Effect of lime and Grape fruit extract as coagulants on chemical composition of Sudanese white soft cheese during storage

Abstract

This study was conducted to determine the effects o coagulant type on the chemical composition of Sudanese white cheese. Three batches of lime acid cheese and grapefruit acid cheese were made. Fresh raw cow's milk (20 liters) was heated to 82ºC for 30 minutes, then salt was added at a rate of 4% and lime juice solution pH 3.2 (50% lime juice+ 50% distilled water) was added at the rate of 5%, and grapefruit juice pH 3.7 was added at a rate of 10%, then the mixture was left until complete coagulation occurred the curd pressed over night(700 gm /pressing), then the cheeses were packed into polyethylene bags 100 gm in each, and stored at 5ºC for a month. Chemical analysis was carried out for the cheese samples at day1, day 7, day 14, day 21 and day30. The results of statistical analysis showed that the fat content of the lime cheese and grapefruit cheese samples stored in refrigerator were high, while the fat contents of the same cheese stored in whey at room temperature were low. The protein content of lime cheese stored in refrigerator (19.45±3.06%), and whey at room temperature (16.28±1.29%) was higher than that of grapefruit cheese (17.03±2.83). Total solids (TS) content of the lime cheese samples stored in whey at room temperature increased significantly (P≤0.001) from 45.51±1.31% at week one to 66.94±1.48% at week four. Ash content of lime cheese samples stored in whey at room temperature (1.10±0.26%) was significantly (P≤0.001) higher than those of lime cheese stored in the refrigerator (0.05±0.01%). Acidity of lime cheese stored in whey increased significantly (P≤0.001). The acidity of lime cheese stored in refrigerator increased from 0.20±0.00% at week one to 0.30±0.00% at week four. Its concluded that the storage period has significant effect on the chemical composition of white cheese made with lime and grape fruit extracts also the method of preservation had clear effect on the chemical composition of the white cheese.

Full text

Int. J. Adv. Res. Biol.Sci. 1(7): (2014): 123–130
123
International Journal of Advanced Research in Biological Sciences
ISSN : 2348-8069
www.ijarbs.com
Research Article
Effect of lime and Grape fruit extract as coagulants on chemical composition of
Sudanese white soft cheese during storage
HashimAhmedElhaseen 1and Omer Ibrahim Ahmed Hamid 2and Osman Ali Osman El Owni3
1,2Sudan University of Science and Technology, College of Animal Production Science and Technology, Department
of Dairy Science and Technology
3Khartoum University, Faculty of Animal production, Department of Dairy Production
*Corresponding author: omerhi30@sustech.edu
Abstract
This study was conducted to determine the effects o coagulant type on the chemical composition of Sudanese white cheese. Thre e
batches of lime acid cheese and grapefruit acid cheese were made. Fresh raw cow’s milk (20 liters) was heated to 82ºC for 30
minutes, then salt was added at a rate of 4% and lime juice solution pH 3.2 (50% lime juice+ 50% distilled water) was added at
the rate of 5%, and grapefruit juice pH 3.7 was added at a rate of 10%, then the mixture was left until complete coagulation
occurred the curd pressed over night(700 gm /pressing), then the cheeses were packed into polyethylene bags 100 gm in each, and
stored at 5ºC for a month. Chemical analysis was carried out for the cheese samples at day1, day 7, day 14, day 21 and day30.
The results of statistical analysis showed that the fat content of the lime cheese and grapefruit cheese samples stored in
refrigerator were high, while the fat contents of the same cheese stored in whey at room temperature were low. The protein
content of lime cheese stored in refrigerator (19.45±3.06%), and whey at room temperature (16.28±1.29%) was higher than that
of grapefruit cheese (17.03±2.83). Total solids (TS) content of the lime cheese samples stored in whey at room temperature
increased significantly (P≤0.001) from 45.51±1.31% at week one to 66.94±1.48% at week four. Ash content of lime cheese
samples stored in whey at room temperature (1.10±0.26%) was significantly (P≤0.001) higher than those of lime cheese stored in
the refrigerator (0.05±0.01%). Acidity of lime cheese stored in whey increased significantly (P≤0.001) . The acidity of lime
cheese stored in refrigerator increased from 0.20±0.00% at week one to 0.30±0.00% at week four. Its concluded that the storag e
period has significant effect on the chemical composition of white cheese made with lime and grape fruit extracts also the method
of preservation had clear effect on the chemical composition of the white cheese.
Keywords: cheese, coagulant, lime, grapefruit, chemical composition
Introduction
Sudanese white soft cheese (Gibna Beyda) is the most
common cheese in Sudan. It has a strong odor and
taste. It is made from raw or pasteurized whole milk,
skim milk or reconstituted milk depending on natural
lactic acid bacteria; no starter is used and coagulated
by rennet enzyme. It salted by adding odium chloride
directly to milk (Abdel Razig, 1996).
Cheese plays an important role in the Sudanese diet,
and many people eat a certain amount of cheese with
at least twice per week in one of their meals, most of
the cheese is consumed either directly or with bread
(Dhoul and Hamid, 2014). Warsama et.al (2006)
reported that Sudanese white soft cheese contained
47.8% total solids, 14.0% fat, 15.9 % protein and 6.2%
ash, and it is locally known in Sudan as (Gibna Bayda)
or Gibbna which is the most famous name and it is
usually stored in containers filled with whey . Natural
cheese should be stored at suitable temperatures to
ensure good quality because a high temperature leads
to evaporation of moisture and growth of unwanted
bacteria and other faults ( Ramakant ,2006).

Int. J. Adv. Res. Biol.Sci. 1(7): (2014): 123–130
124
Gibna Beyda (Sudanese white cheese), like most other
cheeses, is coagulated using rennet as a coagulating
agent. Other types of cheese may also be made using a
combination of heat and acid as coagulating agents. A
type of cheese made by this process is called Queso
Blanco; it is an important domestic white cheese in all
parts of Latin America. Covacevich (1981) Found that
cottage cheese produced by direct chemical
acidification was too low in moisture content and
insufficiently firm in texture unless the direct chemical
acidification was preceded by fermentation of the milk
with starter.
The acid coagulation, which is reached by lactic acid
bacteria, which convert the lactose of the milk into
lactic acid resulting in decrease of the pH of the row
milk from 6.7 to 4.6 and at pH 4.6, the lactic acid
effects the casein which starts coagulating. According
to Abdalla et al., (2001) the factors affecting acid
coagulation are related to dissolve calcium salts,
acidity, temperature, quantity of rennet and the heat
treatment.
The objectives of the research are to study the effect of
direct acidifications using natural acidifiers of
different citrus fruit juices extracted from lemon,
orange and grapefruit on the chemical and
microbiological properties of white cheese.
Materials and Methods
Materials
Three batches of lime acid cheese and grapefruit acid
cheese were made.
The experimental procedures were done in the Dairy
Processing unit of the Department of Dairy
Production. University of Khartoum, during the period
from September 2010 –December 2010.
Source of milk
Fresh raw whole-cow’s milk (20 liters) was obtained
from the farm of College of Animal Production
Science and Technology-Sudan University.
Sources of Lime and Grapefruit
Lime (Citrus aurantifolia) of 3.2 pH and Grapefruit
(Citrus paradisi) of 3.7 pH were purchased from
Khartoum Fruit central Market and then juices were
prepared by mechanical pressing of two dozen of
grapefruit and three dozen of lime .
Graduated measuring cylinder:
This was used to measure the required lime and grape
juices in ml.
Cloth filter:
This cloth filter was used to filter lime and grape
juices and salt.
Salt:
White fine table salt was purchased from local market.
Cheese manufacture:
Fresh raw cow’s milk (20 liters) was heated to 82ºC
for 30 minutes, then salt was added at a rate of 4% and
lime juice solution pH 3.2 (50% lime juice+ 50%
distilled water) was added at the rate of 5%, and
grapefruit juice pH 3.7 was added at a rate of 10%,
then the mixture was left until complete coagulation
occurred (about 15 seconds), then the curd of each
treatment was transferred to a mould lined with cheese
cloth and pressed over night(700 gm /pressing) , it was
removed from the mould and cut into square cubes of
100 gm each, then the cheeses were packed into
polyethylene bags 100 gm in each, and stored at 5ºC
for a month. Chemical analysis was done for the
cheese samples at day 1, 7, 14, 21 and 30 intervals.
Chemical analysis
Titratable acidity, Total solids content, protein
contents, ash and fat contents were determined
according to AOAC (1990)..
Statistical analysis
The SPSS program, version 10 was used. General
linear models were used to estimate the effect of
storage periods on the chemical composition of white
cheese. Least Significance Difference (LSD) was used
for mean separation between the treatments.
Results
Cheese yield
The average yield of acid white soft cheese was
affected by the kind of acidulant. The average yield of

Int. J. Adv. Res. Biol.Sci. 1(7): (2014): 123–130
125
cheese made by lime juice (15%) was lower than the
average yield of grapefruit cheese made by grapefruit
juice (17%). This might be due to the high volume
used in case of using grapefruit juice.
Effect of coagulant type and preservation method
on chemical composition of white cheese:
Table (1) shows the main effects of preservation
method on chemical composition of acid cheeses. Fat,
protein, total solids, ash and titratable acidity content
of lime cheese.
Fat content
The fat content of lime cheese stored in the
refrigerator (15.25±4.14%) showed highly significant
value during storage (Table 1) Similarly the fat
content of lime cheese stored in whey at room
temperature (14.58± 3.68%), showed high
significantly value (P≤0.001), moreover the fat
content of grapefruit cheese stored in refrigerator
(11.67±3.92%) revealed highly significant value than
that of lime cheese stored in whey at room temperature
and refrigerator for different storage periods showed
no significant difference compared to that of grapefruit
cheese stored in whey at room temperature and
refrigerator (Table 1).
Protein content
The protein content of lime cheese (Table 1) stored in
refrigerator (19.45±3.06%), and whey at room
temperature (16.28±1.29%) was higher than the
protein content of grapefruit cheese (17.03±2.83,
13.76±3.15 respectively). Protein content of lime
cheese samples stored in whey at room temperature
decreased significantly (P≤0.001); from 18.13 ±
0.91% at week one to 16.30± 0.35% at week four
(Table 3). However the protein content of the same
cheese stored in the refrigerator increased
to19.40±0.30% at week one to 24.20±0.69% at week
four. It was also decreased significantly (P≤0.001) in
grapefruit cheese stored in whey at room temperature
from 17.07±0.32% at week one to 16.30±0.35% at
week four. Those samples kept in the refrigerator
increased from 15.97±0.23% at week one to 21.67±
0.58% at week four (Table 3). The protein content of
grapefruit cheese stored for one week in the
refrigerator decreased from 15.97 ± 0.23 % to 15.00
±0.00% at week three, compared with week four 21.67
±0.58 %. The protein content of lime cheese
17.07±0.32 preserved in whey for one week decreased
significantly (P≤0.001) to 10.10 ± 0.00 % in week
two, however it increased from 11.57± 1.03 % at week
three to 16.30 ± 0.35 % at week four (Table 3) .
Total solids content
Total solids content of lime cheese (53.55 )
stored in whey at room temperature were higher than
the total solids of grapefruit cheese (36.85 )
stored in whey. However the total solids of grapefruit
cheese stored in refrigerator showed higher values
(Table 1).
Total solids (TS) content of the lime cheese samples
stored in whey at room temperature increased
significantly (P≤0.001) from 45.51±1.31% at week
one to 66.94±1.48% at week four. Then it was also
increased significantly (P≤0.001) in lime cheese
preserved in the refrigerator from 42.77±1.90% at
week one to 43.79±1.64% at week four. However the
TS decreased significantly (P≤0.001) in grapefruit
cheese stored in whey at room temperature
48.01±1.29% at week one compared to week four
30.63±0.87%. Similarly the TS decreased significantly
(P≤0.001) in cheese stored in the refrigerator from
55.12±0.72% at week one to 30.63±0.87% at week
four (Table 4).
Ash content
Ash content of lime cheese stored in whey at room
temperature was , while ash content of
grapefruit cheese stored in refrigerator was
0.42 (Table 1).
Table (5) shows the effect of preservation method and
storage period on ash content of white cheese made by
lime and grapefruit juices. Ash content of lime cheese
samples stored in whey at room temperature
(1.10±0.26%) was significantly (P≤0.001) higher than
those of lime cheese stored in the refrigerator
(0.05±0.01%). Also the ash content of grapefruit
cheese stored in the refrigerator (1.20±0.10) was
significantly (p≤0.001) higher than those of cheese
stored in whey at room temperature (0.47±0.55).

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Acidity
Acidity of lime cheese stored in whey at room
temperature and refrigerator were % and
0.25± 0.05 % respectively, the acidity content of
grapefruit cheese stored in whey at room temperature
and refrigerator were 0.89± 0.19 % and 0.41± 0.33 %
respectively. (Table 1).
Table (6) showed the effect of preservation method
and storage period (week) on acidity level of white
cheese made by lime and grapefruit juices. Acidity of
lime cheese stored in whey increased significantly
(P≤0.001) from 0.80±0.00 % at week one to
1.47±0.12% at week four. The acidity of lime cheese
stored in refrigerator increased from 0.20±0.00% at
week one to 0.30±0.00% at week four. Also the acidity
of grapefruit cheese stored in whey and refrigerator
were increased with the increase of storage period. It
revealed 0.90±0.00% at week one and 1.00±0.00% at
week four for cheese stored in whey, it revealed
0.20±0.00% at week one and 0.37±0.06% at week four
for cheese stored in refrigerator (Table 6).
Discussion
The fat content of the lime cheese and grapefruit
cheese samples stored in refrigerator were high, while
the fat contents of the same cheese stored in whey at
room temperature were low. Generally fat content of
lime cheese was higher than fat content of grapefruit
cheese. Hofi et al. (1976), Nofal et al. (1981) and
Abdel Razig (1996) reported that the high fat content
of the cheese samples stored at room temperature
could be attributed to the loss of degradation products
in the pickling whey. This finding is lower than those
of Babiker (1987) and Kosikowski (1967). However
the present result is in the range stated by SSMO
(2002) that cheese fat content should be between 15-
20 %.
Generally protein content of lime cheese was higher
than protein content of grapefruit cheese; this is
explained by the low TS and high moisture content of
the grapefruit cheese, this result agrees with Abdel
Razig and Babiker (2009) who found that the protein
content of Quso Blanco lime cheese is higher than that
of grapefruit cheese. The lower protein content of lime
cheese and grapefruit cheese were found for samples
stored in whey at room temperature, while the higher
protein content was for cheese samples stored in the
refrigerator. This could be due to inhibition of
proteolytic activities of microorganisms in low storage
temperature (Abdel Razig and Babiker (2009), Hamid
(1998) and Nofal et al., (1981) claimed that the
increase in the crude protein content of the cheese
stored at room temperature could be due to low
moisture content and high acidity in the curd which
inhibited the growth of proteolytic bacteria. The low
crude protein content of the samples stored in the
refrigerator was possibly attributed to absorption of
high level of moisture by the curd (Zaki et al. 1974) .
This result agrees with Alla Gabo (1986), Makki
(1987), SSMO (2002) and Kosikowski (1967) who
reported 22.6%, 23.6-25.6%, 15% (as lowest limit),
and 24.9% respectively. The protein content of lime
cheese stored in the refrigerator and whey was higher
than those of grapefruit cheese stored in the same
conditions.
The total solids content of the grapefruit cheese was
higher than lime cheese. This result disagreed with
Abdel Razig and Babiker (2009) who found that total
solids content of lime cheese is higher than that of
grapefruit cheese. The present finding agreed with El
Owni and Hamid (2008) who found that the total
solids increase with time, it also agreed with Alla
Gabo (1986) who reported 38.85% total solids.
However it disagrees with Nuser (2001) who found
that the total solids decrease with time due to
proteolytic and lipolytic effect of microorganisms. The
high total solids content was in the grapefruit cheese
stored in refrigerator, while the low total solids content
was in the grapefruit cheese stored in whey at room
temperature.
This result agreed with Salama et al., (1983);
Collombo (1992) and Walstra et al. (1999) who
explained the increase in total solids content of the
cheese stored in refrigerator might be due to inhibition
of proteolytic and lipolytic activities of
microorganisms by low storage temperature. Nuser
(2001) and Hayaloglou et al. (2005) found that total
solids decrease during storage period due to
proteolytic and lipolytic effect of microorganisms on
proteins and dissolution of fats into pickling. This
result of the lime cheese and grapefruit cheese
matched with that found by Salama et al., (1983);
Colombo et al. (1992) and Walstra et al. (1999).
During the storage period total solids content of the
cheese was increasing at room temperature, which
were higher that probably might be due to low

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Table 1: Effect of preservation method on chemical composition Mean± SD of lime (Citrus aurantifolia) and
grapefruit (Citrus paradisi) white cheese (Gibna bayda)
Means bearing the same superscripts are not significantly (P<0.05) different
**: Significant level at (p≤0.01); ***: Significant level at (p≤0.001)
Table (2): Fat content of white cheese (Gibna bayda) made by coagulating milk with lime (Citrus aurantifolia) and
grapefruit (Citrus paradisi) juices
Measurements
Lime cheese
Grapefruit cheese
L.S
preservation method
Periods (Week)
Whey/room
Refrigerator
Whey/room
Refrigerator
1
15.0 ±4.36
15.00±5.20
12.33±4.93
12.67±4.62
***
2
14.67±4.62
16.00±5.20
10.67±2.89
13.33 ± 4.04
***
3
14.67±4.62
15.67± 4.62
8.33±3.21
9.33± 4.04
***
4
14.00± 3.46
14.33±4.04
10.67±1.16
11.3 ± 0.04
***
Means bearing the same superscripts are not significantly (P<0.05) different
***: Significant level at (p≤0.001)
Table (3): Protein content of white cheese (Gibna bayda) made by coagulating milk with lime (Citrus aurantifolia)
and grapefruit (Citrus paradisi) juices
storage Period
(Week)
Preservation method
Lime cheese
Grapefruit cheese
L.S
Whey/Room
Refrigerator
Whey/Room
Refrigerator
1
18.13 ± 0.91
19.40 ± 0.30
17.07±0.32
15.97± 0.23
***
2
15.00 ±0.00
16.90 ±0.69
10.10 ± 0.00
15.00±0.00
***
3
15.70 ±0.35
17.30 ± 0.00
11.57 ± 1.03
15.00±0.00
***
4
16.30 ±0.35
24.20 ± 0.69
16.30 ± 0.35
21.67±0.58
***
Means bearing the same superscripts are not significantly (P<0.05) different
Table (4): Total solid content of white cheese (Gibna bayda) made by lime (Citrus aurantifolia) and grapefruit
(Citrus paradisi) juices
Storage period
Treatments
Week)
Lime cheese
Grapefruit cheese
L.S
Whey/Room
Refrigerator
Whey/Room
Refrigerator
1
45.51±1.31
42.77± 1.90
48.01±1.29
55.12± 0.72
***
2
40.91±1.80
45.58 ±0.31
42.24± 1.59
49.13 ± 1.24
***
3
60.86±1.84
45.16 ±0.53
26.51± 1.05
53.96± 0.28
***
4
66.94 ±1.48
43.79 ±1.64
30.63±0.87
30.63± 0.87
***
Means bearing the same superscripts are not significantly (P<0.05) different
***: Significant level at (p≤0.001)
Chemical
composition
Lime cheese
Grapefruit cheese
Whey/room
Refrigerator
Whey/room
Refrigerator
L.S
Fat (%)
14.58±3.68
15.25±4.14
10.50±3.21
11.67±3.92
**
Protein (%)
16.28±1.29
19.45±3.06
13.76±3.15
17.03±2.83
***
Total solids (%)
53.55±11.25
44.32±1.60
36.85±9.09
57.87±9.59
***
Ash (%)
0.64±0.43
0.50±0.01
0.42±0.32
0.74±0.32
***
Acidity (%)
1.09±0.26
0.25±0.05
0.89±0.19
0.41±0.33
***

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128
Table (5): ash content of white cheese (Gibna bayda) made by coagulating milk with lime (Citrus aurantifolia) and
grapefruit (Citrus paradisi) juices
Measurement
Lime cheese
Grapefruit cheese
Preservation method
Periods (Week)
Whey/Room
Refrigerator
Whey/Room
Refrigerator
L.S
1
0.50 ±0.00
0.05± 0.01
0.57±0.06
0.63± 0.25
***
2
0.70±0.01
0.03 ±0.31
0.57±0.06
0.43 ± 0.06
***
3
0.90 ±0.00
0.05± 0.01
0.07±0.03
0.70 ± 0.00
***
4
1.10 ±0.26
0.05± 0.01
0.47±0.55
1.20 ± 0.10
***
Means bearing the same superscripts are not significantly (P<0.05) different
***: Significant level at (p≤0.001)
Table (6): Effect of preservation method and storage period (week) on acidity of white cheese (Gibna bayda) made
by lime (Citrus aurantifolia) and grapefruit (Citrus paradisi) juices
Treatment
Lime cheese
Grapefruit cheese
L.S
Preservation method
Period (Week)
Whey/Room
Refrigerator
Whey/Room
Refrigerator
1
0.80 ±0.00
0.20± 0.00
0.90± 0.00
0.20± 0.00
***
2
1.10±0.00
0.23 ±0.06
1.07±0.06
0.83 ± 0.46
***
3
1.00 ±0.00
0.27± 0.06
0.60±0.00
0.23 ± 0.06
***
4
1.47 ±0.12
0.30± 0.00
1.00±0.00
0.37 ± 0.06
***
***: Significant level at (p≤0.001)
moisture content as a result of high acidity of the
cheese. The result also agreed with that recorded by
Abdel Razig and Babiker (2009) and El Owni and
Hamid (2008) who found that the total solids of
Sudanese white soft cheese increased during storage
period. The variations during storage period in the
grapefruit cheese stored in whey at room temperature,
Table (4) showed the decrease in total solids of
cheese. This was attributed to continuous loss of
moisture. Nuser (2001); Aly and Galal (2002) and
Hayaloglou et al. (2005) attributed the total solids
decrease during the storage period could be due to
proteolytic and lipolytic effect of microorganisms on
proteins and dissolution of fats into pickling.
Ash content of grapefruit cheese is higher than those
of lime cheese. The ash contents of lime cheese
samples stored in whey at room temperature increased
from 0.50±0.00% at week one to 1.10±0.26% at week
four. Then they were constant in cheese stored in
refrigerator during storage period. Also the ash content
of grapefruit cheese samples stored in whey at room
temperature were decreased from 0.57±0.06% at week
one to 0.47±0.55% at week four. The grapefruit cheese
stored at the refrigerator showed ash content of
0.63±0.25%, 0.43± 0.06 %, 0.70±0.00% and
1.20±0.10% from week one to week four (Table 5).
The ash content of the lime cheese samples stored in
whey at room temperature was found to increase
during storage period. However the ash was stable in
cheese stored in refrigerator. This agreed with El Owni
and Hamid (2008) who reported that ash content
increases during storage period due to decrease in
moisture content.
In this study the ash content of lime cheese was higher
in cheese stored in whey at room temperature, while
ash content was lower in cheese stored in refrigerator.
This finding disagreed with Abdalla (1992) and Bilal
(2000) who found that the ash content of the cheese
stored in refrigerator were higher than those stored in
room temperature.

Int. J. Adv. Res. Biol.Sci. 1(7): (2014): 123–130
129
Zaki et al. (1974); Nofal et al. (1981) and Tayar
(1995) justified that the absorption of salt by the curd
at the low storage temperature. Although results of ash
content of grapefruit cheese stored in refrigerator
agreed with Abdalla (1992) and Bilal (2000) . The
result obtained is lower than that of Alla Gabo (1986);
Ali (1987) who reported 5.2% and 6.85, respectively.
On the other, hand the SSMO (2002) stated 5% as the
lowest limit.
In the current study the higher acidity was obtained for
the lime cheese and grapefruit cheese stored in whey
at room temperature, while the lower acidity was in
cheese stored in refrigerator. This finding agreed with
Hamid and El Owni (2007) and Nofal et al. (1981)
who showed that the higher acidity of the cheese
stored at room temperature might be attributed to
increase level of lactic acid, which could be due to the
activation of lactic acid bacteria in room temperature.
The low acidity of cheese stored in refrigerator might
be explained by the fact that low temperature inhibited
growth and activity of lactic acid bacteria, which
consequently lower the rate of acid development (El
Owni and Hamid 2007 and Nofal et al. 1981).
Conclusion
It was noticed from the results the chemical
composition of the lime cheese was better than
grapefruit cheese; quick milk coagulation at high
temperature (82ºC). Using these fruits; the produced
cheese would be of high quality if processing
conditions and handling procedures are satisfactorily
achieved.The study concluded that the average yield
of grapefruit cheese was higher than that of lime
cheese.
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