International Journal of Food Properties

ISSN: 1094-2912 (Print) 1532-2386 (Online) Journal homepage: http://www.tandfonline.com/loi/ljfp20

Biochemical Changes in Dressed Priacanthus

hamrur (Bulls Eye) During Frozen Storage and
Its Effect on Physical and Sensory Quality of Fish
Sausage

B. Umesha Bhatta, R. M. Prabhu, A. Manjunatha Reddy & K. Elavarasan

To cite this article: B. Umesha Bhatta, R. M. Prabhu, A. Manjunatha Reddy & K. Elavarasan
(2015) Biochemical Changes in Dressed Priacanthus hamrur (Bulls Eye) During Frozen Storage
and Its Effect on Physical and Sensory Quality of Fish Sausage, International Journal of Food
Properties, 18:4, 897-908, DOI: 10.1080/10942912.2013.837062

To link to this article: http://dx.doi.org/10.1080/10942912.2013.837062

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Published online: 20 Mar 2014.

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International Journal of Food Properties, 18:897908, 2015
Copyright Taylor & Francis Group, LLC
ISSN: 1094-2912 print/1532-2386 online
DOI: 10.1080/10942912.2013.837062

Biochemical Changes in Dressed Priacanthus hamrur

(Bulls Eye) During Frozen Storage and Its Effect on Physical
and Sensory Quality of Fish Sausage

B. Umesha Bhatta, R. M. Prabhu, A. Manjunatha Reddy, and K. Elavarasan

Department of Fish Processing Technology, Karnataka Veterinary, Animal and Fisheries Sciences
University, Bidar College of Fisheries, Mangalore, Karnataka, India

In the present investigation, biochemical changes in dressed and frozen stored Priacanthus hamrur and
their influence on quality of fish sausage have been studied. During the frozen storage period of five
months, moisture content decreased and led to the increment on other proximate constituents due to
percentage balance. Quality indices such as total volatile base nitrogen and trimethyl amine nitrogen
increased significantly (P < 0.05) and reached 44.80 mg N/100 g and 22.00 mg N/100 g of meat,
respectively. Non-protein nitrogenous contents were found to decrease due to the loss in thaw drip.
Lipid freshness parameters such as peroxide value and free fatty acid value increased and resulted in
reduced extractability of salt soluble proteins. Gel strength of fish sausage decreased significantly (P <
0.05) during the storage period and this was also supported by the folding test. Increments in the express-
ible moisture content clearly indicated the loss of protein functionality, like water holding ability, due
to denaturation during frozen storage. From the sensory evaluation, dressed P. hamrur can be stored
frozen without any cryoprotective and preservative agents up to four months when it is intended for the
preparation of fish sausage and this was also evidence from the changes in biochemical constituents.

Keywords: Priacanthus hamrur, Bulls eye, Quality changes, Frozen storage, Fish sausage, Gel strength.

INTRODUCTION

Due to the over exploitation of resources, deep sea fishes are the emerging fishery resources for
developing countries like India. Priacanthus hamrur is an underutilized deep sea fish widely dis-
tributed in the Indo-Pacific region and identified as one of the potential species available for the
production of surimi and surimi based products. Surimi is a wet concentrate of myofibrillar pro-
teins used as an intermediate product in the development of various fish mince based products. Fish
sausage is a prominent surimi or fish mince based value-added product, which is gaining a wide
fame in recent years. Fish sausage, a hybrid product between kamaboko and meat sausage,[1] is
prepared from fish mince by mixing with additives, spices, followed by stuffing the mix/paste into
synthetic casing and heat processing.

Received 7 February 2013; accepted 18 August 2013.

Address correspondence to R.M. Prabhu, Department of Fish Processing Technology, Karnataka Veterinary, Animal and
Fisheries Sciences University, Bidar College of Fisheries, Mangalore 575001, India. E-mail: rmprabhufpt@gmail.com

897
898 BHATTA ET AL.

Eating quality or consumer preference of fish sausage is determined by the gel forming abil-
ity. Formation of gel involves partial denaturation of proteins with irreversible ordered aggregation
leading to the formation of a three dimensional network with the entrapment of water and added
ingredients.[2] Gelation is one of the most important functional properties that influence the tex-
tural properties of fish products. Gel strength of fish protein gels mainly depends on the quality
of raw material used for the production. Species from the same genus (Priacanthus sp) have been
reported for the comparable gel forming ability compared to fishes such as threadfin bream, croaker,
lizardfish, etc.[3]
The utilization of fish species for the preparation of gelled products like fish sausage calls for
deeper understanding on the biochemical changes in fish during various preservation methods and
their impact on the quality of products. Freezing and frozen storage is a long term preservation
method widely employed in fish processing industry and makes the raw material availability around
the year. Quality changes in fish and shellfishes are evaluated for years in the seafood industry by
measuring the chemical constituents such as trimethylamine (TMA) and total volatile base nitrogen
(TVB-N)[4,5] and limits of these compounds are regulated according to Directive (91/493/EEC)
and Directive (95/149/EEC), respectively. Spoilage due to microbial activity is terminated during
freezing and frozen storage. But the endogenous enzymes and changes in proteins are limiting the
storage life of frozen stored fish.
The gel forming ability of lizard fish, hoki, and silver carp have been reported to decrease during
frozen storage due to the denaturation of myofibrillar proteins.[6,7,8] Denaturation and aggregation of
myofibrillar proteins are associated with the formation of free fatty acids (FFAs) and lipid oxidation
products leading to the loss of functionality and texture.[9,10] Hydrophobic association of oxidized
and hydrolysed lipid based products to the surface of proteins decrease the solubility which is a
major factor determining the functional properties.[11,12] In the present study, changes in the bio-
chemical constituents of dressed P. hamrur during frozen storage and their influence on the physical
and sensory quality of fish sausage have been investigated.

MATERIALS AND METHODS

Preparation of Raw Material for Storage Study

Fresh bulls eye (Priacanthus hamrur) was procured from the local fish landing center, Mangalore,
located along the west coast of India and brought to the laboratory within an hour. During trans-
portation, the raw material was packed in an insulated container with ice in alternative layers. The
fish was thoroughly washed with chilled water prior to processing. The processing was carried out
under hygienic condition.
Fish was dressed to remove scales, head, and entrails. Dressed fish was washed thoroughly
in chilled water, and then packed in polyethylene bags of 2 kg each in a single layer. Around
20 such bags were made and they were frozen in the air blast freezer (Armfield Limited, Ringwood
Hampshire, England) at 35 C and then all the packs were stored in the deep freezer (Blue star,
India) at 20 C for a period of five months. Monthly, about 6 kg (3 2 kg) of dressed fish was
taken out and thawed at 4 C in refrigerator for 68 h. Meat was separated and analyzed for bio-
chemical changes. Fish sausage preparation was carried out using the recipe formulated by Bhatta,
Prabhu, and Reddy.[13] Process flow for the fish sausage preparation and quantity of ingredients used
are given in Flowchart 1 and Table 1, respectively. The study was conducted for the frozen storage
period of five months.
 FROZEN STORAGE EFFECT ON SAUSAGE FROM PRIACANTHUS 899

Minced meat

Mixing with additives (Table1) in a silent cutter

Stuffing the sausage mix in to Krehelon casings

Ringing

Washing

Heat processing

Boiling at 8890C for 60 min

Cooling at 15C for 15 min

Reboiling at 100C for 1min

Wiping and drying under fan

Sausage for quality analysis

FLOWCHART 1 Process flow for the preparation of sausage.

TABLE 1
Composition of ingredients used in fish sausage preparation

Material Weight in grams

Minced fish meat 700

Salt 20
Sugar 15
STPP 2
MSG 2
Color 5ml
Coriander 4
Chili powder 1
Potassium sorbate 2
Starch 90
Chilled water 100200 ml
Oil 5055 ml

Analysis
Proximate composition
The proximate composition namely moisture, total protein, and total ash of fish meat were deter-
mined according to the methods described in AOAC.[14] The moisture content was determined by
the standard hot air oven (100 2 C for 18 h) method. Total nitrogen was determined by Kjeldahl
method of AOAC and crude protein was calculated by multiplying with the factor of 6.25 and
expressed in percentage on wet weight basis. Total ash was determined by heating an incinerated
sample in a muffle furnace (550 C for 10 h) and values were expressed on wet weight basis (AOAC).
900 BHATTA ET AL.

Total lipids were extracted by the procedure of Bligh and Dyer[15] using chloroform and methanol
mixture.

Quality indices
Total volatile base nitrogen (TVB-N) and trimethyl amine nitrogen (TMA-N) were estimated by
the method described by Beatty and Gibbons.[16]

Lipid freshness evaluation

Peroxide value (PV) and FFA value were determined according to the method described by
Jacobs[17] and Takagi, Hayashi, and Itabashi,[18] respectively using chloroform-methanol extract
of lipids prepared from fish meat.

Salt soluble proteins (SSP)

SSP were extracted with 5% (w/v) sodium chloride solution buffered with 0.02 M sodium bicar-
bonate, pH 7.2, using the method of Dyer, Frennch, and Snow.[19] Extraction was carried at 4 C.
Protein content in the extract of SSP was determined by the method of Kjeldhal as described in
AOAC.[14]

Non-protein nitrogen
The non-protein nitrogen content of meat was analyzed according to the method described by
Velankar and Govindan[20] and expressed as mg N/100 g of meat.

Physical Quality Evaluation of Fish Sausage

Gel strength measurement
The gel strength of fish sausage was determined using Okada gelometer (Saitama Keki Seisa
Kuso Co. Ltd., Tokyo, Japan) as described by Suzuki.[21]

Expressible moisture content (EMC)

The EMC of the sausage was measured according to the method described by Okada.[22]

Folding test
The folding test of fish sausage in synthetic casing was carried out according to the method
described by Suzuki.[21] The method involves the use of round shaped cut piece of fish sausage with
5 mm thickness. While testing, the piece is folded along its diameter. Folding test grades are given
based on the place and the extent of damage and is described as follows.

AA: The sausage slice does not break even when it is folded twice in axis of the first fold being
perpendicular to that of the second.
A: the slice is intact without any break in the first fold.
B: crack occurs only on the part of the folded edge of the slice.
C: crack runs through the line of folding.
D: the slice breaks apart completely.
 FROZEN STORAGE EFFECT ON SAUSAGE FROM PRIACANTHUS 901

Sensory Analysis
Fish sausages prepared in a synthetic casing were subjected to sensory evaluation by a team of
trained panelists. The minimum number of ten panelists evaluated the fish sausage. The procedure
for organoleptic evaluation was followed as described by Karthikeyen et al.[23] The attributes that
were evaluated by trained panelists included appearance, color, texture, flavor, taste, and overall
acceptability. The scoring pattern of each attribute is (hedonic scale):
Like extremely9;
Like very much8;
Like moderately7;
Like slightly6;
Neither like nor dislike5;
Dislike slightly4;
Dislike moderately3;
Dislike very much2;
Dislike extremely1.

Statistical Analysis
Analyses were carried out in triplicates for the samples drawn randomly from three different frozen
dressed fish packs and each was packed with 2 kg of dressed fish. Correlation coefficient between the
variables was analyzed using Pearson correlation coefficient and the significance of correlation was
tested using students T test. Data were subjected to ANOVA and Duncan multiple range mean
comparison test to determine the homogeneity of variance and significant differences in biochemical
indices, lipid freshness, and sensory scores as a function of storage days. Statistical analysis was
performed using statistical program SPSS 16 (SPSS.16.0 for windows, SPSS Inc., Chicago, IL,).

RESULTS AND DISCUSSION

Changes in Proximate Composition

Changes in the proximate composition of P. hamrur during the frozen storage period of 150 days
are given in Table 2. Moisture content decreased from 77.28 to 72.87%. The observed decrement
of 5.71% from the initial moisture content can be attributed to the structural changes in the protein

TABLE 2
Changes in proximate composition of meat from bulls eye during frozen storage

Storage period in months Moisture (%) Protein (%) Total lipid (%) Total ash (%)

0 77.28 2.80a 18.95 1.25a 1.27 0.01a 2.42 0.58a

1 76.75 1.17ab 18.82 0.3a 1.5 0.01b 2.35 0.31a
2 75.53 0.71abc 18.92 0.43a 1.93 0.02c 3.23 1.04ab
3 74.57 1.13bcd 18.98 0.5a 2.01 0.28c 3.75 1.33ab
4 73.32 0.23cd 19.55 0.55a 2.24 0.01d 4.04 0.94b
5 72.87 0.41d 19.53 0.45a 2.3 0.02d 4.35 0.2b
Presented
values represent the mean standard deviation;
Mean with different letters in superscripts in the same column indicate that the results are significantly different
(P < 0.05).
902 BHATTA ET AL.

fractions due to freezing and frozen storage. Protein, lipid, and ash contents varied from 18.95 to
19.53%, 1.27 to 2.30%, and 2.42 to 4.35% during the storage period, respectively. There was no sig-
nificant (P > 0.05) increment in the protein content throughout the storage period. But the increment
in the total lipid and total ash were found to be significant compared to the initial value (P < 0.05).
There was a significant correlation between the decrement in the moisture content and increment in
the lipid (r = 0.741; P < 0.01). The similar trend was also observed between moisture and ash values
(r = 0.650; P < 0.01). Moisture content of surimi from pink perch has been reported to decrease
from 78.50 to 76.55% during the frozen storage period of 36 weeks.[24] In the present study, the
lipid and ash contents increased and this can be attributed to the decrement in the moisture content
(due to the percentage balance between the proximate constituents). A similar relationship between
moisture and lipid has been reported for the restructured trout during frozen storage.[25]

Changes in the Chemical Quality Indices

Quality changes of fish during preservation by various methods are commonly evaluated by mea-
suring the TVB-N and TMA-N. Changes in the TVB-N and TMA-N of P. hamrur during 150 days
(5 months) of frozen storage are given in Fig. 1a. The TVB-N and TMA-N contents were observed
to increase significantly from the initial value of 8.40 to 44.80 mg N/100 g and 4.20 to 22.00 mg
N/100 g of meat (P < 0.05), respectively. A positive correlation was observed for TVB-N (r =
0.963; P < 0.01) and TMA-N (r = 0.943; P < 0.01) with storage periods and found to be signifi-
cant. Generally, marine fish with the TVB-N value of 1520 mg N/100 g is considered as good in
PV(milliequivalent of O2/kg of fat)
TVB-N / TMA-N (mg N / 100 g)

50 20
60 PV FFA
A d B

FFA (% as oleic acid)

TVB- N TMA- N e 40 d
50 15
e
40 c
d 30
d 10
30 bc
c c 20 c c
ab
b c a
20 b b b 5
10 a
a b b
10
a
0 0
0 0 2 4 6
0 2 4 6
Storage period (months) Storage period (months)
20
200
C D
SSP (g / 100 g of meat)

NPN (mg N / 100 g)

16 a
a 160 a
ab
12 abc bcd
cd 120 b
d bc
8 c c
80
4 40
0 0
0 2 4 6 0 2 4 6
Storage period (months) Storage period (months)

FIGURE 1 Changes in (A) total volatile base nitrogen and trimethyl amine nitrogen, (B) peroxide value and free
fatty acid value, (C) salt soluble proteins, and (D) non-protein nitrogenous contents of dressed P. hamrur during
frozen storage. Error bars represent the standard deviation from triplicates. Different small letters indicate that the
results are significantly different (P < 0.05).
 FROZEN STORAGE EFFECT ON SAUSAGE FROM PRIACANTHUS 903

quality whereas 50 mg N/100 g is regarded as poor in quality.[26] The acceptable limit of TMA-N
varies from 1015 mg/100 g of meat. In the present investigation, TMA-N value reached 18.90 mg
N/100 g of meat on fourth month of frozen storage, whereas the TVB-N content was within the
acceptable limit (44.80 mg N/100 g of meat) up to the fifth month. It is well established that accumu-
lation of TMA-N and TVB-N during frozen storage is due to the bacterial activity and endogenous
enzyme, TMAOase. Though frozen storage is having detrimental effect on the bacterial prolifera-
tion, enzymes mediated changes were likely to be continued and resulted in the increment of TVB-N
and TMA-N. Siddaiah et al.[8] reported 32.85 mg of TVB-N for 100 g of mince from silver carp
(Hypophthalmichthys molitrix) for the frozen storage period of six months. TMA-N is responsible
for the fishy odor of ice stored or frozen stored fish. In the present research, a negative correlation
was observed for the changes in the sensory score of odor of the prepared sausage with TMA-N (r =
0.467; P < 0.05) and TVB-N (r = 0.600; P < 0.01) and found to be significant. As the marine fish
contains higher amount of TMA compared to fresh water fishes, TMA and TVB-N are considered
as better indices of quality which vary greatly between the species and exhibit greater difference.[27]

Changes in Lipid Freshness

The lipid freshness of fish meat during frozen storage is evaluated by measuring FFAs, PV, and
thiobarbituric acid (TBA) values. FFAs are formed due to the activity of lipolytic enzymes which
are known for their stability during low temperature preservation.[2830] Peroxides formation is the
indication of early oxidation process and formation of TBA reactive substances indicates the sever-
ity of oxidative deterioration of lipids.[25] In the present investigation, lipid freshness was evaluated
by measuring PV and FFA and results are given in Fig. 1b. PV and FFA were observed to increase
significantly (P < 0.05) from 13.03 to 41.25 mEq of O2 /kg of fat and 3.67 to 13.16% of total lipids
as oleic acid, respectively. Mince from silver carp has been studied for the changes in the PV and
FFA during frozen storage for the storage period of 180 days and it was reported that the PV and
FFA increased from 16.93 to 145.54 mmol of O2 /kg of fat and from 7.66 to 30.76% of total lipids
as oleic acid, respectively.[8] A significant positive correlation was observed for PV (r = 0.929; P <
0.01) and FFA (r = 0.929; P < 0.01) with storage period. Similarly, a significant positive correla-
tion was observed between FFA and PV (r = 0.937; P < 0.01). Solubility and textural properties
of proteins were reported to be influenced by FFA and PV. The interaction between the oxidized
products of lipids/FFA with protein molecules lead to the formation of protein aggregates.[31] The
interaction occurs mainly with amino acids of fish proteins by binding to reactive sulfhydryl groups
of cysteins, amino groups of lysine, aspartic acid, tyrosin, methionine, and arginine.[8,3234] A sig-
nificant negative correlation for PV (r = 0.574; P < 0.01) and FFA (r = 0.611; P < 0.01) with
texture score of prepared sausage allotted by sensory panelist was observed. Oxidation of lipid
and also the oxidation of protein were implicated in textural deterioration. Lipid derived reactive
oxygen species, such as peroxy radicals, are potential initiators of protein oxidation. Oxidation of
lipids during frozen storage could be due to the release of oxidative enzymes and pro-oxidants from
various ruptured cellular organells. These processes are commonly linked to a decrease in muscle
protein functionality, leading to increasing expressible moisture and weaker protein gels.[35,36] A
significant negative correlation was recorded for the changes in the flavor and odor with PV and
FFA (P < 0.05).

Changes in SSP and Non-Protein Nitrogen

SSP are the extractable muscle proteins in intermediate or high ionic strength buffer constitute about
55 to 60% of total muscle proteins, or 10% of the weight of meat.[37] Solubility of SSP mainly is
affected during frozen storage due to the formation of protein aggregates.[3] Changes in SSP fraction
of P. hamrur during frozen storage for the period of 150 days is given in the Fig. 1c. SSP decreased
904 BHATTA ET AL.

significantly (P < 0.05) from the initial value of 13.47 to 7.58 g/100 g of meat at the end of frozen
storage. The observed decrement of about 44% from the initial solubility may be due to protein
denaturation/aggregation of proteins mediated by the oxidation of lipids and proteins. During frozen
storage, actin and myosin become gradually less extractable in salt solutions forming high molecular
weight protein aggregates.[38] A decrease in SSP after frozen storage has been reported for many
fish species including red hake mince, farmed rainbow trout, tilapia surimi.[3941] A significant
negative correlation was observed for SSP with the storage days (r = 0.860; P < 0.01), PV (r =
0.714; P < 0.01), FFA (r = 0.760; P < 0.01). SSP are responsible for the textural properties like
gel strength of fish paste product, fish sausage. Similar results have been reported for the protein
solubility of mince from Sardinella longiceps, Hypothalmichthys molitrix, Ilex argentinus, Saurida
micropectoralis.[8,33,42,43]
Measurement of NPN content during frozen storage indicates the proteolytic activity of endoge-
nous muscle proteases which bring the hydrolytic changes in major muscle protein fractions.[44]
Changes in NPN content of meat from P. hamrur during the period of study are given in Fig 1d.
NPN content decreased significantly from 140 to 67 mg N/100 g meat (P < 0.05). The results con-
tradict the observation made by Singh and Balange[24] on changes in NPN of pink perch surimi
during frozen storage, where a sharp increment in NPN from 13.63 to 22.35 mg/100 g of surmi for
the storage period of 36 weeks. A significant inverse relationship was observed between the frozen
storage period and NPN content (r = 0.918; P < 0.01). Non-protein nitrogenous (NPN) con-
stituents include free amino acids, nucleotides, and also the smaller molecular weight peptides like
di- and tri-peptides likely to be generated during frozen storage by the action of muscle proteases.
The decreasing trend in NPN content in the present study could be due to the loss of nitrogenous
constituents in the thaw drip or due to degradation of NPN by enzymes. Similar result has been
reported by Binsi et al.[45] in the study conducted on the physicochemical changes in green muscle
during ice storage.

Changes in the Quality of Fish Sausage Prepared from Frozen Stored Priacanthus hamrur
Every month of storage period, meat from frozen stored dressed P.hamrur was picked, minced,
and mixed with the additives, stuffed, and heat processed. The product, called fish sausage was
studied for the quality attributes by evaluating gel strength, EMC, folding test, and organoleptic
characteristics such as appearance, color, texture, odor, and flavor.

Gel Strength
Quality of mince based products is mainly judged by the attribute called gel strength. In the present
investigation, gel strength of prepared sausages was evaluated using Okada gelometer and is given
in Fig. 2a. Gel strength of fish sausage prepared at zero and the fifth month of frozen stored P.
hamrur was 302.89 and 140.17 g.cm, respectively. There was no significant (P > 0.05) difference
in the gel strength of fish sausage prepared from three, four, and five months of frozen stored P.
hamrur. Similarly, there was no significant difference between the gel strength of fish sausage pre-
pared from the frozen stored fish meat for first two months. However, significantly (P < 0.05) a
higher gel strength was recorded for the fish sausage prepared at zero month (frozen and thawed
immediately). Freezing and storage time have remarkable influence on the quality of food products
prepared from thawed foodstuffs.[46] Gelation of myofibrillar protein is influenced by various fac-
tors, like, nature of myosin, source of muscle protein, concentration of protein, pH, ionic strength,
and rate of heating.[47] Protein solubility is the prime requirement for the formation of three dimen-
sional network with the entrapment of water. In the present investigation, SSP was found to decrease
during the storage period. Prabhu et al.[48] reported a significant decrease in the the kamaboko (gel)
 FROZEN STORAGE EFFECT ON SAUSAGE FROM PRIACANTHUS 905

350 25
a

Expressible water
300 b A cd cd d B
b 20
Gelstrength

250 c
b
200 c c 15

(%)
c a
150
10
100
50 5
0 0
0 1 2 3 4 5 6 0 1 2 3 4 5 6
Storage period (months) Storage period (months)

FIGURE 2 Changes in (A) gel strength and (B) expressible moisture content of fish sausage prepared during frozen
storage of dressed P. hamrur. Error bars represent the standard deviation from triplicates. Different small letters
indicate that the results are significantly different (P < 0.05).

forming ability of fish sausage prepared from lesser sardine mince during frozen storage. A sim-
ilar report has been given for mince from sardine[49] and silver carp.[8] It is more preferable to
store the fish as a whole than fish mince as the latter case is established to be more prone to protein
denaturation in turn affect the gelation process. In the present study, a significant negative correlation
was established between the storage period and gel strength (r = 0.935; P < 0.01). Gel forming
ability of different tropical fish species have been reported to decrease during frozen storage.[3]
Decrease in the gel forming ability of fish muscles during frozen storage was associated with the
freeze denaturation of actomyosin via the aggregation of protein chains, leaving them unavailable
for subsequent gel formation during heat processing. This results in an inferior gel network forma-
tion, causing a lower elasticity with poor water holding capacity in the gel matrix. The formation
of protein aggregates via disulfide bond formation and hydrophobic interaction has been reported
to decrease the gel forming ability of fish muscle proteins during frozen storage.[41,43] Gel strength
was also found to have a positive correlation with texture (r = 0.727; P < 0.01) and SSP (r = 0.868;
P < 0.01).

EMC
Measurement of EMC supports the correlation between the protein denaturation and frozen stor-
age period. In the present investigation, EMC increased from 10.74 to 18.80% (Fig. 2b). A similar
result has been reported for the effect of pre-freezing icing duration on frozen Nile perch.[50] The
water holding capacity of tissue (protein) decreases due to changes brought out in micro-structure
of myofibrillar proteins from a continuous filamentous matrix to a globular matrix by freezing and
frozen storage.[51,52] Decreasing water holding capacity resulted in increasing EMC.[53] EMC of fish
sausage prepared from the mince of silver carp during frozen storage has been reported to increase
significantly. In the present investigation, increment in EMC was found to have significant positive
correlation with storage period (r = 0.884; P < 0.01) and also a significant negative correlation
with SSP (r = 0.643; P < 0.01). An inverse relationship between the gel strength and EMC was
recorded. Moisture present inside the gel in the free form can easily be expelled by pressure. Similar
correlation has been reported between the gel strength and expressible water content of gels from
pelagic fatty fish, Sardinella longiceps.[54]

Folding Test
Folding test of surimi gel is used to grade the quality of surimi in the industry. In the present inves-
tigation, folding test revealed that the quality of fish sausage for first two months were of A grade.
Based on the folding test, fish sausage prepared on two to five months of frozen storage graded B.
906 BHATTA ET AL.

TABLE 3
Changes in sensory scores of sausage prepared from frozen bulls eye during frozen storage

Storage period Overall

in months Appearance Color Texture Taste Flavor Odor acceptability

0 8.09 0.60a 8.55 0.46a 7.65 0.89a 8.06 0.26a 8.08 0.51a 8.36 0.49a 8.03 0.40a
1 8.00 0.43a 8.02 0.82a 7.50 1.29ab 8.07 0.73a 7.84 0.85a 7.89 0.78ab 7.94 0.15a
2 8.00 0.81a 7.97 0.6a 7.40 0.89ab 7.87 1.43a 7.75 1.25a 7.37 0.94ab 7.75 0.33a
3 7.70 0.67a 7.60 0.55ab 7.30 0.67ab 7.10 0.89ab 6.80 0.83ab 7.00 1.00ab 7.20 0.26ab
4 7.40 0.54ab 7.40 0.96ab 5.80 0.83bc 6.40 1.14ab 6.36 1.09ab 6.90 0.54ab 6.76 0.69b
5 6.80 0.28b 6.45 0.25b 5.20 0.88c 5.08 1.18b 5.84 1.20b 6.28 1.05b 6.68 0.82b
Presented
values represent the mean standard deviation;
Mean with different letters in superscripts in the same column indicate that the results are significantly different
(P < 0.05).

From the results obtained it can be inferred that the gel forming ability/elasticity of fish sausage
decreases during frozen storage.

Sensory Evaluation
Changes in sensory attributes of fish sausage prepared during every month of frozen storage for a
period of five months are given in Table 3. There was no significant difference (P > 0.05) in the
scores allotted by the panelists for the sensory attributes like appearance, color, flavor, and odor up
to the storage period of four months. At the end of the fifth month all the above attributes scored
significantly low. Texture of fish sausage prepared from P. hamrur was found not to vary for the
first three months of storage period. However, the sensory scores obtained for overall acceptability
revealed that the sausages were liked slightly by panelists at the end of four months. A significant
negative correlation was recorded between the storage days and the sensory attributes.

CONCLUSION

The study was aimed to evaluate the changes in the biochemical properties of P. hamrur during
frozen storage for the period of five months and their influence on the quality of mince based prod-
uct, sausage. Biochemical indices such as TVB-N, TMA-N, FFA, and PV increased significantly.
Extractability of salt soluble muscle proteins and NPN constituents reduced during the period of
frozen storage. An inverse relationship was observed between EMC and gel strength of meat gels
prepared from dressed and frozen stored P. hamrur. Based on the sensory and physical analysis, fish
sausage prepared from P. hamrur had better quality for the period of four months. The gel forming
quality of fish meat decreased during frozen storage and can be attributed to the denaturation of
proteins by the interaction of proteins with lipid oxidized and hydrolyzed compounds.

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