LWT - Food Science and Technology 90 (2018) 270–276

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LWT - Food Science and Technology

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Spirulina for snack enrichment: Nutritional, physical and sensory evaluations T

a b a
Bárbara Franco Lucas , Michele Greque de Morais , Thaisa Duarte Santos ,
Jorge Alberto Vieira Costaa,∗
a
Laboratory of Biochemical Engineering, College of Chemistry and Food Engineering, Federal University of Rio Grande (FURG), P.O. Box 474, 96203-900, Rio Grande,
RS, Brazil
b
Laboratory of Microbiology and Biochemistry, College of Chemistry and Food Engineering, Federal University of Rio Grande (FURG), P.O. Box 474, 96203-900, Rio
Grande, RS, Brazil

A R T I C L E I N F O A B S T R A C T

Keywords: The aim of this work was to develop snacks enriched with Spirulina and thus improve their nutritional char-
Extruded acteristics. Two formulations were developed: SP added with 2.6% Spirulina sp. LEB 18 and C (0% Spirulina).
Extrusion These snacks were evaluated with respect to the nutritional content (protein, lipids, ash, carbohydrates, car-
Microalga otenoids and in vitro protein digestibility), physical properties (expansion index, bulk density, hardness, water
Nutrition
absorption index, water solubility index, microstructure and color parameters) and sensorial characteristics
Sensory acceptance
(flavor, color, taste, texture, overall acceptance and purchase intention), as well as microbiological analyses. The
addition of Spirulina provided nutritional increase of 22.6% in proteins, 28.1% in lipids and 46.4% in minerals,
without significantly affecting (p > .05) the physical parameters such as expansion index and hardness. The
pigments of such microalga significantly influenced (p < .05) the color parameters evaluated. Microstructure
evaluation of the formulation added with Spirulina showed structures with thin cell walls (18.50 ± 1.50 μm).
Furthermore, this formulation resulted in a sensory acceptance index of 82%. It was concluded that Spirulina can
be used in the concentration of 2.6% resulting in snacks with high nutritional content and sensory acceptance.
Thus, this product can be used as ready-to-eat food by consumers which look for a healthier diet.

1. Introduction
Microalgae have been used in human nutrition for thousands of
years (Spolaore, Joannis-Cassan, Duran, & Isambert, 2006; Vigani et al.,
2015). These microorganisms are capable of accumulating compounds
of high nutritional value, such as essential fatty acids, essential amino
acids, phycocyanin and carotenoids (Spolaore et al., 2006). The mi-
croalga Spirulina has received GRAS certification (Generally Recognized
As Safe) by the Food and Drug Administration (FDA) and had its con-
sumption authorized as food or food supplement (FDA, 2002). Several
studies using Spirulina in foods have been performed. Among the de-
veloped foods, there are cookies (Morais, Miranda, & Costa, 2006),
shake for the elderly (Santos, Freitas, Moreira, Zanfonato, & Costa,
2016) and extruded snacks (Joshi, Bera, & Panesar, 2014).
In recent years, the demand for snacks with improved nutritional
and functional properties has increased (Rathod & Annapure, 2016),
due to the high practicality, convenience and acceptability of these
foods, especially among children (Potter, Stojceska, & Plunkett, 2013).
Usually, the extrudates available on the market have low nutritional
value, with reduced protein and biocompounds concentrations (Anton,
Fulcher, & Arntfield, 2009; Sumargo, Gulati, Weier, Clarke, & Rose,
2016). Thus, the development of snacks enriched with Spirulina sp. LEB
18 can combine the versatility and convenience of extrudates with the
nutritional quality of this microalga.
The extrusion process used in the snacks production has advantages
such as rapid processing, lower cost and greater flexibility, leading to
minimal degradation of nutrients and inactivation of undesirable
compounds (Anton et al., 2009). This process uses high temperature
during short time (HTST) to modify the raw materials, changing me-
chanical and sensorial properties, and improving digestibility (Lazou &
Krokida, 2010; Pastor-Cavada et al., 2011). In the extrudates formula-
tion, physical properties such as expansion, hardness and bulk density
are important parameters in terms of sensorial acceptance. Among the
sensorial attributes, texture plays a major role in the evaluation of ex-
trudates (Pamies, Roudaut, Dacremont, Le Meste, & Mitchell, 2000).
Studies have investigated the influence of different food sources on
the enrichment of snacks (Basto et al., 2016; Cian, Caballero, Sabbag,
González, & Drago, 2014; Morsy, Sharoba, El-Desouky, Bahlol, & Abd El
Mawla, 2014; Potter et al., 2013), however, little information is found
about the effect of Spirulina on the nutritional and sensory quality of

∗
Corresponding author.
E-mail address: jorgealbertovc@terra.com.br (J.A.V. Costa).

https://doi.org/10.1016/j.lwt.2017.12.032
Received 31 August 2017; Received in revised form 3 December 2017; Accepted 12 December 2017
Available online 13 December 2017
0023-6438/ © 2017 Elsevier Ltd. All rights reserved.
B.F. Lucas et al.
these products (Morsy et al., 2014; Joshi et al., 2014). In this context,
the aim of this work was to develop snacks enriched with Spirulina sp.
LEB 18 and thus improve their nutritional characteristics.
2. Materials and methods
2.1. Raw material
Spirulina sp. LEB 18 biomass was produced at the Pilot Plant of the
Laboratory of Biochemical Engineering and isolated from Mangueira
Lagoon (Morais et al., 2008) located in Rio Grande do Sul, Brazil
(33°30′13″S and 53°08 ′59″W). The organic flours used were corn flour
(Ecobio, Coronel Bicaco, Brazil) and rice flour (Volkmann Alimentos
Ltda, Camaquã, Brazil).
The biomass was ground in a ball mill (Model Q298, Quimis, Brazil)
and sieved to granulometry of 0.3 mm. Then, the biomass was vacuum
packed (Model Supervac 400, Sulpack, Brazil) and stored under re-
frigeration conditions (7 °C).
2.2. Sample preparation
Two samples were prepared: C and SP. The control sample (C) was
formulated with a 2:1 ratio of organic rice flour and organic corn flour,
respectively. The sample SP was prepared by addition of 2.6% Spirulina
sp. LEB 18 in the control formulation. This concentration of microalga
was defined in preliminary studies (Lucas, Morais, Santos, & Costa,
2017).
The ingredients were homogenized with filtered water using pla-
netary mixer (Kitchen Aid Professional, Model K45SS, Cameron Park,
Australia). Corrections of deviations in moisture were performed until
the mixture reached 16.2% of moisture (Lucas et al., 2017). Afterwards,
the samples were packed in polypropylene packages and stored in a
refrigerator at 7 °C for 24 h.
2.3. Extrusion
The extrusion trials were performed using a co-rotating twin-screw
extruder (Werner Pfleiderer Co., Model ZSK-30, Ramsey, USA). Results
from preliminary studies (Lucas et al., 2017) were used to select the raw
material levels and the suitable extruder operating conditions. Feed rate
(12.6 kg h−1), screw speed (250 rpm) and temperature of zones 1, 2, 3
and 4 (75, 100, 120 and 143 °C, respectively) were fixed.
The ratio L/D of the screw was 29.07 and a circular die with dia-
meter of 3.8 mm was used. The following screw configuration, which
was composed of transport and mixing elements was used: 2 elements
60/30; 2 elements 42/21; 1 element 28/14; 1 element kneading block
90/5/28; 1 element 21/21; 1 element 28/14; 4 elements 20/10; 1
element kneading block 90/5/28; 1 element 21/21; 1 element 28/14; 5
elements 20/10; 1 element 28/14; 1 element 14/14; 1 element
kneading block 45/5/14 flow inverter; 4 elements 20/10; 1 element
28/14; 1 element 10/10 flow inverter; 3 elements 20/10; 1 kneading
block 45/5/20; 3 elements 20/10. The extruded were dried in a forced
air oven at 85 °C (Tecnal, Model TE-394/2, Piracicaba, Brazil) to obtain
a product with less than 6% moisture content.
2.4. Flavoring
Snacks were flavored in a dragee pan (Siaht, model Siaht 10L,
Jundiaí, Brazil), by aspersion of 5% sunflower oil using pneumatic
spray. Afterwards, a mixture of 2% micronized salt, 0.7% micronized
monosodium glutamate and 2% of onion and parsley flavor was added
by external application on the extrudates. The snacks were homo-
genized until obtaining the entire flavored surface. The ingredients and
the flavor proportion used in the formulation were defined from fla-
voring pre-tests. The extrudates were packed in laminated bi-oriented
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LWT - Food Science and Technology 90 (2018) 270–276
polypropylene bags and stored at 18 °C in 50 g portions.
2.5. Analyses of snacks
2.5.1. Proximate composition
The proximate composition was determined according to the
Association of Official Analytical Chemists (AOAC, 1995): protein
(method 960.52) with conversion factor of 6.25, lipid (method
920.39C), ash (method 923.03) and moisture (method 925.10). Car-
bohydrates were quantified by difference.
2.5.2. Carotenoids
Carotenoids were extracted and total content was determined in
spectrophotometer (UVmin-1240, Shimadzu, Kyoto, Japan) at 450 nm,
according to Rodriguez-Amaya and Kimura (2004).
2.5.3. In vitro protein digestibility
In vitro protein digestibility was determined following Akeson and
Stahmann (1964). The extrudates were hydrolyzed with pepsin and
pancreatin. Results were quantified using standard curve of tyrosine as
reference.
2.5.4. Expansion index
Expansion index (EI) was calculated following Gujska and Khan
(1990) (Equation (1)), where DS is the diameter of the snack (cm) and
DD is the die diameter (cm).
DS
EI =
DD (1)
2.5.5. Bulk density
The bulk density (BD) of snacks was determined by measuring its
dimensions, according to Alvarez-Martinez, Kondury, and Harper
(1988), as described in Equation (2).
4×m
BD (g cm−3) =
π × D2 × L (2)
Where m is the weight of the snack (g), D is the diameter (cm) and L
is the length of the snack (cm).
2.5.6. Hardness
Extruded snacks were analysed using a texturometer (TA-XTplus
Texture Analyzer, Stable micro systems, Surrey, UK). Hardness was
calculated as the maximum force required for the cylindrical probe with
20 mm diameter to penetrate 50% of the sample and was measured in
Newton (N). The test speed was 1 mm s−1 and the distance from the
sample was 25 mm.
2.5.7. Water absorption index (WAI) and water solubility index (WSI)
Water absorption and water solubility indexes were determined
according to Anderson, Conway, Pfeifer, and Griffin (1969).
2.5.8. Color analysis
Color was analysed using a colorimeter (Chroma meter CR-400,
Konica Minolta, Tokyo, Japan) and was expressed by L* [black (0)/
white (100)], a* [green(-60)/red(+60)] and b* [blue(-60)/yellow
(+60)]. The total color difference (ΔE) between samples was calculated
according to Equation (3), where L, a and b are the Hunter Lab values of
the SP sample, and L0, a0 and b0 are the control sample (C) values.
Chroma (C*) and hue angle (h) were calculated according to Equations
(4) and (5), respectively (Konica Minolta, 2007).
ΔE = (L − L0)2 + (a − a0)2 + (b − b0)2 (3)
C∗ = (a∗2 + b∗2 )1/2 (4)
B.F. Lucas et al. LWT - Food Science and Technology 90 (2018) 270–276

b∗
ho = tan−1 ⎛ ⎞
⎝ a∗ ⎠ (5)
2.5.9. Scanning electron microscopy (SEM)
Cross sections of the snacks were cut to a thickness of 4–5 mm.
Samples were sputter coated with gold (Desk-V-Standard, Denton
Vacuum, LLC, Moorestown, N.J, USA). Microstructure was examined in
a scanning electron microscope (JSM-6610LV, JEOL Ltd., Japan) using
a 15 kV accelerating voltage.
2.5.10. Microbiological analyses
Microbiological analyses were carried out to verify the hygienic-
sanitary conditions of the snacks processing. Extrudates were evaluated
with respect to coliforms at 45 °C and Salmonella spp (Goes et al., 2015).
2.5.11. Sensory analysis
Snacks were evaluated in terms of flavor, color, texture, taste and
overall acceptability, using 9-point hedonic scale (1 = dislike extremely
and 9 = like extremely). A purchase intention test was performed using
5-point hedonic scale, where 1 = certainly would not buy and
5 = certainly would buy (Meilgaard, Civille, & Carr, 2007). The ac-
ceptability index (AI) was calculated according to Equation (6), where
X is the mean of the scores obtained for overall acceptance (Queiroz &
Treptow, 2006).
X during extrusion, reducing the extrudates expansion. The SP sample
AI (%) = × 100
9 (6)
The snacks samples were evaluated by 100 panelists, presented in
plate coded, monadic and randomly, accompanied by mineral water
and the product evaluation form. The sensory evaluation was approved
by the Research Ethics Committee of the Federal University of Rio
Grande (approval no 49/2016, Process 23116.002572/2016-05).
2.5.12. Stability during storage (shelf life)
Physical properties (moisture and hardness) and microbiological
attributes (coliforms at 45 °C and Salmonella spp) were evaluated after
12 months of storage, according to the procedures described in items
2.5.1, 2.5.6 and 2.5.10.
2.5.13. Statistical analysis
Data were submitted to variance analysis (ANOVA) and the differ-
ence among the means were determined using Tukey's test, with a
confidence interval of 95% (p < .05) (Montgomery, 2009).
3. Results and discussion
3.1. Nutritional composition of snacks
The nutritional content increase after the microalga addition was
22.6% in protein content, 28.1% in lipid content and 46.4% in mineral
concentration (Table 1). Cian et al. (2014) also obtained increase in the
protein, ash and lipid contents when using red algae to develop ex-
trudates. In this study, the nutritional content found by the authors was
9.60% of protein, 0.57% of ash and 0.31% of lipids, being the protein
increment of 7.6% when compared to the control without algae. In
addition, the protein concentration obtained in the present study was
higher than the concentration found in commercial snacks popular
among children, which represents only 2.7–5.5% according to Potter
et al. (2013).
Studies using microalgal biomass for food enrichment also have
shown improvements in different nutritional parameters (DeMarco,
Steffolani, Martinez, & León, 2014 ; Figueira, Crizel, Silva, & Salas-
Mellado, 2011; Morais et al., 2006). The Spirulina addition resulted in
increase of protein content, phenolic compounds and antioxidant ac-
tivity in pasta (De Marco et al., 2014). Moreover, increase of protein
content in cookies (Morais et al., 2006) and in gluten-free bread
(Figueira et al., 2011).
Digestibility is one of the parameters used to determine protein
quality (Pastor-Cavada et al., 2013). The results achieved showed high
digestible protein content and were similar to those obtained by Pastor-
Cavada et al. (2013) for in vitro protein digestibility in corn (81.8%) and
rice (84.1%) extrudates. Furthermore, Cian et al. (2014) found 91.2%
for protein digestibility of snacks added with red algae, which is con-
sistent with this work.
Lipids concentration is an important factor to be considered in the
process, since according to Basto et al. (2016) lipids can act as lubricant
results demonstrated a 28.1% increase in the lipid content, when
compared to control, and that increase did not significantly affect
(p > .05) the extrudates expansion (Table 2). Lipids from microalgae
are biocompounds of interest mainly because they contain poly-
unsaturated fatty acids (PUFAs), such as eicosapentaenoic (EPA), doc-
osahexaenoic (DHA) and γ-linolenic (GLA). These fatty acids are helpful
in the prevention and treatment of diseases, however the human or-
ganism cannot synthesize them (Matos et al., 2016).
Regarding carotenoids, the content was higher in the SP sample. A
similar trend was also observed by Basto et al. (2016) which extruded a
blend of corn flour and 15% of red peach palm and obtained
9.43 μg g−1 of carotenoids. In the same study, authors also verified that
there was no reduction in carotenoid content due to extrusion (130 °C).
According to the researchers, this result is due to extrusion being a short
shearing-cooking process (Basto et al., 2016). Therefore, that short time
was not sufficient to degrade the pigments, which is consistent with this
research (Table 1).
The increase of minerals was demonstrated from the ash con-
centration in the snacks. Santos et al. (2016) developed a shake for the
elderly and observed similar behavior, with a significant increase in
mineral content after Spirulina addition in the formulation. In this work
authors obtained 3.5% of ash in the shake added with Spirulina sp. LEB
18, which was higher than the content found in the sample without
microalgae (3.1%). Therefore, the present study demonstrated that the
microalga Spirulina sp. LEB 18 contributes to increase the nutrient
content of the snacks.

Table 1
Nutritional composition of control snacks (C) and snacks enriched with 2.6% Spirulina sp. Table 2
LEB 18 (SP). Physical parameters evaluated in the control snacks (C) and in snacks enriched with 2.6%
of Spirulina sp. LEB 18 (SP).
Composition C SP
Parameters C SP
−1 b
Protein (g 100g ) 8.38 ± 0.09 10.27 ± 0.06a
Protein digestibility (%) 88.77 ± 1.10a 89.99 ± 0.45a Expansion index (cm cm−1) 4.61 ± 0.09a 4.57 ± 0.09a
Lipids (g 100g−1) 0.57 ± 0.03b 0.73 ± 0.06a Bulk density (g cm−3) 0.070 ± 0.006b 0.077 ± 0.003a
Ashes (g 100g−1) 0.69 ± 0.01b 1.01 ± 0.01a Hardness (N) 24.26 ± 1.94a 25.43 ± 1.08a
Carbohydrates (g 100g−1) 90.36 87.99 WAI (g g−1) 9.43 ± 0.05a 8.69 ± 0.65a
Carotenoids (μg g−1) 1.57 ± 0.02b 14.68 ± 0.03a WSI (%) 29.82 ± 1.44a 29.98 ± 0.68a

Means ± standard deviation (n = 3). Different letters in the same line mean significant Means ± standard deviation (n = 10). Different letters in the same line mean significant
differences between samples (p < .05). Carbohydrates were calculated by taking the differences between samples (p < .05). WAI: water absorption index; WSI: Water solu-
difference. All results were expressed on a dry basis. bility index.

272
B.F. Lucas et al.
Table 3
Color parameters of control snacks (C) and snacks enriched with 2.6% Spirulina sp. LEB 18
(SP).
Parameters C SP
L* 63.39 ± 0.42a 34.13 ± 1.11b
a* −2.67 ± 0.06b 0.45 ± 0.05a
b* 18.58 ± 0.79a 10.65 ± 1.00b
C* 18.77 ± 0.78a 10.66 ± 0.99b
h 98.19 ± 0.36a 87.58 ± 0.38b
ΔE (*) 30.50 ± 0.98
Means ± standard deviation (n = 10). Different letters in the same line mean significant
differences between samples (p < .05). L*: Lightness; +a*: redness; -a*: greenness; +b*:
yellowness; -b*: blueness; C* = Chroma; h = hue angle; ΔE: Total color difference; (*):
Standard values (L*0, a*0; b*0) used in the calculation of ΔE.
3.2. Physical analysis
Expansion is the most important physical property in snacks (Joshi
et al., 2014). This property is directly related to the microstructure and
consequently to the crunchiness attribute, being a determining factor
for the technological quality of extrudates. When comparing the sam-
ples, it was observed that there was no significant difference in the
expansion index (Table 2), demonstrating that the addition of the mi-
croalga did not reduce such physical quality of the snack. The results
were similar to those reported by Morsy et al. (2014), which ranged
from 3.860 to 4.890 cm cm−1 for snacks made by corn and Spirulina.
The bulk density allows to evaluate the extrusion quality, the pro-
cess parameters and the raw material influence on the technological
quality of the snack. This measure considers expansion in all directions
(Jeyakumari, Das, Bindu, Joshy, & Zynudheen, 2016). In the present
study, the SP sample presented the highest bulk density, demonstrating
that the microalga addition had an influence on this property, probably
due to its high protein content.
According to Sumargo et al. (2016), this occurs due to the hydro-
philicity of certain proteins that tend to compete with the starch by
water, preventing the starch granule hydration and resulting in bulk
density increase. A similar behavior was observed by Lazou and
Krokida (2010) in extrudates, when incorporating lentil flour with high
protein concentration. Moreover, such results were similar to those
obtained by Jeyakumari et al. (2016) for snacks based on rice and corn
added with protein hydrolyzate and shrimp powder (0.045 and
0.082 g cm−3) and lower than those reported by Ding, Ainsworth,
Tucker, and Marson (2005) who obtained 0.10 g cm−3 for rice-based
snacks.
The SP and C samples did not differ statistically in relation to
hardness. This result can be considered positive, indicating that, despite
the addition of Spirulina sp. LEB 18 caused a significant increase in bulk
density, enrichment with the microalga did not affect hardness. The
results are among those cited by Joshi et al. (2014), which obtained
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LWT - Food Science and Technology 90 (2018) 270–276
values of hardness ranging from 16.2 to 43.4 N in corn-based snacks
with Spirulina.
The WAI is used to determine the volume occupied by the starch
after water absorption and retention and can be used as a gelatinization
index (Ding et al., 2005; Joshi et al., 2014). In the present study, al-
though the SP sample has 2.6% Spirulina sp. LEB 18 in substitution of
rice and corn flours, this enrichment did not significantly influence
(p < .05) the WAI of the snacks, when compared to the sample C.
The WSI measures the amount of soluble components released
during the extrusion, and is often used as an indicator of starch de-
gradation (Ding et al., 2005). The samples did not present significantly
difference (p > .05) in relation to the water solubility index (WSI). The
values obtained in this study were close to those reported by Ding et al.
(2005) for rice-based extrudates (21.5–32.7%) and by Rathod and
Annapure (2016) for lentil flour based snacks (up to 28.6%).
3.3. Color
Color is an attribute that improves food quality, contributing to
attract the consumer (Jeyakumari et al., 2016). The Spirulina sp. LEB 18
addition significantly affected (p < .05) all color parameters of the
extrudates (Table 3). Spirulina sp. LEB 18 resulting in reduction of L*,
b* and C*, indicating the darkening of the snacks (Fig. 1). Similar re-
sults were observed by Figueira et al. (2011) in gluten-free bread en-
riched with Spirulina and by Fradique et al. (2010), which compared
pasta added with microalgae (Spirulina and Chlorella) with samples
without microalgae and was observed lower L* values for the samples
containing microalgae.
According to Goes et al. (2015), the lightness reduction is related to
protein content increase. The authors observed reduction in lightness
values when snacks were enriched with fish residues. The L* values
ranged from 61.5 to 71.8 and were lower than the control sample
(79.1).
The hue (h) of the C sample showed results between yellow (+b*)
and green (-a*), which was expected due to the use of rice and corn
flours. The SP sample obtained h between red (+a*) and yellow (+b*),
probably due to microalga pigments like the carotenoids, which re-
mained stable after the extrusion process (Basto et al., 2016). The total
color difference (ΔE) between the samples was considered high, once
according to Jozinović, Šubarić, Ačkar, Babić, and Miličević (2016)
values above 6 demonstrate that the samples are in different groups of
colors. Higher ΔE between the samples is interesting when it is desired
to obtain product with color differentiated of the commercially snacks.
In addition, the color provided by Spirulina addition may be an at-
traction in snacks developed for children.
3.4. Microstructure
The effects caused by Spirulina sp. LEB 18 addition could be
Fig. 1. Control snacks (a) and snacks enriched with 2.6% Spirulina
sp. LEB 18 (b).
B.F. Lucas et al. LWT - Food Science and Technology 90 (2018) 270–276

Fig. 2. Micrographs of control snack with 15× (a), 100×

(b) and 180× magnification (c) and of snack enriched with
2.6% Spirulina sp. LEB 18 with 15× (d), 100× (e) and
180× magnification (f). Fig. 2 (A, D) corresponding to the
air bubbles inside the extrudates. Fig. 2 (B, C, E, F) corre-
sponding to the cell walls structures of the extrudates.

observed using scanning electron microscopy (SEM) (Fig. 2). The snacks
presented cell walls with thickness between 12.67 μm and 20.12 μm.
Fig. 2 shows that the control snack presented homogeneous structure,
with air bubbles of similar sizes and cell walls with thickness mean of
15.42 ± 1.83 μm (Fig. 2a–c). Structures with non-homogeneous
bubble size (Fig. 2d–f) containing thicker cell walls (18.50 ± 1.50 μm)
were associated with the protein addition in the formulation. Spirulina
sp. LEB 18 has a high protein concentration that tends to assimilate part
of the moisture content on the extrusion process, reducing starch hy-
dration as well as gelatinization (Sumargo et al., 2016). This process
causes the formation of smaller air bubbles and non-homogeneous
structure.
Similar results were obtained by Lazou and Krokida (2010), when
they developed corn flour-based snacks with concentrations of 10–50%
of lentil flour with high protein concentration. The authors observed
the thickening of the cell walls of the extrudates, as well as air bubbles
amount reduction formed in the extrusion.
color attribute (p < .05) (Table 4). In general, the snacks received
scores between 7 (like moderately) and 8 (like very much) for the at-
tributes evaluated. The significant difference (p < .05) between the
samples on color sensory evaluation was expected, due to the high total
color difference (ΔE = 30.50) which was verified in the instrumental
analysis (Table 3). Although the SP sample had obtained lower scores
for the color attribute (5.5 ± 2.0), the overall acceptance was not in-
fluenced (7.4 ± 1.0), and did not differ significantly (p > .05) from
the C sample.
Texture is the most important attribute in extrudates evaluation
(Jeyakumari et al., 2016; Pamies et al., 2000). In the present study, the
mean for samples texture was considered high (7.8), since it remained
between (7) like moderately and (8) like very much. This elevated mean
confirm the results obtained in instrumental evaluations of expansion
Table 4
Sensory evaluation of control snacks (C) and snacks enriched with 2.6% Spirulina sp. LEB
18 (SP).

3.5. Sensory analyses Parameter C SP

Flavor 7.2 ± 1.4a 7.2 ± 1.4a

Samples were considered safe for consumption after evaluation of Color 7.6 ± 1.0a 5.5 ± 2.0b
microbiological parameters controlled in extruded foods (Goes et al., Texture 7.5 ± 1.4a 7.8 ± 1.3a
2015). The results were < 3 MPN g−1 for coliforms at 45 °C and ab- Taste 7.6 ± 1.3a 7.7 ± 1.2a
Overall acceptance 7.5 ± 1.2a 7.4 ± 1.0a
sence of Salmonella spp in 25 g of sample, indicating that the snacks
Purchase intention 3.8 ± 0.9a 3.8 ± 0.8a
were processed under adequated hygienic conditions.
The statistical analysis of the sensory evaluation showed that there Means ± standard deviation (n = 100). Different letters in the same line mean sig-
was no significant difference (p > .05) between samples, except for the nificant differences between samples (p < .05).

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B.F. Lucas et al.
Table 5
Physical properties and microbiological attributes of control snacks (C) and snacks enriched with 2.6% Spirulina sp. LEB 18 (SP) after elaboration and after 12 months of storage.
Parameters C (0 days)
−1 b c
Moisture (g 100g ) 4.83 ± 0.08
Hardness (N) 24.26 ± 1.94a
Coliforms at 45 °C (MPN g−1) <3
Salmonella spp Absence in 25 g
Means ± standard deviation (n = 3 for moisture, n = 10 for hardness and n = 3 for microbiological parameters). Different letters in the same line mean significant differences between
samples (p < .05).
index and hardness (Table 2). Moreover, microstructural evaluation
(Fig. 2) also guaranteed the understanding of texture attribute, since
structures with thin cell walls were observed. A similar trend was also
observed by Cian et al. (2014) which obtained high sensory acceptance
and average of 8.3 for texture of snacks added with 3.5% of the red alga
Porphyra columbina, In this study, the purchase intention of the SP
sample also showed acceptable results (3.8 ± 0.8), situated between
“maybe would buy” and “probably would buy”. Furthermore, the ac-
ceptability index (AI) for C and SP samples were 83.3% and 82.2%,
respectively. According to Queiroz and Treptow (2006), for a product to
be accepted with respect to its sensorial characteristics, it is necessary
an acceptance index higher than 70%, which is consistent with our
results. Thus, we can conclude that extrudates with good sensory ac-
ceptability can be produced using organic rice flour, organic corn flour
and Spirulina sp. LEB 18.
3.6. Stability during storage (shelf life)
The shelf life is an important factor in the development of foods
which ensures the product quality for the consumers (Santos et al.,
2016). The evolution of snacks hardness as well as the moisture content
and microbiological attributes with time can be observed in Table 5.
Although the C and SP snacks showed a slight increase in the moisture
content, the results were below 6% even after 1 year of storage. This
result is considered suitable, since the moisture of extrudates with
adequate physical and sensory properties remain between 4 and 7%, as
reported in several studies (Borah, Mahanta, & Kalita, 2016; Cian et al.,
2014; Jeyakumari et al., 2016; Joshi et al., 2014).
Overall, it should be noted that for all samples, moisture variations
did not promote any modification on texture. The snacks did not pre-
sent significant (p > .05) changes in hardness after 12 months of sto-
rage. The results of the microbiological analyses indicated adequate
hygienic sanitary conditions of processing and manipulation of the raw
material during extrusion and flavoring. This is also due to extrusion
being a HTST process, and the low moisture of the final product, which
reduces the risk of microbiological contamination.
4. Conclusion
Spirulina sp. LEB 18 showed potential for enrichment of foods of
practical consumption. The addition of 2.6% of Spirulina sp. LEB 18
resulted in a protein increase of 22.6%, in addition of carotenoids and
minerals, without significantly affecting physical properties (expansion
index, hardness, WAI and WSI). The microstructural analysis revealed
that the snacks enriched with Spirulina sp. LEB 18 presented thin cell
walls (18.5 μm). Sensory evaluation indicated that Spirulina sp. LEB 18
did not influence the sensory characteristics (flavor, texture, taste and
overall acceptance) and also resulted in snacks with high acceptability
index (82%). Furthermore, the snacks presented physical and micro-
biological stability along 12 months of storage. Therefore, the snacks
added with Spirulina are an option for improving the nutritional health
of the population that look forward for food to practical consumption.
275
LWT - Food Science and Technology 90 (2018) 270–276
SP (0 days) C (12 months) SP (12 months)
a
3.89 ± 0.02 5.60 ± 0.03 4.98 ± 0.10b
25.43 ± 1.08a 25.21 ± 1.87a 25.18 ± 0.47a
<3 <3 <3
Absence in 25 g Absence in 25 g Absence in 25 g
Acknowledgements
The authors would like to thank to the Ministry of Science,
Technology, Innovations and Communications (MCTIC) (Project
01200.005005/2014-49), the Coordination for the Improvement of
Higher Education Personnel (CAPES) and the National Council for
Scientific and Technological Development (CNPq) (Project 490469/
2013-2) for financial support, the Inflex Company for donation of
packages, Duas Rodas Company for donation of flavors and CEME-SUL/
FURG (Electron Microscopy Center of Southern) due to the scanning
electron microscopy.
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