CHEMICAL ENGINEERING TRANSACTIONS
VOL. 79, 2020
A publication of
The Italian Association
of Chemical Engineering
Online at www.cetjournal.it
Guest Editors: Enrico Bardone, Antonio Marzocchella, Marco Bravi
Copyright © 2020, AIDIC Servizi S.r.l.
ISBN 978-88-95608-77-8; ISSN 2283-9216
From Haematococcus Pluvialis Microalgae
a Powerful Antioxidant for Cosmetic Applications
Tiziana Marinoa, Angela Iovinea,b, Patrizia Casellab, Maria Martinoc, Simeone
Chianesea, Vincenzo Laroccac, Dino Musmarraa, Antonio Molinob,*
a
Department of Engineering, University of Campania “Luigi Vanvitelli”, Via Roma, 29 - 81031 Aversa, Italy
b
ENEA, Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Territorial and
Production System Sustainability Department, CR Portici Piazzale Enrico Fermi, 1 - 80055 Portici (NA),
Italy*antonio.molino@enea.it
c
ENEA, Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Department of
Sustainability-CR ENEA Trisaia, SS Jonica 106, km 419+500, 75026 Rotondella (MT), Italy
antonio.molino@enea.it
The global cosmetic market is promptly growing, showing a strong boost with the growth of economic well-
being. In this context, the demand of innovative, more and more specific cosmetic ingredients has been the
key for searching alternative, preferably naturally-based, active components. Microalgae represent one of the
most attracting microorganisms and natural deposit for bioactive compounds for their peculiar compositions
and properties. In the cosmetic sector, their efficient application has recently been highlighted by the placing
on the market of different cosmetic preparations. A powerful antioxidant, 550 times more effective than vitamin
E, with enormous potential for cosmetic and pharmaceutical applications, is astaxanthin. Although high-quality
astaxanthin can be obtained from microalgae, particularly from H. pluvialis specie, actually the major part of
the market is occupied by the synthetic form (99%), conferring substantial differences between their price. In
fact, algal-based astaxanthin costs ˃6000 €/Kg, while the synthetic form 600 €/kg. Thus, along with novel
natural product launch, the development of cost-effective technologies able to match the existing ones,
represents the major challenge for the microalgae application. This study aims to determine the feasibility of
microalgal-based astaxanthin production, by exploring both procedural issues and costs evaluation.
1. Introduction
In accordance with the Federal Food, Drug & Cosmetic Act of the US FDA and article L5131-1 of the French
Public Health Code, a cosmetic product is defined as any substance or preparation that is to be rubbed,
poured, sprinkled, or sprayed on, introduced into or applied to external parts of the body, in particular the
epidermis, hair and capillary systems, nails, lips and external genitals, or to the teeth and the mucous
membranes of the oral cavity as the product cleans, perfumes, protects them, modifies their appearance,
keeps them in good condition or helps to reduce body odours (Cosmetics DGCIS 2012). In short, its function
is to assure personal hygiene and beautification.The global cosmetic market is promptly expanding, and it had
a strong growth with the advance of economic well-being. Recently it has been evidenced as the use of
cosmetic in human daily life has become a necessity. In fact, more than 70% of the population considers
cosmetic and personal care products essential for their daily lives (www.cosmeticseurope.eu, 2019).In
particular, facial cosmetics might contribute to increase the perceptions of attractiveness, by improving
appearance. Etcoff et al. (2011) evidenced as the characteristics of a beautiful face can constitute relevant
bio-signals, proposing a potential correlation with judgments of social attributes, proposing a potential
correlation with judgments of social attributes. According to some estimates, the global cosmetics market in
2017 has reached 475 billion euros with a Compound Annual Growth Rate (CAGR) between 7% and 7.14%,
and with a forecast that could raise more than 625 billion euros in 2022 (www.alliedmarketresearch.com,
2019; www.globenewswire.com, 2019).
DOI: 10.3303/CET2079046
Paper Received: 14 September 2019; Revised: 24 January 2020; Accepted: 2 March 2020
Please cite this article as: Marino T., Iovine A., Casella P., Martino M., Chianese S., Larocca V., Musmarra D., Molino A., 2020, From
Haematococcus Pluvialis Microalgae, Chemical Engineering Transactions, 79, 271-276 DOI:10.3303/CET2079046
271
The increasing demand for cosmetics is directly linked to the search of innovative, preferably natural,
functional ingredients to meet the customer requests, that is improving personal hygiene, facial, body skin and
hair health.
Nowadays consumers prefer to use biological and environmentally sustainable cosmetics respect to products
with chemical ingredients fossil derived. There is a crescent demand, for example, for microalgal
biomass/extracts, which could significantly affect the cosmetic market in the near future (Chew et al., 2017;
Molino et al., 2019a, Molino et al., 2018a; Savvidou et al., 2019). This is due to the outstanding microalgae
properties and the possibility to easily modulate their growth driving it towards the production of selected
antioxidant/pigments or other nutrients and secondary metabolites. Until now, the known algal species are
more than 20000 (Christaki et al., 2012), and they are able to mitigate greenhouse gas emissions providing for
about 40–50% global photosynthesis each year (Wang et al., 2015). Both whole algae biomass and algae
extracts are present in some cosmetic formulations, mainly in skin and body creams, lotions, gels, soaps,
shampoos and they show protective actions against UV rays and as anti-aging, conferring hydration and
protection of the skin (Ariede at al., 2017). The extracts mainly derive from coccoid and filamentous
microalgae, Phaeodactylum tricornutum species, Chlorella sp. or from macroalgae like Fucus. Proteins and
peptides are also extracted from Porphyra, Wakame and microalgae such as Arthrosira platensis and
Chlorella. Haematococcus pluvialis, Isochrysis galbana, Nannochloropsis gaditana, Phaodactylum
tricornutum, Tetrasemis chuii and Dunaliella salina are present attracting microorganisms with enormous
potential (Marino et al., 2019; Molino et al., 2018b; Molino et al., 2018c) and their use as ingredient in
cosmetics is allowed at European level since 2006 when 2006/257/CE Commission Decision has been issued
for the establishment of an inventory of cosmetics (www. eur-lex.europa.eu, 2019) . This Regulation
introduced the Cosmetic ingredient database or CosIng (www. eur-lex.europa.eu, 2019) mentioning
ingredients that have been authorized for the production of cosmetics. The CosIng database is available and
can be consulted on the European Commission website by simply entering the necessary information such as
the full name of the selected compound. Among the microalgae extracts, oil and biomass present in the
CosIng database, those of Haematococcus pluvialis, which represents the richest natural xanthophyll
carotenoid astaxanthin (3,3′-dihydroxy-β, β′-carotene-4,4′-dione) source, is one of the most interesting
cosmetic ingredient due to the outstanding metabolite properties. In fact, the red-pink pigment astaxanthin is a
potent anti-inflammatory and has more antioxidant capabilities than vitamin A and vitamin E. The United
States Food and Drug Administration has already accepted the use of astaxanthin as food colorant in animal
feed and the European Commission considers natural astaxanthin as a food dye and the Haematococcus
pluvialis oleoresin as food supplement as evidenced by the Commission Implementing Regulation
2017/2470/EU (Casella et al., 2019). In the cosmetic sector the CosIng reports the following forms: Dunaliella
salina/Haematococcus pluvialis extract, Haematococcus pluvialis extract/oil/powder with the related functions
reported in Table 1.
Table 1: Cosmetic ingredients (EC number, International Nomenclature of Cosmetics (INCI)) of interest for this
work and authorized to be used in cosmetic formulation (CosIng search, 2019)
Name EC number INCI Description Function(s)
Haematococcus pluvialis 919-412-5 Haematococcus pluvialis
extract
Haematococcus pluvialis
Extract is an extract of the
Alga, Haematococcus
pluvialis,
Stephanosphaeraceae
Antioxidant
Unavailable Haematococcus pluvialis Oil Haematococcus pluvialis Oil is
the oil expressed from the
Alga, Haematococcus
pluvialis,
Stephanosphaeraceae
Antioxidant
Skin
conditioning
Unavailable Haematococcus pluvialis
Powder
Haematococcus pluvialis
Powder is the powder derived
from the Alga, Haematococcus
pluvialis,
Stephanosphaeraceae
Antioxidant
unavailable Dunaliella
salina/Haematococcus
pluvialis Extract
Dunaliella
Salina/Haematococcus
pluvialis Extract is the extract
of the alga, Dunaliella salina
and Haematococcus pluvialis
Antioxidant
Skin protecting
272
The aim of this work was to evaluate the economic feasibility of a natural astaxanthin production based on the
cultivation of Haematococcus pluvialis microalgae and the subsequent antioxidant extraction via supercritical
CO2 utilization.
2. Materials and Methods
The research work was performed in the context of VALUEMAG project (Horizon 2020-Grant Agreement No
745695). More precisely the presented algal refinery is composed by three main components: 1) the algae
production system, 2) the biomass pretreatment and 3) CO2 extraction of bioactive compounds. The Block
Flow Diagram of the VALUEMAG bioefinery is represented in Figure 1.
Figure 1. Natural astaxanthin production via Haematococcus pluvialis microalgae cultivation,
dewatering,biomass drying and pretreatment, pigment extraction and carbon dioxide recovery and recycle.
For microalgae cultivation within the VALUEMAG project a specific conical reactor (called SOMAC), for an
optimal cells growth, has been developed. Moreover, the pioneering approach is based on the
microorganisms modification by means the incorporation of superparamagnetic iron oxide nanoparticles which
could enhance the biomass production (Savvidou et al., 2019). The correct cultivation of magnetic modified
microalgae (MAGMA) is assured by their immobilization on the a soft magnetic conical surface of SOMAC
reactor and a thin water layer. After the algal growth phase, biomass is harvested through a dewatering
system which allows at the same time, to recover and recycle water, hence perfectly integrated in the
sustainability context. For this purpose, an hydraulic pressure difference applied across the two sides of a
polysulfone microfiltration membrane permits to obtain a permeate that is reused as a cultivation medium and
a retentate that contains the harvested microalgae biomass (Marino et al., 2019). Algal cells biomass are dried
by using a lyophilizer and after drying process, biomass is mechanically pretreated for enhancing solvent
penetration during the subsequent extraction process of bioactive compounds. The resulting biomass enters in
the supercritical CO2 unit in which gaseous carbon dioxide, after passing through a condenser subcooling
device, is sent by means of a pump to the extractor filled with the microalgal matrix. Downstream of the
extractor, a separator is located to isolate the desired bioactive compounds from the carbon dioxide used
during the process. Finally, the gas is sent again to the condenser subcooling apparatus which allows to
recycle for other extraction step.
On the basis of the above mentioned overall process, a preliminary economic assessment of the astaxanthin
production by Haematococcus pluvialis cultivation has been carried out considering the contributes of the
different steps. The costs, expressed in terms of euros/day, have been calculated by considering the
previously optimized operational conditions related to the astaxanthin extraction.
3. Results and discussion
For an optimal microalgae growth, considering the overall biorefinery described above (Section 2) and taking
into account a productivity of 0.5-1 Kg/day on dry basis, the content range of the various nutrients necessary
for Haematococcus pluvialis cells, have been estimated and the related costs calculated (Table 2).
273
Table 2: Culture medium composition required for Haematococcus pluvialis cultivation
Nutrient Content (g/day) Cost (eurocents/day)
NaNO3 150 ÷ 225 7.50 ÷ 11.25
MgSO4*7H2O 7.50 ÷ 11.25 0.075 ÷ 0.1125
CaCl2*2H2O 3.6 ÷ 5.4 0.122÷0.184
K2HPO4*3H2O 4.0 ÷ 6.0 28.4 ÷ 42.6
C6H8O7 0.6 ÷ 0.9 0.036 ÷ 0.054
C6H8FeNO7 0.6 ÷ 0.9 8.8 ÷ 13.2
H3BO3 0.286 ÷ 0.429 2.12 ÷ 3.18
Na2CO3 2 ÷ 3 0.056 ÷ 0.084
Na2*EDTA 0.10 ÷ 0.15 2.58 ÷ 3.87
Traces including: (CuSO4*5H2O, ZnSO4*7H2O
MnCl2*4H2O, Na2MoO4*2H2O, Co(NO3)2*6H2O)
69.28 ÷ 103.92(x10-3) 6.63 ÷ 9.94
TOTAL 169-253 56.32 ÷ 84.47
Before astaxanthin extraction, a mechanical ball-mill pretreatment (operational conditions as reported by
Molino et al., 2018b,c) stage was performed in order to break down the cellular wall of microalgae strain.
The optimized operational conditions which allowed to recover astaxanthin by using supercritical carbon
dioxide, are summarized in Figure 2.
Figure 2. Optimized operational conditions adopted for extracting astaxanthin from Haematococcus pluvialis
via supercritical CO2 operation (Molino et al., 2018b, c).
Supercritical CO2 pressure of 350 bar, temperature of 50 °C, CO2 flow rate of 0.47 Kg/min are optimized
conditions to obtain 23.58wt% of extract that contains the 48.4 of astaxanthin recovered, 97.2% of β-carotene
recovered as well as 53.3% of lutein respect to the biomass content. Table 3 reports the initial biomass and
the extract composition. The proteins content passed from 25.69% to 55.37% and similarly, the carotenoids
increased from 2.87% to 6.38% and lipids from 2.60 to 8.72% in the biomass and the extract, respectively. On
the contrary, carbohydrates and TDF amount reduced after the extraction operation.
274
Table 3: Haematococcus pluvialis red phase and extract composition
Starting from these results, the calculated energy demand and the costs associated with the VALUEMAG
biorefinery approach, have allowed to estimate that the most expensive operation is represented by the
supercritical fluid extraction, which implies an average cost of 31.33 euros/day, followed by the drying step,
the microalgae growth (mainly due to the use of LED lights), the nutrients and finally the mechanical
pretreatment.
Figure 3. Economic estimate of astaxanthin production from microalgae
The energy consumption mirrors the economic impact. In fact, to perform the supercritical CO2 extraction
about 190.4 kWh/day are required, that is a value higher the sum of the remaining employed steps. However,
the overall biorefinery offer precious advantages for the sustainable astaxanthin production, since it
guarantees to preserve the pigment integrity without any contamination due to possible solvent residues; in
addition, it avoids the application of any downstream purification which might imply additional costs for
astaxanthin recovery prior its placing on the cosmetic market.
4. Conclusions
In this study, the feasibility of astaxanthin production from microalgae has been evaluated in a biorefinery
context which, starting from Haematococcus pluvialis cultivation, allows to obtain the powerful antioxidant by
means of supercritical CO2 extraction. Within this biorefinery concept a dewatering step enables to reduce
algal cultivation costs together with a carbon dioxide recovery and reuse for the environment protection.
3.07
euros/day
0.55
euros/day
7.43
euros/day
0.24
euros/day
31.33
euros/day
Plant for Growth
Nutrients
Drying
Pre-treatment
CO2-Extraction
Biomass Extract
Ash 4.02% 3.61%
Protein 25.69% 55.37%
Carbohydrates 6.30% 0.26%
Total dietary fiber (TDF) 58.52% 25.66%
Carotenoids 2.87% 6.38%
Lipids 2.60% 8.72%
of which FAME: 88.38% 88.84%
Carotenoids composition:
Astaxanthin 69.71% 65.67%
β-carotene 3.45% 6.51%
Lutein 26.84% 27.82%
FAMEs Composition:
SFA 28.14% 26.47%
MUFA 23.66% 26.07%
PUFA 48.20% 47.46%
275
Characterization of the extract after the use of supercritical CO2 revealed that the content of proteins, lipids
and carotenoids increased in comparison to that of the biomass. On the contrary, after the extraction TDFs
amount was lower than initial biomass. A preliminary cost analysis, performed on the basis of previously
optimized operational conditions, have evidenced that, considering the potential productivity of 0.5-1 Kg/day
on dry basis, an average cost of ~42.60 euros/day is required for the efficient astaxanthin recovery. It is
possible to assume that a high impact is due to astaxanthin extraction step. However, the supercritical CO2
utilization constitutes a valid eco-friendly alternative to the traditional techniques. In fact, it guarantees the
production of high-stability algal metabolite and assures CO2 recycle without any dangerous emission in the
environment.
Funding: This research was funded by Bio Based Industries Joint Undertaking under the European Union’s
Horizon 2020 research and innovation program under Grant Agreement No 745695 (VALUEMAG).
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