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Algal Research
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A R T I C L E I N F O A B S T R A C T
Keywords: The Spirulina sp. biomass has been used as an alternative for the production of high added-value functional and
Spirulina sp., nutraceutical foods. This study aimed to develop a ready-made functional sauce enriched with Spirulina sp.
Sauce biomass and evaluate its physicochemical, sensory, antioxidant, and microbiological properties over 45 days of
Enriched
storage. The formulation with 4% Spirulina sp. biomass presented significantly increased protein (34%), fiber
Antioxidant
Functional food
(57%), ash (40%), monounsaturated fatty acid (240%), and mineral (192%) contents than the control. The
storage stability for up to 45 days was noticed in both physicochemical and microbiological parameters. The
phenolic content also doubled, with a 12-fold increase in antioxidant activity, as shown by the IC50. The enriched
sauce presented a high capacity to mask microalgae's characteristic flavor, enhancing the nutritional quality of
the product. Therefore, the sauce enriched with 4% biomass showed the best characteristics among the con
centrations evaluated. It can be concluded that Spirulina sp. biomass can be used as a natural ingredient to
develop a functional sauce with antioxidant and nutritional potential.
* Corresponding author.
E-mail address: leila.rodrigues@ufba.br (L.M.R. Almeida).
https://doi.org/10.1016/j.algal.2021.102387
Received 28 February 2021; Received in revised form 6 June 2021; Accepted 11 June 2021
Available online 30 June 2021
2211-9264/© 2021 Elsevier B.V. All rights reserved.
L.M.R. Almeida et al. Algal Research 58 (2021) 102387
malnutrition, obesity, cardiovascular disease, and cancer, among others. Four sauce formulations, containing 0 (control), 2.0, 3.0, and 4.0%
Thus, Spirulina sp. can be used as an alternative for the production of Spirulina sp. biomass (wt%), tomato (69.00 g), onion (11.00 g), soybean
high added-value functional and nutraceutical foods [1,8–12]. oil (3.60 g), garlic (0.97 g), cumin (0.38 g), sodium chloride (0.31 g),
The global food industry has been widely exploiting microalgal as rosemary (0.24 g), parsley (0.14 g), oregano (0.11 g), and water (14.00
promising sources for the development of new products. The incorpo mL) were prepared. The experimental design is given in Fig. 1.
ration of the biomass of Spirulina sp. in certain foods is considered an The ingredients were ground, mixed in a blender, transferred to a
innovation with high technological and commercial potential since it metal container, and heated for 10 min at 95 ◦ C (except Spirulina sp.
adds nutritional value to products already known by the population biomass). After this step, the sauce was left to rest until it reached a
[13,14]. Compared to Spirulina sp. in commercial powder form, the temperature of 60 ◦ C, and then Spirulina sp. biomass was added and
foods incorporated with biomass are more accepted by the consumers mixed into the sauce.
due to their better sensorial properties [15]. The freshly prepared sauces were poured into 200 mL glass jars with
The novel biomass incorporated food products represent a sustain metal lids, previously sterilized in boiling water (100 ◦ C) for 30 min. The
able trend in the food industry. This trend considers the increasing sauces were pasteurized by heating the jars for 40 s at 98 ◦ C [21] and
consumer demand for healthy and practical foods with a high content of stored in a refrigerator (4–6 ◦ C, 60% of relative humidity).
protein and bioactive compounds and low fat and sugar. In this context, The formulations were evaluated regarding their parameters pH and
ready-made sauces stand out in the market because of their convenience acidity, texture, and sensory acceptance 24 h after preparation. In
and practicality in preparing elaborate dishes and because new products addition to the control formulation, the formulation containing Spirulina
and formulations are continuously launched [16,4]. sp. biomass that presented the best results for all parameters was
Spirulina sp. biomass's main disadvantages are its odor, flavor, and selected for the stability test.
color. These attributes of biomass are percept by most people as un In the stability test, the two formulations (control and a selected
pleasant [1]. However, its incorporation into a sauce can mask these formulation containing Spirulina sp.) were stored for 45 days under
attributes, with the added benefit of enhancing the nutritional and refrigeration (4–6 ◦ C). Both formulations were periodically evaluated (0,
functional quality of the product. 7, 15, 30, and 45 days) for their acidity, pH, water activity, total pro
Some foods such as yogurts and functional beverages have already teins, total lipids, carbohydrates, crude fiber, total ash, color, and anti
been developed of Spirulina sp. biomass supplementation [14,15,17,18]. oxidant properties - 2, 2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), 3-
Despite this, there is a lack of research on the development and char ethylbenzothiazoline-6-sulfonic acid (ABTS), and ferric reducing anti
acterization of biomass-based functional sauces. Besides, it is worth oxidant potential (FRAP).
mentioning that enriched sauces are versatile products that could be The fatty acid profile and the microbiological analysis were evalu
used to prepare other foods. ated at the initial time (zero-day) and after 45 days of storage under
This study aimed to develop a ready-made functional sauce enriched refrigeration (4–6 ◦ C). The minerals were evaluated at the final storage
with Spirulina sp. biomass and evaluate its physicochemical, sensory, time (45th day). Three replicates of all formulations were prepared and
antioxidant, and microbiological properties over 45 days of storage. analyzed in triplicate.
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L.M.R. Almeida et al. Algal Research 58 (2021) 102387
Analysis
performed using a 100-mL beaker with 30 g of sample and a cylinder spoon and randomly encoded with three-digit numbers. As a vehicle for
probe (diameter, 38.1 mm; height, 20 mm (TA4/1000)). The samples tasting the sauce, 25-g portions of pasta were served in disposable
were compressed to 50% of the original thickness in a double cycle at containers. The tasters evaluated the overall acceptance of the formu
0.5 mm⋅s− 1 test speed and 5 g trigger load, over two cycles. The lations using a nine-point hedonic scale, ranging from 1 (dislike
following texture profile parameters were assessed: firmness (N), extremely) to 9 (like extremely), for the following traits: appearance,
cohesiveness and elasticity (mm) [26]. Data were processed using aroma, flavor, texture, and overall impression [23,27].
Texture Expert for Windows 1.20 software (Stable Micro Systems). The tasters were also asked about their intention to purchase the
products, on a scale from 1 (definitely would not buy) to 4 (definitely
2.2.4. Sensory evaluation would buy). The Research Ethics Committee from the Federal University
A team of 40 not trained tasters performed the sensory evaluation of Rio Grande approved the study (process number 1.525.184).
[27]. The group consisted of students and staff of the Federal University
of Bahia (UFBA), of both sexes (80% women, 20% men), aged from 19 to 2.2.5. Fatty acid identification and quantification
45. The session was conducted by first presenting four sauce samples The transesterification of the extracted lipid fraction was performed
corresponding to the formulations with different Spirulina sp. biomass to determine the fatty acid composition, according to Nascimento et al.
concentrations and the control formulation. [28]. The fatty acid methyl esters were separated on a gas chromato
The samples were served on small, white plates with a disposable graph (Clarus 680, Perkin Elmer) equipped with a flame ionization
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L.M.R. Almeida et al. Algal Research 58 (2021) 102387
detector (GC-FID) and an Elite-WAX fused silica capillary column (30 m 2.2.9. Statistical analysis
× 0.32 mm × 0.25 μm). The equipment was set to 250 ◦ C injector The analyses were performed in triplicate and expressed as the mean
temperature and 280 ◦ C detector temperature. values ± standard deviation. Data were subjected to analysis of variance
The programmed column temperature was set to 150 ◦ C for 16 min, (ANOVA), followed by the Tukey's test (p < 0.05) to evaluate the dif
increasing 2 ◦ C per min until 180 ◦ C, remaining at this temperature for ferences between the formulations zero (control), 2, 3 and 4% incor
25 min, increasing 5 ◦ C until 210 ◦ C, and staying at this temperature for porated with Spirulina sp. biomass 24 h after the preparation.
25 min. Helium was used as a carrier gas at 1.0 mL min− 1. Samples were The formulation that presented the best physicochemical parameters
injected in split mode (1:90) with an injection volume of 1 μL. and sensory attributes was also evaluated during 45 days of storage
Fatty acids were identified by comparing the peak retention times of (zero, 7, 15, 30 and 45 days). The Student's t-test was used to assess the
samples with the retention times of fatty acid methyl ester standards differences between the control and the formulation that presented the
(C4–C24, 18919-AMP, Sigma-Aldrich®, USA). The quantification was best physicochemical parameters and sensory attributes at days zero and
accomplished by adding an internal standard (C23:0 Sigma, USA) to the 45 storage. A linear regression was used to verify the correlation be
fatty acid methyl esters. The calculations were performed according to tween dependent variables. The tests were performed using the free
Eq. (2). The values were expressed as mg of fatty acids per g lipid (mg software environment for statistical computing and graphics R Com
g− 1). mander 3.5.1.
( )
mg AFA × WIS × F
Concentration = 1000 × (2) 3. Results and discussion
g AIS × FFA × W
3.1. Preliminary results
AFA = area under the fatty acid methyl ester peak in the sample chro
matogram;WIS = weight of the internal standard added to sample, in mg;
The control formulation had lower pH and higher acidity. Overall,
F = correction factor for the response of each fatty acid methyl ester in
the four formulations significantly differed in both parameters (p <
the ionization detector relative to C23:0;AIS = area under the internal
0.05). The differences increased with the content of Spirulina sp. biomass
standard peak in the sample chromatogram;W = weight of the sample in
incorporated into the sauce (Table 1). The supplementation with Spir
milligrams; andFFA = fatty acid methyl ester to the fatty acid conversion
ulina sp. biomass increased the sauces' pH and decreased the product's
factor.
acidity. These differences were attributed to the biomass' pH, which is
neutral or slightly basic, ranging from 7.0 to 8.0.
2.2.6. Phenolic compounds and antioxidant activity
Similarly to pH, total acidity is also an important parameter for
Each extract's total phenolic content was determined using the Folin-
preserving and accepting a food product. Food decomposition, either by
Ciocalteu method described by Singleton and Rossi [29]. The readings
hydrolysis, oxidation, or fermentation, typically alters the hydrogen ion
were performed at 760 nm (Lambda 35 spectrophotometer, Perkin-
concentration and, consequently, the acidity [34]. The total acidity
Elmer). For quantification, a standard curve (50–250 μg⋅mL− 1) was
values differed among the formulations.
prepared with gallic acid, and the results were expressed in milligrams
Considering that the increase in Spirulina sp. biomass concentration
of gallic acid equivalent per gram of sample (mgGAE⋅g− 1). The radical
reduced the acidity, the formulation with 4% biomass had the lowest
scavenging activity of DPPH (2,2-diphenyl-1-picrylhydrazyl, Sigma-
value. This can be explained by the alkaline character of Spirulina sp.
Aldrich Chemical) was determined according to the method proposed
biomass. The Spirulina sp. biomass presented a buffering effect in the
by Vinson et al. [30], reading the samples at 517 nm.
sauce, reducing the concentration of free hydrogen ions. This is due to
Antioxidant activity (half maximal inhibitory concentration IC50)
some components such as proteins, peptides, and amino acids [35,36].
was expressed as the final concentration in mg⋅mL− 1 sample necessary
Interestingly, the addition of biomass in some food products de
for reducing the concentration of DPPH by 50%. The ferric reducing
creases the pH and increases the total acidity. The biomass addition to
ability of plasma (FRAP) was determined according to the method
fermented products can stimulate some bacteria to produce organic
described by Benzie & Strain [31], reading the samples at 595 nm. A
acids. Thus, explaining the inverse effect in both pH and acidity when
standard curve was constructed with ferrous sulfate (500–2000 μM
compared to those observed in the present study. pH and acidity showed
mL− 1), and the results were expressed as μmol Fe2+g sample− 1.
a significant and inversely proportional correlation (r = − 0.99).
The radical scavenging activity of ABTS•+ (2,2′ -azino-bis (3-ethyl
Thus, demonstrating that the lower the sauce's pH, the higher the
benzothiazoline-6-sulfonic acid; Sigma-Aldrich Chemical) was assessed
total acidity value will be. The negative correlation between pH and
according to the method proposed by Re et al. [32], reading the samples
acidity is consistent since the titratable acidity is related to the amount
at 734 nm. A standard curve was constructed with Trolox (6-hydroxy-
of organic acids present in the sample. The pH is associated with the
2,5,7,8-tetramethylchroman-2-carboxylic acid; Sigma-Aldrich Chemi
concentration of H3O+ ions and this concentration is also proportional to
cal) (100–2000 μM mL− 1), and the results were expressed as μM Trolox g
the concentration of acids.
sample− 1.
Thus, both pH and acidity are important parameters for assessing
food conservation during storage since they can undergo changes caused
2.2.7. Color analysis
by microorganisms and enzymatic activity [37,38]. Barkallah et al. [23]
Color analysis was performed on a digital colorimeter (CIE L*a*b*
have also observed an inversely proportional relationship between the
system; CR-5 benchtop colorimeter, Konica Minolta). Calibration was
performed with a standard white plate according to the manufacturer's
instructions. Results were expressed as L*, (luminosity) ranging from Table 1
0 (black) to 100 (white); a*, ranging from − a* (− 60.0 green) to þa* pH and acidity values for the control sauce and the sauces supplemented with
(+60.0 red); and b*, from − b* (− 60.0 blue) to +b* (+60.0 yellow). different Spirulina sp. biomass concentrations after 24 h of storage.
Parameters Control sauce Spirulina sp. biomass content (%)
2.2.8. Microbiological evaluation 2 3 4
The samples were tested for Salmonella spp. (absence or presence in
pH 3.04 ± 0.01a 3.14 ± 0.01b 3.23 ± 0.02c 4.32 ± 0.02d
25 g), total coliforms, coagulase-positive Staphylococcus, mold, and yeast
Total acidity (%) 3.19 ± 0.09d 3.15 ± 0.02c 2.97 ± 0.59b 1.65 ± 0.01a
according to the Compendium of Methods for the Microbiological Ex
amination of Foods, American Public Health Association (APHA) [33]. Data are expressed as the mean ± standard deviation of triplicates. Different
lowercase letters on the same line indicate a significantly difference between the
sauces analyzed according to the Tukey's test at α = 0.05.
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L.M.R. Almeida et al. Algal Research 58 (2021) 102387
pH and the acidity in the yogurts incorporated with Spirulina sp. presence of sulphur volatile compounds present in onions (such as iso-
biomass. allin, methiin, and propiin) [44] and garlic (allicin) [45] Besides, the
Texture, appearance, and flavor are the three organoleptic attributes cumin (o-cymene and cumic alcohol) [46], rosemary (several mono
that determine consumer acceptability. Texture parameters correlate terpene hydrocarbons) [47], parsley (monoterpenes as myristicin, beta-
with sensory properties and are fundamental food attributes [39]. The phellandrene, and myrcene) [48], and oregano (p-cymene and carva
textural (firmness, cohesiveness, and elasticity) and sensory parameters crol) [49] have also contributed to masking the biomass' undesirable
for the four formulations can be seen in Table 2. odor.
The firmness, cohesiveness, and elasticity values of the sauces con It is worth mention the considerable concentration of spices in the
taining 2%, 3%, and 4% Spirulina sp. biomass significantly differed from sauce and that their odors are very perceptible to the olfactory system.
each other and the control (p < 0.05). Accordingly, supplementation The sauce containing 4% biomass presented the highest flavor score,
with biomass changed the overall texture profile, resulting in increas which did not significantly differ from the control score (p = 0.924).
ingly firm and elastic sauces with increasing microalgal concentration. The addition of the microalga substantially changed the color of the
The sauce containing 4% biomass presented the highest firmness value. products from red to dark green. Therefore, the color was perceived as
Like in the present study, Bolanho et al. [40] also reported higher inadequate by the tasters, as shown in the appearance scores (Table 2).
firmness values for cookie formulations with 5% Spirulina sp. biomass This perception could have been caused because the tasters had not been
than for formulations containing zero and 2% of the same biomass. The trained. Thus, the control sauce scored significantly higher (p < 0.05) in
firmness values increase proportionally with biomass concentration. the appearance attribute than the sauces supplemented with 3 and 4%
This trend can be related to the microalgal's high protein content that Spirulina sp. biomass. Bolanho et al. [40] have also reported a similar
increases protein-protein interactions generating a more rigid and result. The cookies enriched with 5% Spirulina sp. biomass scored lower
cohesive gel network [5]. in the appearance attribute than the control.
The incorporation of Spirulina sp. biomass into the sauces also Despite the color difference, purchase intent did not differ signifi
resulted in significant differences in cohesiveness and elasticity among cantly (p > 0.05) between the control sauce and the sauces supple
all treatments (p < 0.05). The sauce containing 4% Spirulina sp. biomass mented with Spirulina sp. biomass (Table 2). This attribute score's
presented the lowest cohesiveness and highest elasticity. These results reached values near 3.0, classified in the hedonic scale as “probably
can also be correlated with protein content in biomass. Proteins are would buy”. Although the difference was not significant, the sauce
amphiphilic molecules, and they migrate spontaneously to an air-water containing 4% biomass scored the highest (2.95), with 57.5% of the
interface or an oil-water. Once at the interface, it can interact with the tasters indicating they would buy this sauce. Accordingly, aroma, flavor,
neighboring molecules and form an elastic film withstands thermal and and texture showed no significant change after the addition of 2%, 3%,
mechanical motions. However, for cohesiveness, there may have been a and 4% Spirulina sp. biomass, demonstrating general acceptability by
reduction in the chemical bonding resistance that makes up the product tasters.
[41]. The sauce added with 4% biomass showed the highest scores for the
The sensory analysis showed that the incorporation of Spirulina sp. purchase intent and overall impression. Therefore, indicating that the
biomass did not affect the tasters' acceptance of the product. Incorpo appearance did not harm these attributes. Besides, the sauce added with
rating 2, 3, and 4% biomass into the sauces did not significantly affect 4% biomass presented optimal physical characteristics (pH, acidity, and
the aroma, flavor, texture, and overall impression purchase (p > 0.05). texture), ensuring the final product's quality. Higher nutritional values
The undesirable fish flavor and smell from Spirulina sp. biomass have from the formulations with higher content of incorporated biomass are
already been associated with a mixture of several volatile compounds expected due to the proximate composition of Spirulina sp. biomass [20].
[42,43]. The spices' combination aroma used to prepare the sauces in In this study, the sauce containing 4% of Spirulina sp. biomass was
this study was necessary to mask Spirulina sp. biomass's unpleasant chosen as the best final product. This sauce was selected for further
flavor. analyses to evaluate its physicochemical, microbiological, and antioxi
Thus, the undesirable fish aroma of biomass has been masked by the dant properties over 45 days of storage under refrigeration. Barkallah
et al. [23], have also chosen the formulation for evaluation during
Table 2
storage, based on the product's textural properties and sensory
Textural and sensory parameters for the control sauce and the sauces containing acceptability.
different concentrations of Spirulina sp. biomass.
Parameters Control Spirulina sp. biomass content (%)
3.2. Physicochemical characteristics
sauce
2 3 4
No chemical preservative was added to the sauce formulations. The
Firmness (N) 0.40 ± 0.01a 0.40 ± 0.10a 0.43 ± 0.06b 0.45 ± control of pH, acidity, moisture, and water activity is essential for
0.06b ensuring a product's food safety over time. The pH of both the control
Cohesiveness 1.23 ± 0.18b 1.38 ± 0.12c 1.42 ± 0.00 1.09 ±
d
0.03a
sauce and the sauce supplemented with 4% Spirulina sp. biomass grad
Elasticity (nm) 3.39 ± 0.47a 4.31 ± 0.20b 4.66 ± 0.01c 5.75 ± ually decreased during the storage period (Table 3). The formulation
0.01d containing microalgal biomass showed a minor decrease (4.9%) than the
Appearance 7.20 ± 0.90b 6.65 ± 6.05 ± 0.70a 5.92 ± control (8.6%) after 45 days. Throughout the storage period, both sauces
0.80ab 1.01a
had pH values lower than 4.5. Similarly, Barkallah et al. [23] reported
Aroma 6.90 ± 0.90a 6.75 ± 0.90a 6.50 ± 0.72a 6.80 ±
0.83a pH values ranging from 4.04 to 4.53 for yogurt fortified with 0.25%
Flavor 6.62 ± 0.71a 6.10 ± 0.93a 6.35 ± 0.75a 6.82 ± Spirulina sp. biomass over 28 days of storage.
1.00a TA values were higher in the control sauce (90–217%), depending on
Texture 7.00 ± 0.65a 6.87 ± 0.84a 6.75 ± 0.80a 7.00 ± storage time (Table 3). The control's TA values significantly (p < 0.05)
0.90a
Overall 6.87 ± 1.12a 6.42 ± 1.30a 6.40 ± 0.95a 6.90 ±
differed from the sauce containing 4% Spirulina sp. biomass. The results
impression 1.04a show that the control sauce was more acidic than the supplemented
Purchase intent 2.82 ± 0.72a 2.65 ± 0.83a 2.75 ± 0.78a 2.95 ± sauce. The analyzed sauces showed a slight increasing trend in acidity
0.75a over time. This increase may indicate the onset of hydrolysis of fatty
Data are expressed as the mean ± standard deviation of triplicates. Different acids present in the products.
lowercase letters on the same line indicate a significant difference between the As a result of the action of enzymes, the hydrolysis of triacylglycerols
sauces analyzed according to Tukey's test at α = 0.05. promotes the release of free fatty acids. These, in turn, are susceptible to
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L.M.R. Almeida et al. Algal Research 58 (2021) 102387
Table 3 protein, ash, and total fiber than the control (Table 3).
Evaluation of the control sauce's physicochemical characteristics and stability The microalgal's biomass supplementation increased the protein
and the sauce supplemented with 4% Spirulina sp. biomass over 45 days of content by 52% over the control sauce. The high protein increment
storage. noticed can be explained due to the high protein content (around 65%
Parameter/sauce Time (days) wt) present in the Spirulina sp. biomass used in this study [20]. It is
0 7 15 30 45 worth mention that the protein from microalgal biomass is qualitatively
complete because it presents the essential amino acid ratios recom
pH Control 3.04 ± 2.91 ± 2.87 ± 2.85 ± 2.80 ±
0.01dA 0.02cA 0.01bA 0.02aA 0.01aA
mended by the Food and Agriculture Organization (FAO) [51].
4% 4.32 ± 4.26 ± 4.22 ± 4.14 ± 4.12 ± Incorporating 4% Spirulina sp. biomass into sauce adds nutritional
Sauce 0.01cB 0.03bB 0.02bB 0.01aB 0.01aB value to the product, which may help individuals reach the recom
Total acidity Control 3.19 ± 3.72 ± 3.76 ± 3.78 ± 3.87 ± mended daily protein intake. A medium portion (60 g) of the sauce
(%) 0.09aB 0.01bB 0.01abB 0.01abB 0.08cB
enriched with Spirulina sp. biomass contributes 25% to 33% of the rec
4% 1.65 ± 1.70 ± 1.72 ± 1.75 ± 1.78 ±
Sauce 0.01aA 0.02bA 0.02bA 0.07cA 0.01dA ommended protein intake according to the Dietary Reference Intakes
Aw Control 0.99 ± 0.99 ± 0.98 ± 0.98 ± 0.99 ± (DRIs) [52] (15–20%) for a healthy adult.
0,01aB 0,01aB 0,01aB 0.01aB 0.01aB The ash content in the sauce supplemented with Spirulina sp. biomass
4% 0.96 ± 0.95 ± 0.95 ± 0.95 ± 0.94 ± was significantly higher than that in the control sauce. No variations in
Sauce 0,01aA 0,01aA 0,04aA 0.01aA 0.01aA
Moisture (%) Control 86.01 87.63 87.09 86.41 86.13
ashes during storage were observed. The sauce supplemented with 4%
± ± ± ± ± biomass had higher ash content. This can be explained due to the
0.36bB 1.12aB 0.81abB 0.35abB 0.66abB essential minerals such as magnesium, iron, and calcium naturally
4% 83.46 83.55 83.19 83.33 83.40 present in the microalgal biomass. The total fiber content was also
Sauce ± ± ± ± ±
higher in the formulation containing biomass than in the control
0.51aA 0.07aA 0.05aA 0.18aA 0.15aA
Crude protein Control 5.44 ± 5.07 ± 4.93 ± 4.90 ± 4.91 ± formulation.
(%) 0.28aA 0.41aA 0.17aA 0.65ªA 0.45ªA These results corroborate the findings of Bolanho et al. [40], which
4% 8.26 ± 8.03 ± 7.86 ± 7.77 ± 7.81 ± reported higher ash and fiber contents in cookies enriched with 5%
Sauce 0.15aB 0.24aB 0.11aB 0.15aB 0.30aB Spirulina sp. biomass than its control sample. The carbohydrate content
Total lipids Control 2.09 ± 2.08 ± 2.00 ± 2.06 ± 2.04 ±
(%) 0.04aB 0.08aB 0.07aB 0.07ªB 0.03ªB
was higher in the enriched sauce than in the control sauce. Possibly, due
4% 1.95 ± 1.90 ± 2.01 ± 2.01 ± 2.00 ± to the amount of this compound present in the microalgal biomass,
Sauce 0.05aA 0.10aA 0.08aA 0.13aA 0.09aA which is approximately 10% [20]. The minor variations in the carbo
Carbohydrate Control 4.44 ± 3.24 ± 3.93 ± 4.69 ± 4.90 ± hydrate content during the 45 days were attributed to the indirect
(%) 0.03dA 0.02aA 0.01bA 0.22cA 0.03eA
quantification method used in this study.
4% 5.26 ± 5.47 ± 5.91 ± 5.83 ± 5.73 ±
Sauce 0.02aB 0.03aB 0.04bB 0.03bB 0.05bB
Ash Control 0.43 ± 0.42 ± 0.43 ± 0.42 ± 0.40 ± 3.3. Fatty acids composition
(%) 0.01ªA 0.02ªA 0.01ªA 0.01aA 0.04aA
4% 1.07 ± 1.05 ± 1.03 ± 1.06 ± 1.06 ±
The sauce supplemented with 4% biomass presented 14 fatty acids.
Sauce 0.03aB 0.11aB 0.03aB 0.04aB 0.03aB
Crude fiber Control 0.60 ± 0.57 ± 0.57 ± 0.54 ± 0.54 ±
The C18:2 n-6 was the predominant, followed by C18:1 n-9 (Table 4). In
(%) 0.01aA 0.01aA 0.01aA 0.01ªA 0.01ªA the control sauce, C18:2 n-6 was also the major fatty acid derived from
4% 1.15 ± 1.16 ± 1.14 ± 1.13 ± 1.13 ± the soybean oil added to both formulations. Notably, the main lipid
Sauce 0.02aB 0.01aB 0.02aB 0.01aB 0.01aB source in the control sauce was derived from the lipid fraction of soy
Aw: water activity. Data are expressed as the mean ± standard deviation of bean oil, which has high levels of polyunsaturated fatty acids [53],
triplicates. Different lowercase letters on the same line indicate a significant whereas the lipids in the sauce supplemented with Spirulina sp. biomass
difference between the averages for the same sauce (control or 4% sauce) at the also included fatty acids derived from microalgal, such as C16:1 n-7,
different times analyzed by the Tukey test (α = 0.05). Different capital letters in C18:1 n-9, and C18:3 n-3.
the same column indicate a significant difference between the control sauce and The sauce supplemented with 4% of biomass presented richer than
the 4% sauce simultaneously according to Student's t-test at α = 0.05. the control because its formulation contained oleic (C18:1 n-9, 136.73
mg⋅g− 1) and α-linolenic (C18:3 n-3, 43.57 mg⋅g− 1) acids. Daily intake of
oxidative processes, resulting in the formation of compounds respon food sources of omega-3 fatty acids is important for human nutrition.
sible for rancified taste and odor, changing the quality of the product However, industrialization has led to a gradual increase in the con
[50]. Çelekli et al. [17], evaluated the increase in the growth and ac sumption of refined oils with a high linoleic acid (C18:2 n-6) content and
tivity of probiotic bacteria by adding Spirulina sp. biomass to ayran. The to a decrease in the consumption of foods rich in alpha-linolenic acid
authors found that during storage (21 days; 4 ◦ C), the samples added to (C18:3 n-3), resulting in an inadequate n-6/n-3 ratio [54]. Thus, the
Spirulina sp. also had lower acidity levels than the control. sauce enriched with Spirulina sp. biomass is an alternative practical food
The supplemented sauce's moisture and water activity values were that helps increase n-3 intake, thereby providing health benefits because
smaller than those found for the control sauce. The biomass is a dry the sauce supplemented with microalgal biomass contains C18:3 n-3.
powder (less than 10% of moisture) containing a variable content of In the sauce containing 4% biomass, the sum of monounsaturated
total solids. Beyond the high content of proteins, carbohydrates, and fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) prevailed
some lipids, the biomass also has carbonates, bicarbonates, chloride, over the total saturated fatty acids (SFAs). The sauce supplemented with
sulfide, phosphate, and others. The inorganic species comes from the 4% biomass showed an increase of 130.84 mg⋅g− 1 in MUFAs and of
cultivation media and the cellular metabolism of microalgae. approximately 25% in SFAs and PUFAs compared to the control.
The addition of the biomass implies the substitution of a 4% (m m− 1) In contrast to the present study results, Barkallah et al. [23] reported
of high moisture content from the control by a dry content, resulting in a 70.56% increase in SFAs, overshadowing the increase in UFAs
decreased moisture and water activity values observed. Thus, the (29.19%) and MUFAs in the yogurt fortified with Spirulina sp. biomass.
formulation that contains 4% (m m− 1) of this dry powder contains more These differences can be attributed to the Spirulina sp. biomass compo
macronutrients and total solids than the control. As a result, either the sitions, which strongly depend on the strains and the growth conditions
moisture or the water activity tends to decrease. It can be interpreted as used in the production of this microalga [20].
a substitution of 4% of a mass with high moisture content in the control In general, both sauces' storage for 45 days caused no significant
sample by the dried biomass. The supplemented sauce presented higher change (p > 0.05) in the fatty acid composition. Natural antioxidants
6
L.M.R. Almeida et al. Algal Research 58 (2021) 102387
Table 4 Table 5
Fatty acid composition of the control and 4% Spirulina sp. biomass sauces at the Total phenolic content and antioxidant activity of the control sauce and sup
beginning and end of storage. plemented with 4% biomass after 45 days of storage (4–6 ◦ C).
Class Fatty acid Sauce (day 0) Sauce (day 45) Sauce Time (days)
7
L.M.R. Almeida et al. Algal Research 58 (2021) 102387
Table 6 Table 7
Mineral composition of the control sauce and the sauce containing 4% Spirulina Variations in L* (luminosity). a* (red) and b* (yellow) in the control sauce and
sp. biomass with 45 days of storage.a the sauce with 4% Spirulina sp. biomass stored for 45 days at 4–6 ◦ C.
Minerala Control sauce 4% Spirulina sp. sauce Increase (%) Sauce Time (days) Color
8
L.M.R. Almeida et al. Algal Research 58 (2021) 102387
characteristic flavor of the microalgal and had properties that meet the [5] A.M. Pereira, C.R. Lisboa, J.A.V. Costa, High protein ingredients of microalgal
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