LWT - Food Science and Technology 75 (2017) 379e385

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LWT - Food Science and Technology

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(Butia odorata) ice cream: Development,

Probiotic butia
characterization, stability of bioactive compounds, and viability
of Bifidobacterium lactis during storage

,
Claudio Eduardo dos Santos Cruxen, Jessica Fernanda Hoffmann, Giovana Paula Zandona
^
Angela Maria Fiorentini, Cesar Valmor Rombaldi, Fabio Clasen Chaves*
~o do Lea
Agroindustrial Science and Technology Department, Federal University of Pelotas, Campus Capa ~o, s/n, Pelotas, RS, 96010-900, Caixa Postal 354,
Brazil

a r t i c l e i n f o a b s t r a c t

Article history: The addition of fruit with bioactive properties such as Butia odorata, henceforth called butia
, and pro-
Received 3 April 2016 biotic bacteria may improve the nutritional and functional value of ice cream. In this study, butia
ice
Received in revised form cream supplemented with Bifidobacterium lactis (Bl-04) was developed and characterized. Butia
ice
7 September 2016
cream showed 71.2 g 100 g
1 of overrun, a density of 575 g cm
3, and a pH of 4.7. During 90 days of
Accepted 8 September 2016
Available online 13 September 2016
frozen storage (
18  C), a 10% reduction in total phenolic content and a 5% reduction in L-ascorbic acid
content were observed. Carotenoids and flavonoids increased during frozen storage. Probiotic bacteria B.
lactis (Bl-04) remained viable (above 106 CFU g
1) in butia
ice cream during storage, confirming the
Keywords:
probiotic characteristic of the product. Sensory analyses showed that butia
ice cream had high accept-
Processing
Native fruit ability (6.8e7.8) and purchase intent (above 80%). The combination of B. lactis and butia
fruit attributed
Probiotic functional characteristics to the ice cream.
Functional food © 2016 Elsevier Ltd. All rights reserved.
Bifidobacterium lactis

1. Introduction incorporation of ingredients that promote health, flavor, and con-

venience to consumers in well-accepted products (Khurana &
Ice cream is a dairy product containing milk, sweeteners, sta- Kanawjia, 2007). Thus, the combination of fruit containing bioac-
bilizers, emulsifiers, and flavoring agents produced by the mixing of tive properties and the benefits from ingestion of probiotic bacteria
these ingredients, followed by pasteurization and freezing, to can impart a functional food characteristic to ice cream. The
ensure the retention of the product in the frozen or partially frozen development of new products containing native fruits is recom-
state during storage, transport, and delivery to the consumer mended and promoted by the United Nations Food and Agriculture
(Karaman & Kayacier, 2012). However, this dairy-based food is Organization for the indispensable integration of social, economic,
considered poor in bioactive compounds such as carotenoids, and environmental concerns involving food (FAO, 2008).
vitamin C, and phenolic compounds (Sun-Waterhouse, Edmonds,
, is a
Jelly or pindo palm fruit (Butia odorata), also known as butia
Wadhwa, & Wibisono, 2013). Currently, in order to meet con- fruit native to South America containing bioactive properties with
sumer demands, a novel food product must combine functional and potential for commercialization (Barbieri, Costa Gomes, Alercia, &
nutritional properties as well as meet sensory expectations of the Padulosi, 2014; Hoffmann, Barbieri, Rombaldi, & Chaves, 2014). It
consumer. The nutritional and functional value of foods can be has sweet, acidic, fibrous pulp, rich in bioactive compounds
improved by the addition of antioxidants, vitamins, minerals, and including ascorbic acid, carotenoids (provitamin A), and phenolic
bacteria with probiotic potential (Cruz, Antunes, Sousa, Faria, & compounds (Beskow et al., 2015). These bioactive compounds have
Saad, 2009). been associated with several biological effects, including antioxi-
The dairy product market can be further expanded by the dant, antimicrobial, anti-inflammatory, vasodilator, and the
reduction of the development of several chronic diseases (Fraga,
Galleano, Verstraeten, & Oteiza, 2010; Yahia, 2009).
Microorganisms are currently being used to impart functional
* Corresponding author. characteristics to foods. Probiotics are live microorganisms which,
E-mail address: fabio.chaves@ufpel.edu.br (F.C. Chaves).

http://dx.doi.org/10.1016/j.lwt.2016.09.011
0023-6438/© 2016 Elsevier Ltd. All rights reserved.
380 C.E. dos Santos Cruxen et al. / LWT - Food Science and Technology 75 (2017) 379e385

when administered in adequate amounts, confer health benefits to concentrations were evaluated by sensory analysis and for bioac-
the host (FAO, 2001). Some of the beneficial effects promoted by the tive compound content in order to determine the best formulation
ingestion of probiotic bacteria include: antimicrobial activity, to be inoculated with the probiotic bacteria. Bioactive compounds
reduction of lactose intolerance, prevention and reduction of the content and sensory analyses of the ice creams made with the three
development of diarrhea, prevention of inflammatory bowel dis- pulp concentrations were carried out on the 7th day of storage
ease, and a hypocholesterolemic effect (Vasiljevic & Shah, 2008). at
18  C. The methodological procedures for these analyzes are
Bifidobacterium is a genus of anaerobic, gram-positive, catalase- described below in 2.4.3 and 2.4.5.
negative, strictly fermentative, probiotic bacteria. Bifidobacterium
animalis subsp. lactis exhibits elevated oxygen tolerance, and 2.3. Addition of Bifidobacterium lactis (Bl-04) to ice cream made
therefore is widely used in dairy foods (Meile et al., 1997). In order
pulp
with butia
to achieve a therapeutic effect, a minimum intake of a daily dose of
at least 108 to 109 viable cells is required. Such a dose could be After ice cream formulation optimization, B. lactis was added to
achieved with a daily intake of at least 100 g of a product containing the product. Lyophilized cells of B. lactis were resuspended in whole
between 106 and 107 viable cells g
1 (Gomes & Malcata, 1999). milk at a ratio of 1:30 (1 g lyophilized culture to 30 mL of sterile
The objective of this study was to develop and characterize butia
whole milk) and incubated in a shaker (Agimaxx®) at 150 rpm for
ice cream supplemented with B. lactis and to evaluate its physico- 30 min at 37  C. Immediately after the incorporation of air in the ice
chemical characteristics, the stability of bioactive compounds, and cream, 0.1% of the probiotic culture (1012 CFU g
1) was added to
the viability of probiotic bacteria during 90 days of storage make up an initial concentration of bacteria of 109 CFU g
1. The ice
at
18  C. cream was stirred with a spatula for 30 s for proper homogeniza-
tion of the probiotic bacteria. Ice cream samples were placed in
2. Materials and methods 100 mL plastic containers and stored at
18  C until analysis.
Samples of butia
ice cream supplemented with B. lactis were
2.1. Materials evaluated for density, overrun, melting, chemical composition, pH,
acidity, stability of bioactive compounds, and viability of B. lactis
Whole ultra high temperature (UHT) milk (Danby®), cream of during 90 days of storage (
18  C). A control ice cream formulation
milk (Danby®), whole milk powder (Danby®), ice cream emulsifier was prepared as described above without addition of B. lactis.
and stabilizer (Emustab, Selecta®), a mix of guar gum and
carboxymethylcellulose (Super Liga Neutra, Selecta®), and sugar
ice cream supplemented with B. lactis
2.4. Analyses of butia
(Cristal®) were purchased at a local supermarket in Pelotas, RS.
Bifidobacterium lactis (Bl-04) culture was purchased from Danisco®. 2.4.1. Chemical composition, pH, and acidity
Butia
fruit pulp was prepared with fruit picked from a pool of
Water, fat, ash, and protein contents, pH, and acidity were
Butia odorata plants at the Palma Agricultural Center of the Federal determined according to AOAC (2005).
University of Pelotas. After harvest, fruit were washed, sanitized
with sodium hypochlorite (150 mg L
1 for 15 min), de pulped in a
2.4.2. Density, overrun, and melting properties
horizontal depulper, and passed through a sieve with a
The apparent density (AD) was calculated using the following
3.5 mm mesh followed by a 0.5 mm mesh. Sieved pulp was heated
equation:
at 95e100  C for 15 min, cooled, frozen, and stored at
18  C in
sealed polypropylene bags for 12 months until it was used in the ice h .  i
AD ¼ sample weight ðgÞ occupied volume cm3
cream formulations. Stored pulp contained 11.1 g 100 g
1 of total
soluble solids, with a pH of 3.2, acidity of 1.7 g citric acid 100 g
1,
The overrun was calculated according to Daw and Hartel (2015),
14.5 mg b-carotene equivalents 100 g
1, 85.9 mg gallic acid
using the equation:
equivalents 100 g
1, 58.1 mg (
)-epicatechin equivalents 100 g
1,
and 32.3 mg L-ascorbic acid 100 g
1. In order to achieve the desired Overrun ð%Þ ¼ ð½Final volume
initial volume=initial volumeÞ
total soluble solids concentration of 17 g 100 g
1, the pulp was
concentrated by heating and the final pulp characteristics were: pH  100
of 3.5, acidity of 2.4 g citric acid 100 g
1, 21.2 mg b-carotene
equivalents 100 g
1, 133 mg gallic acid equivalents 100 g
1, 89 mg The melting test was performed as described by Muse and
(
)-epicatechin equivalents 100 g
1, 68 mg L-ascorbic acid 100 g
1, Hartel (2004), with adaptations. After 15 days of storage
and 89.4% of DPPH radical inhibition capacity. at
18  C, 80 g of ice cream was placed on screens (4 holes cm
2),
allowed to melt into a graduated cylinder below, and the volume

pulp
2.2. Optimization of ice cream formulation with butia collected was recorded every 5 min. The temperature used for the
experiment was 25  C ± 1  C. The melted volume (mL) was plotted
Three butia
pulp concentrations were used in the ice cream against time (min).
formulation: treatment 1 contained 30 g 100 g
1, treatment 2
contained 40 g 100 g
1, and treatment 3 contained 50 g 100 g
1.
ice cream
2.4.3. Bioactive compounds content in probiotic butia
Initially, 1L of whole milk was heated to 50  C and mixed with Bioactive compounds content was evaluated in butia
ice cream
whole milk powder (10 g 100 g
1). Subsequently, cream of milk upon preparation and after 90 days of frozen storage (
18  C).
(10 g 100 g
1) and sugar (20 g 100 g
1) were incorporated. The mix
of guar gum and carboxymethylcellulose was added (1 g 100 g
1) 2.4.3.1. Preparation of extracts for phenolic, flavonoid, and antioxi-
and heated at 80  C for 5 min. The slurry was incubated for 24 h at dant analyses. Two g of each ice cream sample were homogenized
a pulp (30, 40, or 50 g 100 g
1)
4  C (maturation process). Then, buti
with 20 mL of methanol for 1 min using Ultra Turrax (IKA®, digital
and emulsifier (1 g 100 g
1) were added followed by mixing with T18), and subsequently centrifuged for 10 min at 6700g at 15  C.
an electric mixer (Arno®) for 5 min to homogenize and promote air The supernatant was separated and used as a crude extract to
incorporation. Ice cream was stored in 100 g flasks at
18  C. determine total phenolic content, total flavonoid content, and
Ice creams prepared with the three different pulp antioxidant capacity.
 C.E. dos Santos Cruxen et al. / LWT - Food Science and Technology 75 (2017) 379e385 381

2.4.3.2. Total phenolic content. Total phenolic content was deter- individual booths under fluorescent white light (85% relative hu-
mined using the Folin-Ciocalteu reagent, with absorbance midity, 25  C) with 50 untrained panelists between 10 and 11 a.m.
measured at 725 nm in a spectrophotometer (Jenway, 6700), and The panelists were students, professors, and staff from UFPel,
results were expressed in equivalent mg of gallic acid 100 g
1 fresh 20e45 years old, male and female, all regular ice cream consumers
sample based on a calibration curve using a gallic acid standard tolerant to lactose and willing to participate. Following the regu-
(Sigma-Aldrich) (Beskow et al., 2015). lation of Ethics Committee of the Faculty of Medicine, Federal
University of Pelotas (project number 38484814.0.0000.5317), all
2.4.3.3. Flavonoid content. Total flavonoid content was determined participants were informed of every detail of the scope of the
according to the method of Zhishen, Mengcheng, and Jianming research. Before the analysis, the ice cream was removed from the
(1999). Absorbance was measured at 510 nm and results were freezer and kept at room temperature (25  C) for 10 min.
expressed as mg (
)-epicatechin 100 g
1 fresh sample based on a
calibration curve using an (
)-epicatechin standard (Sigma-
ice cream with different pulp con-
2.4.5.1. Sensory analysis of butia
Aldrich). centrations. Sensory analysis was carried out using the acceptance
testing whereby ice cream samples were evaluated for color, aroma,
2.4.3.4. Antioxidant capacity. Ice cream antioxidant capacity was flavor, texture, and overall quality using a hedonic scale ranging
determined by the method described by Brand-Williams, Cuvelier, from 1 (disliked it very much) to 9 (liked it very much). In the
and Berset (1995) using the free radical 2,2-diphenyl-1-picrylhy- preference test, panelists were requested to list the samples in or-
drazil (DPPH). Absorbance was measured at 517 nm after 24 h of der of preference, assigning the following grade: 3 (most preferred
reaction. Results were expressed as percentage of inhibition of formulation), 2 (intermediate), and 1 (least preferred formulation).
DPPH radical.

ice cream. To perform the
2.4.5.2. Sensory analysis of probiotic butia
2.4.3.5. Carotenoid content. Total carotenoid content was deter- sensory analysis of the final product, 2 L of ice cream was produced
mined by spectrophotometry (AOAC, 2005). Two grams of sample as previously described, supplemented with B. lactis, fractionated
were extracted with 15 mL of an extracting solution (hex- into 50 mL pots and stored at
18  C until the sensory analysis. For
ane:acetone:ethanol:toluene, 10:7:6:7), and vortexed for 1 min. the acceptance testing, ice cream samples were evaluated for color,
Then, 1 mL of KOH in methanol (10% m/v) was added and the flask aroma, flavor, texture, and overall quality using a hedonic scale
placed in a water bath at 56  C for 20 min. The mixture was kept at ranging from 1 (disliked it very much) to 9 (liked it very much).
room temperature in the dark for 1 h. Then 15 mL of hexane was Purchase intent was conducted using an intention scale ranging
added, vortexed for 1 min, and 19 mL of Na2SO4 (10% m/v) was from 1 (would definitely not buy) to 5 (would definitely buy).
added and subsequently vortexed for 1 min. The mixture was kept
at room temperature in the dark for 1 h and the absorbance of the 2.5. Statistical analysis
upper phase was measured at 450 nm. Results were expressed in
equivalent mg of b-carotene 100 g
1 fresh sample based on a The experimental design was completely randomized. The
calibration curve using a b-carotene standard (Sigma-Aldrich). experiment consisted of two replicates, and all analyses were per-
formed in triplicate. For pulp concentration optimization, a uni-
2.4.3.6. L-ascorbic acid content. For the extraction of L-ascorbic acid, variate analysis comparing three treatments was used. To examine
15 mL of extraction solution (3% w/v metaphosphoric acid and 8% v/ the effect of frozen storage on probiotic ice cream, a univariate
v acetic acid in ultrapure water) was added to 2 g of the sample. The analysis comparing 0 and 90 days of frozen storage was used. Data
mixture was homogenized and centrifuged at 6700g for 15 min. were analyzed for normality by the Shapiro-Wilk's test, homosce-
The supernatant was filtered using a nylon syringe filter (0.45 mM). dasticity by Hartley's test, and examined for violations of as-
Ten microliters of the extract were injected into the HPLC (Shi- sumptions by inspection of plots of standardized residuals. Then
madzu, Prominence). L-ascorbic acid was separated using a C18 data were subjected to analysis of variance (p  0.05), and in case of
guard column (2.0  4 mm) and a C18 column (Luna; 2.0  150 mm, statistical significance, confidence intervals (95%), Tukey's test
100 Å, 3 mm) (Phenomenex, Torrance, CA, USA), with formic acid in (p  0.05), or LSD test (p  0.05) were applied.
water (0.1% v/v eluent A) and methanol (eluent B) used as mobile
phases at a flow rate of 0.2 mL min
1 and column temperature of 3. Results and discussion
30  C. An isocratic elution with 1% B was used for a 10 min total run
time. The PDA detector was set to scan in the 210e370 nm range 3.1. Formulation optimization
(Vinci, Rot, Mele, & Ruggieri, 1995). Results were expressed in mg of

1
ice cream formulations with pulp concentrations of
L-ascorbic acid 100 g fresh sample based on a calibration curve Three butia
using an L-ascorbic acid standard (Sigma-Aldrich). 30, 40, and 50% were subjected to sensory analyses (preference and
acceptance tests). The panelists did not show a preference for any
2.4.4. Viability of B. lactis specific formulation nor did they differentiate among the three
The viability of B. lactis was determined immediately after ice products based on overall quality attributes, which were consid-
cream preparation, after 2, 8, and 24 h of frozen storage, and after 7, ered high for all samples (Table 1). When looking at each attribute
15, 30, 60, and 90 days of frozen storage. For viable cell count separately, aroma and flavor did not differ among treatments, while
determination, 10 g of sample was homogenized in 90 mL of 0.1% color was graded distinctly higher in the formulations with 40 and
peptone water (Oxoid®). Then, serial decimal dilutions were per- 50% buti
a pulp. Texture had the highest acceptability in the
formed and 0.1 mL of the suspension was inoculated on Petri plates formulation with 30% pulp. Panelists commented that they ex-
containing Man Rogosa Sharpe - MRS agar (Acumedia®). The plates pected a more pronounced fruit aroma but did not experience it. In
were placed under anaerobic conditions and incubated for 48 h at a commercial product, this attribute will likely need to be improved
37  C. to fulfill consumer expectation. Regarding color, the higher the pulp
concentration, the more intense the yellow color of the product.
2.4.5. Sensory analysis The texture of the products made with both 40 and 50% butia
pulp
Sensory analysis was carried out in a laboratory environment in was affected by the pulp water content, which lead to the formation
382 C.E. dos Santos Cruxen et al. / LWT - Food Science and Technology 75 (2017) 379e385

Table 1 Senaka Ranadheera, Evans, Adams, & Baines, 2013). Buti
a ice

ice cream with different pulp
Mean panelist acceptability and preference of butia cream contained less protein than ice cream in other studies, but
concentrations.
this was expected given the high concentration (40 g 100 g
1) of
Attributesa Pulp concentration low protein-containing pulp that was added to the product.
30% 40% 50% Nonetheless, the amount of protein present was sufficient to
contribute to the structural and textural properties of the product
Color 6.8 b 7.6 a 7.8 a
Aroma 5.9 a 6.3 a 6.3 a including emulsification, aeration, water retention, and viscosity
Flavor 7.0 a 6.8 a 7.0 a

(Cheng, Ma, Li, Yan, & Cui, 2015). The carbohydrates present in butia
Texture 7.4 a 6.8 ab 6.5 b ice cream (19.0 g 100 g
1), which mainly originated from the
Overall quality 8.8 a 8.7 a 8.8 a
lactose and sucrose added to the formulation, contributed to the
Preferenceb 2.1 a 1.9 a 2.0 a
desired sweet taste and assisted in fixing aromas and providing
Means followed by the same letter within a row are not significantly different by flavor intensity (Karaman et al., 2014). The non-fat solids (protein,
Tukey's test (p  0.05).
a
Rated from 1 to 9 (1 e disliked very much, 9 e liked very much).
lactose, and minerals) play an important role in palatability by
b
Ranked from 1 to 3 (1 e least preferred, 3 e most preferred). influencing taste during the residence time in the mouth, as well as
helping to lower the freezing point and increase the viscosity of the
product (Cheng et al., 2015).
of large ice crystals that were perceived in the sensory analysis.
Karaman et al. (2014) also used hedonic scales to evaluate ice cream 3.2.2. Overrun, density, and meltdown tests
enriched with persimmon puree at concentrations varying up to The overrun is an important characteristic of the product, since
40% (w/w). The authors found that concentrations higher than 24% the presence of air in ice cream gives it a light and pleasant texture,
has lower overall acceptance. and influences the physical characteristics such as hardness and
To assist in the choice of formulation that would receive the B. meltdown (Sofjan & Hartel, 2004). In this study, overrun and
lactis treatment, the concentration of the bioactive compounds density were 71.3 ± 4.9 g 100 g
1 and 575 ± 25 g cm
3, respectively.
present in the butia
ice cream prepared with different pulp con- Magarin ~ os, Selaive, Costa, Flores, and Pizarro (2007) obtained
centrations was measured after seven days of cold storage (Table 2). overrun values ranging from 106 to 108 g 100 g
1 and a density of
Ice cream formulations with 40 and 50% butia
pulp had higher 1076.7 ± 28.9 g cm
3 in ice cream formulations supplemented with
concentrations of flavonoids and phenolic compounds than the ice probiotic bacteria. Variation in the overrun can be attributed to
cream with 30% pulp. Ice cream made with 50% pulp had the several factors such as fat, protein, emulsifier, and non-fat solids
highest water content (70%). Therefore, given the high acceptability contents (Sun-Waterhouse et al., 2013). The freezing process may
by tasters, the concentration of bioactive compounds, and the pulp also have influenced the final product, because discontinuous
water content, ice cream made with 40% pulp was chosen to be freezing, used in the present work, is less efficient for air incorpo-
supplemented with the probiotic B. lactis (Bl-04). ration when compared to continuous freezing (Akalin & Erişir,
2008).

ice cream supplemented with B. lactis
3.2. Analysis of butia The melting of ice cream can be influenced by many variables,
such as overrun, emulsifying characteristics, and concentration of
3.2.1. Proximate composition, pH, and acidity lipids and proteins (Sofjan & Hartel, 2004). Butia
ice cream sup-

ice cream supplemented with B. lactis had the following
Butia plemented with B. lactis began melting after 15 min and reached
composition: 65.6 ± 1.3 g 100 g
1 water, 1.1 ± 0.03 g 100 g
1 pro- complete melting (80 g) after 60 min (Fig. 1). While no information
tein, 0.98 ± 0.03 g 100 g
1 ash, 13.6 ± 1.0 g 100 g
1 lipids, on buti
a (or related fruit) ice cream is available currently, ice cream
19.0 g ± 1.3100 g
1 carbohydrates, and 20.8 ± 1.3 g 100 g
1 non-fat made with goat's milk started melting at 5 min and was completely
solids. Total caloric value was 200 ± 5.2 kcal 100 g
1, pH 4.7 ± 0.01, melted (100 g) at 40 min (da Silva et al., 2014), while green kiwi ice
and acidity 0.07 ± 0.002 g 100 g
1 lactic acid. cream (120 g) started melting after at 20 min and complete melting
The composition of ice cream may vary, but it typically contains was observed at 145 min (Sun-Waterhouse et al., 2013).
from 8 to 20 g 100 g
1 lipids, 8e15 g 100 g
1 of non-fat solids,
13e20 g 100 g
1 of carbohydrates, and 55e65 g 100 g
1 of water
pulp upon processing
3.2.3. Bioactive compounds stability in butia
(Arbuckle, 1986). The lipids present in ice cream are important for
ice cream after frozen storage
and in probiotic butia
the development of desirable characteristics including smooth Flavonoids are phenolic compounds that play an important role
texture, viscosity, and flavor (Ohmes, Marshall, & Heymann, 1998). in the sensory and functional quality of foods (Linderborg,
Several studies have shown that the amount of protein in ice cream Laaksonen, Kallio, & Yang, 2011). Total phenolic compound and
typically ranges from 2.5 to 4.5 g 100 g
1 (da Silva, Bezerra, dos flavonoid contents were higher in the ice cream product than in the
Santos, & Correia, 2014; Erkaya, Dag demir, & Sengül, 2012; pulp; however, after 90 days of storage at
18  C, phenolic content

Table 2

ice cream prepared with three different pulp concentrations.
Bioactive compounds content in butia

Pulp (%) Water (%) Flavonoida Phenolicb Carotenoidc L-ascorbic acidd Scavenging of DPPHe

30 62 c 61.8 ± 2.4 b 48.9 ± 1.2 c 6.9 ± 1.7 a 19.1 ± 2.3 a 17.6 ± 1.1 b
40 68 b 71.5 ± 2.6 a 60.2 ± 2.7 b 8.8 ± 2.2 a 25.0 ± 4.1 a 20.5 ± 1.4 ab
50 70 a 70.7 ± 4.7 a 66.6 ± 2.6 a 8.3 ± 1.8 a 22.9 ± 1.9 a 23.8 ± 1.7 a

Results expressed as mean ± standard deviation.

Means followed by the same letter within a column are not significantly different by Tukey's test (p  0.05).
a
mg (
)-epicatechin equivalents 100 g
1 fresh weight (fw).
b
mg gallic acid equivalents 100 g
1 fw.
c
mg b-carotene equivalents 100 g
1 fw.
d
mg L-ascorbic acid 100 g
1 fw.
e
Expressed as % inhibition.
 C.E. dos Santos Cruxen et al. / LWT - Food Science and Technology 75 (2017) 379e385 383

80 Bruhn, & Barrett, 2007). In this study, carotenoid content was

higher after processing and 90 days of storage at
18  C (Table 3).
Different studies have reported the increase of carotenoid content
upon processing and storage. For example, lycopene was shown to
60 be 27% higher upon tomato processing (Re et al., 2002). An increase
in b-carotene (102%) was observed in carrots under frozen storage
Melted volume

(Guerra-Vargas, Jaramillo-Flores, Dorantes-Alvarez, & Hernandez-

(mL/80mL)

40
Sanchez, 2001). These increases can be attributed to isomeriza-
tion or higher carotenoid extractability from tissues (Rickman,
Bruhn et al., 2007). The average daily intake of carotenoids from
natural sources is estimated to be about 2 mg per day (Rickman,
20 Bruhn et al., 2007). Therefore, Buti

a ice cream can be considered
to be a significant source of carotenoids.

3.2.4. Viability of B. lactis in butia
ice cream

0
0 5 10 15 20 25 30 35 40 45 50 55 60 No significant reduction in the concentration of B. lactis was
observed within the first 24 h of frozen storage (Fig. 2A). Usually the
Time (minutes)
largest decrease in microorganisms occurs within the first few

ice cream supplemented with Bifidobacterium lactis (Bl-
Fig. 1. Melting curve for butia hours of processing and cold storage due to the formation of ice
04) at 25  C and 85% relative humidity. Error bars represent ± standard deviation. crystals, thermal shock, and mechanical action by the air incorpo-
ration process (Turgut & Cakmakci, 2009). da Silva et al. (2014)
in buti
a ice cream supplemented with B. lactis was reduced by 10% found that ice cream supplemented with B. animalis subsp. lactis
while flavonoids increased by 31% (Table 3). Similarly, an accu- showed a reduction of more than 1.0 Log CFU g
1 in the first 2 h of
mulation of anthocyanins was observed in strawberries and rasp- storage at
18  C. In this study, B. lactis remained viable with counts
berries when fruit were stored at 20  C (Kalt, 2005). The reduction exceeding 106e107 Log CFU g
1 during ninety days of storage
in phenolic content during frozen storage may have occurred due to at
18  C, confirming the probiotic potential of this food product
oxidation and chemical degradation (Rickman, Barrett, & Bruhn, (Fig. 2B).
2007). On the other hand, Gahler, Otto, and Bohm (2003) found a The viability of probiotic bacteria added to a food matrix, ice
44% gain in total phenolics during the baking of tomatoes and up to cream in particular, depends on various factors such as the type of
a 64% increase in total phenolics during the cooking of tomato culture that is being added, the product acidity, overrun, and
sauce. Re, Bramley, and Rice-Evans (2002) also observed a 60% in- presence of cryoprotectors. Acidic foods are usually poor carriers of
crease in chlorogenic acid content in tomatoes upon processing. bacteria of the genus Bifidobacterium (Charteris, Kelly, Morelli, &
Phenolic compound content is highly dependent on the commodity Collins, 2002). Butia
ice cream, while acidic, did not affect the
and processing method (Rickman, Barrett et al., 2007). A possible viability of B. lactis. A possible explanation is that at freezing tem-
explanation for the increase seen in phenolic compound and peratures, the low molecular motion of the system limited the
flavonoid contents is that the elevated temperatures during pro- interaction between acidic molecules and bacterial cells. In this
cessing may have made the compounds more available, or dehy- study, air incorporation did not affect the viability of B. lactis,
dration may have concentrated the compounds. possibly due to the fact that the addition of bacterial cells occurred
L-Ascorbic acid (AA) content is considered a marker to estimate
after air incorporation and was immediately followed by freezing,
nutrient loss in food products due to its high susceptibility to thereby avoiding mechanical damage. There is evidence that some
degradation by oxidation (Rickman, Barrett et al., 2007). In this species such as B. lactis can withstand the presence of oxygen
study, no significant differences were observed in L-ascorbic acid (Meile et al., 1997). Shin and Park, (1997) report that aero tolerance
content after processing and storage (Table 3). Antioxidant capacity of some Bifidobacterium species can be obtained by detoxification of
of hydrophobic extracts is generally highly correlated with phenolic H2O2 by NADH oxidative enzymes. Moreover, it is also possible that
compounds and L-ascorbic acid content (Yahia, 2009). In this study, the presence of carbohydrates (lactose and sucrose) and protein
butia
ice cream supplemented with probiotic bacteria had a low (casein) acted as cryoprotectants and preserved cells during frozen
antioxidant capacity against DPPH radical and was further reduced storage (Magarin ~ os et al., 2007; Vasiljevic & Shah, 2008). No B. lactis
upon both processing and storage (Tables 2 and 3). Butia
ice cream was detected in the control ice cream without bacterial inoculation.
was not found to be a relevant source of phenolic, flavonoid, and L-
ascorbic acid contents. 3.2.5. Sensory analysis of probiotic ice cream
Carotenoids are fat-soluble compounds and are more stable Taste score is an appropriate way to demonstrate the sensory
under storage, yet may suffer degradation due to light, tempera- characteristics of a product or to compare formulations. Soukoulis,
ture, and amount of available oxygen during processing (Rickman, Lyroni, and Tzia (2010), for example, evaluated consumer

Table 3

pulp upon ice cream making (processing) and after 90 days of storage at
18  C.
Variation (%) of bioactive content in butia

Flavonoid content Phenolic content Carotenoid content L-ascorbic acid content Antioxidant capacity

Processinga þ100* þ13* þ4 ns
8 ns
43*

Storagec þ31*
10* þ16*
5 ns
7*

*significant difference by LSD test at p  0.05.

ns e not significant.
a
Processing variation % ¼ (pulp content e ice cream contentb)  100.
b
60% dilution was considered given that ice cream was prepared with 40% of pulp.
c
Storage variation % ¼ (ice cream content at the beginning of the storage e ice cream content at the end of storage)  100.
384 C.E. dos Santos Cruxen et al. / LWT - Food Science and Technology 75 (2017) 379e385

10.0 10.0
A CI 0.05 = 0.21 B CI 0.05 = 0.11

9.5 9.5
log.CFU/g

log.CFU/g
9.0 9.0

8.5 8.5

8.0 8.0
0 2 8 24 7 15 30 60 90

Time (hours) Time (days)

ice cream during the first 24 h of storage (A) and over 90 days of storage at
18  C (B). Vertical bars represent ± standard deviation.
Fig. 2. Bifidobacterium lactis viability in butia
CI ¼ confidence interval (95%).

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