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International
Journal of Nutrition and Food Sciences
2016; 5(5): 361-366
http://www.sciencepublishinggroup.com/j/ijnfs
doi: 10.11648/j.ijnfs.20160505.17
ISSN: 2327-2694 (Print); ISSN: 2327-2716 (Online)
Probiotics,Organoleptic and Physicochemical Properties of
Vegetable Milk Based Bio-ice cream Supplemented with
Skimmed Milk Powder
Mohamed Omer Elsamani
1, 2
1
Department of Basic Medical Sciences, University of Al-Baha, Baha City, Kingdom Saudi Arabia
2
Department of Food Sciences &Technology, University of Omdurman Islamic, Khartoum, Sudan
Emailaddress:
mhmdomer6@gmail.com
*
Correspondingauthor (M. O. Elsamani)
Tocitethisarticle:
Mohamed Omer Elsamani. Probiotics, Organoleptic and Physicochemical Properties of Vegetable Milk Based Bio-ice cream Supplemented
with Skimmed Milk Powder. International Journal of Nutrition and Food Sciences. Vol. 5, No. 5, 2016, pp. 361-366.
doi: 10.11648/j.ijnfs.20160505.17
Received: August 9, 2016; Accepted: August 18, 2016; Published: October 17, 2016
Abstract:
The aim of the present study was to evaluate non fermented probiotic ice cream made from vegetable (lupine
and peanuts) milk, and to evaluate its probiotics, organoleptic and physicochemical characteristics. Ice cream samples were
produced from blends of vegetable milk and skimmed milk powder together with the added Probiotics bacteria of
Lactobacillusacidophilus and Bifidobacterium. lactis. The changes of Bio–ice cream qualities and survival of added
microbes were evaluated at 0, 1, and 30 days. The viable of L. acidophilus decreased by 1.62 and 2.05 log cycles in lupine
and peanuts milk ice creams throughout 30 days storage respectively. The counts of B. lactis reduced by 1.32 and 2.22 log
cycles in lupine and peanuts ice creams respectively. Regardless of ice cream color, incorporation of vegetable milk ice
cream significantly (P ≤ 0.05) enhanced the taste, texture, flavor, and overall acceptability of ice cream. The highest total
solids and protein were found in ice creams containing peanuts milk and lowest fat and total acceptability were found in ice
creams containing lupine milk. The produce of ice cream with vegetable milks developed the growth and viability of B.
lactis and L. acidophilus.
Keywords:
Bio- ice cream, Lupine, Peanut Milk, Probiotic, Bacteria
1. Introduction
Consumer is looking for the functional foods can offer
more health benefits. Full-fat milk is rich in saturated fatty
acids, which has been shown to significantly elevate total,
Low-density lipoprotein (LDL), and high-density
lipoprotein (HDL)-cholesterol) [4] and [3] which has long
been associated with increased risk of cardiovascular
disease. Functional food that contains viable probiotics in
particular addresses the improvement in intestinal micro
flora activation of the immune system, reduction in serum
cholesterol and inhibition of potential pathogens [12].
However, these probiotics must be present in adequate
amount (10
6
–10
7
cfu/g) at the time of consumption in order
to render it being effective [13].Fermented yogurt and milk
can effectively be used to deliver probiotic bacteria. The
former unfortunately may cause loss of viable probiotic due
to pH reduction and accumulation of organic acids as a
result of fermentation [14]. In this regard ice cream, due to
its neutral pH, may be used to deliver the probiotics [15].
Partial replacement cow milk with vegetable milks such as
soy milk could improve the pH of probiotic ice cream
resulting increased the survival of probiotics [16]. On the
other hand, Ice cream is a delicious, wholesome, nutritious
frozen dairy product, which is extensively consumed in
different parts of the world. Ice cream has nutritional
significance but encompasses no therapeutic properties [17].
Ice cream is traditionally made from cow s milk and thus
contains about 16% (w/w) lactose [18]. The demand for
alternatives to cows' milk is growing due to problems with
362 Mohamed Omer Elsamani: Probiotics, Organoleptic and Physicochemical Properties of Vegetable Milk
Based Bio-ice cream Supplemented with Skimmed Milk Powder
its lactose (Lactose intolerance) and cholesterol contents
and desire for vegetarian alternatives. Ice cream can be
made a functional food by adding fruits, protein rich
ingredients and also using vegetable milk such as coconut
and soy milks and the addition of probiotics. Moreover, the
lupine and peanut milks contain unique nutrient
compositions in peanut and lupine milks can support the
growth and survival of the lactic acid bacteria in ice cream
and thus could improve the health benefits and nutritional
components and health benefits of probiotic ice creams.
In particular, lupine proteins are receiving attention in
terms of health benefits, particularly in relation to a number
of conditions now known as ‘metabolic syndrome’ which
includes a cluster of factors such as, obesity, high blood
pressure, insulin resistance and elevated blood cholesterol
(5).
Lupine enriched foods have the potential to: beneficially
influence satiety (appetite suppression) and energy balance
[10], beneficially influence glycaemic control [6], improve
blood lipids [7], reduce hypertension [8] and improve bowel
health [9]. Partial replacement of cow milk with vegetable
milks may affect the physical properties of ice creams.
Fortification of yoghurt ice cream with soy protein has been
shown to improve the textural quality of the product
including firmness and viscosity [19]. Lecithin in the soy
also acts as emulsifiers which increase the viscosity and
stability of ice cream, refined texture and extend the
melting time [20]. Abdullah et al. [21] improved the quality
of ice cream by using different ratios of skim milk in
soymilk blend and found that large quantity of skim milk in
soymilk decline beany flavour of soy beans and increased
quality of ice cream. Fatemeh and Other authors [22]
Effects of Vegetable Milk on Survival of Probiotics and
Rheological and Physicochemical Properties of Bio-ice
cream, studied the Physicochemical and Rheological
properties of soy and coconut milk ice cream by adding soy
oil and sugar contents and found the addition of sugar and
replacing skim milk powder with in Vegetable milk ice
cream increased ice cream mix viscosity and reduced
melting rate of ice cream. The present study was carried out
to investigate the effect of lupine and peanut milks on the
survival of probiotic in ice cream during storage and the
physicochemical and organoleptic properties of non
fermented Bio-ice cream. There has been a shortage of milk
production. Thus, the supplementation and/or substitution
of cow milk with lupine and peanut milk would improve the
yield and nutritional quality of ice cream at a comparatively
lower cost. Up to now, no work has been published on
producing ice cream using cow milk supplemented with
Vegetable milk (lupine and peanut milk). Therefore, the aim
of the present study was to investigate the effect of
incorporation of Vegetable milk on physicochemical,
microbiological and organoleptic characteristics of Bio-ice
cream.
2. Materials and Methods
2.1. Bacteria Strain
Lactobacillus acidophilus and Bifidobacteriumbifidum
were obtained as pure freeze-dried probiotic culture from
CHR-Hansen (Horsholm, Denmark).
2.2. Preparation of Starter Cultures
Each one gram of strain was cultured in 100 mL of
sterilized skimmed milk (10 w/v) , simplified by the
addition of 0.05% (w/v) L-Cys-HCl, 1% (w/v) yeast extract
and 2% (w/v) glucose. The incubation was carried out under
aerobic condition in a water bath (40°C) until pH was
reached to 5.0 [23].
2.3. Preparation of Vegetable Milk
Lupine and peanuts milk were extracted from the seeds
by the methods according to [1: 2].
2.4. Ice Cream Production
Fresh pasteurized and homogenized milks (lupine and
peanuts milk), skimmed milk powder, butter, sugar, and
vanilla was purchased from local super market. Stabilizer
(Danisco AS, Copenhagen, Denmark) containing cellulose
gum, guar gum, mono- and diacyl- glycerol’s of fatty acid,
was used as stabilizer. Ice cream mix formulated according
to some properties of mixture milks (Table 1) such that it
had 38.7%–40.8% total solids and 10 - 10.5% fat for a total
batch of 100kg (Table 5).
The vegetable milk (lupine and peanuts milk) was
initially heated to 50°C followed by the other additives. The
mixture was mixed at 65°C in two stages by a homogenizer
with 16000 rpm for 5 min followed by pasteurization (10
min heating at 80°C followed by cooling to 4°C). The
mixture was aged overnight at 4°C prior to the addition of
each probiotic culture (4% w/w) followed by immediate
freezing in batch ice cream maker (1.5 L, Baumatic
Gelato1SS). The frozen mixture was packed in 100 mL
plastic cups and then stored in a freezer (−18°C). Three
different batches of ice cream per treatment were
manufactured.
2.5. Physicochemical Analysis
The vegetable Milk and ice creams were analyzed for pH
by a digital pH meter and for titratable acid (TA) by
titrating a sample (10g) with NaOH (0.1N)using
phenolphthalein (0.1w/v) as an indicator. The dry matters
were measured by drying samples at100±1°C for 3.5h using
an air oven [24]. The protein and fat contents of milk and
ice creams were measured by the Kjeldahl and Soxhlet
methods respectively [24]. All chemical analyses were
carried out in triplicate.
International Journal of Nutrition and Food Sciences 2016; 5(5): 361-366 363
Table 1. Physicochemical compositions of lupine and peanuts milk
Parameters pH T.S(g/100g T.A(%lactic acid) M.C(g/100g) Protein(g/100g) Fat(g/100g)
Peanuts Milk 7.03±0.10 14.67±0.10 0.10±0.10 85.33±0.12 5.40±0.12 5.00±0.12
Lupine Milk 6.30±0.10 12.14±0.07 0.60±0.01 87.86±0.12 5.10±0.12 3.30±0.12
Values are means of triplicate samples (±SD).
T.S: Total solids
T.A: Titritable acidity
M.C: Moisture content
Table 2. The content of components used in Bio-ice cream made from vegetable milk (%by weight).
Ice cream mixture Milk Skimmed Milk Powder Sugar butter water Stabilizer Vanillin
Lupine 50.0 10.0 15.0 4.4 20.0 0.5 0.1
Peanuts 50.0 10.0 15.0 4.4 20.0 0.5 0.1
2.6. Bacteriological Analysis
The viability of probiotics was measured immediately after inoculating the probiotic cultures and after 1 and 30 days of
frozen storage at −20°C. The samples (10 g) were decimally diluted with sterile peptone water (1 g L-1; Merck). One mL
aliquot dilutions were pour plated in triplicate on MRS agar for L. acidophilus and MRS agar supplemented with 0.05%
(w/v) L-Cys-HCI (Merck) for B. lactis. The plates were incubated at 38±1°C for 72 h under aerobic condition with 5% CO
2
(v/v) for L. acidophilus and anaerobic condition (Anaerocult A) for B. lactis. The bacterial viability was represented as
survival rate [23].
Table 3. Probiotic bacteria count of Bio- ice creams with different milks.
Bio-ice creams Mixture(0days)(Log10cfu/g)
Icecream1days(Log10cfu/g)
Icecream30days(Log10cfu/g)
Survival Rate (%)
L.L 8.87±0.12 8.85±0.15 7.25±0.15 97.57±0.15
L.B 8.59±0.12 8.57±0.15 7.27±0.15 98.85±0.15
P.L 9.58±0.12 9.46±0.15 7.53±0.15 98.87±0.15
P.B 9.87±0.12 9.74±0.15 7.65±0.15 97.85±0.15
Values are means of triplicate samples (±SD).
L.L: ice creams inoculated with Lactobacullusacidophilus made using lupine milk or P.L: peanut milk
LB: ice creams inoculated with Bifidobacteriumbifidum made using lupine milk or P.B: peanut milk
2.7. Organoleptical Analysis
The organoleptically characteristics of the ice creams were evaluated following the IDF standards [11]. A trained panel of 12
members, composed of adult male (4,age ranged from 25 to 35) and female (8,age ranged from 25 to 40), was assigned to
determine the quality of the ice creams (color, flavor, taste, texture and overall acceptability). Members were asked to score1-5
hedonic scale (1 = poor, 2 = acceptable, 3 = good, 4 = very good and 5 = excellent). The samples were randomized and
presented using tag for each one. To determine the differences in judges ‘response, the means cores were analyzed by Duncan’s
multiple range tests.
Table 4. Organoleptic property scores of ice creams with different milks.
Bio-ice creams Color Texture Taste flavor Overall acceptability
L.L 3.22±0.03
a
3.18±0.03
a
3.36±0.03
b
3.38±0.05
a
3.32±0.01
a
L.B 3.32±0.05
a
3.12±0.02
b
3.32±0.03
b
3.32±0.05
b
3.30±0.05
a
P.L 3.22±0.05
a
3.33±0.05
b
3.45±0.03
a
3.23±0.05
c
3.35±0.05
a
P.B 3.23±0.03
a
3.32±0.03
b
3.37±0.05
b
3.22±0.05
c
3.33±0.05
a
Values with different letters in the same column are significantly (P<0.05) different
L.L: Ice creams inoculated with L.acidophilus made using lupine milk
P.L: Ice creams inoculated with L.acidophilus made using peanut milk
LB: ice creams inoculated with Bifidobacteriumbifidum made using lupine milk
P.B: ice creams inoculated with Bifidobacteriumbifidum made using peanut milk
364 Mohamed Omer Elsamani: Probiotics, Organoleptic and Physicochemical Properties of Vegetable Milk
Based Bio-ice cream Supplemented with Skimmed Milk Powder
Table 5. The physicochemical compositions of vegetable milk and probiotic ice creams, made using lupine and peanut milks.
Vegetable milk/
Bio-ice creams T.S(g/100g) pH T.A(%lactic acid) Protein(g/100g) Fat(g/100g)
L.M 12.0±0.1 7.00±0.5 0.72±0.2 4.40±0.5 3.30±0.1
P.M 14.67±0.1 7.03±0.3 0.91±0.1 5.20±0.5 5.00±0.3
L.L 39.0±0.3 7.11±0.1 0.72±0.4 2.13±0.1 10.0±0.2
L.B 38.7±0.1 7.15±0.1 0.70±0.3 2.21±0.3 10.3±0.1
P.L 39.9±0.2 6.71±0.1 0.12±0.1 2.8±±0.1 10.3±0.1
P.B 40.8±0.1 6.74±0.1 0.11±0.1 2.32±0.1 10.5±0.2
Values are means of triplicate samples (±SD).
L.M: lupine Milk
P.M: Peanut Milk
L.L: Ice creams inoculated with L.acidophilus made using lupine milk
P.L: Ice creams inoculated with L.acidophilus made using peanut milk
LB: ice creams inoculated with Bifidobacteriumbifidum made using lupine milk
P.B: ice creams inoculated with Bifidobacteriumbifidum made using peanut milk
2.8. Statistical Analysis
The viability of the probiotic micro-organisms were
evaluated at three times of storage (0, 1 and 30 days), in
triplicates and the results were expressed as mean ±S.E.M
(standard mean error) values. The statistical analysis was
performed using Minitab programmer (1998). Three separate
samples were analyzed and mean values were calculated. The
data were assessed by analysis of variance (ANOVA) and
followed by Duncan’s multiple range method for mean
comparison. The mean values and the standard error were
calculated from the data obtained with triplicate trials. The
criterion for statistical significance was (P<0.05)[25].
3. Results and Discussion
3.1. Viability of Probiotic Bactrian Ice Cream
Table3: shows the changes in bacterial counts in non-
fermented ice creams made using vegetable milks. The
survival rate of probiotic bacteria in ice creams after 30 days
tend to be higher in B.bifidum (7.65 and 7.27 logcfug
-1
for
peanut and lupine milk ice cream respectively) than in the
presence of Lactobacullus acidophilus (7.53 and 7.25
logcfug
-1
peanut and lupine milk ice cream respectively)
(Table3). These finding agreed with Fatemeh et al. [27] they
found the survival rate of probiotic bacteria in ice creams
after 30 days tend to be higher in B. bifidum (7.767 and 7.371
log cfug
—
1
for soy and coconut ice cream respectively) than
in L. acidophilus (7.847 and 6.870 log cfug
—
1
for soy and
coconut ice cream respectively) .However, the decline in
viable bacterial counts due to freezing is associated with
freeze injury on these cells. In addition the mechanical
stresses associated with the mixing and freezing process
which incorporates oxygen in to the mixture may be
responsible in further reduction in bacterial count [23]. The
survival of both probiotics in ice cream was high (P<0.01) in
the presence of lupine and peanuts milk. This could be
explained by the high pH of lupine and peanuts ice creams
which are known to be conducive to probiotic survival since
these organisms are susceptible to inactivation when stored in
acidic conditions [28]. As a result probably, these proteins
can cover probiotics as a capsule. The high survival rate of L.
Acidophilus cells during the frozen storage in other studies
was attributed to the protection provided to the cells by the
solid ingredients and the high fat content of the ice cream in
the form of emulsion [29].
Heenan et al.,[16] demonstrated that the survival of
probiotics increased in the frozen soy dessert due to the
prevailing neutral pH. The highest survival of both probiotics
was in lupine milk ice cream. It is probably due to the lupine
milk proteins which provide physical protection against
freezing damage by encapsulating probiotics by form stable
net work looks like a gel structure [26].
3.2. Organoleptic Property
Table 4: shows the organoleptic property scores in non
fermented ice creams made using vegetable milks (Table4).
The presence of either L. Acidophilus or B.bifidum had no
significant influence on the Color and Overall acceptability
properties of both probiotics ice creams. The highest (P≤0.05)
scores of overall acceptability was (3.35) seen in peanut milk
ice cream than the lowest core (3.30) seen in lupine milk ice
cream. It is probably due to the lupine milk beany flavors,
which could be explained by the soy milk woody or beany
off flavors [21].
3.3. Physicochemical Parameters of Milks and Probiotic Ice
Creams Made Using Lupine and Peanut Milk
Table5: The physicochemical compositions of the
probiotic ice creams, lupine and peanut milks are shown in
Table5. The highest (7.15) pH value and lowest (0.70%)
titritable acidity (TA) and fat content (10%) were in ice
creams containing lupine milk. The protein contents varied
between 2.51% to 2.32% in ice creams inoculated with
B.bifidum made using lupine and peanut milk, respectively.
The highest (40.8%) total solids, fat (10.5%) and protein
(40.8%) were found in ice creams containing peanuts milk
and lowest (10.0%) fat content and total acceptability (3.30)
International Journal of Nutrition and Food Sciences 2016; 5(5): 361-366 365
score were found in ice creams containing lupine milk. The
total solids content of 40.8g/100g obtained for sample
containing peanut milk is higher (P≤0.05) than other samples
(39.0g/100g and 38.8g/100g) for sample containing lupine
milk. This may be due the higher content of protein and fat of
peanut milk. Lupine milk based ice creams showed lower
(P≤0.05) fat content than ice cream made with peanut milk.
This may be due to that lupine milk has generally low fat
(3.30g/100g) content compared to that of peanuts milk
(5.0g/100g).
4. Conclusions
Probiotic Food is being one of the largest markets of
functional foods represent a vast growth potential for the
food industry and may be explored through the development
of innovative ingredients, processes, and products. This
research proves that the rear serious confirmations those
supporting develop of new lupine and peanut ice cream
formulation and application of growing probiotic culture
have important role in design of various functional products
including vegetable milk ice cream. Stability during storage
is important characteristics that are proved observing
Probiotic characteristics of produced samples of vegetable
milk ice cream. Moreover, incorporation of lupine and
peanuts milk in ice cream manufacturing resulted in cost
saving and improvement of the nutritional value and
organoleptic quality.
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