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

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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

the human health (Santos, de Freitas, Moreira, Zanfonato, & Costa,
Microalgae are microscopic photosynthetic organisms that are 2015).
found in marine and freshwater environments (Chu, 2012). For centu- Spirulina is a photosynthetic cyanobacterium that is produced com-
ries they have been exploited as food and animal feed, especially for mercially for alimentary use, as a dietary supplement and food additive.
aquaculture (Spolaore, Joannis-Cassan, Duran, & Isambert, 2006; Furthermore, Spirulina possesses proven biological functionality such as
Vigani et al., 2015b; Yamaguchi, 1997). However, recent achievements antiviral, anti-inflammatory and antioxidant activity (Chu, Lim,
have renewed the interest in them, since studies have pointed out the Radhakrishnan, & Lim, 2010; Wu, Ho, Shieh, & Lu, 2005). Therefore, it
ability of obtaining various high-value molecules from microalgae, is widely cultivated to produce biologically active food additives able
such as carotenoids, chlorophylls, proteins (López et al., 2010; Pasquet to treat several diseases, including diabetes and obesity (Anitha &
et al., 2011; Spolaore et al., 2006; Vaz, Moreira, de Morais, & Costa, Chandralekha, 2010), arthritis (Kumar, Singh, Patro, & Patro, 2009), ane-
2016). Thus, the microalgal use in diversified sectors, including pharma- mia (Selmi et al., 2011), cardiovascular diseases (Deng & Chow, 2010),
ceutical, energy and food industries, has become especially attractive, allergies (Vo, Ngo, & Kim, 2012), tumors and cancer (Singh, Kate, &
with the latter sector benefiting greatly from this advancement Banerjee, 2005). Moreover, functional compounds from Spirulina are
(Chacon-Lee & Gonzalez-Marino, 2010; Enzing, Ploeg, Barbosa, & widely used in food industry as colorants (Martelli, Folli, Visai, Daglia,
Sijtsma, 2014; Vigani et al., 2015a). Microalgae are able to enhance the & Ferrari, 2014) and have been suggested also as emulsifiers (Batista,
Raymundo, Sousa, & Empis, 2006).
⁎ Corresponding author. Currently, the annual production of Spirulina exceeds 3000 tons on a
E-mail address: spcheng@central.ntua.gr (S. Papadaki). 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 food chain towards the reduction of its overall environmental footprint.
chemical composition (proteins, polyunsaturated fatty acids and vita- These interventions may include, among others, (a) source material re-
mins). Dried spirulina contains about 60% proteins, as well as, many pig- duction, (b) process upgrading, (c) reuse of waste material and (d)
ments which may be beneficial, including beta carotene and zeaxanthin, recycling or composting practices (Kyriakopoulou, Papadaki, &
plus phycobiliproteins, such as C-phycocyanin and allophycocyanin Krokida, 2013, 2015; Schau et al., 2008). In addition, LCA as a tool can
(Campanella, Crescentini, & Avino, 1999; Oliveira, Rosa, Moraes, & lead to the development of a cross-disciplinary agenda of research,
Pinto, 2009; Tang & Suter, 2011). Among those the most important linking together agricultural, environmental, social, and health concerns
functional ingredients are phycobiliproteins, a pigment-protein com- under the principle of sustainability and also applying them to daily
plex which is used into various food products to enhance their nutri- consumption practices (Schau et al., 2008).
tional qualities acting as food colorant, antioxidant and emulsifier, and In this study, the environmental performance of the isolation of phy-
can sufficiently replace or reduce the use of synthetic additives. C-phy- cocyanin from Spirulina platensis microalga using UAE and polar sol-
cocyanin is the main pigment and reaches 20% in dry weight of the cell vents such as water, ethanol or phosphate buffer was evaluated using
protein, depending on cultivation conditions (Cuellar-Bermudez et al., the Life Cycle Assessment methodology. The analysis was carried out,
2015), and lately has been approved by the Food and Drug Administra- using the Ecoinvent 2 database provided with the SimaPro 7 software
tion as a natural blue food colorant (Fda, 2014). (Pre' Consultants, 2014; SimaPro Manual PRe Consultants, 2008).The
For the effective recovery of phycocyanin, the nutrient should be ex- system examined included the open pond cultivation of the microalgae,
tracted from the microalgae biomass. Dehydration is considered an es- the hot air drying step and their extraction processing for the recovery
sential step pre-treatment step due to the high moisture content of of extracts equivalent to 1 kg of phycocyanin.
the material, however it may lead to the decrease of its phycocyanin
content. The drying of Spirulina constitutes approximately 30% of the 2. Life cycle assessment methodology
total production cost (Oliveira, Duarte, Moraes, Crexi, & Pinto, 2010).
The most commonly used method is the spray drying technique, in According to International Standards Organization (ISO) 14,000 se-
which the product is obtained in powder form, but usually, the powder ries, the technical framework for LCA methodology consists of four
does not satisfy all the criteria required for food use (Desmorieux & phases: (1) the goal and scope definition; (2) the inventory analysis;
Decaen, 2005). Desmorieux et al. reported that apart from high opera- (3) the impact assessment; and (4) the interpretation (ISO, 2006). De-
tion cost, the dried product obtained by spray drying did not have the fining the goal and scope involves the description of the purpose, the
same aspect and color as the low-cost drying methods (Desmorieux & functional unit (FU), the system boundaries, the data quality, assump-
Decaen, 2005). Greenhouse drying and oven-drying have the potential tions and simplifications. The life cycle inventory involves collecting
to approach the advantages derived from spray drying in terms of qual- data for each unit process regarding all relevant inputs and outputs of
ity and bioavailability at a lower cost (Tiburcio, Galvez, Cruz, & Gavino, energy and mass flows, as well as data on emissions to air, water and
2007). The development of an optimized drying process would there- soil. The life cycle impact assessment phase evaluates the potential en-
fore encourage farmers to grow Spirulina, as this will give them the as- vironmental impacts of the examined system, product and processes,
surance that they can produce dried Spirulina of marketable quality. while in the interpretation phase the results of the inventory analysis
The crucial step for the efficient phycocyanin recovery is the selec- and the environmental impacts are combined together.
tion of the optimum combination of extraction technique and solvent
system. The increasing legislative restrictions on the use of organic sol- 3. Goal and scope definition
vents in foods coupled to their negative effects on the functional prop-
erties of compounds are a challenge for an effective and eco-friendly The objective of this study was to compare the environmental as-
recovery process (Chemat et al., 2017; Chemat, Vian, & Cravotto, pects and impacts associated with different extraction methods for the
2012). Nowadays, Ultrasound Assisted Extraction (UAE) due to its recovery of phycocyanin from the microalga Spirulina platensis and to
high efficiency and low solvent consumption tends to replaced conven- identify the “hot spots” of the evaluated technologies as a way to poten-
tional extraction methods (Dey & Rathod, 2013; Patist & Bates, 2008). tially improve their environmental performance. This study includes the
The higher yield obtained in UAE is of major interest from an industrial analysis of different production stages, such as the cultivation, the har-
point of view, since the technology is an “add on” step to the existing vesting, the drying treatment and the extraction of the microalgal bio-
process with minimum alteration, application in aqueous extraction mass till the recovery of phycocyanin rich extracts. To evaluate the
where organic solvents can be replaced with generally recognized as impact of the different processes, an inventory registration of the mate-
safe solvents, shortening the extraction time (Vilkhu, Mawson, rial and energy flows, as well as, the emissions that occur over these
Simons, & Bates, 2008). The use of ultrasonic for extraction purposes processes was established.
in high-cost raw materials is an economical alternative to traditional ex-
traction processes, which is an industry demand for a sustainable 3.1. Functional unit
development.
The environmental assessment of industrial processes, such as the To remove performance variation and provide a fair comparison be-
phycocyanin recovery is based on Life Cycle Assessment (LCA) proce- tween the extracts recovered by the proposed production lines, the
dure. LCA is a tool that is being used to evaluate the environmental as- functional unit was defined as 1 kg of phycocyanin extracted from
pects associated with the entire life cycle of a product or process. It dried Spirulina platensis microalga to be used as a natural colorant and
monitors the materials, energy and wastes involved in each phase of bioactive component in food, nutraceuticals and cosmetic applications.
the product's life cycle, from raw materials extraction to final disposal, In general, the functional unit defines what the LCA study is measuring
by compiling an inventory of elementary flows. These flows are then and provides a reference to which the inputs and outputs can be related.
assigned to environmental impact categories according to the sub- A direct comparison of different materials is not in accordance with ISO
stances' ability to contribute to different key environmental issues 14040 as their properties may vary and lead to unambiguous definition
(JRC European Commission, 2010). The results of the analysis will regarding their common function (Kyriakopoulou et al., 2015).
allow identifying the phases/processes and the relevant resource flows
along the entire chain with the most significant environmental impacts 3.2. Product system and system boundaries
(Andersson, Ohlsson, & Olsson, 1994; Schau, Fet, & Declarations, 2008).
This will lead to the suggestion of effective alternative interventions (ei- The system model is based on a fictional microalgae farming estab-
ther technologies or management practices) for the upgrading of the 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 environmental factors and the high running costs due to the large salt-
their simple technology and low energy, as well as, the fact that they water volumes (Prieto et al., 2011). The biomass productivity in such
are inexpensive and well researched on different strain cultivations systems can reach up to 15 g m− 2 d−1 (Jiménez et al., 2003; De
(Jorquera, Kiperstok, Sales, Embiruçu, & Ghirardi, 2010). The simplicity Bhowmick et al., 2014), however it is difficult to maintaining optimal
of raceway ponds lays on their construction, since they are shallow cultivation parameters (Guterman et al., 1990; Zhang et al., 2015).
open basins with a considerate length constructed using a concrete Following, the cultivation, a series of dewatering and drying process-
shell lined with polyvinyl chloride (PVC) with various dimensions es are applied in order to concentrate the microalgae biomass. Accord-
(Aitken & Antizar-Ladislao, 2012). On the contrary to photo-bioreactors, ing to Grierson et al. (2013), a concentration of the microalgae
which present high energy and cost intensity production and operation, cultivation by a factor of 18.4 can be achieved by flocculation is applied,
their energy demand is limited, especially when utilizing natural sun- followed by air floatation (Grierson et al., 2013). An additional concen-
light, decreasing their environmental impact. Open pond systems have tration can be achieved by centrifugation leading to approximately
been used for the production of Spirulina biomass (Chanawongse, Lee, 28.6% solids content and a recycling of the growth medium (Grierson
Bunnag, & Tanticharoen, 1994; Radmann, Reinehr, & Costa, 2007). et al., 2013). The moisture content of the Spirulina platensis biomass
Moreover, the warm and sunny climate of the Aegean and Mediterra- after centrifugation is around 88% (Olguín, Galicia, Camacho, Mercado,
nean regions are ideal for outdoor cultivation, while algae can be also & Pérez, 1997; Stramarkou, Papadaki, Kyriakopoulou, & Krokida,
grown in greenhouses utilizing the sunlight. Therefore, to place the 2016). Combining the cultivation and handling specifications of Spiru-
microalga cultivation for this study the Greek island of Chios was lina with input and output data for open pond microalga cultivations
selected. provided by the literature (Campbell, Beer, & Batten, 2011; Collet et
In this LCA analysis, cradle-to-gate systems of phycocyanin rich ex- al., 2011), the mass and energy balances of the life cycle inventory of
tract production were considered, broken down into two stages; the ac- the cultivation and harvesting for the production of 1 kg wet biomass
quisition and treatment of the different raw material prior to extraction were constructed. This inventory analysis refers to a fictional open
(cultivation, harvesting, drying, etc.) and the extraction process for the pond cultivation system utilizing sunlight and sea water, along with
recovery of phycocyanin which can be used as additive in food, nutra- several nutrients for the cultivation of Spirulina biomass. Energy is re-
ceutical or cosmetic products. As in previous studies energy, water, car- quired for pumping and recirculating the water, for pumping the carbon
bon dioxide and nutrient requirements are treated as system inputs, dioxide needed for the cultivation and for the dewatering of the biomass
while solar radiation provided for the cultivation stage is not modeled by centrifugation. In Table 1 are presented the inputs and outputs of the
in the process as it is considered a natural energy source inventory for the production of 1 kg wet Spirulina biomass.
(Kyriakopoulou et al., 2015). Regarding the open pond installation,
site leveling works for capital infrastructure, including plumbing, 4.2. Pretreatment of Spirulina platensis and phycocyanin recovery
pumps, sheds, processing plant and machinery is not taken into consid-
eration, due to the assumed low attribution of these elements, while the Phycobiliprotein recovery involves the cell rupture of the microalga
land occupation and the energy demands for the biomass production and the release of these proteins from within the cell (Moraes, Sala,
were incorporated in the analysis. The energy consumption of each Cerveira, & Kalil, 2011). Traditionally, the recovery is achieved with
component employed in the production process (pumps, centrifuge, the use of polar solvents such as water, ethanol and buffer (Mahadev,
drier, ultrasound extractor) was calculated based on their specification 2005). The microalgal biomass is suspended and the pigments are
(considering commercial equipment) and application time. leached in the selected solvent (Prabuthas, Majumdar, Srivastav, &
Mishra, 2011). However, the wet biomass is immediately susceptible
4. Life cycle inventory to bacterial decomposition and degradation due to its nutritional com-
position. Therefore, dried biomass treatment gained popularity since it
Spirulina platensis has been widely used as a food supplement due to leads to effective storage and handling and at the same time retain the
its high protein content and nutritional value. In order to acquire the de- bioactive content (Oliveira et al., 2010).
sired biomass several steps including cultivation of the microalgae in The microalgae paste collected is dried in an industrial rotary dryer,
appropriate media, the harvesting of the biomass and the conditions as the one describe in the study of Show, Lee, Tay, Lee, and Chang
of post-harvest treatment (drying temperature, storage conditions,
etc.). Table 1
Life cycle inventory of the cultivation and harvest of the production of 1 kg wet Spirulina
4.1. Cultivation, harvest of microalgae 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 In this study phycocyanin recovery from both wet and dried Spiru-
powder ready for extraction. According to Show et al. (2015), wet slurry lina biomass is examined and the specifications of the extraction pro-
containing 30% solids of microalgae is treated at 120 °C for about 10 s in cesses are presented in Table 2. The selection of the conditions was
a pilot drum-dryer consuming about 52 kWh. In their assessment on en- made based on previous experimental work in which the comparison
ergy requirement for drying algae with a water content of approximate- on the yield of phycocyanin was made after treating the samples in se-
ly 94% to 4%, they report that for the production of 1 kg of dry algae lected extraction time, ultrasound power and temperatures
product 18.2 kg of water need to be evaporated. The energy demand (Stramarkou et al., 2016). Taking into consideration the experimental
for this process was 15.7 Mcal, leading to 1.15 Mcal/kg of evaporated yielding the mass and energy balances of each extraction line case
water (or 4.815 MJ/kg water) (Show et al., 2015). In the case of the cen- study were prepared for the production of 1 kg of phycocyanin in the
trifuged Spirulina biomass with an initial moisture content of 88% to form of liquid extract (Table 3).
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 5. Environmental impact assessment and interpretation
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. The environmental impacts associated with the recovery of bioac-
As the water content is a determining factor on the energy consump- tive, phycobilin-rich extracts from Spirulina platensis were quantified
tion, the dewatering and centrifugation steps prior to drying are consid- using the problem-oriented approach, CML 2 baseline 2000 v2.04
ered essential to reduce the amount of water entering the rotary drier. (Center for Environmental Studies, University of Leiden) method pro-
Adjusting not only the initial moisture content (to lower levels), but vided by the SimaPro LCA tool (de Bruijn, van Duin, & Huijbregts,
also the moisture content of the dried microalga biomass (to acceptable 2002; European Commission—Joint Research Centre—Institute for
but not extremely low levels) can affect the energy requirement, the Environment and Sustainability, 2010; Guinée et al., 2002). The selected
processes economic viability and the environmental performance. method includes a series of impact categories, namely Ozone layer de-
Nowadays, for the disruption of Spirulina's cell walls from both dried pletion, Human toxicity, Fresh water aquatic ecotoxicity, Marine aquatic
or wet biomass, sonication is suggested since it enhances the bioactive ecotoxicity, Terrestrial ecotoxicity, Photochemical oxidation, Global
compound recover leading to shorter treatment times (Dey & Rathod, warming (indicator of Carbon footprint), Acidification, Abiotic depletion
2013; Hadiyanto, 2016). UAE is considered an inexpensive extraction and Eutrophication, and has been previously used for the evaluation of
techniques which involves the use of ultrasound of 20 kHz to 40 kHz; microalgae cultivation and different extraction processes for the recov-
which increases the permeability of cell walls and produces cavitation ery of bioactive compounds (Collet et al., 2014; Lardon, Hélias, Sialve,
(the formation, growth, and implosive collapse of bubbles in a liquid) Steyer, & Bernard, 2009; Rodríguez-Meizoso et al., 2012). The impacts
(Drosou, Kyriakopoulou, Bimpilas, Tsimogiannis, & Krokida, 2015; in this method are quantified with appropriate physical units based on
Kyriakopoulou et al., 2015). The cavitation effect helps in the cell wall the life cycle inventory data according to physically based formulae
disruption allowing the solvent to penetrate into the biomass and in- (Guinée et al., 2002).
creasing the contact surface area between the solvent and compounds In this study a life cycle assessment from cradle to gate was per-
of interest, resulting into increased mass transfer also through good formed including the production of wet and dried Spirulina platensis
mixing (Naveena, Armshaw, & Tony Pembroke, 2015). Therefore, UAE biomass and the extraction of the biomass for the production of 1 kg
provides increased yielding and leaching rate, along with the advantage of phycocyanin in the form of extract. Therefore, the environmental per-
of reduced solvent volumes and temperatures, which is crucial for the formance of the phycocyanin recovery process lines involves the foot-
recovery of thermolabile compounds (Dey & Rathod, 2013). prints of all the necessary steps; the cultivation (raceways ponds and
For the recovery of phycocyanin from wet and dried Spirulina harvesting), the drying and the final biomass treatment using different
platensis, the combination of ultrasound with solvents such as water, solvents. Each step was found to contribute to the environmental foot-
ethanol and phosphate buffer (pH 7) is suggested (Agustini, Suzery, print according to its energy and resources consumption.
Sutrisnanto, & Ma'ruf, 2015; Kumar, Dhar, Pabbi, Kumar, & Walia, Specifically, for the cultivation step the most important contribu-
2014; Stramarkou et al., 2016). The selection of solvents is based on tions to the energy demand come from the electricity required to circu-
the nature of phycobilins, which are covalently bound to a water-solu- late the culture and the embodied energy in pond construction. These
ble protein that aggregates on the surface of the photosynthetic mem- energy fractions were reported to be 22% to 79% and 8% to 70%, respec-
brane located in the cytoplasm or in the stroma of the chloroplast tively (Slade & Bauen, 2013). In addition, the embodied energy in the
(Cuellar-Bermudez et al., 2015). The most efficient pigment extraction fertilizers, such as nitrogen and phosphorus, is reported that it may con-
procedures combine mechanical and chemical methods that lead to tribute to the energy demand by 6% to 40% (Slade & Bauen, 2013). De-
protein release (Sobiechowska-Sasim, Stoń-Egiert, & Kosakowska, spite these energy demands, the cultivation stage yields to some net
2014). However, each combination of solvent and extraction conditions benefit in relation to global warming impact due to the intake of CO2
leads to different yielding as it was presented in our previous study from microalgae, presenting a slightly reduced overall carbon footprint.
(Stramarkou et al., 2016). The difference in the yield on the compounds However, the electricity usage, also for the centrifugation, and fertilizer
of interest, which can be a result of the material selected or the recovery consumption contribute not only the global warming potential but also
process examined, has shown among different studies than it can affect to the acidification and abiotic depletion potential.
the environmental performance of the production process On the other hand, the recycling of harvest water during dewatering
(Kyriakopoulou et al., 2015; Papadaki, Kyriakopoulou, & Krokida, and centrifugation reduces the nutrient use and controls the fertilizer
2016; Silva et al., 2015). usage, which leads to the reduction of the natural resources depletion

Table 2
Specification of extraction conditions examined.

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

Spirulina platensis Wet paste Wet paste Wet paste Dry powder Dry powder Dry powder
Solvent Water Ethanol Buffer Water Ethanol Buffer
Solid liquid ratio (kg d.b.:L) 1:20 1:20 1:20 1:20 1:20 1:20
Conditionsa 30 °C, 5 min, 30 °C, 5 min, 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 25 kHz, 450 W 25 kHz, 450 W
a
The extraction conditions are derived from the research of Stramarkou et al., 2016.
 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 the phycocyanin yield, pretreatment steps of high energy demand
(Clarens, Resurreccion, White, & Colosi, 2010; Lardon et al., 2009). Eu- such as wet biomass drying influence insignificantly the overall envi-
trophication potential is reduced due to the avoidance of nitrate and ronmental performance. Even though drying is considered an energy
phosphate leaching to the ground water, and ammonia and nitrous ox- demanding step, especially due to the high moisture content of Spirulina
ides to the air (Kyriakopoulou et al., 2015). platensis, the effect of the treatment on the phycocyanin content or its
Although the dewatering steps together with drying have been re- availability in the biomass showed opposite results on the overall envi-
ported as the most energy and cost intensive steps in algal biomass pro- ronmental performance.
duction due to the low concentration of microalgae in the culture Examining the treatment processes for the extraction of phycocyanin
medium (Soomro et al., 2016; Uduman, Qi, Danquah, Forde, & from wet biomass using water, ethanol and buffer as solvent (Table 4),
Hoadley, 2010), dewatering and centrifuge operation are considered significant differences on several impact categories were observed.
an important step for condensing the microalga paste prior to extraction Among the three Ultrasound Assisted Extraction processes, UAE 2
or drying (Grierson et al., 2013). Therefore, the selection of energy effi- using ethanol as solvent exhibited the lowest environmental impact in
cient systems is important to result to lower energy consumption and categories of carbon footprint, ozon layer depletion and human toxicity
lower emissions (Weschler, Barr, Harper, & Landis, 2014). However, even though an organic solvent was used. The positive effect of ethanol
the selection of the drying processing can significantly affect the quality on the recovery of phycobiloproteins affects the environmental perfor-
of the final dried microalga powder and thus the extractability of the mance positively, since less biomass is needed to be cultivated, harvest-
targeted bioactive compounds (Papadaki, Kyriakopoulou, Stramarkou, ed and extracted for the recovery of the desired amount of phycocyanin.
Tzovenis, & Krokida, 2017). In this study an industrially applied hot air However, in some cases the effect of ethanol overcomes the positive ef-
drying method was selected for all the production lines. Although the fect of the high yielding, leading to slightly worsen impact, especially in
dried powder used in all extraction processes was considered of the the photochemical oxidation and the ozone layer depletion potential.
same quality, the experimental extraction results for each process pre- The combination of buffer as solvent with dried Spirulina platensis
sented significant differences, affecting greatly the environmental per- biomass leads to significantly higher phycocyanin yields, contributing
formances of phycocyanin production. the lowest on the overall environmental performance, despite the fact
As far as the extraction of both wet and dried biomass is concerned, that an extremely energy consuming step is added prior to extraction.
the environmental impact is primarily affected by the energy demand of The treatment of dried biomass with ethanol exhibits the highest foot-
the ultrasound extractor and the nature of the solvent system used. The prints due to the low yielding and the high material (biomass and sol-
energy demand according to the experimental data was 54 MJ/kg dry vent) consumption (Table 5).
biomass treated, while the corresponding energy demands for other ul- The carbon footprint of the two preferable extraction procedures for
trasound extractors can reach up to 148 MJ/kg biomass treated wet (UAE 2) and dried biomass (UAE 6) is 2.06E + 03 and 1.18E + 03 kg
depended on the time of the treatment, the scale and the specifications CO2 eq, respectively. This difference of the latter from the former is
of the machinery used (Ferreira, Dias, Silva, & Costa, 2016). However, fo- around 43% and shows once more that the yielding is the crucial factor
cusing further on the recovery of bioactive compound the crucial factor for the environmental performance of phycocyanin recovery. The dry-
for the overall environmental performance of the recovery process is ing step prior to extraction permits the highest penetration of the sol-
the achieved yield. If the overall footprint of the process is allocated to 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

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