC, G' values of fillets inj ected with TPP and NaCl + TPP solutions

were significantly lower (P < 0.05) than those of control samples (Figure 4-58). However, tilapia

fillets inj ected with NaCl solution showed no significant difference (P > 0.05) at 5% inj section

level compared to control, but had a significantly lower (P < 0.05) G' at 10 and 15% inj section

levels compared to control. After cooling at 5.0oC (Figure 4-59) the G' values for fillets inj ected

with TPP and NaCl + TPP solutions were not significantly different (P > 0.05) than control. At

10 and 15% inj section levels, fish fillets inj ected with NaCl solutions showed no significant

difference (P > 0.05) in G' after cooling when compare to its control sample. Increase in protein

concentration of trout myofibrils induced an increase in G' thus resulting is stronger gel

formation (62). Another reason for weak gel might be due to higher pH of muscle. Similar study

was performed by Venugopal et al. (63) in which homogenate of shark meat was converted into

gel by lowering the pH to 3.5 using acetic acid. The gel formed by shark meat treated with acetic

acid was stronger than control. Gelation was associated with reduction in myosin heavy chain









and sulfhydryl groups. When TPP were added to chicken surimi in the presence ofNaCl the final

G' value was greater then without TPP (64). TPP in the presence of NaCl aids in solubilization of

actomyosin in meat systems (65), and also reduces the thermal stability of denaturation

temperatures of action and myosin (66). Wang and Smith (67) reported that when chicken breast

muscle myosin is heated to 55oC and cooled to 25oC, hydrogen bonded structures were formed,

which contributed to the stability and elasticity of myosin gel networks. The gel strength of

myosin or actomyosin gels increased as protein concentration increased (68). Increasing protein

concentration decreased the average intermolecular distance between protein molecules,

increasing the potential for protein-protein interactions (69). Final G' of chicken surimi increased

as pH decreased from 6.4 to 6.0 (64). Young et al. (70) reported that the rigidity of bovine

myofibrillar protein gels was greater at pH 6.0 than 8.0 at 85oC. Wang et al. (71) showed that the

gel strength of chicken breast muscle was greater at pH 5.5 than 7.5 at 80oC. As pH decreases

towards the isoelectric point, intermolecular repulsive forces decrease, favoring protein-protein

interactions and hence formation of a gel network (64).

It can be concluded that fillets inj ected with protein isolate solution containing TPP and

NaCl showed lower cook losses compared to fillets injected with solution containing TPP and

NaCl (without PI). The drip losses for fillets inj ected with PI and PI + NaCl were higher then

those inj ected with NaCl solution. The gel forming ability (G') of those fillets inj ected with TPP

and TPP + NaCl solutions were similar to those inj ected with protein isolate solution containing

TPP and TPP + NaC1, but G' value was lower than those inj ected with NaCl solution or PI

containing NaC1. There was no difference in overall texture and color of fillets inj ected with PI

solutions and with no PI.











SNaCl
9 TPP
O NaCl + TPP


50000 -
45000
40000
35000
30000
25000
20000
15000 -
10000
5000
0


15% Injection


Control 5% Injection 10% Injection


Figure 4-57. Gel formation of uncooked tilapia fillets at 9.2oC inj ected with NaC1, TPP and NaCl
+ TPP solutions to obtain 5, 10 and 15% injection levels.

SNaCl
9 TPP

40000m a NaCl + TPP


35000 b M

30000

25000

20000

15000 _

10000

5000


O


Control 5% Injection 10% Injection


Figure 4-58. Gel formation of uncooked tilapia fillets (at 80. 1oC) inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.


d d



15% Injection










SNaCl
9 TPP
180000 NaCI + TPP
160000
140000
120000 ~ b
S100000- b b b b
0 b b

60000
40000
20000


Control 5% Injection 10% Injection 15% Injection



Figure 4-59. Gel formation of uncooked tilapia fillets (at 5.0oC) inj ected with NaC1, TPP and
NaCl + TPP solutions to obtain 5, 10 and 15% injection levels.

Comparison between Fillets Injected With Solutions Containing Protein Isolate (PI) and
Without PI

The pH of uncooked control fillets was 6.4 6.5 (Table 1), which increased significantly

(P < 0.05) to pH 6.7 after injection with PI containing TPP. However, there was no significant

difference (P > 0.05) in pH when fillets were inj ected with PI solutions without TPP. Similar

pattern was observed in cooked fillets (Table 2) injected with PI solutions. The pH of control

cooked fillets was 6.6 6.7, which increased significantly (P < 0.05) to pH 6.85 with PI

solutions containing TPP. However, there was no significant difference (P > 0.05) in pH when

fillets were inj ected with PI solutions without TPP. The pH of cooked and uncooked fillets

(Table 3, 4) inj section with TPP and TPP + NaCl solutions (without PI) also significantly

increased (P < 0.05) but there was no significant difference (P > 0.05) in pH when fillets were

inj ected with solutions without TPP.









Our study showed -0.5% drip losses and -10% cook losses of control samples after storage

at 4oC for 1 day followed by cooking. Cook losses based on green weight decreased -5 to -6%

with inj section of PI solutions. In fact, cook losses were improved by +3% to +6% with inj section

TPP containing PI solutions (Table 7). When compared with fillets inj ected with solutions with

PI. The cook losses increased to -14% with inj section solution containing NaC1. Addition of

phosphates in inj section solution decreased cook losses to -0.5% (Table 1 1).

Inj ected weight cook losses of tilapia fillets inj ected with PI solution with TPP were -6% to

-10%, however increased from -12% to -18% with inj section of PI without TPP (Table 8). Fillets

inj section with solutions NaCl (no PI) showed inj ected weight cook losses from -15% to -25%.

Where as those fillets inj ected with solution with TPP and TPP + NaCl (no PI) were -5% to -

12% (Table 12).

Drip losses based on inj ected weight were -0.4% to -0.9% with inj section of PI solution

with TPP. However, drip losses reduced to -3% with PI solution without TPP (Table 6). When

compared with solution without PI. The drip losses were not only up to -2.8 with TPP solutions,

but increased to -8.7 with only NaCl solutions (Table 10).

Phosphates are effective in altering a decline in postmortem pH and improves the quality

of pork (40). Researchers have attributed the increase of WHC in meats treated with phosphates

to changes in pH and ionic strength (17). Others have suggested that phosphates induced

solubilization of actomyosin which increased with increasing pH (72). Another study showed

that addition of 0.5% tripolyphosphate to pork sausages formulated with or without carrageenan

increased hardness and reduced cooking losses(73). Similar change in texture upon addition of

phosphates was also reported (74). They reported an increase in firmness of turkey frankfurters

that contained 0.4% phosphate at either 1.5% or 2.0% salt. Meat pH greatly affected the water-










holding capacity of cooked pork sausages. The lower the pH, the greater was the drip thaw loss

in all treatments (75). Salt (NaC1) contributes to water and fat biding in meat products. Its

reduction in meat increase cooking loss and weaken the texture (76). It appeared that the strength

and water-holding capacity of protein gels were not always related. The SDS-PAGE analyses

were performed on the exudates of cooked samples inj ected with PI solutions. Figure 4-60,

showed no difference in protein composition among the exudates of tilapia fillets inj ected with

PI solutions containing TPP, TPP + NaCl (at 15% injection level) and control samples. Similar

results were observed among the other treatments. These analyses showed no difference (DATA

NOT SHOWN) in protein composition of exudates of tilapia fillets after inj ected with different

solutions (without PI).

Table 4-1. Uncooked fish fillets inj ected with different PI solutions (pH study).
Samples PI + TPP PI + NaCI + TPP PI PI + NaCI
Control 6.39 10.02 6.48 10.06 6.48 10.07 6.52 10.05
5% Injection 6.68 10.03 6.72 10.06 6.49 10.08 6.61 10.13
10% Injection 6.72 10.02 6.74 10.03 6.41 10.03 6.63 10.13
15% Injection 6.70 10.01 6.73 10.03 6.40 10.06 6.55 10.02

Table 4-2. Cooked fish fillets inj ected with different PI solutions (pH study).
Samples PI +TPP PI +NaCI +TPP PI PI +NaCI
Control 6.59 10.05 6.66 10.06 6.67 0.02 6.76 10.10
5% Injection 6.85 10.10 6.80 10.02 6.69 0.05 6.84 10.11
10% Injection 6.79 10.02 6.78 10.04 6.79 0.06 6.77 10.11
15% Injection 6.85 10.05 6.82 10.03 6.78 0.08 6.87 10.13

Table 4-3. Uncooked fish fillets inj ected with different inj section solutions without PI (pH study).
Samples NaCI TPP NaCI + TPP
Control 6.44 10.09 6.41 10.01 6.33 10.02
5% Injection 6.41 10.06 6.77 10.05 6.63 10.09
10% Injection 6.33 10.03 6.76 10.02 6.64 10.02
15% Injection 6.59 10.03 6.68 10.01 6.64 10.03

Table 4-4. Cooked fish fillets inj ected with different inj section solutions without PI (pH study).
Samples NaCI TPP NaCI +TPP
Control 6.64 10.03 6.72 10.00 6.52 10.08
5% Injection 6.51 10.04 6.84 10.05 6.70 10.02
10% Injection 6.57 10.04 6.76 10.07 6.72 10.01
15% Injection 6.38 10.05 6.76 10.15 6.73 10.01














Table 4-5. Green weight drip losses of fish fillets inj ected with different PI solutions.
Samples PI + TPP PI + NaCI + TPP PI PI + NaCI
Control -0.4 10.0 -0.6 10.3 -0.7 10.2 -0.6 10.0
5% Injection 5.0 10.7 5.4 10.6 4.2 10.3 4.3 1.
10% Injection 9.2 10.2 10.0 10.6 6.9 1217.7 10.5
15% Injection 15.4 101 14.2 10.2 10.8 1128.4 10.2

Table 4-6. Inj ected weight drip losses of fish fillets injected with different PI solutions.
Samples PI + TPP PI + NaCI + TPP PI PI + NaCI
5% Injection -0.8 10.3 -0.7 10.3 -1.9 110-1.6 1.
10% Injection -0.9 10.2 -1.0 10.0 -3.6 1. -3.0 10.7
15% Injection -0.4 10.4 -0.9 10.3 -3.9 10.7 -5.2 10.3

Table 4-7. Green weight cook losses of fish fillets inj ected with different PI solutions.
Samples PI + TPP PI + NaCI + TPP PI PI + NaCI
Control -11.5 114 -10.6 10.9 -10.5 117-9.6 1.
5% Injection -1.8 10.9 -1.0 10.6 -6.7 10.6 -9.5 1.
10% Injection -0.4 1143.2 10.9 -6.2 10.6 -4.9 1.
15% Injection 3.3 12.5 5.9 10.4 -6.3 115-5.7 1.


Table 4-8. Inj ected weight cook losses of fish fillets inj ected with different PI solutions.
Samples PI + TPP PI + NaCI + TPP PI PI + NaCI
5% Injection -7.2 10.5 -6.7 10.2 -12.2 10.8 -14.6 1.
10% Injection -9.6 110-7.1 10.5 -15.4 10.7 -14.4 1.
15% Injection -10.8 12.5 -8.2 101 -18.7 114 -17.6 1.

Table 4-9. Green weight drip losses of fish fillets inj ected with different inj section solutions
without PI.
Samples NaCI TPP NaCI + TPP
Control -0.5 10.3 -0.8 10.6 -0.6 10.3
5% Injection 1.7 10.8 3.7 10.4 3.8 10.7
10% Injection 1.1 1186.6 10.9 8.1 10.9
15% Injection 4.4 11011.3 11511.2 10.8


Table 4-10. Inj ected weight drip losses of fish fillets inj ected with different injection solutions
without PI.
Samples NaCI TPP NaCI + TPP
5% Injection -3.8 10.7 -1.6 101-1.4 10.6
10% Injection -7.6 116-2.3 101-2.0 1.
15% Injection -8.7 113-2.4 10.5 -2.8 10.3














Table 4-1 1. Green weight cook losses of Eish fillets inj ected with different inj section solutions
without PI.
Samples NaCI TPP NaCI + TPP
Control -9.7 10.6 -9.6 113-9.1 11.1
5% Injection -10.4 12.7 -0.8 10.7 -2.3 1.
10% Injection -12.5 119-0.3 10.7 0.8 1.
15% Injection -14.2 13.3 3.0 1190.4 11.1


Table 4-12. Inj ected weight cook losses of Eish fillets injected with different inj section solutions
without PI.
Samples NaCI TPP NaCI + TPP
5% Injection -15.2 12.8 -5.9 10.4 -7.2 10.2
10% Injection -20.0 117-8.6 10.2 -8.6 11.1
15% Injection -24.9 13.2 -9.7 10.7 -12.2 10.7


Myosin HC I lh


r

--)/-
i


-- -


205 KDa
116 KDa
97 KDa
66 KDa
55 KDa
45 KDa
36 KDa
29 KDa
24 KDa
20 KDa
14.2 KDa


ii --- rrrrrr rrr*r ~-n~


Actin ao ie





Myosin


Pl + TTP


Control


Pl + TTP + NaCI Standard


Figure 4-60. Protein composition of exudates of tilapia fillets inj ected with PI solutions after
cooking performed my SDS-PAGE electrophoresis.










CONCLUSION


This study demonstrated the effect of tilapia protein isolate on water-holding capacity and

quality of tilapia fillets. Tilapia protein isolate (PI) solutions with TPP improved cook yield and

reduced drip losses compared to PI solutions without TPP. PI solutions showed no significant

change in the texture and color of uncooked and cooked tilapia fillets. However, gel forming

ability (G') of tilapia fillets inj ected with PI solution with TPP was decreased. This is probably

due to higher amount of moisture retained with tilapia fillets.

These results were compared with tilapia fillets injected with TPP, NaC1, and TPP +

NaCl solutions without PI. PI solutions with TPP showed higher cook yield and reduced drip loss

compared to inj section solution with TPP. However, PI solutions without TPP showed lower cook

yield and higher drip loss compared to TPP solution. Inj section solution with NaCl showed lowest

cook yield and highest drip loss compared to all inj section treatments.










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BIOGRAPHICAL SKETCH

Saqib Hussain was born in Rawalpindi, Pakistan. He received his bachelor' s in Food

Technology from University of Arid Agriculture, Pakistan in September 2001. After his

internship in PEPSI as an undergraduate student, he came to United States in October 2001. He

started his master' s degree in Spring 2005 under the supervision of Dr. Hordur G. Kristinsson.

Saqib is a member of Institute of Food Technologists (IFT), Chicago, IL and participated in the

annual IFT meeting held in Orlando, FL in July 2006. While at the University of Florida he was

joined Phi Tau Sigma, the honorary society of food scientists.