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Fortification of biscuits with fish protein

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World Journal of Dairy & Food Sciences 9 (2): 242-249, 2014
ISSN 1817-308X
© IDOSI Publications, 2014
DOI: 10.5829/idosi.wjdfs.2014.9.2.1142

Fortification of Biscuits with Fish Protein Concentrate

1 G.F. Mohamed, 2 A.M. Sulieman, 1N.G. Soliman and 2S.S. Bassiuny

1Department of Food Technology, National Research Centre, Dokki, Giza, Egypt

2Department of Food Science, Zagazig University, Zagazig, Egypt

Abstract: This study was aimed to evaluate the impact of the fortification of biscuits with carp fish protein
concentrate (CFPC) and shark fish protein concentrate (SFPC). The results show that the protein and total ash
content increases by increasing the fortification level of both types of fish protein concentrate compared to the
control sample. The crude fiber and carbohydrate of biscuit samples decreased by increasing the fortification
levels compared to the control sample. Total Essential amino acids of biscuits were 2.99, 3.39 and 3.77% for
biscuit control; biscuits fortified with 3% CFPC and 3% SFPC respectively. Besides, the corresponding value of
Non-Essential amino acids were 4.22, 4.93 and 6.24 for biscuit control, biscuits fortified with 3% CFPC and 3%
SFPC respectively. Acid value (AV), Peroxide value (PV) and Thiobarbituric acid number (TBA) increased
gradually up to the end of the storage time in all samples. Biscuits prepared by fortification with CFPC and SFPC
had a lower AV than control sample. From the obtained judging results, it could be observed that the fortification
of CFPC and SFPC at levels 1, 2 and 3% into wheat flour blend used in biscuit making didn't cause any
significant (at p 0.05) deleterious effect on overall acceptability of produced biscuits which exhibited the
hedonic scale of very good or excellent judging scores (7.5-9.5) and better acceptability.

Key words: Fish protein concentrate Biscuits quality Sensory quality Nutritional quality

INTRODUCTION tonnes) in 2012 [2]. Mean while, the carp species are
generally low in the food chain and have not been utilized
Fish is one of the most important sources of animal widely due to a lot of inter- muscular bones [3]. The
protein in the range of 17-20% and other elements for the protein content of sharks forms approximately 22% of the
maintenance of a healthy body [1]. Farmed common carp flesh. Shark protein can be considered as a good source of
production was nearly 14 percent of the total global essential amino acids and can serve as a cheap substitute to
freshwater aquaculture production in 2002 (33 138 962 fulfill several amino acid deficiencies in protein-poor
tonnes). Common carp production increased by an average diets. In sharks, the largest fraction of this NPN consists of
global rate of 9.5 percent/yr between 1985 (681 319 urea, which occurs not only in the liver (as in land
tonnes) and 2002. In the past decade (1993-2002) this has animals), but all over the body [4, 5]. Fish Protein
increased to 10.4 percent/yr. This is greater than the Concentrate (FPC) has been defined as the product
expansion rate of farmed grass carp (10.1 percent/yr), resulting by removing of water and oil from fish, thus
silver carp (8.8 percent/yr) and bighead carp production increasing the concentration of the protein, that it is an
(7.2 percent/yr), but less than that for tilapias (11.8 excellent source of highly digestible amino acids and other
percent/yr) during this decade. In Europe, common carp nutrient materials. The quality of the materials used to
production was 144 602 tonnes in 2002. This represents a prepare FPC will influence the quality of the produced
substantial reduction from peak production of over 402 material [6, 7]. Bakery products in the Middle East
000 tonnes in 1990, caused by changes in Eastern Europe. especially in the Arab countries consider an important
However, European production appears to be gradually source of nutrients i.e. carbohydrate, protein, minerals and
increasing again; the 1993-2002 trough was 125 274 vitamins. Most bakeries products can easily be enriched
tonnes in 1997. Common carp production increased from and fortified with proteins, vitamins and minerals to meet
(3 228 240 tonnes) in 2009 to (3 791 912 specific needs of the target groups of the

Corresponding Author: Gamal Fouad Mohamed, Food Technology Department,

National Research Centre, Dokki, Giza, Egypt.
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 World J. Dairy & Food Sci., 9 (2): 242-249, 2014

population [8]. Good eating quality makes biscuits

attractive for protein fortification and other nutritional
improvements. Development of fortified biscuits or other
composite flour bakery products is the latest trend in
bakery industry. In the same time, most of the effort
directed towards improving the diets of people is centered
on the improvement of the protein content and quality of
cereals by the addition of protein-rich foods, protein
concentrates and/or essentials amino acids. Also, it was
found that 6% FPC is the best level for supplementation of
biscuits [9] and 10% FPC increased nutritional value and
percent of satisfaction values for some common foods
[10]. The overall acceptability of salt biscuits was
improved by the supplementation with 5% fish protein
concentrate [11]. The present study was carried out to
determine the possibility of producing high nutritional
biscuits enriched with fish protein concentrate and the
effect of storage on the quality of product.

MATERIALS AND METHODS

Biscuits ingredients (wheat flour (72% extract) - salt

– sugar – shortening fat – vanilla – baking powder – eggs
– milk) were purchased from local market, Cairo, Egypt.
Fish protein concentrate was prepared from carp and shark Fig. 1: Flow diagram of Marine beef (fish protein
fish at the laboratory, National Research Center (NRC), concentrate) production.
Dokki, Giza, Egypt as the following in Fig. 1.
homogenized with a hand blender (Braun, Kronberg,
Dough Characteristics: Wheat flour of 72% extraction Germany) for a few seconds. Then, add sugar and
was well mixed with CFPC and SFPC to produce shortening and mixed for 20 sec. at speed 3, half of the
individual mixtures containing 0, 1.0, 2.0 and 3.0% flour and all other ingredients were then included and
fortification levels to manufacture of biscuits (on flour mixed for 20 sec. The remaining flour was added and
weight basis). The rheological properties of dough was mixed for 140 sec to give a total mixing time of 3.0 min.
determined by Farinograph (Model Type No: 81010 (31, following a rest time of 20 min, then the dough was
50 and 63 rpm), Brabender, OHG, Duisburg, 1979, sheeted for 5 mm thickening with a diameter of 50 mm.
Germany) in department of food technology National Biscuits formed manually and baked at 180°C for 20 min.
Research Center (NRC) according to the standard methods in a thermostatically controlled oven with air fan (Shel
(AACC) [12]. Parameters measured were water Lab 1370 FX, USA). The biscuits cooled at room
absorption, dough development time, dough stability, temperature for 75.0 min.
weakening and mixing tolerance index (MTI).
Physical Characteristics of Biscuits
Preparation and Evaluation of Biscuits: Biscuits Baking Quality of Biscuits: Diameter (w) was measured by
preparation was carried according to the method described Boclase (HL 474938, STECO, Germany). Also, volume
by American Association of Cereal Chemists (AACC) (v) and thickness (T) of biscuits were determined
[12]. The ingredients were weighed at the formula: 100 g according to standard methods. The spread ratio W/T was
of wheat flour of 72% extraction and CFPC and SFPC calculated. Percent spread ratio was calculated according
fortification at levels (0, 1.0, 2.0 and 3.0%); 50.0 g of fine to standard methods described by American Association of
sugar; 1.0 egg ( 50.0 g); 28.0 g of shortening; 0.93 g of Cereal Chemists (AACC) [12] by dividing the average
salt; 1.11 g of sodium bicarbonate; 1.0 g of vanilla and 15 value of diameter (w) by the average value of thickness
ml whole milk. The eggs were initially (T) of biscuits.

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 World J. Dairy & Food Sci., 9 (2): 242-249, 2014

Color Analysis: The color was measured by using a Statistical Analysis: The obtained results were
Hunter Lab. Model D25 color and difference Meter [13]. statistically analyzed using SPSS statistical package
This color assessment system is based on the Hunter L*, (Version 9.05) according to McClave and Benson [19]
a* and b* coordinates. L* representing lightness and analysis of variance (ANOVA), Duncan’s multiple range
darkness, + a redness, + b yellowness and – b blueness test and least significant difference (LSD) was chosen to
with white Tile of Hunter Lab color standard: (L=92.56, determine the significant difference among various
a=-0.87 and b= -0.15). treatments. Differences considered significant at p<0.05.

Major Chemical Composition: Moisture content, crude RESULTS AND DISCUSSION

protein, crude fiber, fat content and total ash were
determined according to the method described by AOAC Fish protein concentrate (FPC) is a low cost animal
[14]. Total carbohydrates were calculated by difference. protein with high quality, so it can be used as a protein
supplement to increase nutritive value of foods [20, 21].
Amino Acids Composition: Amino acids composition Table 1 shows the chemical composition of wheat flour
was determined with an automatic amino acid analyzer and carp fish protein concentrate and shark fish protein
(LKB 4151 plus, Biochrom Ltd., Cambridge, UK) concentrate. From data presented in Table 1 it's clear that,
according to Bidlingmeyer et al. [15]. The degree of the contents of moisture, protein, fat content, crude fiber
variability in different groups of amino acids (according to and total ash were 11.2%, 12.00%, 1.56%, 1.02% and
type of side chain) was expressed as percentages of total 0.86% in wheat flour, respectively. Similar results were
amino acids in each species and the ratio of essential obtained by Yassen et al. [22], who reported that the
amino acid was determined. chemical composition (% on dry weight basis) of wheat
flour (72%) were moisture 11.59%, protein 10.86%, fat
Lipid Autoxidation: The acid value (AV), peroxide value 1.56% and ash 0.60%. Also, the results showed that the
(PV) and thiobarbituric acid value (TBA) were the carp fish protein concentrate contained 9.10% moisture,
parameters used for the assessment of lipid auto-oxidation. 88.70% protein, 0.04% fat and 2.10% ash and the shark
These three parameters were determined in the oil fish protein concentrate contained 8.55% moisture,
extracted from the samples of grounded biscuits using n- 89.12% protein, 0.01% fat and 2.20% ash content. These
hexane [16]. The extracted oil was kept in a tightly closed values were in agreement with Bolly [23], who reported
dark bottle in a deep freezer at (–20°C) for subsequent that the moisture content of (fish protein concentrate)
analysis. The acid and peroxide values were determined marine beef in wet base is < 10% for good quality marine
according to the methods of AOAC [14]. Thiobarbituric beef, protein content of marine beef was 60-89%, fat
acid value was determined as described by Kirk and content was 0-3.71% and ash content was 2-3.5%. From
Sawyer [17]. Stability of biscuit lipids were followed these results, it could be concluded that the fish protein
periodically at intervals of 1 month during storage for 3 concentrate (FPC) are high protein content by removing
months at ambient temperature. maximum moisture and fat from fish and much lower in
fat by removing with ethanol or isopropanol.
Sensory Evaluation: Organoleptic properties of the
biscuits were evaluated in National Research Center Rheological Properties of Dough Samples as Affected
(NRC) department of food technology by 10 panelists by Fortification FPC: Rheological properties of dough
according to Ogunjobi and Ogunwolu [18]. Five coded can be used as quality indicators for cereal products [24].
samples of biscuit were presented to each of the panelist This fact has contributed to use rheological testing for
and they were asked to assess the biscuits for color, following the changes in dough systems and mechanical
flavour, taste, texture, appearance and overall acceptability properties. Farinograph parameters such as water
using 10-point hedonic scale (0-2 = Dislike very much, 3- absorption, arrival time, dough development time, dough
4.9 = Dislike moderately, 5-5.9 = Dislike slightly, 6-6.9 = stability, weakening and mixing tolerance index were
Neither like nor dislike, 7-7.9 = Like slightly, 8-8.9 = Like different between control and blends congaing CFPC or
moderately, 9-9.9 = Like, 10 = Like very much) for the SFPC at different levels are showed in Table 2. The water
assessment. absorption was found in the range of 63.0% to 64.5% and

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 World J. Dairy & Food Sci., 9 (2): 242-249, 2014

Table 1: Chemical composition of wheat flour, carp fish protein concentrate (CFPC) and shark fish protein concentrate (SFPC)
Samples
---------------------------------------------------------------------------------------------------------------------
Compounds* Wheat flour CFPC SFPC
Moisture content (%) 11.20 9.10 8.55
Crude protein (N% x 6.25) 12.00 88.70 89.12
Crude fat (%) 1.56 0.04 0.01
Ash content (%) 0.86 2.10 2.20
*On wet weight basis

Table 2: Effect of fortification of wheat flour with FPC on rheological properties of dough by farinograph
Carp fish protein concentrate (CFPC) Shark fish protein concentrate (SFPC)
-------------------------------------------------- ----------------------------------------------------
Parameters Control 1% 2% 3% 1% 2% 3%
Water absorption (%) 63.5 63.0 63.5 64.5 64.0 64.5 65.0
Arrival time (min) 1.2 1.3 0.5 1.6 1.2 2.2 0.6
Dough development time (min) 1.75 2.00 1.50 2.00 1.75 3.50 1.20
Dough stability (min) 4.7 5.9 10.5 6.9 3.0 5.0 3.9
Weakening (BU) 55 40 20 30 110 30 30
Mixing tolerance index (BU) 40 30 10 20 60 30 20
Where: BU=Brabender units.

Table 3: Chemical composition of biscuit fortified with different levels of FPC

Carp fish protein concentrate (CFPC) Shark fish protein concentrate (SFPC)
--------------------------------------------------------- ---------------------------------------------------------------
Chemical composition of samples Control 1% 2% 3% 1% 2% 3%
Moisture 3.56±0.12a 3.68±0.05b 3.80 ±0.03a 4.00 ±0.01b 3.90±0.14a 4.20 ±0.02b 4.26 ±0.01a
Protein 9.51±0.03b 10.22±0.08a 11.10±0.01b 12.00±0.05b 10.24±0.11a 11.21±0.06a 12.11±0.03b
Crude fiber 0.60±0.03b 0.59±0.01a 0.58±0.14a 0.57±0.05b 0.59±0.03b 0.58±0.04a 0.57±0.14a
Total ash 1.80±0.08a 1.80±0.03b 1.84±0.01a 1.88±0.02b 1.82±0.11b 1.86±0.06a 1.88±0.05b
Fat content 21.30±0.06a 21.31±0.11a 21.32±0.03b 21.33±0.08a 21.30±0.01b 21.31±0.04a 21.32±0.08a
Carbohydrate* 63.23±0.11a 62.40±0.08a 60.36±0.03a 58.22±0.05b 62.15±0.02b 59.84±0.06a 58.86±0.04a
The values are mean ± S.D of three independent determinations. The values with different superscripts in a column differ significantly (p 0.05). * Calculated
by difference method

Table 4: color quality of biscuits from different formulas

Color characteristics value
----------------------------------------------------------------------------------------------------------------------------------------------------------
Upper surface of biscuit Bottom surface of biscuit
------------------------------------------------------------------------- -------------------------------------------------------------------------
Fortification treatment L* a* b* L* a* b*
Control 60.52±0.05 de 11.25±0.06a 25.52±0.14b 65.41±0.03a 10.52±0.11c 26.00±0.14abc
1% SFPC 58.82±0.08d 12.68±0.14a 22.10±0.05 a 65.71±0.08bcd 11.01±0.03ab 25.72±0.11cd
2% SFPC 57.50±0.02a 12.99±0.16 c 25.14±0.06a 66.21±0.11bc 11.54±0.02a 26.24±0.03ab
3% SFPC 56.73±0.07c 13.32±0.07a 25.74±0.08a 67.45±0.11bc 11.80±0.05 a 26.99±0.07bcd
1% CFPC 58.71±0.11c 12.39±0.03a 21.93±0.11c 65.58±0.06cd 10.88±0.07b 25.58±0.09d
2% CFPC 57.32±0.08c 12.75±0.09c 22.99±0.06a 66.05±0.02c 11.32±0.08a 25.02±0.12c
3% CFPC 56.59±0.02a 13.14±0.11c 25.56±0.08a 67.36±0.05 c 11.65±0.06a 26.71±0.16c
*Means with the same superscripts at the same column are not significant at (p<0.05), L+ = lightness (100 = white; 0 = black), a+ = redness to a- = green; b+
= yellowness to b- =blue.

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 World J. Dairy & Food Sci., 9 (2): 242-249, 2014

Table 5: Amino acids composition (%) of biscuit control and fortified with 3% of FPC
Essential amino acids Biscuit control 3% CFPC 3% SFPC Nonessential amino acids Biscuit control 3% CFPC 3% SFPC
Lysine 0.21 0.31 0.39 Aspartic 0.45 0.58 0.68
Therionine 0.23 0.29 0.34 Glutamic 1.84 2.02 2.09
Valine 0.40 0.41 0.50 Arginine 0.31 0.40 0.59
Methionine 0.14 0.15 0.19 Serine 0.38 0.43 0.51
Isolucine 0.29 0.31 0.38 Proline 0.71 0.76 1.02
Leucine 0.55 0.61 0.69 Glycine 0.26 0.38 0.79
Phenylalanine 0.51 0.56 0.56 Alanine 0.27 0.36 0.56
Tyrosine 0.28 0.32 0.32 -- -- -- --
Histidine 0.20 0.22 0.24 -- -- -- --
Cystine 0.18 0.21 0.16 -- -- -- --
Total EAAs 2.99 3.39 3.77 Total NAAs 4.22 4.93 6.24
Total AAs -- -- -- -- 7.21 8.32 10.01
EAAs: Essential Amino acids, NAAs: Non- Essential Amino acids, AAs: Amino acids

64.0% to 65.0% in blends containing CFPC and blends Amino Acids Composition (%) of Biscuit Fortified with
containing SFPC. However, the blends containing SFPC 3% of FPC: Table 5 shows the amino acids compositions
showed higher water absorption due to more protein. (%) for biscuits (control) and fortified with 3% of CFPC
Compared to the control, water absorption was increased and SFPC. Total Essential amino acids were 2.99, 3.39
by the addition of CFPC or SFPC. Similarly, the dough and 3.77%, respectively. Besides, the corresponding value
development time was higher in all blends compared to of non-essential amino acids were 4.22, 4.93 and 6.24,
control. The addition of CFPC or SFPC increased the respectively. From the obtained results, it could be
stability of dough blends compared to the controls at levels illustrated that the CFPC and SFPC composed of all
1% and 2%. While weakening and mixing tolerance index essential amino acids which were higher than control.
decreased by addition CFPC or SFPC at different levels. Therefore, FPC had a good nutritional protein quality.
These results are in agreement with those reported by These results are nearly in agreement with those obtained
Abou-Zaid and Elbandy [25]. by Ibrahim [26].

Chemical Composition of BiscuitFortified with Chemical Quality Attributes: The least stable macro-
components in food are the lipids. The biscuits become
Different Levels of FPC: The chemical composition for
unacceptable and are rejected by the panelists as a result of
different samples of biscuits fortified with CFPC and
rancidity development. To prolong the shelf-life of such
SFPC at levels 0, 1, 2 and 3% are showed in Table 3. It
foods and prevent the occurrence of rancidity, the presence
was observed that, the protein and total ash content
of antioxidants is required. The changes in fat quality
increases by increasing the fortification level of both types
parameters, i.e. AV, PV and TBA of the biscuits fortified
of FPC compared to the control sample. On the other hand,
with SFPC and CFPC were followed throughout the
crude fiber and carbohydrate of biscuit samples decreased storage period for 3 months at room temperature and the
by increasing the fortification levels compared to the results obtained in Table 6. From the table, it could be
control sample. These results are in agreement with those noticed that AV, PV and TBA increased gradually up to
obtained by Ibrahim [26]. the end of the storage time in all samples. Biscuits fortified
with SFPC and CFPC had a lower AV than control.
Physical Characteristics of Biscuits Similar findings were reported by Jeyasanta et al. [27]
Color Values of Biscuits: The data presented in Table 4 since they showed that peroxide values were within the
shows the color attributes of the biscuits. In regard to the acceptable limit of 10-20 mEq/kg of fat throughout the
surface color, it was cleared that the addition of CFPC and storage period. The Thiobarbituric acid value also was
SFPC decreased the color characteristics as CFPC and well with in the acceptable limit which was less than 3 mg
SFPC levels increased. The lightness (L*) value of crust malonaldehyde kg-1 of sample which indicates good
surface decreased, but the redness (a*) value increased. quality fishery products. This indicates that the level of
These results also showed that, the highest increasing in quality indicators such as hydrolysis of lipid (FFA),
redness (a*) and yellowness (b*) found in biscuit samples primary and secondary lipid oxidation does not increase
fortified with FPC at levels 2 and 3% from the control. during storage. This is because there was low fat content
These results were in agreement with those reported by in the sample and also there was increase in extraction of
Abou-Zaid and Elbandy [25]. total lipids at the time of processing.

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 World J. Dairy & Food Sci., 9 (2): 242-249, 2014

Table 6: Effect of storage period on the chemical attributes of the stored biscuit
Time of storage (month)
----------------------------------------------------------------------------------------------------------------------------- ----------------------------------------------
0 1 2 3
------------------------------------- ------------------------------------ --------------------------------- --------------------------------------
Treatments AV PV TBA AV PV TBA AV PV TBA AV PV TBA
Control 0.62 3.20 0.010 0.92 5.20 0.021 1.15 7.5 0.030 1.46 9.36 0.092
1% SFPC 0.63 3.22 0.011 0.93 7.00 0.027 0.96 7.75 0.031 1.11 9.21 0.090
2% SFPC 0.64 3.24 0.012 0.94 7.30 0.028 0.94 7.66 0.032 1.10 9.21 0.092
3% SFPC 0.65 3.26 0.014 0.92 7.40 0.029 0.93 7.50 0.034 1.12 9.00 0.095
1% CFPC 0.64 3.23 0.013 0.92 7.10 0.028 1.11 8.23 0.032 1.30 9.40 0.092
2% CFPC 0.65 3.25 0.015 0.93 7.20 0.029 1.07 8.25 0.035 1.33 9.36 0.094
3% CFPC 0.67 3.27 0.017 0.94 7.30 0.030 1.11 8.21 0.038 1.29 9.33 0.096
*AV: acid value; PV: peroxide value; TBA: thiobarbituric acid.

Table 7: Sensory characteristics of produced biscuits as affected by the fortification of SFPC and CFPC
Samples Appearance (10) Color (10) Texture (10) Taste (10) Flavor (10) Overall acceptability (10)
Control 9.0±0.63a 9.5±0.57a 10.0±0.95a 10.0±0.85a 9.0±0.58a 9.6±0.77a
1% SFPC 8.5±1.25a 9.5±1.60a 8.6±0.71b 8.5±0.70b 9.0±0.95a 9.3±0.79a
2% SFPC 8.0±0.76b 9.5±0.95a 7.5±0.63c 7.0±1.22c 8.5±0.67b 9.4±0.92a
3% SFPC 7.0±0.88c 9.3±1.00a 7.5±0.67d 8.5±1.52d 8.0±0.79c 9.2±1.30a
1% CFPC 9.0±0.60a 9.4±0.74a 10.0±0.75a 9.0±1.06a 9.0±0.75a 9.5±0.70a
2% CFPC 8.8±1.14a 9.5±0.96a 7.5±1.52c 7.0±1.06c 9.0±0.84a 9.4±1.03a
3% CFPC 8.5±1.23a 9.2±0.96a 6.5±0.21d 6.0±1.03cd 8.4±0.67b 9.4±1.34a
L.S.D 0.05* 0.45 0.51 Ns Ns 0.6 Ns**
*Means with the same superscripts at the same column are significant at (p<0.05), **Ns: not significant

Sensory Characteristics of Produced Biscuits as

Affected by the Fortification of SFPC and CFPC: The
effect of fortification of SFPC and CFPC at different
levels (0, 1, 2 and 3%) on the judging scores of sensory
quality characteristics; flavor, color, taste, appearance,
texture and overall acceptability of produced biscuit was
studied and the obtained results are shown in Table 7 and
Fig. 2. From the obtained judging results, it could be
observed that the fortification of SFPC and CFPC at levels
1, 2 and 3% into wheat flour blend used in biscuit making
didn't cause any significant (at p 0.05) effect on overall
acceptability of produced biscuits which exhibited the
hedonic scale of very good or excellent judging scores
(7.5-9.5) and better acceptability. These results are in
agreement with those reported by Abou-Zaid and Elbandy
[25].

CONCLUSION

In conclusion the present work provides that raw carp

and shark fish are generally considered as a good source to
produce FPC. The FPC made from carp and shark fish had
high nutritional value and it was more effective for
fortification biscuits whereas, it maximizing the amount of
Fig. 2: Biscuits fortification with carp and shark fish protein content. Moreover, the level used (1, 2 and 3%)
protein concentrate. didn't affect on the appearance and other

247
 World J. Dairy & Food Sci., 9 (2): 242-249, 2014

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