Download Supplementation of Some Fruit Nectars with Technological Barley

Journal of Life Sciences and Technologies Vol. 1, No. 1, March 2013
Supplementation of Some Fruit Nectars with
Technological Barley Preparations as Prebiotic
Sources
Abd-El Tawab S. Barakat, Sherein I. Abd-Elmoez*, Mona F. Masoud and Manal M. Hagag
Food Technology Research Institute; * National Research Center
Email: [email protected]

antimicrobial substances which inhibit the growth of
pathogenic bacteria in the intestine. Also, [6] evaluated
the in vivo prebiotic potential of barley β- glucan and
concluded that it induced a strong bifidogenic effect. It is
reported that daily intake of a cake containing barley βglucan is well tolerated and demonstrated significant
bifidogenic properties in older healthy volunteers. [7]
reported that barley is gaining renewed interest as an
ingredient for production of functional foods due to its
high contents of bioactive compounds such as β- glucans,
tocopherols and tocotrienols. The US Food and Drug
Administration has allowed whole-grain barley products
that can supply β-glucan at levels of 0.75 g per serving or
3 g per a day to carry a claim that they reduce the risk of
coronary heart disease [8]. Also, [9] cited that oat and
barley β-glucan were studied as functional ingredients in
bakery systems to deliver health benefits. A number of
studies have evaluated the effect of barley β- glucans in
traditional food products such as bread, pasta, cereal
flakes, and others on postprandial glycemia in both
healthy [10] and overweight subjects [11]. On the other
hand, [12] concluded that beta glucan consumed with
orange juice is more effective in lowering total and LDL
cholesterol concentrations and the ratio of total to HDL
cholesterol than is the same preparation administered in
bread and cookies. Also, [13] mentioned that a barley βglucan resulted in significant reductions in postprandial
glycemia and insulinemia in healthy individuals, when
incorporated into a beverage. However, a barley β-glucan
concentrate had low effect on glycemic response when
incorporated into solid food matrix. There is
epidemiological evidence linking a diet rich in fruits and
vegetables with reduced incidences of coronary heart
disease, cancer, and various chronic diseases [14]. The
Kame Project carried out with Japanese- Americans
between 1992 and 2001 found that subjects with a higher
intake of fruit and vegetable juices had a substantially
reduced incidence of Alzheimer’s disease [15]. One glass
of fruit juice is an important source of fluids and can
provide vitamin C, folate, potassium and antioxidants.
[16] reported that the demand for the strawberry is
increasing worldwide because of its delicious flavor and
attractive color. Nutritionally, strawberry contains low
calorie carbohydrate and a potential source of vitamin C,
fibers and provides more vitamin C than oranges. Mango
(Mangifera indica L.), as an emerging tropical export
Abstract—Prebiotics are being implicated in gut health
maintenance, colitis prevention, cancer inhibition,
immunepotentiaton, cholesterol removal, reduction of
cardiovascular disease and prevention of obesity and
constipation. It was found that barley had a great positive
effect on viability and enhanced the growth of
bifidobacterium group. This search acts as a trial to
produce some functional nectars using the technological
preparations of barley Talbina as prebiotic sources.
Strawberry, guava and mango nectars were supplemented
with classic and instant barley Talbina by 5% and 7%,
respectively, as prebiotic sources. The results revealed the
capability of the classic and instant Talbina to enhance the
bacterial growth of highly beneficial bacteria as
Bifidobacterium langium, Lactobacillus acidophilus and L.
palantarum. Data showed that addition of both instant and
classic Talbina increased protein, carbohydrates, fiber, βglucan, viscosity, TSS and pH and slightly decreased acidity
of nectars. The results recommend that addition of the
classic or instant Talbina to the nectars did not affect the
sensory properties of these fruit nectars.
Index Terms—Barley, classic talbina, instant talbina,
nectars, prebiotics.
I. INTRODUCTION
Prebiotics are generally defined as non-digestible
polysaccharides and oligosaccharides which promote the
growth of beneficial lactic acid bacteria in the colon and
exert antagonism to Salmonella sp. or Escherichia coli,
limiting their proliferation [1]. Prebiotics are being
implicated in starter culture formulation, gut health
maintenance, colitis prevention, cancer inhibition,
immunepotentiaton, cholesterol removal, reduction of
cardiovascular disease and prevention of obesity and
constipation [2]. [3] reported that barley grain provides
essential vitamins and minerals that can promote growth
of probiotic and intestinal bacteria. [4] found that fecal
bifidobacterium was significantly increased by addition
of barley. Moreover, [5] reported that addition of barley
extract or malt had a great positive effect on viability and
enhanced the growth of L. acidophilus and L. plantarum.
Lactic acid bacteria and Bifidobacteria are natural
components of gastrointestinal microbiota. They produce
Manuscript received September 15, 2012; revised December 30,
2012
©2013 Engineering and Technology Publishing
doi: 10.12720/jolst.1.1.38-43
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Journal of Life Sciences and Technologies Vol. 1, No. 1, March 2013
growth of L. acidophilus and MRS broth only for L.
Plantarum [25]. The antimicrobial activity of these tested
probiotic strains was carried out against two of the most
pathogenic food poisoning bacterial strains which are
highly contagious to human and animal health;
Eshereshia coli O157 and St. aureus. The experiments
were carried out using three different groups of probiotic
preparations; Group 1 used as negative control; pure
culture of the probiotic stains without Talbina. Group 2
probiotic strains with the addition of 20% classic Talbina.
Group 3 probiotic strains with the addition of 20% of
instant Talbina. For determination of the role of Talbina
for enhancement of the bacterial growth, Serial dilutions
up to 1010 of the three probiotic strains were prepared.
Total bacterial counts of these strains were estimated
after anaerobic incubation for 24 and 48 hrs.
In vitro antibacterial activity of probiotic bacteria: In
vitro antibacterial activity of the three tested probiotic
strains in the previously mentioned groups was evaluated
using agar well diffusion test on Muller Hinton agar [26].
These plates were prepared and wells were drilled out
using Pasture pipettes. The plates were inoculated with
pathogenic bacterial strains prepared in conc. equivalent
with 0.5 MacFerland, and then 50 µl aliquots of cell free
cultures supernatant of probiotic strains grown in MRS
broth were suspended in the agar wells. Plates were
incubated at 37˚C/24 hrs under microaerophillic
condition. The experiment was carried out in duplicate.
The diameters of inhibition zones around wells were
measured in mm using a ruler.
Chemical composition: Moisture, ash, total dietary
fiber, lipids and protein were determined according to the
methods recommended by the [27]. Total carbohydrates
were calculated by difference according to the following
equation: Total carbohydrates = 100 – (% crude protein
+ % crude fat + % ash). Total calories were calculated
using the equation mentioned by [28]. Where, energy
(calories) = 4 (carbohydrate + protein) + 9 (fat). Betaglucan was extracted from barley flour and instant
Talbina powders using the method of [29].
Physical properties: Total soluble solids were
measured at 25 °C with Abbe refrectometer Model 1T
according to [27] and expressed as Brix. The pH value
was measured with a pH meter model Consort pH meter
P107. Total acidity was determined by titration with
NaOH 0.1 N solution using phenolphthalein as indicator
according to [27]. Rheological properties: Viscosity,
Torque (%) and Shear stress (D/ cm2) measurement was
carried out by the Brookfield Digital Viscometer Model
DV lll at rotational speeds 30 rpm. A temperaturecontrolled water bath was used to regulate the
temperature of the samples at 30 oC. The Brookfield
spindles UL adapter and Brookfield small sample adapter
were used according to [30] with some modifications.
Sensory evaluation: A panel of ten judges selected
from staff of Food Technology Research Institute
evaluated the product fortnightly for color, taste, odor and
overall acceptability by the method of [31] using a scale
from 1 to 9, where 1 represented extremely disliked and 9
represent extremely liked. Initial experiments showed that
crop is produced in about 90 countries in the world with a
production of over 25.1 million tons [17]. The fruits of
mango (Mangifera indica L.) have a pleasant taste and
aroma and soluble polysaccharides (pectic substances),
cell wall material (cellulose) and uronic acids [18]. As
stated in the Australian Guide to Healthy Eating [19],
fruit juice can count towards a serve of fruit a day. Some
fruit juices available on the market are also fortified with
dietary fiber, calcium, folate and vitamin A. Consumption
of natural juice with additional benefits was highly
appreciated by consumers [20]. This search acts as a trial
to produce some functional nectars using the
technological preparations of barley as prebiotic sources.
II. MATERIALS AND METHODS
Naked barley (Hordeum vulgare variety Giza l30)
was obtained from Barley Research Department, Field
Crop Research Institute, Agriculture Research Center,
Giza, Egypt. Strawberry (Fragariq sp.), guava, mango
fruits (Mangifera indica L.) and sucrose were purchased
from local market.
Preparation of the classic and instant talbina: Clean
barley grains were milled in stone mill to produce whole
barley flour (control). Whole barley flour was added to
water at room temperature by 1:5 (w/v) and stirred to
produce barley flour solution. Barley flour solution was
boiled for 10 min. with continuous stirring. The cooked
barley flour solution is then named "classic Talbina
syrup" [21]. A portion of the previous Talbina was spray
dried at 280ºc for 2min. in Misr coffee Co. 10th of
Ramadan City, Egypt, (patent No. 24427/2007) to
produce the instant Talbina powder [22].
Preparation of fruits purees: Fruits were washed with
running water, mango fruits were hand peeled and cut.
The fruits were mechanically extracted by using
Moulinex blender (Blender Mixer, type: 741). The purees
were strained by 0.023 inch screen avoid coarse pulp
particles and to have only fine particles of almost
colloidal consistency.
Preparation of fruits nectars: The previous purees
(strawberry, guava or mango) were added to water by
25%. Sucrose was added by 12%. Classic or instant
Talbina was added by 5%, 7% and 9% (w/v).
Microbiological evaluation: Pour-plate method was
employed. Aliquots of 1 ml from the suitable dilutions of
barley flour and instant Talbina powder were transferred
to petri dishes. Specific medium was added and mixed
thoroughly. Three replicates were prepared from each
dilution. Colony counts were obtained after three days of
incubation at 30°C. Total bacteria, spore form,
Thermophilic on Thornton's medium, total fungi on
Martin’s medium, actinomycetes on Jensen's medium and
yeast on PDA (Potato-Dextrose Agar) medium [23].
Coliform on Macconoky Agar medium.
Probiotic bacterial growth and bacterial colony count
of the instant Talbina: The probiotic strains;
Bifidobacterium langium, Lactobacillus acidophilus and
Lactobacillus plantarum were cultured in De Man,
Rogosa, Sharpe (MRS) broth with Cystiene (0.3 %) for
the growth of Bif. lungium, with bile salts (0.05%) for the
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Journal of Life Sciences and Technologies Vol. 1, No. 1, March 2013
addition of the classic Talbina up to 5% or instant Talbina
up to 7% produced the acceptable nectars.
Statistical analysis: Data of sensory evaluation of the
studied cakes and biscuits were subjected to analysis of
variance and least significant difference (L.S.D) at 0.05
level according to the method described by [32].
2) Prebiotic activity of the classic and instant talbina:
Prebiotic activity of the classic and instant Talbina on
probiotic strains against E. coli O157 and S. aureus is
shown in Table Π. The supernatant of Bif. lungium
(group 1) showed more antibacterial effect against S.
aureus and E. coli O157 with an inhibition zones equal
1.3 mm and 1.2 mm, respectively. While it showed the
same activity (1.2 mm) in group 2 with E. coli O157 after
24 and 48 hrs, respectively. However the activity of
Group 2 bifidobacterium against S. aureus after 24 hrs
increased to 1.3 mm then decreased to 1.1 mm after 48
hrs. The least antibacterial activity of Bifidobacterium
was shown in group 3, as the inhibitory zone against E.
coli: O157 was 1.2 mm and 1.1 mm for S. aureus after 48
hrs. These results were in agreement with [34]. They
found that Bifidobacterium sp. showed considerable
inhibitory activity against both S. aureus and E. coli. The
supernatant of L acidophilus as well showed great
hindrance of both tested strains in group 1 (1.4 mm)
while its probiotic activity decreased in group 2 and no
hindrance, was shown in Group 3. L. palantarum showed
no hindrance against pathogenic bacteria in all groups as
shown Table Π. In contrast to our results, L. palantarum
showed a great inhibitory effect on both E. coli O 157
and Salmonella sp. when examined on MRS medium [35].
These results showed also that the increased bacterial
count of the probiotic strains might hinder the ability of
release of the antibacterial compounds from the probiotic
bacteria in vitro or it might prevent the ability of heaving
pure supernatant carrying the probiotic released
components which have great antibacterial activity as the
supernatant will contain probiotic bacterial cells which
did not precipitate after centrifugation due to over
bacterial growth. [36] demonstrated that feed mice with
probiotic B. thermacidophilum can reduce the severity of
E. coli: O157 infection. Also, [35] reported that L.
palantarum showed a great inhibitory effect against both
E. coli: O157 and Salmonella sp. when examined on
MRS medium.
A. Results and Discussion
1) Effect of the classic and instant talbina on some
probiotic bacteria:
Effect of the classic and instant Talbina on some
probiotic bacteria is shown in Table Ι. The results
indicated that total counts of tested probiotic strains were
highly affected with the addition of Talbina. As shown in
Table Ι colony counts of Bifidobacterium sp. of group (1)
(negative control without Talbina) were 180 and 250
colony forming unit (cfu) after 24 and 48 hrs,
respectively. Whereas L. acidophilus showed no growth
after 24 or 48 hrs, the growth of L. palantarum, was
observed only after 48 hrs and reached 173 cfu and was
absent after 24 hrs. On the other hand, group (2)
(containing 20% classic Talbina) showed high growth
activity with counts of 210 and 280 for Bifidobacterium
sp, 18 and 36 for L. acidophilus and 230 and 280 for L.
palantarum after 24 and 48 hrs respectively. Moreover,
the total counts of Group (3) (containing 20% Instant
Talbina) were the highest as reached more than 300 (cfu)
for Bifidobacterium sp, 28 and 57 for L. acidophilus and
more than 300 cfu for L. palantarum after 24 and 48 hrs
respectively. These results indicated the capability of the
classic and instant Talbina to enhance the bacterial
growth of highly beneficial bacteria as Bif. langium, L.
acidophilus and L. palantarum. Also, results revealed that
the instant Talbina is more effective than classic Talbina
in enhancing the bacterial growth. Similar results were
reported by [33]. They found that fecal bifidobacterium
increased significantly by addition of barley. Moreover,
addition of barley extract or malt had a great positive
effect on viability and enhancing the growth of L.
acidophilus and L. plantarum [4].
TABLE II. PREBIOTIC ACTIVITY OF THE CLASSIC AND INSTANT TALBINA.
TABLE I. EFFECT OF THE CLASSIC AND INSTANT TALBINA ON SOME
PROBIOTIC BACTERIA.
Probiotic strains
Probiotic strains
10
Bacterial colony count (10 )
Zone of inhibition in mm
E. coli O157
After
After
24 hrs
48 hrs
S. aureus
After
After
24 hrs
48 hrs
After 24 hours
After 48 hours
180
250
Bifidobacterium
1.2
1.2
1.3
1.3
L. acidophilus
No growth
No growth
L. acidophilus
1.4
1.4
1.4
1.4
L. palantarum
No growth
173
L. palantarum
0.0
0.0
0.0
0.0
Group 1
Bifidobacterium
Group 1
Group 2
Group 2
Bifidobacterium
210
280
Bifidobacterium
1.2
1.2
1.3
1.1
L. acidophilus
18
36
L. acidophilus
0.0
1.1
1.4
0.0
L. palantarum
230
280
L. palantarum
0.0
0.0
0.0
0.0
Bifidobacterium
Group 3
Bifidobacterium
L. acidophilus
L. palantarum
Group 3
> 300
> 300
0.0
1.2
0.0
1.1
28
57
L. acidophilus
0.0
0.0
0.0
0.0
> 300
L. palantarum
0.0
0.0
0.0
0.0
> 300
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Journal of Life Sciences and Technologies Vol. 1, No. 1, March 2013
3) Effect of classic and instant talbina on the chemical
properties of nectars:
Effect of instant and classic Talbina on the chemical
properties of strawberry, guava and mango nectars is
shown in Table Ш. Data showed that addition of both
instant and classic Talbina with 5%, 7% and 9%
increased protein, carbohydrates, fiber, β- glucan and
energy content of the produced nectars because of the
high content of both instant and classic Talbina of these
components.
TABLE Ш. EFFECT OF INSTANT AND CLASSIC TALBINA ON THE CHEMICAL PROPERTIES OF NECTARS.
Nectar of:
Strawberry with:
Control
5% classic Talbina
7% instant Talbina
Guava with:
Control
5% classic Talbina
7% instant Talbina
Mango with:
Control
5% classic Talbina
7% instant Talbina
Protein
(%)
Carbohydr
ates (%)
Fiber
(%)
Ash
(%)
0.1
0.6
0.8
11.7
15.0
16.6
0.5
1.5
1.6
0.10
0.24
0.28
0.00
0.38
0.56
0.1
0.6
0.8
11.5
15.1
16.5
1.0
1.8
2.1
0.20
0.34
0.38
0.00
0.38
0.56
0.3
0.8
1.1
12.5
16.1
17.8
2.0
2.8
3.2
0.22
0.35
0.41
0.00
0.40
0.57
4) Effect of the classic and instant talbina on the
physical properties of nectars:
Effect of instant and classic Talbina on the physical
properties of strawberry, guava and mango nectars is
shown in Table ΙV. Data showed that TSS of strawberry,
guava and mango nectars was 12.12, 13.15 and 15.11,
respectively. Data showed also that pH of strawberry,
guava and mango nectars was 3.5, 3.5 and 4.9,
respectively. Data showed also that acidity of strawberry,
guava and mango nectars was 1.43, 1.43 and 0.33,
respectively. Data cleared also that addition of both
classic and instant Talbina with 5% and 7% slightly
increased TSS and pH and decreased acidity of nectars.
Data obvious also that the effect of both instant and
classic Talbina on the physical properties of nectars was
similar.
Effect of the classic and instant Talbina powder on the
sensory properties of juices of strawberry, guava and
mango is shown in Table V. Data cleared that addition of
the classic and instant Talbina with 5% and 7% did not
affect the color of strawberry, guava or mango nectars.
Data showed also that addition of the classic Talbina with
5% and instant Talbina with 5% and 7% did not affect
taste, odor and acceptability of strawberry, guava or
mango nectars. From data presented in Table V, it could
be noticed that addition of the classic Talbina up to 5% or
instant Talbina up to 7% did not affect the sensory
properties of strawberry, guava or mango nectars.
TABLE V. EFFECT OF THE INSTANT TALBINA POWDER ON THE
SENSORY PROPERTIES OF NECTARS.
TSS
pH
5% classic Talbina
7% instant Talbina
Taste
(9)
Odor
(9)
Acceptability
(9)
Control
8.5a
8.7 a
8.5 a
8.6 a
5% classic Talbina
8.3 a
8.5 a
8.3 a
8.4 a
7% instant Talbina
8.4 a
8.7 a
8.5 a
8.6 a
8.6 a
8.7 a
8.8 a
8.6 a
8.6
a
8.5
a
8.5
a
8.5 a
8.5
a
8.7
a
8.5
a
8.6 a
Strawberry with:
Acidity (%)
Guava with:
Strawberry with:
Control
Color
(9)
Juice of:
TABLE IV. EFFECT OF THE CLASSIC AND INSTANT TALBINA ON THE
PHYSICAL PROPERTIES OF NECTARS.
Nectar of:
ß-glucan
(%)
12.12
12.21
12.23
3.5
3.6
3.6
Control
1.43
5% classic Talbina
1.31
7% instant Talbina
1.31
Mango with:
Guava with:
Control
13.15
5% classic Talbina
13.16
7% instant Talbina
13.16
3.5
3.7
3.6
1.31
Mango with:
15.11
4.9
0.33
5% classic Talbina
15.21
5.0
0.31
7% instant Talbina
15.21
5.0
0.31
8.7 a
8.5 a
8.6 a
8.5
a
8.5
a
8.3
a
8.5 a
7% Instant Talbina
8.6
a
8.7
a
8.5
a
8.6 a
L. S. D.
0.5
5% Classic Talbina
1.25
Control
8.7 a
Control
1.43
0.8
0.5
0.7
6) Effect of the classic and instant talbina on the
rheological properties of nectars:
Effect of instant and classic Talbina on the rheological
properties of strawberry, guava and mango nectars is
shown in Table VI. Data showed that the torque of
strawberry, guava and mango nectars was 0.6%, 0.8%
5) Effect of the classic and instant talbina on the
sensory properties of nectars:
41
Journal of Life Sciences and Technologies Vol. 1, No. 1, March 2013
[7]
and 0.4%, respectively. Data showed also that addition of
both classic and instant Talbina with 5% and 7%
increased the torque of nectars. Data cleared also that
classic Talbina increased the torque of nectars more than
the instant. Data in Table VI showed also that viscosity of
strawberry, guava and mango nectars was 4, 5 and 2.5 cP,
respectively. Data cleared also that addition of both
classic and instant Talbina with 5% and 7% increased the
viscosity of nectars. Data showed also that classic Talbina
increased the viscosity of nectars more than the instant.
Data in Table VI cleared also that shear stress of
strawberry, guava and mango nectars was 0.0, 0.0 and 2.0
D/ cm2, respectively. Data cleared also that addition of
both classic and instant Talbina 5% and 7% to strawberry
and guava nectars did not affect the shear stress of the
produced nectars. Data cleared also that addition of both
classic and instant Talbina 5% and 7% to mango nectars
increased the shear stress of the produced nectars and the
classic Talbina increased it more than the instant.
[8]
[9]
[10]
[11]
[12]
[13]
TABLE VI: EFFECT OF THE CLASSIC AND INSTANT TALBINA ON THE
RHEOLOGICAL PROPERTIES OF NECTARS.
[14]
Nectar of:
Torque
(%)
Viscosity
(cP)
Shear stress
(D/ cm2)
0.6
0.0
[15]
Strawberry with:
Control
5% classic Talbina
1.8
4.00
6.90
7% instant Talbina
1.8
8.50
0.8
0.0
0.0
[16]
0.0
[17]
[18]
Guava with:
Control
5% classic Talbina
2.2
5.00
9.70
7% instant Talbina
2.4
10.5
0.0
0.0
Control
0.4
2.50
2.00
5% classic Talbina
1.9
8.37
14.0
7% instant Talbina
2.0
11.4
16.20
Mango with:
[19]
[20]
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Abdeltawab S. Barakat: was born in Egypt, in
1970. B. Sci. Food Sci., Zagazig University, 1992;
M. Sci. Nutrition and Food Science, Menoufia
Univ., 1998 and Ph. D. Nutrition and Food
Science, Zagazig Univ., 2003.
He is an Associate Professor at the Food
Technology Research Institute, a specialist in
human nutrition and cereal science and has a
patented in creating a new method to produce an
instant barley drink.
Author’s
Dr.
Barakatformal
a member of the Egyptian Society of Food Science and
photo
Technology.
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