Food Hydrocolloids 107 (2020) 105893

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Food Hydrocolloids
journal homepage: http://www.elsevier.com/locate/foodhyd

3D printed functional cookies fortified with Arthrospira platensis: Evaluation

of its antioxidant potential and physical-chemical characterization
Marta V. Vieira a, Sara M. Oliveira a, Isabel R. Amado a, Luiz H. Fasolin b, c, Antonio A. Vicente b,
Lorenzo M. Pastrana a, Pablo Fucin ~os a, *
a
International Iberian Nanotechnology Laboratory, Department of Life Sciences, Av. Mestre Jos�e Veiga s/n, 4715-330, Braga, Portugal
b
CEB – Centre of Biological Engineering, Universidade do Minho. Campus de Gualtar, 4710-057, Braga, Portugal
c
Department of Food Engineering, School of Food Engineering, University of Campinas, UNICAMP, 13083-862, Campinas, SP, Brazil

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

Keywords: In the last few decades, consumers’ growing attention to the close relationship between health and nutrition is
Arthrospira platensis emerging as a new trend, mostly regarding the incorporation of natural ingredients into food. Among those
Encapsulation ingredients, microalgae are considered as innovative and promising compounds, rich in valuable nutrients and
Food-ink
bioactive molecules. In the present work, 3D printed cookies were fortified with the microalga Arthrospira pla­
Functional food
tensis aiming at developing a new functional food with antioxidant properties. A. platensis antioxidants were
3D printing
recovered using ultrasound-assisted extraction in hydroalcoholic solutions. Ethanol/water and biomass/solvent
ratios were optimised through a Design of Experiments (DOE) approach, using the antioxidant activity (ORAC
and ABTS) and total phenolic content (TPC) as response variables. The highest ORAC, ABTS and TPC values were
observed in the extract obtained with 0% ethanol and 2.0% biomass; thus, this extract was chosen to be
incorporated into a printable cookie dough. Three different incorporation approaches were followed: (1) dried
biomass, (2) freeze-dried antioxidant extract and (3) antioxidant extract encapsulated into alginate microbeads
to enhance the stability to heat, light, and oxygen during baking and further storage. All dough formulations
presented shape fidelity with the 3D model. The cookies had aw values low enough to be microbiologically stable,
and the texture remained constant after 30 days of storage. Moreover, the extract encapsulation promoted an
improvement in the ORAC value and colour stability when compared to all other formulations, revealing the
potential of A. platensis for the development of a functional 3D food-ink.

1. Introduction the increasing cost of the health care and the steady boost of life ex­
pectancy (Betoret, Betoret, Vidal, & Fito, 2011; Lopez-Rubio, Gavara, &
Many nutrition concepts have changed during the past few decades, Lagaron, 2006; Plaza, Herrero, Cifuentes, & Iba �nez, 2009; Sun, Zhou,
and the food industry has made a significant effort to follow them and Yan, Huang, & Lin-ya, 2018).
adapt their products to these changes. Traditionally, the primary role of Functional food is a natural or processed food that contains known
diet was to provide enough nutrients to meet metabolic requirements biologically-active compounds which, when in defined quantitative and
while giving consumers a feeling of satisfaction and well-being. Nowa­ qualitative amounts, provide a clinically proven and documented health
days, however, it is established that beyond meeting nutritional needs, benefit; and, hence, a useful tool for the prevention, management and
the diet may modulate various bodily functions and may play detri­ treatment of diseases. There is a wide range of compounds that have
mental or beneficial roles in some diseases (Bigliardi & Galati, 2013; already been incorporated into functional foods, with particular atten­
Roberfroid, 2000). In this regard, it is possible to observe an increasing tion being given to ingredients from natural resources (Day, Seymour,
consumer’s health consciousness and demand for healthy foods - facts Pitts, Konczak, & Lundin, 2009; Herrero, Martín-Alvarez,
� Senora
�ns,
that are stimulating innovation and new product development in the Cifuentes, & Iba�nez, 2005).
food industry. This trend is also responsible for an ever-increasing Microalgae can be considered an innovative and promising food
worldwide interest in functional food, which also can be explained by ingredient, rich in nutrients such as high-value proteins, long-chain

* Corresponding author.
E-mail address: [email protected] (P. Fuci~
nos).

https://doi.org/10.1016/j.foodhyd.2020.105893
Received 30 September 2019; Received in revised form 24 March 2020; Accepted 26 March 2020
Available online 31 March 2020
0268-005X/© 2020 Elsevier Ltd. All rights reserved.
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

polyunsaturated fatty acids, carotenoids, vitamins, minerals, and grade.

phenolic compounds, as well as other bioactive molecules (Gouveia,
Marques, Sousa, Moura, & Bandarra, 2010). Among them, Arthrospira
2.2. Optimization of A. platensis antioxidants extraction
platensis is one of the main species exploited by the food and nutrition
industries, being traditionally used as food by different cultures. This
A. platensis antioxidants were recovered using ultrasound-assisted
microorganism is a blue-green filamentous prokaryotic cyanobacterium
extraction in hydroalcoholic solutions. The influence of the ethanol/
well known for its unique composition, comprising not only up to 70% of
water and biomass/solvent ratios were assessed through a Design of
protein containing all the essential amino acids, but also poly­
Experiments (DoE) approach, using the antioxidant activity (ORAC and
saccharides, vitamin B12, C, E, and γ-linolenic acid (GLA). Furthermore,
ABTS) and total phenolic content (TPC) as response variables. The lower
it is a source of potent antioxidants, such as carotenoids, polyphenols
and upper limits for the independent variables were based on previously
and phycobiliproteins -a group of photosynthetic pigments majority
reported conditions for extracting antioxidants from Arthrospira spp.
represented by C-phycocyanin, which are related to numerous reported
(El-Baz, El-Senousy, El-Sayed, & Kamel, 2013; Oh, Ahn, Do, & Lee, 2011;
pharmacological properties; including anticancer, antidiabetes, hep­
Silva et al., 2017; Syarina, Karthivashan, Abas, Arulselvan, & Fakurazi,
atoprotective and anti-inflammatory (Czerwonka et al., 2018; Da Silva,
2015). Table 1 shows the coded variables and their real values for
Fernandes, Barros, Fernandes, & Jos�e, 2019; Hu, Fan, Qi, & Zhang,
A. platensis antioxidant extraction. The obtained extracts were analysed
2019; Plaza, Herrero, Cifuentes, & Iba �n
~ ez, 2009; Soni, Sudhakar, &
as described in Section 2.3. The extract with higher antioxidant activity
Rana, 2017).
was freeze-dried for further encapsulation and incorporation into the
The incorporation of microalgae biomass into traditional foods (e.g.
cookie doughs.
breakfast cereals, bread, pasta, cookies, gelled desserts, and beverages),
which are primarily consumed on a daily basis, has been researched and
several products have already been launched in the market (Gouveia 2.3. Antioxidant activity and total phenolic content
et al., 2010; Lafarga, 2019). In particular, cookies are considered a
convenient dense snack food, offering a valuable supplementation The Oxygen Radical Absorbance Capacity (ORAC) of the extracts was
vehicle for nutritional improvement as they are widely accepted and performed in 96-well microplates, based on the method proposed by Ou,
consumed by all age groups. There is a trend for research and innovation Hampsch-Woodill, and Prior (2001) and further modified by D� avalos,
in this market segment, which promotes the inclusion of healthy in­ Go�mez-Cordov� es, and Bartolom� e (2004). In brief, 20 μL of different
gredients into cookies, such as antioxidants, vitamins, minerals, proteins concentrations of the extracts were added to 120 μL of a 116.67 nmol.
and fibers (Batista et al., 2017; Nogueira & Steel, 2018; Saponjac
� et al., L 1 fluorescein solution prepared in 75 mmol.L-1 phosphate buffer at pH
2016). 7.4. The mixture was incubated for 15 min at 37 � C and, subsequently,
Besides the change in consumer’s attitudes towards a healthier diet, 60 μL of 40 mmol.L-1 2,20 -azobis(2-methylpropionamidine)-dihydro­
it is noteworthy that food ingredients and their nutritional needs vary chloride (AAPH) were rapidly added using the automatic reagent
among individuals, especially children, elderly and athletes (Tan, Toh, injector of the plate reader (Biotek Synergy H1). A blank (Fluorescein þ
Wong, & Li, 2018). This context motivates a growing market for AAPH) prepared with 20 μL of phosphate buffer instead of the extracts
personalized healthy nutrition, which aims to tailor food and diets was also analysed, and Trolox was used as standard. Fluorescence was
specifically based on an individual’s health condition. In light of this, recorded every 5 min after AAPH addition (excitation wavelength 485
three dimensional (3D) food printing has gained increasing attention for nm, emission wavelength 520 nm) for 120 min. Results were calculated
its distinctive potential to create complex geometric structures, enabling based on the differences in areas under the fluorescein decay curve
mass customisation while having economic and environmental benefits. between the blank and the samples and were expressed as μmol.L 1 of
The main advantage of this emerging technology is being able to Trolox equivalents/g sample.
personalize food by tailoring nutrition in a novel multi-flavoured, col­ The spectrophotometric analysis of ABTS radical scavenging activity
oured and textured structure, allowing the incorporation of a broad was conducted according to the method of Re et al. (1999). Firstly, an
range of ingredients (Dankar, Haddarah, Omar, Sepulcre, & Pujola �, ABTS solution was prepared by mixing 7 mmol.L 1 ABTS (2,20 -azino-bis
2018; Liu et al., 2018a; P�erez, Nykvist, Brøgger, Larsena, & Falkeborg, (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt with 2.45
2019; Sun et al., 2018). mmol.L 1 potassium persulfate, allowing this mixture to stand at room
Considering the above mentioned, this study aimed at developing 3D temperature for 12–16 h in the dark. Subsequently, the ABTS solution
printed functional cookies fortified with antioxidants extracted from was diluted with Milli-Q water to obtain an absorbance of 0.70 � 0.02 at
A. platensis, to create a new functional food based on an innovative 3D 734 nm. In a 96-well microplate, 10 μL of the sample was added with
food-ink. Due to the inherent instability of C-phycocyanin, carotenoids 200 μL of the ABTS solution, and after 6 min of reaction, the absorbance
and other antioxidant compounds present in this microalga, the was measured at 734 nm. The scavenging capacity percentages (%
encapsulation of its extract in alginate microbeads was proposed as a RadScav) were calculated using Eq. (1). Results were expressed as Tro­
way of improving the cookies stability to heat, light, and oxygen during lox Equivalent Antioxidant Capacity (TEAC) in mmol.L 1 of Trolox per g
the baking and further storage. Parameters such as colour, texture, water of A. platensis biomass (mmol.L 1 TEAC/g).
activity and antioxidant potential were investigated and compared with
% RadScav ¼ 1-(Abss-Absc/Absb)*100 (1)
the freeze-dried extract and whole biomass incorporation into the cookie
dough. Where Abss, Absb and Absc are the absorbance of the sample, blank and

2. Materials and methods Table 1

Full Factorial 2k Design of Experiments for A. platensis antioxidants extraction,
2.1. Materials with two factors and two central points. Real values in parentheses.
Run X1 (Ethanol/Total solvent ratio) X2 (Microalgae mass/Volume of solvent)
Arthrospira platensis biomass was obtained commercially in a
1 1 (100%) 1 (12%)
specialized store (Braga, Portugal). Potassium phosphate dibasic and 2 1 (100%) 1 (2%)
potassium di-hydrogen phosphate were purchased from Fisher Bio­ 3 1 (0%) 1 (12%)
reagents (Pittsburgh, USA) and AppliChem (Darmstadt, Germany), 4 1 (0%) 1 (2%)
respectively. All other reagents were purchased from Sigma-Aldrich (St. 5 0 (50%) 0 (7%)
6 0 (50%) 0 (7%)
Louis, MO, USA). All solvents and reagents used were of analytical

2
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

negative control, respectively. handled gently to avoid structural damage, and they were allowed to
The total phenolic content (TPC) of the extracts was determined rest for 3 min before analysis. Temperature-sweep profiles were per­
according to the method of Singleton, Orthofer, and Lamuela-Ravento �s formed in the range of 25 � C–150 � C at 5 � C/min and 1 Hz. Complex
(1999), using the Folin–Ciocalteu reagent (FCR) and gallic acid as a modulus (G*) and tan δ were evaluated.
standard. Initially, 0.5 mL of sample was mixed with 0.1 mL of Creep-recovery assays were carried out at 25 � C by applying constant
Folin-Ciocalteu reagent and vigorously stirred. After 5 min, 0.5 mL of a stress (55 Pa) for 360 s on the dough and allowing strain recovery for
7.0% sodium carbonate solution was added to alkalinize the medium, 600 s after load removal. The strain was obtained as a function of time,
and the mixture was allowed to react for 1 h at room temperature. The and the data were represented by creep compliance: JðtÞ ​ ðPa 1 Þ ¼ γ=σ,
absorbance was measured spectrophotometrically at a wavelength of where γ and σ are the strain and constant shear stress during the creep
760 nm, and the results were expressed as mg of gallic acid equivalent test, respectively. The creep compliance data of the dough samples were
(GAE) per g of A. platensis biomass (mg GAE/g). fitted with Burger’s model for creep and recovery stages (Eqs. (2) and
(3), respectively).
2.4. Preparation of the cookie dough � � t�� t
JðtÞc ¼ J0 þ Jm 1 exp þ (2)
λ η0
Control cookies were prepared according to the formulation reported
� � t��
by Kim et al. (2019), using wheat flour, butter, powdered sugar, milk JðtÞr ¼ Jmax J0 Jm 1 exp (3)
and xanthan gum. A. platensis incorporation was done by replacing an λ
equivalent amount of wheat flour following three different approaches:
where J0 (Pa 1), Jm (Pa 1), and Jmax (Pa 1) represent the instanta­
(1) direct addition of 2.0% whole A. platensis dried biomass, (2) incor­
neous, viscoelastic, and maximum creep compliance values, respec­
poration of the freeze-dried antioxidant extract obtained from the same
tively; t (s) and λ (s) are the phase and average retardation time,
amount of biomass and (3) incorporation of an equivalent amount of
respectively; and η0 is the viscosity coefficient (Pa.s). The relative elastic
antioxidant extract encapsulated into alginate microbeads. The dough
portion (%) was determined by the ratio between the equilibrium
formulations are presented in Table 2.
compliance and the maximum compliance.
2.4.1. A. platensis extract encapsulation
2.5.2. Texture measurement
Freeze-dried A. platensis extract was encapsulated within alginate
The dough firmness was assessed by a uniaxial compression assay in
microbeads through vibrational extrusion technique using the Büchi
a Texture Analyser TA-HD plus Stable MicroSystem (Godalming, Surrey,
Encapsulator B-395 Pro® (Büchi Labortechnik AG, Flawil, Switzerland).
UK). Cylindrical dough samples (12 mm diameter, 10 mm height) were
Briefly, a 2.0% (w/v) sodium alginate aqueous solution was prepared,
compressed at a speed of 1 mm/s, with trigger force of 5 g up to 90%
and the freeze-dried extract (8% w/v) was added under stirring. The
strain level. Results were expressed by the peak force in the force-time
parameters selected were chosen based on the manufacturer’s recom­
graph (N.s). Measurements were repeated five times for each
mended conditions for air-flow configurations: inner nozzle size of 150
formulation.
μm and outer nozzle of 600 μm, frequency 2000 Hz, electrode 1200 V,
amplitude 2, airflow 0.8 mbar and flow rate of 1.5 mL/min.
The beads formed were collected into a 0.1 mol.L 1 calcium chloride 2.6. 3D printing and post-processing
solution and stirred at 500 rpm. After all the alginate solution was
dispensed, the beads were left at a lower agitation rate (200 rpm) for 2 h The cookies were produced using a 3D food printer (Focus, Byflow,
to complete the hardening process. The resulting A. platensis-calcium Netherlands) equipped with a paste printing head and a 1.6 mm aperture
alginate microbeads were retrieved by filtration using a 100-μm strainer nozzle. A cylinder shape (27.6 mm diameter and 6.72 mm height) was
and rinsed with distilled water. Before weighting for cookie dough sliced (Slic3r software) to micro-extrude 6 layers, each one with 1.12
incorporation, the water excess was removed off the microbeads with mm thickness, through a nozzle moving at a speed of 10 mm s 1. During
filter paper. printing, flow rate and Z-offset were adjusted to obtain the adequate
dough weight and shape (diameter and thickness) according to the 3D
2.5. Dough characterization model. Each formulation was printed at least in triplicate.
The printed cookies were baked at 150 � C for 25 min. Then,
2.5.1. Rheological analyses depending on the type of experiment, the cookies were analysed in
Oscillatory dynamic measurements and creep-recovery tests were triplicate (using three cookies) or in quintuplicate (using five cookies).
carried out in an HR-1 rheometer (TA Instruments, USA) equipped with Measurements of height and diameter were performed in three repli­
a stainless steel parallel plate geometry (40 mm diameter, 1000 μm gap) cates, before and after post-processing, aiming to evaluate shape fidelity.
within the linear viscoelasticity domain, in duplicate. The samples were This parameter was based on the differences between the cookies
theoretical and measured dimensions, as described in Eq. (4). The effect
of baking on the cookie’s shape was determined as the percentual
Table 2
variation of the dimensions before and after baking (Eq. (5)).
Cookies dough formulations fortified with different A. platensis forms.
Ingredients Control (g/ Biomass (g/ Free Extract Encapsulated ðMeasured dimension * 100Þ
Shape fidelity ð%Þ ¼ (4)
100 g) 100 g) (g/100 g) Extract (g/100 g) Theoretical dimension
Wheat flour 40 38 39.2 30
Butter 25 25 25 25
½ðBaked cookie dimension Raw cookie dimensionÞ*100�
Variation ð%Þ ¼
Powdered 22 22 22 22 Raw cookie dimension
sugar (5)
Milk 13 13 13 13
Xanthan 0.5 0.5 0.5 0.5
gum 2.7. Cookies physical-chemical characterization
A. platensis 0 2 0.8a 10b
a
Amount of freeze-dried extract present in 2.0% of A. platensis biomass. All cookies were analysed in terms of colour variation, water activity,
b
Incorporation of 10% of A. platensis alginate microbeads with an amount of texture and antioxidant potential, 24 h after baking and after 30 days of
extract equal to the free extract formulation. storage at room temperature, protected from light.

3
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

2.7.1. Colour analysis agitation at low frequency. The enhanced extraction obtained by ul­
The colour of cookies samples was measured using a colourimeter trasounds is mostly attributed to the acoustic cavitation produced in the
(CR-400; Konica Minolta, Inc., Tokyo, Japan). The first colour mea­ solvent by the passage of an ultrasound wave. Moreover, UAE also exerts
surement was acquired 24 h after baking to ensure and appropriate a mechanical effect, allowing greater penetration of solvent into the cell
cooling before the readings. The results were expressed in terms of L*, wall, increasing the contact surface area between the solid and liquid
lightness (values from 0 to 100%); a*, redness to greenness (60 to 60, phase. As a result, the solute quickly diffuses from the solid phase to the
respectively); b*, yellowness to blueness (60 to 60, respectively), ac­ solvent, increasing bioactive recovery when compared to conventional
cording to the CIELab system. The total colour difference (ΔE*) between methods (Haque, Dutta, Thimmanagari, & Chiang, 2016; Kurd &
sample cookies along storage time (30 days), as well as between raw Samavati, 2015; Liu, Wei, & Liao, 2013; Zou, Jia, Li, Wang, & Wu,
dough and cooked samples, was determined using L*, a* and b* average 2013).
values, according to equation (6). The measurements were conducted Various parameters play a significant role in optimizing the experi­
using a white standard (L* ¼ 93.90, a* ¼ 0.3158, b* ¼ 0.3321), under mental conditions for the development of an extraction method.
artificial fluorescent light at room temperature. Three replicates were Extraction time, temperature and the solid-to-liquid ratio are generally
analysed for each formulation, with five measurement locations per considered to be the most critical factors that affect bioactive recovery.
cookie, including the centre and its surrounding (Batista et al., 2017). The choice of an extracting solvent is also a crucial step towards
� �1 extraction optimization; different solvents will yield different extract
ΔE* ¼ ðΔL* Þ2 þ ðΔa* Þ2 þ ðΔb* Þ2 2 (6) amounts and composition. In the present study, water and ethanol were
employed as extraction solvents for the microalga A. platensis consid­
2.7.2. Texture analysis ering food application safety (Chaiklahan et al., 2013; Zou et al., 2013).
The cookies’ texture was evaluated using a Texture Analyser TA- The recovery of antioxidant compounds from A. platensis biomass
HDplus Stable MicroSystem (Godalming, Surrey, UK) in penetration was optimised through a Design of Experiments (DoE) approach, using a
mode, with a 2 mm cylindrical stainless probe, a target distance of 4 mm 2k full factorial design with 2 factors. The effect of ethanol/water and
and speed test of 0.5 mm s 1. The resistance to penetration (or hardness) biomass/solvent ratios on the antioxidant activity (ORAC and ABTS) and
was measured by the peak force in the force-time graph (N.s). Mea­ total phenolic content (TPC) were analysed as response variables (see
surements were repeated five times for each formulation sample (one Table 1). All the response curves exhibited an excellent fitting (r2 ¼
measurement per cookie). 0.99) and statistical significance. The response surfaces after 30 min and
1 h of ultrasound treatment are represented in Fig. 1. It is possible to
2.7.3. Water activity determination notice that the phenolic content increased proportionally with the
The cookies water activity (aw) was determined using an Aqualab 4 amount of microalga. Additionally, increasing the treatment time to 1 h
TE Water Activity Meter (Meter Group, Inc., Pullman, USA) at 25 � 0.5 promoted a higher phenolic content, which was also higher with
�
C. Measurements were repeated three times for each formulation as a increasing A. platensis content. Nevertheless, the recovery of phenolic
crushed powder. compounds was more efficient at 0% ethanol independently of the time.
The ABTS exhibited a similar trend; although in this case, the extraction
2.7.4. Antioxidant activity time was crucial, being the antioxidant activity at 1 h more than 2.5-fold
The antioxidant activity of the cookies was evaluated following the higher than at 30 min. The ORAC assay corroborated the ABTS results,
methods described in section 2.3 in triplicate; however, with results pointing out that to obtain an extract with high antioxidant activity, the
expressed per gram of cookie instead. For the extraction of cookies ultrasound treatment should be performed during 1 h with 2.0%
antioxidant fraction, aliquots of 0.5 g of the control, free and encapsu­ biomass and 0% ethanol. After freeze-drying the liquid extract obtained
lated extract cookies, previously milled with a mortar and pestle, were in this condition, 0.4 g of dry antioxidant extract was obtained per gram
mixed with 2.5 mL of 75 mmol.L-1 phosphate buffer at pH 7.4 on a of biomass. This extract was used for the cookie formulation
vortex for 1 min; and followed by centrifugation at 9000 rpm for 10 min. experiments.
This process was repeated twice, and supernatants were combined and
filtered through a 0.45 μm syringe filter. Antioxidants from cookies 3.2. Cookie dough characterization
containing A. platensis biomass were recovered by ultrasound-assisted
extraction for 1 h, using 5 mL of 75 mmol.L-1 phosphate buffer at pH Cookies quality is influenced by several factors, such as the quality
7.4 as the solvent. and amount of ingredients used, processing conditions and moulding of
the dough, as well as baking and cooling of the cookies. Among those
2.8. Statistical analysis factors, dough rheology is of considerable importance in cookies
manufacture as it influences the dough machinability and the final
Statistical analysis of the experimental data was performed through sensorial characteristics. Doughs with extreme degrees of firmness or
the t-test or analysis of variance (one way ANOVA), followed by Tukey’s softness will not process satisfactorily on the dough forming equipment
Post Hoc test at a significance level of 95% (p < 0.05), using the software and will not yield adequate products (Manohar & Rao, 2002).
GraphPad Prism 5.0. All results were presented as mean � standard In this work, cookies were shaped through a 3D food printer, where
deviation. Design of Experiments (DoE) and its statistical analysis were the main physical properties involved can be divided into two cate­
performed using Statsoft Inc. Statistica™ (version 13). gories: firstly the ones that affect the extruding process, which includes
the flow behaviour and viscous modulus (G00 ) of the dough; and sec­
3. Results and discussion ondly, the factors which influence the ability to support the three
dimensional structure of the printed products or to maintain its shape
3.1. Optimization of A. platensis antioxidants extraction and structure, such as the elastic modulus (G’), gel strength, among
others (Yang, Zhang, Prakash, & Liu, 2018).
For a practical application in the food industry, antioxidants should Initially, cookies doughs were characterized through a creep-
be first extracted; however, the extraction process efficiency may affect recovery test and texture analysis. During the creep-recovery assay, a
its availability (Wardhani, V� asquez, & Pandiella, 2010). Recently, stress is applied for a specific interval, it is then removed, and the re­
ultrasonic-assisted extraction (UAE) has been widely employed for the covery is monitored for another period. This property provides infor­
recovery of target compounds from many natural products due to its mation about the ability of the sheared and micro-extruded food-ink to
facilitated mass transfer between immiscible phases, through super recover; therefore, the faster the recovery, the higher shape fidelity

4
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

Fig. 1. Response surfaces of the biomass and ethanol concentrations combined effect in A. platensis’ antioxidant activity and TPC.

should be expected. Likewise, less strain during the test indicates a variation (J) as a function of time, which is the ratio of the deformation γ
stronger ability of the material to maintain the shape and structure of to the applied stress τ. Dough deformation could be used to characterize
printed products; which, however, will also require higher extrusion its strength, which means the harder the dough, the higher the amount
rates (Yang, Zhang, Fang, & Liu, 2019). of energy required to achieve the same deformation when compared
Fig. 2a shows the creep–recovery curves expressed by a compliance with a softer dough. Accordingly, if a material has a high compliance, it

Fig. 2. Dough rheology analyses. (a) Creep-recovery curves and temperature-sweep profiles represented by G* (b) and tan δ (c) against temperature.

5
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

will present low rigidity and high deformation or strain; and, conse­ bind with water molecules through hydrogen bonds, suppressing the
quently, better extrudability (Ahmed, 2015). The cookie dough incor­ water absorption and the gluten protein swelling. Consequently, the free
porated with encapsulated extract showed higher maximal strain or extract dough displayed a lower network strength, comparable to the
compliance with the applied stress, while the one prepared with one with encapsulated extract (Liu, Liang, Saeed, Lan, & Qin, 2019).
microalga biomass led to a more stiff dough (Table 3). The replacement The dough properties during the cooking process were evaluated
of a small amount of flour by microalga biomass resulted in the inclusion through a temperature-sweep analysis and results revealed that the
of a complex biological ingredient, rich in proteins and polysaccharides. obtained profiles were consistent with that of the creep-recovery test.
These molecules have an important role in the water absorption process, Fig. 2b and c show the viscoelastic properties (complex modulus and tan
which promote the increase of dough firmness (Bolanho et al., 2014; δ) against temperature increase. G* reflects both contribution of elastic
Gouveia, Batista, Miranda, Empis, & Raymundo, 2007; Gouveia et al., (G0 ) and viscous (G00 ) moduli; while tan δ, which character prevail, is
2008). On the other hand, the encapsulated antioxidant extract dough defined as G’’/G’. In the mechanical spectra, tan δ was lower than 1 (G’
led to an unstructured network due to the water molecules present > G”) for all temperatures, indicating that all doughs behave as a gel-
among the calcium-alginate microbeads, resulting in a reduction of its like material. The dough incorporated with A. platensis biomass
viscosity. Furthermore, the addition of the free antioxidant extract did exhibited higher G* values, as the biomass composition promotes a more
not change the dough behaviour, matching with the one observed for the structured network. Oppositely, the encapsulated extract dough pre­
control cookies. sented lower G* values due to its reduced viscosity. Nonetheless, tan δ
The creep and recovery stages were fitted to Burger’s model (Eqs. (2) spectra did not show great differences, as the proportion between elastic
and (3), respectively) and results are shown in Table 3. The instanta­ and viscous moduli was similar for all formulations.
neous elastic compliance (J0C), viscoelastic compliance (J1C) and the Moreover, throughout the temperature-sweep profile it is also
steady viscosity (μ0) corroborated with Fig. 2a. Biomass cookie dough possible to detect a continuous decrease in G* up to 40 � C, which may be
revealed high μ0 and lower compliances, which means that the higher related to the melting of the butter that accounts for a significant part
the steady-state viscosity, the higher the resistance to deformation. The (25 g/100 g) of the dough, as equally observed by Kim et al. (2019).
opposite behaviour was observed for the encapsulated extract dough, Afterwards, the modulus remains almost constant until 105–110 � C,
which presented higher compliances and lower steady-state viscosity; when it starts to increase. Nevertheless, this phenomenon occurred at a
therefore, less resistance to deformation. lower temperature for cookies incorporated with encapsulated extract
Contrastingly, retardation time (λ) did not show the same behaviour, (~90 � C). When the temperature rose to above 120 � C, a further
and only the biomass and the encapsulated extract doughs were signif­ decrease of G’ and G” was detected. It is inferred that the pyrolytic
icantly different (p < 0.05). However, this parameter agrees with the
compliance results. The retardation time (λ) mainly indicates the time
required for the sample strain to decay to the initial value at 1/e, and the
less stiff dough (encapsulated extract) showed higher (λ). The recovery
phase exhibited similar behaviour, with elastic compliance (J0C),
viscoelastic compliance (J1C) and the steady viscosity (μ0) following the
same tendency. More stiff doughs take longer to deform, and also to
recover their structure. Nevertheless, despite the differences, the elastic
portion of the dough did not differ significantly (p < 0.05) among all
formulations.
Concerning the texture analysis, all doughs incorporated with
different forms of A. platensis presented significant differences (p < 0.05)
when compared to the control. As it can be observed in Fig. 3, the
biomass cookie dough showed higher hardness, which agrees with the
creep-recovery results. Nevertheless, the dough incorporated with the
free extract, which had similar rheological behaviour as the control,
Fig. 3. Texture analysis of the cookie doughs incorporated with different forms
exhibited a low hardness value. This decrease of hardness could be
of A. platensis. Results are presented as mean � standard deviation. The term
attributed to the reduced flour mass and its substitution for components
“ns” denotes a not statistically significant difference.
of the freeze-dried antioxidant extract. Those constituents may further

Table 3
Creep-recovery analysis of the cookies doughs incorporated with different forms of A. platensis. Results are presented as mean � standard deviation.
Dough Creep

Compliance (1/Pa) Retardation Coefficient of Max. Strain (%) Max. r2

time (s) viscosity (Pa.s) Compliance (1/Pa)
J0C J1C
λ1C η0
b b
Control 0.0037 � 0.0002 0.0256 � 0.0009 29.46ab � 3.26 12169a � 11 3.09b � 0.06 0.056b � 0.001 0.99
Biomass 0.0017a � 0.0003 0.0111a � 0.0018 31.60b � 1.15 24343b � 4073 1.45a � 0.24 0.026a � 0.004 0.99
Free Extract 0.0039b � 0.0003 0.0265b � 0.0023 28.75ab � 1.56 11827a � 27 3.18b � 0.14 0.058b � 0.003 0.99
Encapsulated Ext. 0.0073c � 0.0007 0.0484c � 0.0037 22.29a � 1.48 7118a � 741 5.54c � 0.50 0.109c � 0.002 0.99

Dough Recovery
Compliance (1/Pa) Retardation Equilibrium Equilibrium Relative elastic r2
J0C J1C time (s) strain (%) compliance (1/Pa) portion (%)
λ1R

Control 0.0055b � 0.0002 0.0169b � 0.0011 87.00b � 6.40 1.82a � 0.01 0.0330a � 0.0003 58.90a � 1.60 0.90
Biomass 0.0033a � 0.0006 0.0109a � 0.0018 102.70b � 5.20 0.65a � 0.11 0.012a � 0.002 44.60a � 0.20 0.93
Free Extract 0.0062b � 0.0002 0.0179b � 0.0000 85.50b � 2.70 1.80a � 0.15 0.033a � 0.003 56.70a � 2.20 0.95
Encapsulated Ext. 0.0115c � 0.0004 0.0294c � 0.0004 64.60a � 3.20 3.18b � 0.55 0.058b � 0.010 53.30a � 10.50 0.94

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M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

decomposition and leaching of the amylose in flour starch granule were Fig. 5b. Upon baking, the cookie thickness varied from 3.37% for the
induced, which may have caused the corresponding decrease (Kim et al., biomass cookies, until 18.22% for the encapsulated extract cookies. The
2019). increase in this dimension is related to the gas production from the water
vaporization and a higher dough elasticity, which explains the fact that
3.3. 3D printing and post-processing the cookies incorporated with fresh alginate microbeads have an
enlargement superior to the other formulations, corroborating the re­
3D food printing is a digital manufacturing technology, which is used sults obtained in the rheology analyses (Chevallier, Della Valle, Colonna,
to fabricate three-dimensional structures in a layer-by-layer manner, Broyart, & Trystram, 2002).
using liquid-, gel-, or powder-type food materials as a printing medium; Oppositely, the diameter of the cookies had a smaller variation after
it includes three steps: modelling, 3D printing and post-processing. A the post-processing treatment, as it can also be observed in Fig. 5b.
range of 3D printing methods have been utilized for food printing, such Except for the encapsulated extract cookies, which suffered a shrinkage
as selective laser sintering/hot air sintering, hot-melt extrusion/room of around 7% probably due to the high water loss, all the other formu­
temperature extrusion, binder jetting, and inkjet printing (Holland, lations suffered a positive influence of the temperature during baking.
Tuck, & Foster, 2018; Portanguen, Tournayre, Sicard, Astruc, & Mirade, The heat promotes the melting of fat, which confers plasticity and an
2019; Sun, Zhou, Huang, Fuh, & Hong, 2015). Among them, ease flow, resulting in an initial spreading followed by a width retraction
extrusion-based 3D food printing is the most widely adopted method, at the end of the process (Walker, Seetharaman, & Goldstein, 2012).
which consists of a material being extruded through a nozzle moving in
x-, y- and z-direction, building up a structure layer-by-layer (Kim et al., 3.4. Cookies physical-chemical characterization
2019; Sun et al., 2018).
For a successful 3D printing step, it is required a material which can A number of parameters can be scrutinized from baked cookies that
be smoothly extruded through the nozzle and, at the same time, can are of crucial importance to determine their adequacy. Colour is an
support the weight of the subsequent printed layers without deforma­ attribute which impacts food quality, contributing to consumer’s
tion. In this context, the knowledge of the material’s rheological and attraction to a product. The incorporation of A. platensis in different
mechanical profiles is imperative to achieve proper extrudability and forms into cookie doughs stimulated a decrease in luminosity when
structure stability during the process (Liu, Zhang, Bhandari, & Yang, compared to the control, which was more prominent in the biomass and
2018b; Wang, Zhang, Bhandari, & Yang, 2018; Yang et al., 2018). free extract cookies (Fig. 6). Biomass cookies were distinguished by the
Furthermore, 3D food printing is not only affected by the physico­ microalga green tonality, showing negative values of a*; while the free
chemical properties of the ingredients used, but also by the process extract cookies tended to the blue colour, represented by the C-phyco­
parameters, such as nozzle moving speed, extrusion rate, nozzle diam­ cyanin distinctive pigmentation (Lucas, Morais, Santos, & Costa, 2018).
eter, and layer and nozzle heights. This correlation between the food Table 4 presents the total colour differences (ΔE*) between the raw
formula attributes and the operational conditions influences the printing dough and baked cookies, as well as the differences for each formulation
precision and, thus, is a key factor in the end-product quality (Dankar over 30 days of storage time. A. platensis cookies showed significantly
et al., 2018; He, Zhang, & Fang, 2019; P� erez, Nykvist, Brøgger, Larsen, & colour variation upon baking, with ΔE* varying from 17.47 to 25.50.
Falkeborg, 2019). This outcome may be explained primarily by the browning of cookie
Lastly, the printed food pieces may require a further post-deposition surface (lightness decrease and colour parameter increase), possibly due
cooking process (e.g. baking and boiling), which involves different to formation of Maillard reaction products (MRP) through the interac­
levels of heat penetration in the food matrix; resulting in texture mod­ tion of reducing sugars with proteins, but also possibly owing to starch
ifications and, possibly, misshapen structures (Dankar et al., 2018; Sun dextrinization and sugar caramelization. Moreover, changes in tonality
et al., 2018). Cookie dough is a material that unavoidably requires may be related to pigment loss upon exposure to high temperatures
post-processing; however, it rapidly deforms after baking. Kim et al. (Batista et al., 2017; Chevallier et al., 2002).
(2019) have studied the addition of hydrocolloids as a structuring agent, On the other hand, along the conservation time, all cookie formu­
aiming to suppress product deformation in the high-temperature envi­ lations presented a low ΔE*, principally the encapsulated extract cookie.
ronment of the post-processing step, whereas maintaining the in­ According to Mokrzycki and Tatol (2011), ΔE* values between 1 and 2
gredients and product characteristics of the desired cookie. Xanthan indicate that only experienced observers can notice a colour difference,
gum in the concentration of 0.5% was reported to promote the best and values between 2 and 3.5 suggest that an inexperienced observer is
shape accuracy of the selected 3D printed model after the baking pro­ also able to see the difference. Hence, it is possible to conclude that the
cess; therefore, it was chosen to develop the functional cookies of this encapsulation of A. platensis extract improved the cookie colour stability
work. during the 30 days of storage time, when compared to other
3D food printing is a tool which allows the creation of unique, formulations.
innovative, products that other methods cannot emulate. Among Another essential physical stability factor, which gives an identity to
numerous applications, this technology is becoming popular due to the a food product, is the texture (Carter, Galloway, Campbell, & Carter,
possibility to design foods with appealing forms, new textures and 2015). In this work, the cookie’s texture was evaluated through a
personalized nutritional values, where several raw materials can be penetration test, and the results are represented in Fig. 7. As it has been
blended according to individual’s physical and nutritional status. stated for the cookie dough texture analysis (see section 3.2), the addi­
Additionally, this 3D printing flexibility enables the use of alternative tion of A. platensis biomass promoted an increase in the cookie hardness.
ingredients in food processing, such as microalgae, insects and fungi; in Inversely, the incorporation of microalgae antioxidant extract in fresh
the production of tasty, healthy and tailored foods (An, Guo, Zhang, & alginate microbeads resulted in a softer texture, which it was expected
Zhong, 2019; Godoi, Prakash, & Bhandari, 2016; Lille, Nurmela, Nor­ considering the water molecules present among the microparticles.
dlund, Metsa €-Kortelainen, & Sozer, 2018; Severini, Azzollini, Albenzio, Finally, over the 30 days of storage time, there was no significant dif­
& Derossi, 2018; Voon, An, Wong, Zhang, & Chua, 2019). ference (p > 0.05) in the cookie texture for all the developed
Fig. 4 shows the 3D printed cookies incorporated with different formulations.
forms of the microalga A. platensis right after the 3D printing step and The incorporation of A. platensis into cookies by conventional
after the post-processing treatment. methods has been described in the literature by a few authors, pro­
All 3D printed cookies presented shape fidelity in the range of 100 � moting different effects on the final product texture. Onacik-Gür, Zbi­
5.0%, demonstrating dimensional consistency with the 3D model kowska, and Majewska (2018) evaluated the addition of 1%, 2% and 3%
(Fig. 5a). The effect of the cooking process over the cookies is showed in of microalga powder and reported a decrease in the cookies hardness

7
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

Fig. 4. 3D printed cookies incorporated with different forms of A. platensis. First row: after the 3D printing step; second row: after the baking process.

Fig. 5. (a) Shape fidelity of the raw 3D printed cookies and (b) Effect of the cooking process on the 3D cookies measures. Results are presented as mean � standard
deviation. The term “ns” denotes a not statistically significant difference.

with the increase of microalga concentration. On the other hand, Mar­ 3.4.1. Antioxidant activity
cinkowska-Lesiak, Onopiuk, Zalewska, Ciepłoch, and Barotti (2017) The microalga A. platensis is recognized to have notable free radical
incorporated different powder amounts of the same species into short­ scavenging properties and antioxidant activity, due to the presence of
bread biscuits, which promoted an increase in the cookie hardness natural pigments and other bioactive compounds in its composition. Its
directly proportional to the addition of microalga powder; therefore, light-harvesting protein-pigment complexes called phycobilisomes are
corroborating with the results found for the 3D printed cookies of this composed by phycobiliproteins, where C-phycocyanin and allophyco­
study. cyanin are considered the most important ones. Moreover, this micro­
Concomitantly with the food texture, water activity (aw) is also a alga contains phenolic compounds and a spectrum of natural mixed
significant parameter regarding the conservation of low moisture carotene and xanthophyll phytopigments that, together with phycocy­
cookies, particularly for the maintenance of a crispy texture. Further­ anin, seem to be related to its distinguished antioxidant activity (Batista
more, the food physical-chemical and microbiological stability depend et al., 2017; Zaid, Hammad, & Sharaf, 2015).
greatly on the water content and its interaction with food ingredients. The 3D printed cookies developed in this work had their antioxidant
Water activity is a measure of the availability of water molecules to enter capacity evaluated through the ORAC and ABTS assays after the post-
into microbial, enzymatic or chemical reactions. Therefore, this processing step and after 30 days of storage (Fig. 8). After the storage
parameter has been used to assess the potential microbial growth and period, the encapsulated extract cookie exhibited a significantly higher
chemical stability of foods after manufacture. It is established that (p < 0.05) ORAC value compared to all other formulations, showing its
bacteria do not grow at aw values of 0.80 or below, while the limit for improvement against processing and environmental factors. On the
mould and yeast growth is 0.6 (Hough. Buera, Chirife, & Moro, 2001; other hand, no significant difference between the formulations was
Khouryieh & Aramouni, 2012). As Fig. 7 shows, all cookie formulations found for ABTS assay. One possible explanation for this discrepancy
presented aw values below 0.3 throughout the 30 days of storage, indi­ could be due to the differences in the mechanism of action of those
cating high microbiological stability. antioxidant analyses. The ORAC assay measures the affinity of anti­
oxidative compounds to neutralize the free radicals over a period of
time, accounting for any potential lag phases in antioxidant activity

8
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

Fig. 6. Colour parameters L*, a* and b* for the raw cookie doughs and for the baked cookies after 24 h and 30 days of storage. a) Raw dough; b) baked cookies after
24 h and c) baked cookies after 30 days. Results are presented as mean � standard deviation. For results with the same letter, the difference between the means is not
statistically significant.

rather than providing a measurement of only fast acting antioxidants;

Table 4
whereas the ABTS assay neutralize free radicals at a particular point of
Total colour variation (ΔE*) between cooked and raw cookie samples and colour
time without accounting for slow-acting antioxidants (Nayak, Liu, &
stability along conservation time.
Tang, 2015).
Total colour difference (ΔE*) Raw vs. Baked 24 h vs. 30 days Additionally, it is noticeable that antioxidant capacity was also found
Control 9.71 2.85 for the cookies with no incorporation of A. platensis (control). Cookies
Biomass 25.50 2.43 are usually prepared with reducing sugars and a protein source, which
Free Extract 25.29 2.12
leads to the formation of MRPs. Those compounds are one of the main
Encapsulated Extract 17.47 1.30
responsible for the browning process characteristic of many foods, and it

Fig. 7. Texture (a) and water activity (b) analyses of the 3D printed cookies incorporated with different A. platensis forms over 30 days of storage time. Results are
presented as mean � standard deviation. The label “*” denotes a statistically significant difference.

9
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

Fig. 8. Antioxidant activity of the 3D printed cookies incorporated with different A. platensis forms over 30 days of storage time. Results are presented as mean �
standard deviation. The label “*” denotes a difference statistically significant with respect to other formulations at the same sampling time (30 days).

has been associated to an increase of their antioxidant potential Methodology, Investigation. Antonio A. Vicente: Supervision, Writing -
(Nooshkam, Varidi, & Bashash, 2019; Yilmaz & Toledo, 2005). review & editing. Lorenzo M. Pastrana: Writing - review & editing,
According to Manzocco, Calligaris, Mastrocola, Nicoli and Lerici Supervision, Funding acquisition. Pablo Fucin ~ os: Conceptualization,
(2000), it can be stated that in the development of the Maillard reaction, Writing - review & editing, Supervision, Funding acquisition.
there is a positive correlation between colour and antioxidant proper­
ties. This correlation was found in foods where Maillard reaction was the Acknowledgement
sole or the prevalent event related to the antioxidant activity. Such
circumstance generally occurs in food products with no or low content of This work was funded by the European Union INTERREG Atlantic
naturally occurring antioxidants; which means that eventual changes in Area Programme and the European Regional Development Fund (ERDF)
the antioxidant capacity upon processing are only due to the formation through the project “Enhance Microalgae: High added-value industrial
of heat-induced antioxidants. Given that, the antioxidant capacity found opportunities for microalgae in the Atlantic Area” (Ref. EAPA_338/
for the control cookies could be explained by the formation of MRPs, as 2016).
antioxidant compounds are absent in its composition.
References
4. Conclusions
Ahmed, E. M. (2015). Hydrogel: Preparation, characterization, and applications: A
review. Journal of Advanced Research, 6, 105–121. https://doi.org/10.1016/j.
The microalga A. platensis was used as a source of antioxidants in the jare.2013.07.006.
development of 3D printed cookies, based on functional food-inks. The An, Y. J., Guo, C. F., Zhang, M., & Zhong, Z. P. (2019). Investigation on characteristics of
antioxidant extraction was optimised through a DoE approach. Optimal 3D printing using Nostoc sphaeroides biomass. Journal of the Science of Food and
Agriculture, 99, 639–646. https://doi.org/10.1002/jsfa.9226.
conditions were 1 h extraction, with 0% ethanol and 2.0% biomass. All Batista, A. P., Niccolai, A., Fradinho, P., Fragoso, S., Bursic, I., Rodolfi, L., et al. (2017).
cookie dough formulations were suitable for extrusion, forming a ho­ Microalgae biomass as an alternative ingredient in cookies: Sensory, physical and
mogenous filament with a diameter close to the nozzle aperture, and chemical properties, antioxidant activity and in vitro digestibility. Algal Research, 26,
161–171. https://doi.org/10.1016/j.algal.2017.07.017.
presenting dimensional consistency with the 3D model after the post- Betoret, E., Betoret, N., Vidal, D., & Fito, P. (2011). Functional foods development:
deposition step. Furthermore, the fortification with A. platensis resul­ Trends and technologies. Trends in Food Science & Technology, 22, 498–508. https://
ted in 3D printed cookies with an innovative appearance. The encap­ doi.org/10.1016/j.tifs.2011.05.004.
Bigliardi, B., & Galati, F. (2013). Innovation trends in the food industry: The case of
sulation of the antioxidant extract was capable of improving the
functional foods. Trends in Food Science & Technology, 31, 118–129. https://doi.org/
antioxidant activity and colour stability along the storage time when 10.1016/j.tifs.2013.03.006.
compared to all other formulations. Therefore, cookies developed in a Bolanho, B. C., Egea, M. B., J� acome, A. L. M., Campos, I., De Carvalho, J. C. M., &
3D printer could be considered a promising alternative for the incor­ Danesi, E. D. G. (2014). Antioxidant and nutritional potential of cookies enriched
with Spirulina platensis and sources of fibre. Journal of Food and Nutrition Research,
poration of new ingredients, such as microalgae, to obtain a novel 53, 171–179.
functional food with antioxidant properties. Carter, B. P., Galloway, M. T., Campbell, G. S., & Carter, A. H. (2015). The critical water
activity from dynamic dewpoint isotherms as an indicator of crispness in low
moisture cookies. Journal of Food Measurement and Characterization, 9, 463–470.
Declaration of competing interest https://doi.org/10.1007/s11694-015-9254-3.
Chaiklahan, R., Chirasuwan, N., Triratana, P., Loha, V., Tia, S., & Bunnag, B. (2013).
The authors declare no conflict of interest. Polysaccharide extraction from Spirulina sp. and its antioxidant capacity.
International Journal of Biological Macromolecules, 58, 73–78. https://doi.org/
10.1016/j.ijbiomac.2013.03.046.
CRediT authorship contribution statement Chevallier, S., Della Valle, G., Colonna, P., Broyart, B., & Trystram, G. (2002). Structural
and chemical modifications of short dough during baking. Journal of Cereal Science,
35, 1–10. https://doi.org/10.1006/jcrs.2001.0388.
Marta V. Vieira: Conceptualization, Methodology, Investigation, Czerwonka, A., Kaławaj, K., Sławi� nska-Brych, A., Lemieszek, M. K., Bartnik, M., et al.
Formal analysis, Visualization, Writing - original draft. Sara M. Oli­ (2018). Anticancer effect of the water extract of a commercial Spirulina (Arthrospira
veira: Conceptualization, Methodology, Investigation, Formal analysis, platensis) product on the human lung cancer A549 cell line. Biomedicine &
Pharmacotherapy, 106, 292–302. https://doi.org/10.1016/j.biopha.2018.06.116.
Visualization, Writing - original draft. Isabel R. Amado: Methodology, Da Silva, S. C., Fernandes, I. P., Barros, L., Fernandes, A., Jos�e, M., et al. (2019). Spray-
Investigation, Writing - review & editing. Luiz H. Fasolin: dried Spirulina platensis as an effective ingredient to improve yogurt formulations:

10
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

Testing different encapsulating solutions. Journal of Functional Foods, 60, 103427. Manohar, R. S., & Rao, P. H. (2002). Interrelationship between rheological
https://doi.org/10.1016/j.jff.2019.103427. characteristics of dough and quality of biscuits; use of elastic recovery of dough to
Dankar, I., Haddarah, A., Omar, F. E. L., Sepulcre, F., & Pujol� a, M. (2018). 3D printing predict biscuit quality. Food Research International, 35, 807–813. https://doi.org/
technology: The new era for food customization and elaboration. Trends in Food 10.1016/S0963-9969(02)00083-2.
Science & Technology, 75, 231–242. https://doi.org/10.1016/j.tifs.2018.03.018. Manzocco, L., Calligaris, S., Mastrocola, D., Nicoli, M. C., & Lerici, C. R. (2000). Review
D�avalos, A., G�omez-Cordov�es, C., & Bartolom�e, B. (2004). Extending applicability of the of non-enzymatic browning and antioxidant capacity in processed foods. Trends in
oxygen radical absorbance capacity (ORAC-Fluorescein) assay. Journal of Agricultural Food Science & Technology, 11, 340–346. https://doi.org/10.1016/S0924-2244(01)
and Food Chemistry, 52, 48–54. https://doi.org/10.1021/jf0305231. 00014-0.
Day, L., Seymour, R. B., Pitts, K. F., Konczak, I., & Lundin, L. (2009). Incorporation of Marcinkowska-Lesiak, M., Onopiuk, A., Zalewska, M., Ciepłoch, A., & Barotti, L. (2017).
functional ingredients into foods. Trends in Food Science & Technology, 20, 388–395. The effect of different level of Spirulina powder on the chosen quality parameters of
https://doi.org/10.1016/j.tifs.2008.05.002. shortbread biscuits. Journal of Food Processing and Preservation, 42, 1–10. https://doi.
El-Baz, F. K., El-Senousy, W. M., El-Sayed, A. B., & Kamel, M. M. (2013). In vitro antiviral org/10.1111/jfpp.13561.
and antimicrobial activities of Spirulina platensis extract. Journal of Applied Mokrzycki, W. S., & Tatol, M. (2011). Color difference ΔE - a survey. Machine Graphics
Pharmaceutical Science, 3, 52–56. https://doi.org/10.7324/JAPS.2013.31209. and Vision, 20, 383–411.
Godoi, F. C., Prakash, S., & Bhandari, B. R. (2016). 3d printing technologies applied for Nayak, B., Liu, R. H., & Tang, J. (2015). Effect of processing on phenolic antioxidants of
food design: Status and prospects. Journal of Food Engineering, 179, 44–54. https:// fruits, vegetables, and grains—a review. Critical Reviews in Food Science and Nutrition,
doi.org/10.1016/j.jfoodeng.2016.01.025. 55, 887–919. https://doi.org/10.1080/10408398.2011.654142.
Gouveia, L., Batista, A. P., Miranda, A., Empis, J., & Raymundo, A. (2007). Chlorella Nogueira, A. de C., & Steel, C. J. (2018). Protein enrichment of biscuits: A review. Food
vulgaris biomass used as colouring source in traditional butter cookies. Innovative Reviews International, 34, 796–809. https://doi.org/10.1080/
Food Science & Emerging Technologies, 8, 433–436. https://doi.org/10.1016/j. 87559129.2018.1441299.
ifset.2007.03.026. Nooshkam, M., Varidi, M., & Bashash, M. (2019). The Maillard reaction products as food-
Gouveia, L., Coutinho, C., Mendonça, E., Batista, A. P., Sousa, I., et al. (2008). Functional born antioxidant and antibrowning agents in model and real food systems. Food
biscuits with PUFA-ω3from Isochrysis galbana. Journal of the Science of Food and Chemistry, 275, 644–660. https://doi.org/10.1016/j.foodchem.2018.09.083.
Agriculture, 88, 891–896. https://doi.org/10.1002/jsfa. Oh, S. H., Ahn, J., Do, H. K., & Lee, H. Y. (2011). The effect of ultrasonificated extracts of
Gouveia, L., Marques, A. E., Sousa, J. M., Moura, P., & Bandarra, N. M. (2010). Spirulina maxima on the anticancer activity. Marine Biotechnology, 13, 205–214.
Microalgae – source of natural bioactive molecules as functional ingredients. Food https://doi.org/10.1007/s10126-010-9282-2.
Science & Technology Bulletin: Functional Foods, 7, 21–37. https://doi.org/10.1616/ Onacik-Gür, S., Zbikowska, A., & Majewska, B. (2018). Effect of Spirulina (Spirulina
1476-2137.15884. platensis) addition on textural and quality properties of cookies. Italian Journal of
Haque, F., Dutta, A., Thimmanagari, M., & Chiang, Y. W. (2016). Intensified green Food Science, 30, 1–12. https://doi.org/10.14674/IJFS-702.
production of astaxanthin from Haematococcus pluvialis. Food and Bioproducts Ou, B., Hampsch-Woodill, M., & Prior, R. L. (2001). Development and validation of an
Processing, 99, 1–11. https://doi.org/10.1016/j.fbp.2016.03.002. improved oxygen radical absorbance capacity assay using fluorescein as the
Herrero, M., Martín-Alvarez,
� P. J., Se~
nor�ans, F. J., Cifuentes, A., & Ib�
an~ ez, E. (2005). fluorescent probe. Journal of Agricultural and Food Chemistry, 49, 4619–4626.
Optimization of accelerated solvent extraction of antioxidants from Spirulina platensis https://doi.org/10.1021/jf010586o.
microalga. Food Chemistry, 93, 417–423. https://doi.org/10.1016/j. P�erez, B., Nykvist, H., Brøgger, A. F., Larsen, M. B., & Falkeborg, M. F. (2019). Impact of
foodchem.2004.09.037. macronutrients printability and 3D-printer parameters on 3D-food printing: A
He, C., Zhang, M., & Fang, Z. (2019). 3D printing of food: Pre-treatment and post- review. Food Chemistry, 287, 249–257. https://doi.org/10.1016/j.
treatment of materials. Critical Reviews in Food Science and Nutrition, 1–14. https:// foodchem.2019.02.090.
doi.org/10.1080/10408398.2019.1641065, 0. Plaza, M., Herrero, M., Cifuentes, A., & Ib� an
~ ez, E. (2009). Innovative natural functional
Holland, S., Tuck, C., & Foster, T. (2018). Selective recrystallization of cellulose ingredients from microalgae. Journal of Agricultural and Food Chemistry, 57,
composite powders and microstructure creation through 3D binder jetting. 7159–7170. https://doi.org/10.1021/jf901070g.
Carbohydrate Polymers, 200, 229–238. https://doi.org/10.1016/j. Portanguen, S., Tournayre, P., Sicard, J., Astruc, T., & Mirade, P. S. (2019). Toward the
carbpol.2018.07.064. design of functional foods and biobased products by 3D printing: A review. Trends in
Hough, G., Buera, M. D. P., Chirife, J., & Moro, O. (2001). Sensory texture of commercial Food Science & Technology, 86, 188–198. https://doi.org/10.1016/j.
biscuits as a function of water activity. Journal of Texture Studies, 32, 57–74. https:// tifs.2019.02.023.
doi.org/10.1111/j.1745-4603.2001.tb01034.x. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., et al. (1999). Antioxidant
Hu, S., Fan, X., Qi, P., & Zhang, X. (2019). Identification of anti-diabetes peptides from activity applying an improved ABTS radical cation decolorization assay. Free Radical
Spirulina platensis. Journal of Functional Foods, 56, 333–341. https://doi.org/ Biology and Medicine, 26, 1231–1237. https://doi.org/10.1016/S0891-5849(98)
10.1016/j.jff.2019.03.024. 00315-3.
Khouryieh, H., & Aramouni, F. (2012). Physical and sensory characteristics of cookies Roberfroid, M. B. (2000). Concepts and strategy of functional food science: The European
prepared with flaxseed flour. Journal of the Science of Food and Agriculture, 92, perspective. American Journal of Clinical Nutrition, 71, 1660S–4S.Crossref Partial Au1
2366–2372. https://doi.org/10.1002/jsfa.5642. ATL STL Year Vol Fpage Lpage.
Kim, H. W., Lee, I. J., Park, S. M., Lee, J. H., Nguyen, M. H., & Park, H. J. (2019). Effect of Saponjac,
� V. T., Cetkovi�
� c, G., Canadanovi�
� c-Brunet, J., Pajin, B., Djilas, S., et al. (2016).
hydrocolloid addition on dimensional stability in post-processing of 3D printable Sour cherry pomace extract encapsulated in whey and soy proteins: Incorporation in
cookie dough. LWT - Food Science and Technology (Lebensmittel-Wissenschaft und cookies. Food Chemistry, 207, 27–33. https://doi.org/10.1016/j.
-Technologie), 101, 69–75. https://doi.org/10.1016/j.lwt.2018.11.019. foodchem.2016.03.082.
Kurd, F., & Samavati, V. (2015). Water soluble polysaccharides from Spirulina platensis: Severini, C., Azzollini, D., Albenzio, M., & Derossi, A. (2018). On printability, quality and
Extraction and in vitro anti-cancer activity. International Journal of Biological nutritional properties of 3D printed cereal based snacks enriched with edible insects.
Macromolecules, 74, 498–506. https://doi.org/10.1016/j.ijbiomac.2015.01.005. Food Research International, 106, 666–676. https://doi.org/10.1016/j.
Lafarga, T. (2019). Effect of microalgal biomass incorporation into foods: Nutritional and foodres.2018.01.034.
sensorial attributes of the end products. Algal Research, 41, 101566. https://doi.org/ Silva, A. de S., Moreira, L. M., De Magalh~ aes, W. T., Farias, W. R. L., Rocha, M. V. P., &
10.1016/j.algal.2019.101566. Bastos, A. K. P. (2017). Extraction of biomolecules from Spirulina platensis using non-
Lille, M., Nurmela, A., Nordlund, E., Mets€ a-Kortelainen, S., & Sozer, N. (2018). conventional processes and harmless solvents. Journal of Environmental Chemical
Applicability of protein and fiber-rich food materials in extrusion-based 3D printing. Engineering, 5, 2101–2106. https://doi.org/10.1016/j.jece.2017.04.008.
Journal of Food Engineering, 220, 20–27. https://doi.org/10.1016/j. Singleton, V. L., Orthofer, R., & Lamuela-Ravent� os, R. M. (1999). Analysis of total
jfoodeng.2017.04.034. phenols and other oxidation substrates and antioxidants by mean of folin-ciocalteu
Liu, Y., Liang, X., Saeed, A., Lan, W., & Qin, W. (2019). Properties of 3D printed dough reagent. Methods in Enzymology, 299, 152–178. https://doi.org/10.1016/S0076-
and optimization of printing parameters. Innovative Food Science & Emerging 6879(99)99017-1.
Technologies, 54, 9–18. https://doi.org/10.1016/j.ifset.2019.03.008. Soni, A. R., Sudhakar, K., & Rana, R. S. (2017). Spirulina- from growth to nutritional
Liu, Y., Liu, D., Wei, G., Ma, Y., Bhandari, B., & Zhou, P. (2018a). 3D printed milk protein product: A review. Trends in Food Science & Technology, 69, 157–171. https://doi.
food simulant: Improving the printing performance of milk protein concentration by org/10.1016/j.tifs.2017.09.010.
incorporating whey protein isolate. Innovative Food Science & Emerging Technologies, Sun, J., Zhou, W., Huang, D., Fuh, J. Y. H., & Hong, G. S. (2015). An overview of 3D
49, 116–126. https://doi.org/10.1016/j.ifset.2018.07.018. printing technologies for food fabrication. Food and Bioprocess Technology, 8,
Liu, Y., Wei, S., & Liao, M. (2013). Optimization of ultrasonic extraction of phenolic 1605–1615. https://doi.org/10.1007/s11947-015-1528-6.
compounds from Euryale ferox seed shells using response surface methodology. Sun, J., Zhou, W., Yan, L., Huang, D., & Lin-ya, L. (2018). Extrusion-based food printing
Industrial Crops and Products, 49, 837–843. https://doi.org/10.1016/j. for digitalized food design and nutrition control. Journal of Food Engineering, 220,
indcrop.2013.07.023. 1–11. https://doi.org/10.1016/j.jfoodeng.2017.02.028.
Liu, Z., Zhang, M., Bhandari, B., & Yang, C. (2018b). Impact of rheological properties of Syarina, P. N. A., Karthivashan, G., Abas, F., Arulselvan, P., & Fakurazi, S. (2015). Wound
mashed potatoes on 3D printing. Journal of Food Engineering, 220, 76–82. https://doi. healing potential of Spirulina platensis extracts on human dermal fibroblast cells.
org/10.1016/j.jfoodeng.2017.04.017. EXCLI Journal, 14, 385–393. https://doi.org/10.17179/excli2014-697.
Lopez-Rubio, A., Gavara, R., & Lagaron, J. M. (2006). Bioactive packaging: Turning foods Tan, C., Toh, W. Y., Wong, G., & Li, L. (2018). Extrusion-based 3D food printing
into healthier foods through biomaterials. Trends in Food Science & Technology, 17, -Materials and machines. International Journal of Bioprinting, 4(2), 1–13. https://doi.
567–575. https://doi.org/10.1016/j.tifs.2006.04.012. org/10.18063/IJB.v4i2.143.
Lucas, B. F., Morais, M. G., Santos, T. D., & Costa, J. A. V. (2018). Spirulina for snack Voon, S. L., An, J., Wong, G., Zhang, Y., & Chua, C. K. (2019). 3D food printing: A
enrichment: Nutritional, physical and sensory evaluations. Lebensmittel-Wissenschaft categorised review of inks and their development. Virtual and Physical Prototyping,
und -Technologie- Food Science and Technology, 90, 270–276. https://doi.org/ 14, 203–218. https://doi.org/10.1080/17452759.2019.1603508.
10.1016/j.lwt.2017.12.032.

11
M.V. Vieira et al. Food Hydrocolloids 107 (2020) 105893

Walker, S., Seetharaman, K., & Goldstein, A. (2012). Characterizing physicochemical Yang, F., Zhang, M., Prakash, S., & Liu, Y. (2018). Physical properties of 3D printed
changes of cookies baked in a commercial oven. Food Research International, 48, baking dough as affected by different compositions. Innovative Food Science &
249–256. https://doi.org/10.1016/j.foodres.2012.04.003. Emerging Technologies, 49, 202–2010. https://doi.org/10.1016/j.ifset.2018.01.001.
Wang, L., Zhang, M., Bhandari, B., & Yang, C. (2018). Investigation on fish surimi gel as Yilmaz, Y., & Toledo, R. (2005). Antioxidant activity of water-soluble Maillard reaction
promising food material for 3D printing. Journal of Food Engineering, 220, 101–108. products. Food Chemistry, 93, 273–278. https://doi.org/10.1016/j.
https://doi.org/10.1016/j.jfoodeng.2017.02.029. foodchem.2004.09.043.
Wardhani, D. H., V�azquez, J. A., & Pandiella, S. S. (2010). Optimisation of antioxidants Zaid, A. A. A., Hammad, D. M., & Sharaf, E. M. (2015). Antioxidant and anticancer
extraction from soybeans fermented by Aspergillus oryzae. Food Chemistry, 118, activity of Spirulina platensis water extracts. International Journal of Pharmacology, 11,
731–739. https://doi.org/10.1016/j.foodchem.2009.05.057. 846–851. https://doi.org/10.3923/ijp.2015.846.851.
Yang, F., Zhang, M., Fang, Z., & Liu, Y. (2019). Impact of processing parameters and post- Zou, T. B., Jia, Q., Li, H. W., Wang, C. X., & Wu, H. F. (2013). Response surface
treatment on the shape accuracy of 3D-printed baking dough. International Journal of methodology for ultrasound-assisted extraction of astaxanthin from Haematococcus
Food Science and Technology, 54, 68–74. https://doi.org/10.1111/ijfs.13904. pluvialis. Marine Drugs, 11, 1644–1655. https://doi.org/10.3390/md11051644.

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