Innovative Food Science and Emerging Technologies 44 (2017) 217–223

Contents lists available at ScienceDirect

Innovative Food Science and Emerging Technologies

journal homepage: www.elsevier.com/locate/ifset

Environmental impact of phycocyanin recovery from Spirulina

platensis cyanobacterium
Sofia Papadaki a,⁎, Konstantina Kyriakopoulou a, Ioannis Tzovenis b, Magdalini Krokida a
a
School of Chemical Engineering, National Technical University of Athens, Athens, Greece
b
Biology Department, National & Kapodistrian University of Athens, Athens, Greece

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

Article history: Multifunctional extracts from Spirulina platensis are suggested as food additives, due to their high content in func-
Received 8 November 2016 tional ingredients and specifically phycocyanin. The recovery of phycocyanin from the microalgal biomass is per-
Received in revised form 6 February 2017 formed by using ultrasounds and polar solvents such water, ethanol or buffer. The application of drying
Accepted 23 February 2017
pretreatment in combination with the use of different solvents presents variation in the yields, affecting the ac-
Available online 24 February 2017
tual recovery of the protein and hence the environmental impact of the production of 1 kg phycocyanin. Life cycle
Keywords:
analysis on the recovery techniques for the isolation of the desired phycocyanin was performed in order to eval-
Environmental footprint uate the selected extraction processes' sustainability. Drying exhibited increased environmental footprint due to
Extraction of bioactive compounds the energy demand, while at the same time affecting not only the yielding but also the quality of the extracts. The
Microalgae cultivation process use of aqueous solvents can lead to an environmental and efficient extraction, replacing organic solvent systems
Life cycle assessment sufficiently.
Pigments Industrial relevance: Phycocyanin is a pigment-protein complex which is used into various food products to en-
hance their nutritional qualities acting as food colorant, antioxidant and emulsifier, which can sufficiently replace
or reduce the use of synthetic additives. For the effective recovery of phycocyanin, the nutrient should be extract-
ed from the microalgae biomass of Spirulina platensis. The steps to achieve that include the cultivation and har-
vesting of the microalgae, the drying of the biomass if necessary and the extraction process. However, these
steps are resource and energy demanding processes which can affect the environmental footprint and the cost
of the final product. Looking for more efficient practices combinations of materials (wet or dried biomass) and
solvents (water, buffer and ethanol), which are currently used industrially, were examined in order to evaluate
and suggest the most sustainable production line for phycocyanin.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction nutritional content of conventional food and hence, to positively affect

Microalgae are microscopic photosynthetic organisms that are
found in marine and freshwater environments (Chu, 2012). For centu-
ries they have been exploited as food and animal feed, especially for
aquaculture (Spolaore, Joannis-Cassan, Duran, & Isambert, 2006;
Vigani et al., 2015b; Yamaguchi, 1997). However, recent achievements
have renewed the interest in them, since studies have pointed out the
ability of obtaining various high-value molecules from microalgae,
such as carotenoids, chlorophylls, proteins (López et al., 2010; Pasquet
et al., 2011; Spolaore et al., 2006; Vaz, Moreira, de Morais, & Costa,
2016). Thus, the microalgal use in diversified sectors, including pharma-
ceutical, energy and food industries, has become especially attractive,
with the latter sector benefiting greatly from this advancement
(Chacon-Lee & Gonzalez-Marino, 2010; Enzing, Ploeg, Barbosa, &
Sijtsma, 2014; Vigani et al., 2015a). Microalgae are able to enhance the
⁎ Corresponding author.
E-mail address: spcheng@central.ntua.gr (S. Papadaki).
the human health (Santos, de Freitas, Moreira, Zanfonato, & Costa,
2015).
Spirulina is a photosynthetic cyanobacterium that is produced com-
mercially for alimentary use, as a dietary supplement and food additive.
Furthermore, Spirulina possesses proven biological functionality such as
antiviral, anti-inflammatory and antioxidant activity (Chu, Lim,
Radhakrishnan, & Lim, 2010; Wu, Ho, Shieh, & Lu, 2005). Therefore, it
is widely cultivated to produce biologically active food additives able
to treat several diseases, including diabetes and obesity (Anitha &
Chandralekha, 2010), arthritis (Kumar, Singh, Patro, & Patro, 2009), ane-
mia (Selmi et al., 2011), cardiovascular diseases (Deng & Chow, 2010),
allergies (Vo, Ngo, & Kim, 2012), tumors and cancer (Singh, Kate, &
Banerjee, 2005). Moreover, functional compounds from Spirulina are
widely used in food industry as colorants (Martelli, Folli, Visai, Daglia,
& Ferrari, 2014) and have been suggested also as emulsifiers (Batista,
Raymundo, Sousa, & Empis, 2006).
Currently, the annual production of Spirulina exceeds 3000 tons on a
dry weight basis and is realized by many companies in different

http://dx.doi.org/10.1016/j.ifset.2017.02.014
1466-8564/© 2017 Elsevier Ltd. All rights reserved.
218 S. Papadaki et al. / Innovative Food Science and Emerging Technologies 44 (2017) 217–223
countries. The extensive production of Spirulina is due to its original
chemical composition (proteins, polyunsaturated fatty acids and vita-
mins). Dried spirulina contains about 60% proteins, as well as, many pig-
ments which may be beneficial, including beta carotene and zeaxanthin,
plus phycobiliproteins, such as C-phycocyanin and allophycocyanin
(Campanella, Crescentini, & Avino, 1999; Oliveira, Rosa, Moraes, &
Pinto, 2009; Tang & Suter, 2011). Among those the most important
functional ingredients are phycobiliproteins, a pigment-protein com-
plex which is used into various food products to enhance their nutri-
tional qualities acting as food colorant, antioxidant and emulsifier, and
can sufficiently replace or reduce the use of synthetic additives. C-phy-
cocyanin is the main pigment and reaches 20% in dry weight of the cell
protein, depending on cultivation conditions (Cuellar-Bermudez et al.,
2015), and lately has been approved by the Food and Drug Administra-
tion as a natural blue food colorant (Fda, 2014).
For the effective recovery of phycocyanin, the nutrient should be ex-
tracted from the microalgae biomass. Dehydration is considered an es-
sential step pre-treatment step due to the high moisture content of
the material, however it may lead to the decrease of its phycocyanin
content. The drying of Spirulina constitutes approximately 30% of the
total production cost (Oliveira, Duarte, Moraes, Crexi, & Pinto, 2010).
The most commonly used method is the spray drying technique, in
which the product is obtained in powder form, but usually, the powder
does not satisfy all the criteria required for food use (Desmorieux &
Decaen, 2005). Desmorieux et al. reported that apart from high opera-
tion cost, the dried product obtained by spray drying did not have the
same aspect and color as the low-cost drying methods (Desmorieux &
Decaen, 2005). Greenhouse drying and oven-drying have the potential
to approach the advantages derived from spray drying in terms of qual-
ity and bioavailability at a lower cost (Tiburcio, Galvez, Cruz, & Gavino,
2007). The development of an optimized drying process would there-
fore encourage farmers to grow Spirulina, as this will give them the as-
surance that they can produce dried Spirulina of marketable quality.
The crucial step for the efficient phycocyanin recovery is the selec-
tion of the optimum combination of extraction technique and solvent
system. The increasing legislative restrictions on the use of organic sol-
vents in foods coupled to their negative effects on the functional prop-
erties of compounds are a challenge for an effective and eco-friendly
recovery process (Chemat et al., 2017; Chemat, Vian, & Cravotto,
2012). Nowadays, Ultrasound Assisted Extraction (UAE) due to its
high efficiency and low solvent consumption tends to replaced conven-
tional extraction methods (Dey & Rathod, 2013; Patist & Bates, 2008).
The higher yield obtained in UAE is of major interest from an industrial
point of view, since the technology is an “add on” step to the existing
process with minimum alteration, application in aqueous extraction
where organic solvents can be replaced with generally recognized as
safe solvents, shortening the extraction time (Vilkhu, Mawson,
Simons, & Bates, 2008). The use of ultrasonic for extraction purposes
in high-cost raw materials is an economical alternative to traditional ex-
traction processes, which is an industry demand for a sustainable
development.
The environmental assessment of industrial processes, such as the
phycocyanin recovery is based on Life Cycle Assessment (LCA) proce-
dure. LCA is a tool that is being used to evaluate the environmental as-
pects associated with the entire life cycle of a product or process. It
monitors the materials, energy and wastes involved in each phase of
the product's life cycle, from raw materials extraction to final disposal,
by compiling an inventory of elementary flows. These flows are then
assigned to environmental impact categories according to the sub-
stances' ability to contribute to different key environmental issues
(JRC European Commission, 2010). The results of the analysis will
allow identifying the phases/processes and the relevant resource flows
along the entire chain with the most significant environmental impacts
(Andersson, Ohlsson, & Olsson, 1994; Schau, Fet, & Declarations, 2008).
This will lead to the suggestion of effective alternative interventions (ei-
ther technologies or management practices) for the upgrading of the
food chain towards the reduction of its overall environmental footprint.
These interventions may include, among others, (a) source material re-
duction, (b) process upgrading, (c) reuse of waste material and (d)
recycling or composting practices (Kyriakopoulou, Papadaki, &
Krokida, 2013, 2015; Schau et al., 2008). In addition, LCA as a tool can
lead to the development of a cross-disciplinary agenda of research,
linking together agricultural, environmental, social, and health concerns
under the principle of sustainability and also applying them to daily
consumption practices (Schau et al., 2008).
In this study, the environmental performance of the isolation of phy-
cocyanin from Spirulina platensis microalga using UAE and polar sol-
vents such as water, ethanol or phosphate buffer was evaluated using
the Life Cycle Assessment methodology. The analysis was carried out,
using the Ecoinvent 2 database provided with the SimaPro 7 software
(Pre' Consultants, 2014; SimaPro Manual PRe Consultants, 2008).The
system examined included the open pond cultivation of the microalgae,
the hot air drying step and their extraction processing for the recovery
of extracts equivalent to 1 kg of phycocyanin.
2. Life cycle assessment methodology
According to International Standards Organization (ISO) 14,000 se-
ries, the technical framework for LCA methodology consists of four
phases: (1) the goal and scope definition; (2) the inventory analysis;
(3) the impact assessment; and (4) the interpretation (ISO, 2006). De-
fining the goal and scope involves the description of the purpose, the
functional unit (FU), the system boundaries, the data quality, assump-
tions and simplifications. The life cycle inventory involves collecting
data for each unit process regarding all relevant inputs and outputs of
energy and mass flows, as well as data on emissions to air, water and
soil. The life cycle impact assessment phase evaluates the potential en-
vironmental impacts of the examined system, product and processes,
while in the interpretation phase the results of the inventory analysis
and the environmental impacts are combined together.
3. Goal and scope definition
The objective of this study was to compare the environmental as-
pects and impacts associated with different extraction methods for the
recovery of phycocyanin from the microalga Spirulina platensis and to
identify the “hot spots” of the evaluated technologies as a way to poten-
tially improve their environmental performance. This study includes the
analysis of different production stages, such as the cultivation, the har-
vesting, the drying treatment and the extraction of the microalgal bio-
mass till the recovery of phycocyanin rich extracts. To evaluate the
impact of the different processes, an inventory registration of the mate-
rial and energy flows, as well as, the emissions that occur over these
processes was established.
3.1. Functional unit
To remove performance variation and provide a fair comparison be-
tween the extracts recovered by the proposed production lines, the
functional unit was defined as 1 kg of phycocyanin extracted from
dried Spirulina platensis microalga to be used as a natural colorant and
bioactive component in food, nutraceuticals and cosmetic applications.
In general, the functional unit defines what the LCA study is measuring
and provides a reference to which the inputs and outputs can be related.
A direct comparison of different materials is not in accordance with ISO
14040 as their properties may vary and lead to unambiguous definition
regarding their common function (Kyriakopoulou et al., 2015).
3.2. Product system and system boundaries
The system model is based on a fictional microalgae farming estab-
lishment using open pond cultivation systems. The open pond
 S. Papadaki et al. / Innovative Food Science and Emerging Technologies 44 (2017) 217–223 219

(raceway) infrastructure was selected over photo-bioreactors due to
their simple technology and low energy, as well as, the fact that they
are inexpensive and well researched on different strain cultivations
(Jorquera, Kiperstok, Sales, Embiruçu, & Ghirardi, 2010). The simplicity
of raceway ponds lays on their construction, since they are shallow
open basins with a considerate length constructed using a concrete
shell lined with polyvinyl chloride (PVC) with various dimensions
(Aitken & Antizar-Ladislao, 2012). On the contrary to photo-bioreactors,
which present high energy and cost intensity production and operation,
their energy demand is limited, especially when utilizing natural sun-
light, decreasing their environmental impact. Open pond systems have
been used for the production of Spirulina biomass (Chanawongse, Lee,
Bunnag, & Tanticharoen, 1994; Radmann, Reinehr, & Costa, 2007).
Moreover, the warm and sunny climate of the Aegean and Mediterra-
nean regions are ideal for outdoor cultivation, while algae can be also
grown in greenhouses utilizing the sunlight. Therefore, to place the
microalga cultivation for this study the Greek island of Chios was
selected.
In this LCA analysis, cradle-to-gate systems of phycocyanin rich ex-
tract production were considered, broken down into two stages; the ac-
quisition and treatment of the different raw material prior to extraction
(cultivation, harvesting, drying, etc.) and the extraction process for the
recovery of phycocyanin which can be used as additive in food, nutra-
ceutical or cosmetic products. As in previous studies energy, water, car-
bon dioxide and nutrient requirements are treated as system inputs,
while solar radiation provided for the cultivation stage is not modeled
in the process as it is considered a natural energy source
(Kyriakopoulou et al., 2015). Regarding the open pond installation,
site leveling works for capital infrastructure, including plumbing,
pumps, sheds, processing plant and machinery is not taken into consid-
eration, due to the assumed low attribution of these elements, while the
land occupation and the energy demands for the biomass production
were incorporated in the analysis. The energy consumption of each
component employed in the production process (pumps, centrifuge,
drier, ultrasound extractor) was calculated based on their specification
(considering commercial equipment) and application time.
4. Life cycle inventory
Spirulina platensis has been widely used as a food supplement due to
its high protein content and nutritional value. In order to acquire the de-
sired biomass several steps including cultivation of the microalgae in
appropriate media, the harvesting of the biomass and the conditions
of post-harvest treatment (drying temperature, storage conditions,
etc.).
4.1. Cultivation, harvest of microalgae
environmental factors and the high running costs due to the large salt-
water volumes (Prieto et al., 2011). The biomass productivity in such
systems can reach up to 15 g m− 2 d−1 (Jiménez et al., 2003; De
Bhowmick et al., 2014), however it is difficult to maintaining optimal
cultivation parameters (Guterman et al., 1990; Zhang et al., 2015).
Following, the cultivation, a series of dewatering and drying process-
es are applied in order to concentrate the microalgae biomass. Accord-
ing to Grierson et al. (2013), a concentration of the microalgae
cultivation by a factor of 18.4 can be achieved by flocculation is applied,
followed by air floatation (Grierson et al., 2013). An additional concen-
tration can be achieved by centrifugation leading to approximately
28.6% solids content and a recycling of the growth medium (Grierson
et al., 2013). The moisture content of the Spirulina platensis biomass
after centrifugation is around 88% (Olguín, Galicia, Camacho, Mercado,
& Pérez, 1997; Stramarkou, Papadaki, Kyriakopoulou, & Krokida,
2016). Combining the cultivation and handling specifications of Spiru-
lina with input and output data for open pond microalga cultivations
provided by the literature (Campbell, Beer, & Batten, 2011; Collet et
al., 2011), the mass and energy balances of the life cycle inventory of
the cultivation and harvesting for the production of 1 kg wet biomass
were constructed. This inventory analysis refers to a fictional open
pond cultivation system utilizing sunlight and sea water, along with
several nutrients for the cultivation of Spirulina biomass. Energy is re-
quired for pumping and recirculating the water, for pumping the carbon
dioxide needed for the cultivation and for the dewatering of the biomass
by centrifugation. In Table 1 are presented the inputs and outputs of the
inventory for the production of 1 kg wet Spirulina biomass.
4.2. Pretreatment of Spirulina platensis and phycocyanin recovery
Phycobiliprotein recovery involves the cell rupture of the microalga
and the release of these proteins from within the cell (Moraes, Sala,
Cerveira, & Kalil, 2011). Traditionally, the recovery is achieved with
the use of polar solvents such as water, ethanol and buffer (Mahadev,
2005). The microalgal biomass is suspended and the pigments are
leached in the selected solvent (Prabuthas, Majumdar, Srivastav, &
Mishra, 2011). However, the wet biomass is immediately susceptible
to bacterial decomposition and degradation due to its nutritional com-
position. Therefore, dried biomass treatment gained popularity since it
leads to effective storage and handling and at the same time retain the
bioactive content (Oliveira et al., 2010).
The microalgae paste collected is dried in an industrial rotary dryer,
as the one describe in the study of Show, Lee, Tay, Lee, and Chang
Table 1
Life cycle inventory of the cultivation and harvest of the production of 1 kg wet Spirulina
platensis biomass at an open pond installation occupying 1 ha.

Inventory Amount Reference

Industrially applied microalgae cultivation systems include shallow
Inputs
big ponds, tanks, circular ponds and raceway ponds, however according Water, salt, ocean 595.6757 kg Campbell et al. (2011)
to FAO the commercial cultivation of Spirulina platensis is done outdoors Carbon dioxide 1.4286 kg Campbell et al. (2011)
in open-air cultures (Habib et al., 2008). The microalga thrives in alka- Nitrogen fertilizer 0.0069 kg Campbell et al. (2011)
line environment and this preference prevents external contamination, Phosphorus fertilizer 0.0047 kg Campbell et al. (2011)
Iron sulfate b0.0001 kg Campbell et al. (2011)
suggesting its suitability for environmental applications (Wuang, Khin, Occupation land open pond 1 ha
Chua, & Luo, 2016). Electricity pumping water 0.4659 MJ Collet et al. (2011)
For the cultivation phase of this study shallow ponds incorporated Electricity pumping CO2 0.1031 MJ Collet et al. (2011)
with low energy-consuming paddlewheels for gas/liquid mixing and Electricity paddlewheel 0.6090 MJ Collet et al. (2011)
Electricity centrifugation 0.1279 MJ Collet et al. (2011)
circulation, called raceway ponds, are the available infrastructures.
Traction 0.0035 MJ Campbell et al. (2011)
Guillard F/2 commercial media, is selected for the cultivation, diluted
in the manufacturer's recommended dilutions (1:1000) in tap water Outputs
Spirulina platensis 1.0000 kg
with 30 g/L marine salt (Wuang et al., 2016). According to Jorquera et
Emissions to air
al. (2010), temperature regulation is performed during liquid evapora- Carbon dioxide 0.0193 kg Campbell et al. (2011)
tion, since the culture medium is directly exposed to the atmosphere Nitrogen b0.0001 kg Campbell et al. (2011)
(Jorquera et al., 2010). Even though open-air cultures are easy to con- Emissions to water
struct and operate, they present several limitations such as large area Water 397.6834 kg Campbell et al. (2011)
Salts 0.6541 kg Campbell et al. (2011)
requirement, contamination problems, impractical control of some
220 S. Papadaki et al. / Innovative Food Science and Emerging Technologies 44 (2017) 217–223
(2015), and the dried product at approximately 4% moisture is turned to
powder ready for extraction. According to Show et al. (2015), wet slurry
containing 30% solids of microalgae is treated at 120 °C for about 10 s in
a pilot drum-dryer consuming about 52 kWh. In their assessment on en-
ergy requirement for drying algae with a water content of approximate-
ly 94% to 4%, they report that for the production of 1 kg of dry algae
product 18.2 kg of water need to be evaporated. The energy demand
for this process was 15.7 Mcal, leading to 1.15 Mcal/kg of evaporated
water (or 4.815 MJ/kg water) (Show et al., 2015). In the case of the cen-
trifuged Spirulina biomass with an initial moisture content of 88% to
produce 1 kg of dry material with 4% moisture about 8 kg of wet bio-
mass need to be treated. By the end of the drying process 7 kg of
water will be evaporated using 8.05 Mcal (34.704 MJ) and the final
dry product of 1 kg will contain about 0.04 kg of water.
As the water content is a determining factor on the energy consump-
tion, the dewatering and centrifugation steps prior to drying are consid-
ered essential to reduce the amount of water entering the rotary drier.
Adjusting not only the initial moisture content (to lower levels), but
also the moisture content of the dried microalga biomass (to acceptable
but not extremely low levels) can affect the energy requirement, the
processes economic viability and the environmental performance.
Nowadays, for the disruption of Spirulina's cell walls from both dried
or wet biomass, sonication is suggested since it enhances the bioactive
compound recover leading to shorter treatment times (Dey & Rathod,
2013; Hadiyanto, 2016). UAE is considered an inexpensive extraction
techniques which involves the use of ultrasound of 20 kHz to 40 kHz;
which increases the permeability of cell walls and produces cavitation
(the formation, growth, and implosive collapse of bubbles in a liquid)
(Drosou, Kyriakopoulou, Bimpilas, Tsimogiannis, & Krokida, 2015;
Kyriakopoulou et al., 2015). The cavitation effect helps in the cell wall
disruption allowing the solvent to penetrate into the biomass and in-
creasing the contact surface area between the solvent and compounds
of interest, resulting into increased mass transfer also through good
mixing (Naveena, Armshaw, & Tony Pembroke, 2015). Therefore, UAE
provides increased yielding and leaching rate, along with the advantage
of reduced solvent volumes and temperatures, which is crucial for the
recovery of thermolabile compounds (Dey & Rathod, 2013).
For the recovery of phycocyanin from wet and dried Spirulina
platensis, the combination of ultrasound with solvents such as water,
ethanol and phosphate buffer (pH 7) is suggested (Agustini, Suzery,
Sutrisnanto, & Ma'ruf, 2015; Kumar, Dhar, Pabbi, Kumar, & Walia,
2014; Stramarkou et al., 2016). The selection of solvents is based on
the nature of phycobilins, which are covalently bound to a water-solu-
ble protein that aggregates on the surface of the photosynthetic mem-
brane located in the cytoplasm or in the stroma of the chloroplast
(Cuellar-Bermudez et al., 2015). The most efficient pigment extraction
procedures combine mechanical and chemical methods that lead to
protein release (Sobiechowska-Sasim, Stoń-Egiert, & Kosakowska,
2014). However, each combination of solvent and extraction conditions
leads to different yielding as it was presented in our previous study
(Stramarkou et al., 2016). The difference in the yield on the compounds
of interest, which can be a result of the material selected or the recovery
process examined, has shown among different studies than it can affect
the environmental performance of the production process
(Kyriakopoulou et al., 2015; Papadaki, Kyriakopoulou, & Krokida,
2016; Silva et al., 2015).
Table 2
Specification of extraction conditions examined.
Specs UAE 1 UAE 2
Spirulina platensis Wet paste Wet paste
Solvent Water Ethanol
Solid liquid ratio (kg d.b.:L) 1:20 1:20
Conditionsa 30 °C, 5 min, 30 °C, 5 min,
25 kHz, 450 W 25 kHz, 450 W
a
The extraction conditions are derived from the research of Stramarkou et al., 2016.
In this study phycocyanin recovery from both wet and dried Spiru-
lina biomass is examined and the specifications of the extraction pro-
cesses are presented in Table 2. The selection of the conditions was
made based on previous experimental work in which the comparison
on the yield of phycocyanin was made after treating the samples in se-
lected extraction time, ultrasound power and temperatures
(Stramarkou et al., 2016). Taking into consideration the experimental
yielding the mass and energy balances of each extraction line case
study were prepared for the production of 1 kg of phycocyanin in the
form of liquid extract (Table 3).
5. Environmental impact assessment and interpretation
The environmental impacts associated with the recovery of bioac-
tive, phycobilin-rich extracts from Spirulina platensis were quantified
using the problem-oriented approach, CML 2 baseline 2000 v2.04
(Center for Environmental Studies, University of Leiden) method pro-
vided by the SimaPro LCA tool (de Bruijn, van Duin, & Huijbregts,
2002; European Commission—Joint Research Centre—Institute for
Environment and Sustainability, 2010; Guinée et al., 2002). The selected
method includes a series of impact categories, namely Ozone layer de-
pletion, Human toxicity, Fresh water aquatic ecotoxicity, Marine aquatic
ecotoxicity, Terrestrial ecotoxicity, Photochemical oxidation, Global
warming (indicator of Carbon footprint), Acidification, Abiotic depletion
and Eutrophication, and has been previously used for the evaluation of
microalgae cultivation and different extraction processes for the recov-
ery of bioactive compounds (Collet et al., 2014; Lardon, Hélias, Sialve,
Steyer, & Bernard, 2009; Rodríguez-Meizoso et al., 2012). The impacts
in this method are quantified with appropriate physical units based on
the life cycle inventory data according to physically based formulae
(Guinée et al., 2002).
In this study a life cycle assessment from cradle to gate was per-
formed including the production of wet and dried Spirulina platensis
biomass and the extraction of the biomass for the production of 1 kg
of phycocyanin in the form of extract. Therefore, the environmental per-
formance of the phycocyanin recovery process lines involves the foot-
prints of all the necessary steps; the cultivation (raceways ponds and
harvesting), the drying and the final biomass treatment using different
solvents. Each step was found to contribute to the environmental foot-
print according to its energy and resources consumption.
Specifically, for the cultivation step the most important contribu-
tions to the energy demand come from the electricity required to circu-
late the culture and the embodied energy in pond construction. These
energy fractions were reported to be 22% to 79% and 8% to 70%, respec-
tively (Slade & Bauen, 2013). In addition, the embodied energy in the
fertilizers, such as nitrogen and phosphorus, is reported that it may con-
tribute to the energy demand by 6% to 40% (Slade & Bauen, 2013). De-
spite these energy demands, the cultivation stage yields to some net
benefit in relation to global warming impact due to the intake of CO2
from microalgae, presenting a slightly reduced overall carbon footprint.
However, the electricity usage, also for the centrifugation, and fertilizer
consumption contribute not only the global warming potential but also
to the acidification and abiotic depletion potential.
On the other hand, the recycling of harvest water during dewatering
and centrifugation reduces the nutrient use and controls the fertilizer
usage, which leads to the reduction of the natural resources depletion
UAE 3 UAE 4 UAE 5 UAE 6
Wet paste Dry powder Dry powder Dry powder
Buffer Water Ethanol Buffer
1:20 1:20 1:20 1:20
30 °C, 5 min, 30 °C, 5 min, 30 °C, 5 min, 30 °C, 5 min,
25 kHz, 450 W 25 kHz, 450 W 25 kHz, 450 W 25 kHz, 450 W
 S. Papadaki et al. / Innovative Food Science and Emerging Technologies 44 (2017) 217–223 221

Table 3
Life cycle inventory of the recovery processes for the extraction of phycocyanin in the form of liquid extract.a

UAE 1 UAE 2 UAE 3 UAE 4 UAE 5 UAE 6

Experimental data
Phycocyanin yield (%)a 0.46 2.71 0.94 3.22 0.02 3.41

Inputs LCI
Material (kg d.b.) 217.39 36.90 106.38 31.06 5000.00 29.33
Equivalent in wet biomass (kg) 1739.13 295.20 851.06 248.45 40,000.00 234.60
Water evaporated through drying (kg water) Non Non Non 217.39 35,000.00 205.28
Energy for drying (MJ) – – – 1046.74 168,525.00 988.42
Solvent (L) 4347.83 738.01 2127.66 621.12 100,000.00 586.51
Electricity for extraction (MJ) 11,739.13 1992.62 5744.68 1677.02 270,000.00 1583.58

Output LCI
Phycocyanin content (kg) 1 1 1 1 1 1
a
The yield is referred to the consistency of the final extracts' recovery of phycocyanin (Stramarkou et al., 2016) according to which the mass and energy balances are demonstrated in
the table.

and the environmental impacts during nutrients production and use
(Clarens, Resurreccion, White, & Colosi, 2010; Lardon et al., 2009). Eu-
trophication potential is reduced due to the avoidance of nitrate and
phosphate leaching to the ground water, and ammonia and nitrous ox-
ides to the air (Kyriakopoulou et al., 2015).
Although the dewatering steps together with drying have been re-
ported as the most energy and cost intensive steps in algal biomass pro-
duction due to the low concentration of microalgae in the culture
medium (Soomro et al., 2016; Uduman, Qi, Danquah, Forde, &
Hoadley, 2010), dewatering and centrifuge operation are considered
an important step for condensing the microalga paste prior to extraction
or drying (Grierson et al., 2013). Therefore, the selection of energy effi-
cient systems is important to result to lower energy consumption and
lower emissions (Weschler, Barr, Harper, & Landis, 2014). However,
the selection of the drying processing can significantly affect the quality
of the final dried microalga powder and thus the extractability of the
targeted bioactive compounds (Papadaki, Kyriakopoulou, Stramarkou,
Tzovenis, & Krokida, 2017). In this study an industrially applied hot air
drying method was selected for all the production lines. Although the
dried powder used in all extraction processes was considered of the
same quality, the experimental extraction results for each process pre-
sented significant differences, affecting greatly the environmental per-
formances of phycocyanin production.
As far as the extraction of both wet and dried biomass is concerned,
the environmental impact is primarily affected by the energy demand of
the ultrasound extractor and the nature of the solvent system used. The
energy demand according to the experimental data was 54 MJ/kg dry
biomass treated, while the corresponding energy demands for other ul-
trasound extractors can reach up to 148 MJ/kg biomass treated
depended on the time of the treatment, the scale and the specifications
of the machinery used (Ferreira, Dias, Silva, & Costa, 2016). However, fo-
cusing further on the recovery of bioactive compound the crucial factor
for the overall environmental performance of the recovery process is
the achieved yield. If the overall footprint of the process is allocated to
the phycocyanin yield, pretreatment steps of high energy demand
such as wet biomass drying influence insignificantly the overall envi-
ronmental performance. Even though drying is considered an energy
demanding step, especially due to the high moisture content of Spirulina
platensis, the effect of the treatment on the phycocyanin content or its
availability in the biomass showed opposite results on the overall envi-
ronmental performance.
Examining the treatment processes for the extraction of phycocyanin
from wet biomass using water, ethanol and buffer as solvent (Table 4),
significant differences on several impact categories were observed.
Among the three Ultrasound Assisted Extraction processes, UAE 2
using ethanol as solvent exhibited the lowest environmental impact in
categories of carbon footprint, ozon layer depletion and human toxicity
even though an organic solvent was used. The positive effect of ethanol
on the recovery of phycobiloproteins affects the environmental perfor-
mance positively, since less biomass is needed to be cultivated, harvest-
ed and extracted for the recovery of the desired amount of phycocyanin.
However, in some cases the effect of ethanol overcomes the positive ef-
fect of the high yielding, leading to slightly worsen impact, especially in
the photochemical oxidation and the ozone layer depletion potential.
The combination of buffer as solvent with dried Spirulina platensis
biomass leads to significantly higher phycocyanin yields, contributing
the lowest on the overall environmental performance, despite the fact
that an extremely energy consuming step is added prior to extraction.
The treatment of dried biomass with ethanol exhibits the highest foot-
prints due to the low yielding and the high material (biomass and sol-
vent) consumption (Table 5).
The carbon footprint of the two preferable extraction procedures for
wet (UAE 2) and dried biomass (UAE 6) is 2.06E + 03 and 1.18E + 03 kg
CO2 eq, respectively. This difference of the latter from the former is
around 43% and shows once more that the yielding is the crucial factor
for the environmental performance of phycocyanin recovery. The dry-
ing step prior to extraction permits the highest penetration of the sol-
vent resulting to higher extraction rates, leading to lower biomass

Table 4 Table 5
Environmental footprint of processing wet Spirulina biomass for the recovery of 1 kg Environmental footprint of processing dried Spirulina biomass for the recovery of 1 kg
phycocyanin. phycocyanin.

Impact category UAE 1 UAE 2 UAE 3 Impact category UAE 4 UAE 5 UAE 6

Abiotic depletion kg Sb eq 5.49E + 01 2.52E + 01 2.69E + 01 Abiotic depletion kg Sb eq 1.06E + 01 3.86E + 03 1.00E + 01
Acidification kg SO2 eq 4.27E + 01 9.92E + 00 2.09E + 01 Acidification kg SO2 eq 7.94E + 00 1.64E + 03 7.50E + 00
Eutrophication kg PO−3
4 eq 1.51E + 00 1.30E + 00 7.37E − 01 Eutrophication kg PO−3
4 eq 2.76E − 01 1.86E + 02 2.61E − 01
Global warming (GWP100) kg CO2 eq 6.72E + 03 2.06E + 03 3.29E + 03 Global warming (GWP100) kg CO2 eq 1.25E + 03 3.26E + 05 1.18E + 03
Ozone layer depletion kg CFC-11 eq 4.68E − 04 1.06E − 04 2.29E − 04 Ozone layer depletion kg CFC-11 eq 7.49E − 05 1.57E − 02 7.07E − 05
Human toxicity kg 1,4-DB eq 3.74E + 03 7.95E + 02 1.83E + 03 Human toxicity kg 1,4-DB eq 6.31E + 02 1.23E + 05 5.96E + 02
Fresh water aquatic ecotox. kg 1,4-DB eq 9.52E + 02 1.91E + 02 4.66E + 02 Fresh water aquatic ecotox. kg 1,4-DB eq 2.04E + 02 3.69E + 04 1.93E + 02
Marine aquatic ecotoxicity kg 1,4-DB eq 2.42E + 06 4.54E + 05 1.18E + 06 Marine aquatic ecotoxicity kg 1,4-DB eq 5.20E + 05 8.96E + 07 4.91E + 05
Terrestrial ecotoxicity kg 1,4-DB eq 3.49E + 01 6.85E + 00 1.71E + 01 Terrestrial ecotoxicity kg 1,4-DB eq 6.63E + 00 1.19E + 03 6.26E + 00
Photochemical oxidation kg C2H4 1.93E + 00 1.40E + 00 9.44E − 01 Photochemical oxidation kg C2H4 3.48E − 01 2.02E + 02 3.29E − 01
222 S. Papadaki et al. / Innovative Food Science and Emerging Technologies 44 (2017) 217–223
requirements and energy consumption for the extraction process
(Drosou et al., 2015). If the daily consumption of phycobiloproteins is
taken into consideration, functional food products containing small
amounts of phycobilin will be burdened by the environmental footprint
of the recovery process in an extent corresponding on the mass of the
bioactive incorporated in the product. Therefore, with such low burdens
a promising pathway for the use of cyanobacteria by industries to pro-
duce cell biomass for the production of high value products lies ahead
(Mazard, Penesyan, Ostrowski, Paulsen, & Egan, 2016). Modifying
cyanobacteria to utilize waste CO2 can yield industrial- and food-
grade sugars and a nutritional range of products, such as amino acids,
nutrients and vitamins, making them ideal alternatives to plants for car-
bohydrate feedstocks (Hays & Ducat, 2015).
6. Conclusions
Spirulina platensis is a natural rich source of bioactive compounds
which can be recovered under several conditions and steps. The treat-
ment steps employed involve the cultivation, harvesting, drying and ex-
traction of the microalga cells. The environmental performance of each
process is affected by the usage of vast amounts of biomass, consumables
and energy for the recovery of 1 kg of phycocyanin thus extraction pro-
cesses enhancing the yielding of the process such as UAE are considered
essential. UAE's low cost, short time and medium environmental impact
and in combination with the right pretreatment and solvent can be even
greener. Comparing wet and dried biomass treatment, wet biomass in
combination with organic solvents such as ethanol exhibited high selec-
tivity leading to low environmental impact but still higher than that of
dried biomass. In the case of dried biomass extraction, although drying
is an energy consuming steps the combination with solvents such as
buffer lead to extracts richer phycocyanin leading to unpredicted lower
environmental impact. In both cases, the extraction techniques exam-
ined showed their potential for the recovery of phycocyanin rich extracts
which can be used in nutraceutical, cosmetic and food industries.
Abbreviations
GWP Global Warming Potential
LCA life cycle analysis/assessment
LCI life cycle inventory
LCIA life cycle impact assessment
UAE Ultrasound Assisted Extraction
Acknowledgments
The authors would like to thank Mrs. M. Stramarkou for her contribu-
tion and the information regarding the extraction of Spirulina platensis for
the recovery of bioactive compounds and especially phycocyanin.
References
Agustini, T. W., Suzery, M., Sutrisnanto, D., & Ma'ruf, W. F. (2015). Comparative study of
bioactive substances extracted from fresh and dried Spirulina sp. Procedia
Environmental Sciences, 23, 282–289 Ictcred 2014 10.1016/j.proenv.2015.01.042
Aitken, D., & Antizar-Ladislao, B. (2012). Achieving a green solution: Limitations and focus
points for sustainable algal fuels. Energy. http://dx.doi.org/10.3390/en5051613.
Andersson, K., Ohlsson, T., & Olsson, P. (1994). Life cycle assessment (LCA) of food prod-
ucts and production systems. Trends in Food Science and Technology, 5(5), 134–138.
http://dx.doi.org/10.1016/0924-2244(94)90118-X.
Anitha, L., & Chandralekha, K. (2010). Effect of supplementation of spirulina on blood glu-
cose, glycosylated hemoglobin and lipid profile of male non-insulin dependent dia-
betics. Asian Journal of Experimental Biological Sciences, 1(11), 36–46.
Batista, A. P., Raymundo, A., Sousa, I., & Empis, J. (2006). Rheological characterization of
coloured oil-in-water food emulsions with lutein and phycocyanin added to the oil
and aqueous phases. Food Hydrocolloids, 20(1), 44–52. http://dx.doi.org/10.1016/j.
foodhyd.2005.02.009.
Campanella, L., Crescentini, G., & Avino, P. (1999). Chemical composition and nutritional
evaluation of some natural and commercial food products based on spirulina.
Analusis, 27, 533–540. http://dx.doi.org/10.1051/analusis:1999130.
Campbell, P. K., Beer, T., & Batten, D. (2011). Bioresource technology life cycle assessment
of biodiesel production from microalgae in ponds. Bioresource Technology, 102(1),
50–56. http://dx.doi.org/10.1016/j.biortech.2010.06.048.
Chacon-Lee, T. L., & Gonzalez-Marino, G. E. (2010). Microalgae for “healthy”
foods—Possibilities and challenges. Comprehensive Reviews in Food Science and Food
Safety, 9, 655–675. http://dx.doi.org/10.1111/j.1541-4337.2010.00132.x.
Chanawongse, L., Lee, Y. K., Bunnag, B., & Tanticharoen, M. (1994). Productivity of the cy-
anobacterium Spirulina platensis in cultures using sunlight. Bioresource Technology,
48(2), 143–148. http://dx.doi.org/10.1016/0960-8524(94)90201-1.
Chemat, F., Rombaut, N., Sicaire, A. G., Meullemiestre, A., Fabiano-Tixier, A. S., & Abert-
Vian, M. (2017). Ultrasound assisted extraction of food and natural products. Mech-
anisms, techniques, combinations, protocols and applications. A review. Ultrasonics
Sonochemistry. http://dx.doi.org/10.1016/j.ultsonch.2016.06.035.
Chemat, F., Vian, M. A., & Cravotto, G. (2012). Green extraction of natural products: Con-
cept and principles. International Journal of Molecular Sciences, 13(7), 8615–8627.
http://dx.doi.org/10.3390/ijms13078615.
Chu, W. (2012). Biotechnological applications of microalgae. International E-Journal of
Science, Medicine & Education, 6, 24–37.
Chu, W. -L., Lim, Y. -W., Radhakrishnan, A. K., & Lim, P. -E. (2010). Protective effect of
aqueous extract from Spirulina platensis against cell death induced by free radicals.
BMC Complementary and Alternative Medicine, 10, 53. http://dx.doi.org/10.1186/
1472-6882-10-53.
Clarens, A. F., Resurreccion, E. P., White, M. A., & Colosi, L. M. (2010). Environmental life
cycle comparison of algae to other bioenergy feedstocks. Environmental Science and
Technology, 44(5), 1813–1819. http://dx.doi.org/10.1021/es902838n.
Collet, P., Hélias Arnaud, A., Lardon, L., Ras, M., Goy, R. A., & Steyer, J. P. (2011). Life-cycle
assessment of microalgae culture coupled to biogas production. Bioresource
Technology, 102(1), 207–214. http://dx.doi.org/10.1016/j.biortech.2010.06.154.
Collet, P., Lardon, L., Hélias, A., Bricout, S., Lombaert-Valot, I., Perrier, B., ... Bernard, O.
(2014). Biodiesel from microalgae—Life cycle assessment and recommendations for
potential improvements. Renewable Energy, 71, 525–533. http://dx.doi.org/10.1016/
j.renene.2014.06.009.
Cuellar-Bermudez, S. P., Aguilar-Hernandez, I., Cardenas-Chavez, D. L., Ornelas-Soto, N.,
Romero-Ogawa, M. A., & Parra-Saldivar, R. (2015). Extraction and purification of high-
value metabolites from microalgae: Essential lipids, astaxanthin and phycobiliproteins.
Microbial Biotechnology. http://dx.doi.org/10.1111/1751-7915.12167.
De Bhowmick, G., Subramanian, G., & Mishra, Ramkrishna Sen S. (2014). Raceway pond
cultivation of a marine microalga of Indian origin for biomass and lipid production:
A case study. Algal Research, 6(B), 201–209.
de Bruijn, H., van Duin, R., & Huijbregts, M. A. J. (2002). Handbook on life cycle assess-
ment. In J. B. Guinee, M. Gorree, R. Heijungs, G. Huppes, R. Kleijn, A. de Koning, &
H. A. Udo de Haes (Eds.), Operational guide to the ISO standards. Vol. 7. Dordrecht:
Springer Netherlands. http://dx.doi.org/10.1007/0-306-48055-7.
Deng, R., & Chow, T. J. (2010). Hypolipidemic, antioxidant, and antiinflammatory activities
of microalgae spirulina. Cardiovascular Therapeutics. http://dx.doi.org/10.1111/j.
1755-5922.2010.00200.x.
Desmorieux, H., & Decaen, N. (2005). Convective drying of spirulina in thin layer. Journal
of Food Engineering, 66(4), 497–503. http://dx.doi.org/10.1016/j.jfoodeng.2004.04.
021.
Dey, S., & Rathod, V. K. (2013). Ultrasound assisted extraction of β-carotene from Spiru-
lina platensis. Ultrasonics Sonochemistry, 20(1), 271–276. http://dx.doi.org/10.1016/
j.ultsonch.2012.05.010.
Drosou, C., Kyriakopoulou, K., Bimpilas, A., Tsimogiannis, D., & Krokida, M. (2015). A com-
parative study on different extraction techniques to recover red grape pomace poly-
phenols from vinification byproducts. Industrial Crops and Products. http://dx.doi.org/
10.1016/j.indcrop.2015.05.063.
Enzing, C., Ploeg, M., Barbosa, M., & Sijtsma, L. (2014). Microalgae-based products for the
food and feed sector: An outlook for Europe. JRC scientific and policy reports. European
Comission. http://dx.doi.org/10.2791/3339.
European Commission—Joint Research Centre—Institute for Environment and
Sustainability (2010v). International Reference Life Cycle Data System (ILCD) handbook:
Analysing of existing environmental impact assessment methodologies for use in life cycle
assessment. 115.European Commission. http://dx.doi.org/10.2788/38479.
Fda (2014). Generally Recognized as Safe (GRAS). 2(February). (pp. 81536). U.S. Food
and Drug Administration, 81536. http://dx.doi.org/10.1016/B978-0-12-
386454-3.00848-4.
Ferreira, A. F., Dias, A. P. S., Silva, C. M., & Costa, M. (2016). Effect of low frequency ultra-
sound on microalgae solvent extraction: Analysis of products, energy consumption
and emissions. Algal Research, 14, 9–16. http://dx.doi.org/10.1016/j.algal.2015.12.015.
Grierson, S., Strezov, V., & Bengtsson, J. (2013). Life cycle assessment of a microalgae bio-
mass cultivation, bio-oil extraction and pyrolysis processing regime. Algal Research,
2(3), 299–311.
Guinée, J. B., Heijungs, R., Huppes, G., Kleijn, R., de Koning, A., van Oers, L., & Gorrée, M.
(2002). Life cycle assessment. Operational guide to the ISO standards. I: LCA in perspec-
tive. IIa: Guide. IIb: Operational annex. III: Scientific background. The Netherlands: Minis-
try of …. http://dx.doi.org/10.1007/BF02978784.
Guterman, H., Vonshak, A., & Ben-Yaakov, S. (1990). A macromodel for outdoor algal mass
production. Biotechnology and Bioengineering, 35(8), 809–819.
Habib, M. A. B., Parvin, M., Huntington, T. C., & Hasan, M. R. (2008). A review on culture,
production and use of spirulina as food for humans and feeds for domestic animals
and fish. FAO Fisheries and Aquaculture Circular. No. 1034. Rome: FAO 33p.
Hadiyanto, S. H. (2016). Response surface optimization of ultrasound assisted extraction
(UAE) of phycocyanin from microalgae Spirulina platensis. Emirates Journal of Food
and Agriculture, 28(4), 227–234. http://dx.doi.org/10.9755/ejfa.2015-05-193.
Hays, S. G., & Ducat, D. C. (2015). Engineering cyanobacteria as photosynthetic feedstock
factories. Photosynthesis Research. http://dx.doi.org/10.1007/s11120-014-9980-0.
 S. Papadaki et al. / Innovative Food Science and Emerging Technologies 44 (2017) 217–223
International Organization for Standardization (ISO) (2006). Environmental management -
Life cycle assessment - Principles and framework. ISO 14040:2006 (Second Edition )
2006-06. Geneva.
Jiménez, C., Cossío, B. R., & Niell, F. X. (2003). Relationship between physicochemical var-
iables and productivity in open ponds for the production of Spirulina: a predictive
model of algal yield. Aquaculture, 221, 331–345.
Jorquera, O., Kiperstok, A., Sales, E. A., Embiruçu, M., & Ghirardi, M. L. (2010). Comparative
energy life-cycle analyses of microalgal biomass production in open ponds and
photobioreactors. Bioresource Technology, 101(4), 1406–1413. http://dx.doi.org/10.
1016/j.biortech.2009.09.038.
JRC European Commission (2010). Analysing of existing environmental impact assess-
ment methodologies for use in life cycle assessment. International Reference Life
Cycle Data System (ILCD) Handbook.
Kumar, D., Dhar, D. W., Pabbi, S., Kumar, N., & Walia, S. (2014). Extraction and purification
of C-phycocyanin from Spirulina platensis (CCC540). Indian Journal of Plant
Physiology, 19(2), 184–188. http://dx.doi.org/10.1007/s40502-014-0094-7.
Kumar, N., Singh, S., Patro, N., & Patro, I. (2009). Evaluation of protective efficacy of Spiru-
lina platensis against collagen-induced arthritis in rats. Inflammopharmacology, 17(3),
181–190. http://dx.doi.org/10.1007/s10787-009-0004-1.
Kyriakopoulou, K., Papadaki, S., & Krokida, M. (2013). Natural aspect of sustainable
food and cosmetics manufacturing. Green design, materials and manufacturing
processes—Proceedings of the 2nd international conference on Sustainable
Intelligent Manufacturing, SIM 2013 (pp. 197–201) Retrieved from https://
www.scopus.com/inward/record.uri?eid=2-s2.0-84880098008&partnerID=
40&md5=5de51f97d0aab468eb6f89e6f04d53b1
Kyriakopoulou, K., Papadaki, S., & Krokida, M. (2015). Life cycle analysis of β-carotene ex-
traction techniques. Journal of Food Engineering, 167, 51–58. http://dx.doi.org/10.
1016/j.jfoodeng.2015.03.008.
Lardon, L., Hélias, A., Sialve, B., Steyer, J. P., & Bernard, O. (2009). Life-cycle assessment of
biodiesel production from microalgae. Environmental Science and Technology, 43(17),
6475–6481. http://dx.doi.org/10.1021/es900705j.
López, C. V. G., del Carmen Cerón García, M., Fernández, F. G. A., Bustos, C. S., Chisti, Y., &
Sevilla, J. M. F. (2010). Protein measurements of microalgal and cyanobacterial bio-
mass. Bioresource Technology, 101(19), 7587–7591. http://dx.doi.org/10.1016/j.
biortech.2010.04.077.
Mahadev, J. D. J. (2005). An improved and efficient method for the extraction of phycocy-
anin from Spirulina sp. International Journal of Food Engineering, 1(5), 1–11. http://dx.
doi.org/10.2202/1556-3758.1037.
Martelli, G., Folli, C., Visai, L., Daglia, M., & Ferrari, D. (2014). Thermal stability improvement
of blue colorant C-phycocyanin from Spirulina platensis for food industry applications.
Process Biochemistry, 49(1), 154–159. http://dx.doi.org/10.1016/j.procbio.2013.10.008.
Mazard, S., Penesyan, A., Ostrowski, M., Paulsen, I. T., & Egan, S. (2016). Tiny microbes
with a big impact: The role of cyanobacteria and their metabolites in shaping our fu-
ture. Marine Drugs. http://dx.doi.org/10.3390/md14050097.
Moraes, C. C., Sala, L., Cerveira, G. P., & Kalil, S. J. (2011). C-phycocyanin extraction from
Spirulina platensis wet biomass. Brazilian Journal of Chemical Engineering, 28(1),
45–49. http://dx.doi.org/10.1590/S0104-66322011000100006.
Naveena, B., Armshaw, P., & Tony Pembroke, J. (2015). Ultrasonic intensification as a tool
for enhanced microbial biofuel yields. Biotechnology for Biofuels, 8, 140. http://dx.doi.
org/10.1186/s13068-015-0321-0.
Olguín, E. J., Galicia, S., Camacho, R., Mercado, G., & Pérez, T. J. (1997). Production of Spi-
rulina sp. in sea water supplemented with anaerobic effluents in outdoor raceways
under temperate climatic conditions. Applied Microbiology and Biotechnology, 48(2),
242–247. http://dx.doi.org/10.1007/s002530051045.
Oliveira, E. G., Duarte, J. H., Moraes, K., Crexi, V. T., & Pinto, L. A. A. (2010). Optimisation of
Spirulina platensis convective drying: Evaluation of phycocyanin loss and lipid oxida-
tion. International Journal of Food Science and Technology, 45(8), 1572–1578. http://dx.
doi.org/10.1111/j.1365-2621.2010.02299.x.
Oliveira, E. G., Rosa, G. S., Moraes, M. A., & Pinto, L. A. A. (2009). Characterization of thin
layer drying of Spirulina platensis utilizing perpendicular air flow. Bioresource
Technology, 100(3), 1297–1303. http://dx.doi.org/10.1016/j.biortech.2008.05.052.
Papadaki, S. G., Kyriakopoulou, K. E., & Krokida, M. K. (2016). Life cycle analysis of
microalgae extraction techniques. Chemical Engineering Transactions, 52,
1039–1044. http://dx.doi.org/10.3303/CET1652174.
Papadaki, S., Kyriakopoulou, K., Stramarkou, M., Tzovenis, I., & Krokida, M. (2017). Envi-
ronmental assessment of industrially applied drying technologies for the treatment
of Spirulina platensis. IOSR Journal of Environmental Science, Toxicology and Food
Technology, 11(1), 41–46. http://dx.doi.org/10.9790/2402-1101014146.
Pasquet, V., Farhat, F., Piot, J., Baptiste, J., Kaas, R., Patrice, T., ... Picot, L. (2011). Study on the
microalgal pigments extraction process: Performance of microwave assisted extraction.
Process Biochemistry, 46(1), 59–67. http://dx.doi.org/10.1016/j.procbio.2010.07.009.
Patist, A., & Bates, D. (2008). Ultrasonic innovations in the food industry: From the labo-
ratory to commercial production. Innovative Food Science & Emerging Technologies,
9(2), 147–154. http://dx.doi.org/10.1016/j.ifset.2007.07.004.
Prabuthas, P., Majumdar, S., Srivastav, P. P., & Mishra, H. N. (2011). Standardization of
rapid and economical method for neutraceuticals extraction from algae. Ultrasound,
2(May), 93–96.
Pre' Consultants (2014). SimaPro database manual. PRe', 1–48. http://dx.doi.org/10.1017/
CBO9781107415324.004.
Prieto, A., Pedro, Cañavate J., & García-González, M. (2011). Assessment of carotenoid pro-
duction by Dunaliella salina in different culture systems and operation regimes.
Journal of Biotechnology, 151(2), 180–185.
Radmann, E. M., Reinehr, C. O., & Costa, J. A. V. (2007). Optimization of the repeated batch
cultivation of microalga Spirulina platensis in open raceway ponds. Aquaculture,
265(1–4), 118–126. http://dx.doi.org/10.1016/j.aquaculture.2007.02.001.
223
Rodríguez-Meizoso, I., Castro-Puyana, M., Börjesson, P., Mendiola, J. A., Turner, C., &
Ibáñez, E. (2012). Life cycle assessment of green pilot-scale extraction processes to
obtain potent antioxidants from rosemary leaves. Journal of Supercritical Fluids, 72,
205–212. http://dx.doi.org/10.1016/j.supflu.2012.09.005.
Santos, T. D., de Freitas, B. C. B., Moreira, J. B., Zanfonato, K., & Costa, J. A. V. (2015). Devel-
opment of powdered food with the addition of Spirulina for food supplementation of
the elderly population. Innovative Food Science and Emerging Technologies. http://dx.
doi.org/10.1016/j.ifset.2016.07.016.
Schau, E. M., Fet, A. M., & Declarations, E. P. (2008). LCA studies of food products as back-
ground for environmental product declarations. The International Journal of Life Cycle
Assessment, 13, 255–264. http://dx.doi.org/10.1065/lca2007.12.372.
Selmi, C., Leung, P. S. C., Fischer, L., German, B., Yang, C. -Y., Kenny, T. P., ... Gershwin, M. E.
(2011). The effects of Spirulina on anemia and immune function in senior citizens.
Cellular & Molecular Immunology, 8(3), 248–254. http://dx.doi.org/10.1038/cmi.2010.76.
Show, K. Y., Lee, D. J., Tay, J. H., Lee, T. M., & Chang, J. S. (2015). Microalgal drying and cell
disruption—Recent advances. Bioresource Technology, 184, 258–266. http://dx.doi.
org/10.1016/j.biortech.2014.10.139.
Silva, A. G., Carter, R., Merss, F. L. M., Corrêa, D. O., Vargas, J. V. C., Mariano, A. B., ... Scherer, M.
D. (2015). Life cycle assessment of biomass production in microalgae compact
photobioreactors. GCB Bioenergy, 7(2), 184–194. http://dx.doi.org/10.1111/gcbb.12120.
SimaPro Manual PRe Consultants (2008). Introduction to LCA with SimaPro 7. the Nether-
lands: PRé Consultants, 1–88 Version 10.1016/j.soncn.2012.11.001
Singh, S., Kate, B. N., & Banerjee, U. C. (2005). Bioactive compounds from cyanobacteria
and microalgae: An overview. Critical Reviews in Biotechnology, 25(3), 73–95. http://
dx.doi.org/10.1080/07388550500248498.
Slade, R., & Bauen, A. (2013). Micro-algae cultivation for biofuels: Cost, energy balance,
environmental impacts and future prospects. Biomass and Bioenergy, 53(0), 29–38.
http://dx.doi.org/10.1016/j.biombioe.2012.12.019.
Sobiechowska-Sasim, M., Stoń-Egiert, J., & Kosakowska, A. (2014). Quantitative analysis of
extracted phycobilin pigments in cyanobacteria—An assessment of spectrophotomet-
ric and spectrofluorometric methods. Journal of Applied Phycology, 26(5), 2065–2074.
http://dx.doi.org/10.1007/s10811-014-0244-3.
Soomro, R. R., Ndikubwimana, T., Zeng, X., Lu, Y., Lin, L., & Danquah, M. K. (2016). Develop-
ment of a two-stage microalgae dewatering process—A life cycle assessment approach.
Frontiers in Plant Science, 7(February), 1–12. http://dx.doi.org/10.3389/fpls.2016.00113.
Spolaore, P., Joannis-Cassan, C., Duran, E., & Isambert, A. (2006). Commercial applications
of microalgae. Journal of Bioscience and Bioengineering, 101(2), 87–96. http://dx.doi.
org/10.1263/jbb.101.87.
Stramarkou, M., Papadaki, S., Kyriakopoulou, K., & Krokida, M. (2016). Recovery of func-
tional pigments from four different species of microalgae. IOSR Journal of
Environmental Science, Toxicology and Food Technology, 10(9), 26–30. http://dx.doi.
org/10.9790/2402-1009022630.
Tang, G., & Suter, P. M. (2011). Vitamin A, nutrition, and health values of algae: Spirulina,
Chlorella, and Dunaliella. Journal of Pharmacy and Nutrition Sciences, 1(2), 111–118.
http://dx.doi.org/10.6000/1927-5951.2011.01.02.04.
Tiburcio, P. C., Galvez, F. C. F., Cruz, L. J., & Gavino, V. C. (2007). Optimization of low-cost
drying methods to minimize lipid peroxidation in Spirulina platensis grown in the
Philippines. Journal of Applied Phycology, 19(6), 719–726. http://dx.doi.org/10.1007/
s10811-007-9215-2.
Uduman, N., Qi, Y., Danquah, M. K., Forde, G. M., & Hoadley, A. (2010). Dewatering of
microalgal cultures: A major bottleneck to algae-based fuels. Journal of Renewable
and Sustainable Energy. http://dx.doi.org/10.1063/1.3294480.
Vaz, B. d. S., Moreira, J. B., de Morais, M. G., & Costa, J. A. V. (2016). Microalgae as a new
source of bioactive compounds in food supplements. Current Opinion in Food
Science, 7(2016), 73–77. http://dx.doi.org/10.1016/j.cofs.2015.12.006.
Vigani, M., Parisi, C., Rodríguez-Cerezo, E., Barbosa, M. J., Sijtsma, L., Ploeg, M., & Enzing, C.
(2015a). Food and feed products from micro-algae: Market opportunities and chal-
lenges for the EU. Trends in Food Science and Technology. http://dx.doi.org/10.1016/j.
tifs.2014.12.004.
Vigani, M., Parisi, C., Rodríguez-Cerezo, E., Barbosa, M. J., Sijtsma, L., Ploeg, M., & Enzing, C.
(2015b). Food and feed products from micro-algae: Market opportunities and chal-
lenges for the EU. Trends in Food Science & Technology, 42(1), 81–92. http://dx.doi.
org/10.1016/j.tifs.2014.12.004.
Vilkhu, K., Mawson, R., Simons, L., & Bates, D. (2008). Applications and opportunities for
ultrasound assisted extraction in the food industry—A review. Innovative Food
Science & Emerging Technologies, 9(2), 161–169. http://dx.doi.org/10.1016/j.ifset.
2007.04.014.
Vo, T. S., Ngo, D. H., & Kim, S. K. (2012). Marine algae as a potential pharmaceutical source
for anti-allergic therapeutics. Process Biochemistry, 47(3), 386–394. http://dx.doi.org/
10.1016/j.procbio.2011.12.014.
Weschler, M. K., Barr, W. J., Harper, W. F., & Landis, A. E. (2014). Process energy comparison
for the production and harvesting of algal biomass as a biofuel feedstock. Bioresource
Technology, 153, 108–115. http://dx.doi.org/10.1016/j.biortech.2013.11.008.
Wu, L. C., Ho, J. A. A., Shieh, M. C., & Lu, I. W. (2005). Antioxidant and antiproliferative ac-
tivities of spirulina and Chlorella water extracts. Journal of Agricultural and Food
Chemistry, 53(10), 4207–4212. http://dx.doi.org/10.1021/jf0479517.
Wuang, S. C., Khin, M. C., Chua, P. Q. D., & Luo, Y. D. (2016). Use of spirulina biomass pro-
duced from treatment of aquaculture wastewater as agricultural fertilizers. Algal
Research, 15, 59–64. http://dx.doi.org/10.1016/j.algal.2016.02.009.
Yamaguchi, K. (1997). Recent advances in microalgal bioscience in Japan, with special ref-
erence to utilization of biomass and metabolites: A review. Journal of Applied
Phycology, 8, 487–502. http://dx.doi.org/10.1007/BF02186327.
Zhang, L., Chen, L., Wang, J., Chen, Y., Gao, X., Zhang, Z., & Liu, T. (2015). Attached cultiva-
tion for improving the biomass productivity of Spirulina platensis. Bioresource
Technology, 181, 136–142. http://dx.doi.org/10.1016/j.biortech.2015.01.025.