Journal of Biotechnology 215 (2015) 13–19

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Journal of Biotechnology
journal homepage: www.elsevier.com/locate/jbiotec

Antioxidant activity of some Moroccan marine microalgae: Pufa

profiles, carotenoids and phenolic content
Amal Maadane a,b , Nawal Merghoub a,∗,1 , Tarik Ainane a , Hicham El Arroussi a ,
Redouane Benhima a , Saaid Amzazi b , Youssef Bakri b , Imane Wahby a,1
a
Green Biotechnology Center, MAScIR (Moroccan Foundation for Advanced Science, Innovation & Research) Rabat Design Center, Rabat, Morocco
b
Laboratory of Biochemistry–Immunology, Sciences Faculty–University Mohammed V of Rabat, Morocco

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

Article history: In order to promote Moroccan natural resources, this study aims to evaluate the potential of microalgae
Received 5 November 2014 isolated from Moroccan coastlines, as new source of natural antioxidants. Different extracts (ethanolic,
Received in revised form 4 June 2015 ethanol/water and aqueous) obtained from 9 microalgae strains were screened for their in vitro antioxi-
Accepted 16 June 2015
dant activity using DPPH free radical-scavenging assay. The highest antioxidant potentials were obtained
Available online 22 June 2015
in Dunalliela sp., Tetraselmis sp. and Nannochloropsis gaditana extracts. The obtained results indicate that
ethanol extract of all microalgae strains exhibit higher antioxidant activity, when compared to water and
Keywords:
ethanol/water extracts. Therefore, total phenolic and carotenoid content measurement were performed
Microalgae
Antioxidant activity
in active ethanol extracts. The PUFA profiles of ethanol extracts were also determined by GC/MS analysis.
Phenolic content The studied microalgae strains displayed high PUFA content ranging from 12.9 to 76.9 %, total carotenoids
Carotenoids content content varied from 1.9 and 10.8 mg/g of extract and total polyphenol content varied from 8.1 to 32.0 mg
PUFA profiles Gallic acid Equivalent/g of extract weight. The correlation between the antioxidant capacities and the
phenolic content and the carotenoids content were found to be insignificant, indicating that these com-
pounds might not be major contributor to the antioxidant activity of these microalgae. The microalgae
extracts exerting the high antioxidant activity are potential new source of natural antioxidants.
© 2015 Elsevier B.V. All rights reserved.

1. Introduction interest as a source of high value products. The possibility of cul-

Microalgae constitute an important source for biomolecules
with a wide range of applications that can be harnessed for com-
mercial use. They can produce proteins, lipids, vitamins, pigments
and other molecules exploited for health, food and feed additives,
cosmetics and for energy production (De Jesus Raposo et al., 2013;
Barra et al., 2014). Recently microalgae have gained an increasing
Abbreviations: DPPH, 2, 2-diphenyl-1-picrylhydrazyl; FAME, fatty acid methyl
esters; PUFA, polyunsaturated fatty acids; MUFA, monounsaturated fatty acids;
SFA, saturated acids; FRAP, ferric reducing antioxidant potential; TEAC, trolox
equivalent antioxidant capacity; BHT, butylatedhydroxytoluene; BHA, butylated-
hydroxyanisole; GAE/EW, gallic acid equivalent mg of extract weight; ROS, reactive
oxygen species.
∗ Corresponding author at: Green Biotechnology–MASCIR (Moroccan foundation
for Advanced Science, Innovation and Research) Rabat Design Center, Rue Mohamed
Al Jazouli. Madinat Al Irfane–Rabat, 10100, Morocco. Fax: +212 530 27 98 27.
E-mail address: n.merghoub@mascir.com (N. Merghoub).
1
These authors jointly supervised this work
turing them in non arable lands as well as metabolism modulation
following some stresses application make microalgae an attractive
feedstock for industrial production. There is wide range of com-
pounds that could be potentially employed as antioxidant agents
like: pigments including carotenoids, phenolic compounds, sul-
fated polysaccharides and long-chain polyunsaturated fatty acids
(Zaho et al., 2004; Plaza et al., 2009; Goh et al., 2010; Hajimahmoodi
et al., 2009; Goiris et al., 2012).
There is a growing worldwide interest toward finding new,
safe and powerful antioxidants from natural sources in order to
minimize oxidative damage to living cells and prevent oxidative
deterioration in commercialized products such as food, pharma-
ceuticals or cosmetics. Antioxidants compounds exert their action
through different mechanisms including prevention of chain ini-
tiation, chelating of transition metal ion catalysts, decomposition
of peroxidases, prevention of continued hydrogen abstraction and
radical scavenging (Valko et al., 2006).
There is a continuing balance between oxidation and antioxida-
tion reactions necessary for proper physiological function, taking

http://dx.doi.org/10.1016/j.jbiotec.2015.06.400
0168-1656/© 2015 Elsevier B.V. All rights reserved.
14 A. Maadane et al. / Journal of Biotechnology 215 (2015) 13–19
into account the oxidative stress and the critical roles of oxi-
dants in cell signaling, defense against infection, and tissue repair
(Kancheva and Kasaikina, 2013; Wahlqvist, 2013). Furthermore,
natural antioxidants are generally more suitable for human con-
sumption than synthetic ones such as butylated hydroxytoluene
(BHT) or butylated hydroxyanisole (BHA), whose carcinogenic
effects have been previously reported (Namiki, 1990; Pokorny,
1991). Therefore, these harmful effects increased the research of
new natural sources to replace the synthetic antioxidants (Li et al.,
2007; Rodriguez-Garcia and Guil-Guerrero 2008; Catarina Guedes
et al., 2013). Several previous works have been interested on the
evaluation of antioxidant activity of the microalgae species. Nev-
ertheless, to our knowledge, this study is the first to evaluate
antioxidant potential of Moroccan microalgae strains; and investi-
gate the correlation between the antioxidant capacity and phenolic,
carotenoids, and PUFA content.
In the present contribution we screened nine Moroccan marine
microalgae for antioxidant activity. Extraction conditions were
optimized to obtain extracts with maximal antioxidant potential.
The antioxidant activity was assessed by the DPPH (1,1-diphenyl-2-
picryl-hydrazil) radical bleaching method. Active ethanolic extracts
were characterized by determination of PUFA profile using GC/MS.
Total phenolic and carotenoids content measurements were also
performed.
2. Materials and methods
2.1. Microalgae strains and culture conditions
Nine marine microalgae (Nannochlorpsis gaditana, Dunaliella sp.,
Dunaliella salina, Phaedactylium tricornutum, Isochrysis sp., Navicula
sp., Chaeotoceros sp., Chlorella sp. and Tetraselmis sp.) belonging to
MAScIR’s microalgae collection, were selected for this study. Tax-
onomical identification was performed by sequencing partial 18S
region of the ribosomal RNA gene.
Microalgae were grown in sterile natural seawater enriched
with f/2 medium nutrients (Guillard and Ryther, 1962), with addi-
tion of silicate for the diatoms. Batch cultures were performed in
5 L flasks agitated by air bubbling at 25 ◦ C, under continuous illu-
mination with 150 ␮mol m−2 s−1 . The growth was monitored by
measuring the optical density at 680 nm, every two days, using f/2
medium as the blank. The biomass of each strain was harvested at
the stationary phase by centrifugation at (2547 RCF for 10 min) and
then the wet biomass were freeze-dried and stored at −20◦ until
extraction.
2.2. Extraction procedure
Three different extractions were performed to obtain three
types of extracts with different polarities, using (ethanol, water,
ethanol/water). For each extraction, 1 g of dried biomass was
extracted with 100 mL ethanol or ethanol/water (1/1, v/v) for 3 h at
room temperature, in darkness. The extraction was repeated twice
and all extracts were combined. Ethanolic and hydro-alcoholic
extract were filtered with a Whatman’s filter paper (grade 1) and
concentrated under reduced pressure in a rotary evaporator. Water
extracts were performed by ultra-sonication (20 kHz, 400 W, Bran-
son – P/S) of microalgal biomass in ultrapure water (2 g/50 mL H2 O)
for 15 min, on ice. The aqueous extract were then filtered with a
Whatman’s filter paper (grade 1) and lyophilized. All extracts were
stored at –20 ◦ C until use.
2.3. DPPH radical scavenging assay
The DPPH radical-scavenging activity was determined following
the method of Fukumoto and Mazza (2000). This method is simple,
rapid and convenient, independent of sample polarity for screen-
ing of many samples for radical scavenging activity (Marxen et al.,
2007). The stock solution of the extract was prepared in methanol at
the concentration of 20 mg/mL. Twenty microliter of each extracts
was added to 180 ␮L of DPPH radical solution (60 ␮M) in 96-well
plates, in order to obtain final concentrations of 50, 100, 200, 500,
1000 ␮g/mL. The samples were shaken. Due to the colored extracts
that can absorb at 520 nm, it was necessary to prepare a control (i.e.
sample blank) which consisted of 20 ␮L of each sample solution
added to 180 ␮l of methanol. Then the absorbance of samples was
measured at 30, 60 and 120 min at 520 nm, with methanol as the
blank, using spectrophotometer (Multiscan, Thermo-Fisher Scien-
tific). Ascorbic acid (Vit C) and BHT were used as standards. The IC50
(Inhibition Concentration at 50%) was used to compare the radical
scavenging activity. The DPPH radical activity was calculated using
the following equation:
Scavengingeffect(%) = [1-(Asample -Asampleblank )/Acontrol ] × 100.
A sample : is the absorbance of methanolic DPPH solution with the
presence of all of the extract samples and standard.A sampleblank : is
the absorbance of extracts sample in methanol (without DPPH in
order to subtract the absorbance of colored extracts).A control : is the
absorbance of the methanolic DPPH solution.
2.4. Carotenoids content measurement
In this study, determination of carotenoids content was car-
ried out spectrophotometrically according to Lichtenthaler and
Buschmann (2001); Miazek and Ledakowicz, (2013) methods.
Aliquots of the extracts were prepared at concentration of 1 mg/mL
in ethanol. The absorbance of samples was measured at 470, 648
and 664 nm by spectrophotometer (ULTROSPEC 3100 PRO, Thermo-
Scientific). The ethanol was used as a blank. The pigment content
(chlorophyll a, chlorophyll b and total carotenoids) was calcu-
lated using the Lichtenthaler equations previously reported: (Chla:
Chlorophyll a, Chlb: Chlorophyll b).
Chla = 13.36 × A664 − 5.19A648
Chlb = 27.43 × A648 – 8.12A664
Carotenoidstotal = (1000 × A470 − 1.63 × chla − 104.96 × chlb)/221
2.5. Determination of total phenolic compounds
Total soluble phenolic compounds in extracts were deter-
mined with Folin-Ciocalteu reagent according to the method of
Singleton and Rossi (1965), using gallic acid as a standard. Briefly,
in 10 mL volumetric flask, aliquots of 0.1 mL of extracts sam-
ples (1 mg/mL) and standards for gallic acid were mixed with
2.4 mL of deionized water, 2 mL of 2% sodium carbonate (Na2 CO3 ),
and 0.5 mL of Folin–Ciocalteau reagent and adjusted to 10 mL
with distilled water. After incubation at room temperature for
60 min, the absorbance of the reaction mixtures was measured at
750 nm against deionized water as a blank using spectrophotome-
ter (ULTROSPEC 3100 PRO, Thermo-Scientific). The standard curve
was established with gallic acid solutions (0–200 ␮g/mL), the total
concentration of phenolic compounds in the extract was deter-
mined as mg of gallic acid equivalent (GAE)/ mg of extract weight
(EW) by using this equation (linear regression):
Absorbance = 0.003 × total phenols [␮g GAE/mgof dry extract](R2 = 0.997)
 A. Maadane et al. / Journal of Biotechnology 215 (2015) 13–19 15

Fig. 1. Antioxidant activity of the 9 microalgae extracts (ethanol, ethanol/water (1/1) and water extracts (1 mg/ml)), performed using DPPH radical scavenger assay. BHT and
Vit C are used as positive control at concentration of 100 ␮g/ml. Values represent mean of three independent experiments.

2.6. FAME’s analysis Table 1

DPPH radical scavenger activities of the microalgae ethanolic extracts. IC50 values
were calculated (extracts concentration of 1 mg/ml during 1 h). BHT and Vit C were
The fatty acid methyl esters (FAMEs) profile of the selected used as positive control.
microalgae strains was determined after basic transesterification.
Microalgae strains IC50 (␮g/mL)
The reaction was catalyzed by 1.5% NaOH (w/w) in methanol
1:20 at 80 ◦ C during 5 h. FAMEs profile was characterized by gas Tetraselmis sp. 247 ± 0.01
chromatography (GC) (Agilent 7890A Series GC) coupled to mass Dunaliella Salina 283 ± 0.09
Nannochloropsis gaditana. 365 ± 0.05
spectrometry (MS) equipped with multimode injector and 5 MS
Chlorella sp. 382 ± 0.03
column (30 m × 250 um × 0,25 ␮m) and electron impact ioniza- Dunaliella sp. 348 ± 0.04
tion. Two ␮l of FAMEs solubilized in chloroform were injected at Navicula sp. 347 ± 0.02
chromatography splitless mode using helium at 1.5 mL/min as car- Phaeodactylum tricornutum 432 ± 0.01
Chetoceros sp. 395 ± 0.01
rier gas. The ion source and quadruple temperatures were 230 ◦ C
Isochrisis sp. 464 ± 0.01
and 150 ◦ C respectively. Detection was done using full scan mode BHT 6.2 ± 0.002
between 35 and 500 m/s with gain factor 5 and identification Vit C 2.5 ± 0.007
was performed using NIST 2011 MS Library and known standards Means ± standard deviation of three experiments measurements.
Supelco 37Component FAME Mix (47885-U Sigma–Aldrich). The
oven temperature program was started at 70 ◦ C and maintained
1 min, increased at 20 ◦ C/min until 120 ◦ C, then, held one minute
various degrees. However, the nine tested ethanol extracts dis-
before to be increased until 200 ◦ C by 30 ◦ C/min and held one
played significant higher antioxidant activities than ethanol/water
minute then, increased at 250 ◦ C at 10 ◦ C/min and held one minute,
(1/1) and water extracts. The lower IC50 value means the more pow-
then increased until 270 ◦ C at 5 ◦ C/min and finally kept constant for
erful antioxidant capacity. The highest radical scavenging activity
5 min. FAME composition was calculated as percentage of the total
was obtained with ethanol extracts of Tetraselmis sp, exhibiting
FAME presents in the sample, determined from the peak areas.
the lowest IC50 value (247 ± 0.01 ␮g/mL) (significantly different
at p < 0.05 (one-way ANOVA). Ethanol extracts of Dunaliella salina
2.7. Statistical analysis and Navicula sp. presented a similar antioxidant potential with
the IC50 values of 283 ± 0.09 ␮g/mL and 347 ± 0.02 ␮g/mL, respec-
Three replicates of each sample were used for statistical analy- tively (Table 1). The scavenging effect of standard on the DPPH
sis and the values were reported as value ± the standard deviation. radical decreased in the order of VitC > BHT, with IC50 values of
One-way ANOVA test were applied to evaluate significant differ- 6.3 ± 0.002 ␮g/mL and 2.5 ± 0.007 ␮g/Ml respectively.
ences (P < 0.05). To study the relationship between antioxidant Antioxidant activity was measured at different times (30, 60, and
activity and total carotenoids or total phenolic content, linear 120 min), A recent study reported that the reaction time and DPPH
regression analysis were assessed using SPSS (SPSS 17.0 Inc., USA). concentration influence antioxidant activity and kinetic parame-
ters of bioactive molecules and plant extracts in the reaction with
3. Results and discussion the DPPH radical (Angela et al., 2014; Marinova and Batchvarov,
2011). The kinetics of the reaction is different among extracts.
3.1. DPPH radical scavenging activity In samples with an intermediate and slow kinetic behavior, the
antioxidant activity measured after 2 h of reaction is significantly
Antioxidant activity of extracts was determined by DPPH rad- higher than that measured after 30 min. Results are presented in
ical scavenger assay. DPPH radical scavenging percentages are Fig. 2. The ethanol extractsof three microalgae strains (Tetraselmis
presented in Fig. 1. According to the obtained results; all tested sp., Dunaliella sp. and Nannochloropsis gaditana) exhibited signifi-
microalgae extracts possessed the ability of scavenging DPPH at cant time-dependent radical scavenging activity (p < 0.05, one-way
16 A. Maadane et al. / Journal of Biotechnology 215 (2015) 13–19

Fig. 2. Antioxidant activity of the 9 microalgae extracts, at different time (30, 60, 120 min), performed by DPPH radical scavenger assay. BHT and Vit C are used as positive
control at concentration of 100 ␮g/mL. Values represent mean ± standard deviation of three independent experiments.

Table 2 (EW)), followed by Isochrysis sp.(6.1 ± 1.7 mg/g EW), Phaeodactylum

Carotenoids and phenolic content of ethanol extract of different microalgae strains.
tricornutum (6.3 mg/g EW) and Tetraselmis sp. (4.5 ± 0.9 mg/g EW)
Microalgae Carotenoids content Phenolic content whereas lowest carotenoids contents were found in Navicula sp.
Strains (mg/g DE) (mg GAE/g EW) and Chlorella sp.(0.374 ± 0.41 and 0.3 ± 0.4 mg/g EW), respectively.
Tetraselmis sp. 4.6 ± 0.90 25.5 ± 1.50 These results are similar to the finding of Ahmed et al. (2014) which
Dunaliella salina 3.8 ± 1.30 19.3 ± 0.70 reported carotenoids content of some species extracts from sub-
Dunaliella sp. 10.8 ± 2.02 14.0 ± 0.43 tropical coastal and brackish water such as Tetraselmis sp.(5.8 mg/g
Navicula sp. 0.4 ± 0.04 19.7 ± 0.33
DW), Dunaliella tertiolecta (1.1 mg/g DW) and Isochrysis sp.(5.0 mg/g
Chlorella sp. 0.3 ± 0.04 8.1 ± 0.16
Nannochloropsisgaditana. 3.0 ± 0.24 32.0 ± 0.57 DW). Goiris et al., (2012), reported also similar carotenoids content
Isochrysis sp. 6.1 ± 1.70 13.4 ± 0.16 in Phaeodactylum tricornutum and Nannochloropsis sp. (6.1 mg/g
Phaeodactylumtricornutum 6.3 ± 0.10 16.8 ± 0.33 DW and 2.2 mg/g DW, respectively).
Chaetoceros sp. 1.9 ± 0.2 11.9 ± 0.28

3.3. Phenolic content

ANOVA). No significant difference between the antioxidant activity
of the other microalgae strains extracts at different reaction time
(30, 60, and 120 min). There are a number of reports on antioxi-
dant capacity of some species belonging to the genera of Dunaliella
(Herrero et al., 2006), Navicula (Hemalatha et al., 2013), Isochry-
sis and Tetraselmis (Custódio et al., 2014), Chlorella (Li et al., 2007;
Hajimahmoodi et al., 2009) and Phaeodactylum (Guzmán et al.,
2001; Catarina Guedes et al., 2013).
It is well known that the yield of chemical extraction depends on
the temperature and extraction time, the type of solvents, and the
chemical composition of the sample. The antioxidant compounds
of microalgae could have different polarities, thus the antioxidant
capacity of microalgae are strongly influenced by the extracting
solvent (Li et al., 2007; Goh et al., 2010; López et al., 2011; Ahmed
et al., 2014). Uma et al. (2011) shows that the methanolic extracts
displayed greater potential in different antioxidant assays when
compared to ethanol and acetone extracts of two green microalgae.
3.2. Carotenoids content
Carotenoids play an important role in quenching reactive oxy-
gen species (ROS) generated during photosynthesis, especially
singlet oxygen. In order to investigate the effect of carotenoids on
the antioxidant activity of studied microalgae, total carotenoids
content of ethanolic extract was determined. As shown in
Table 2, ethanol extract of Dunaliella sp. displayed the high-
est total carotenoid content (10.8 ± 2.02 mg/g Extract Weight
The total phenolic content of ethanolic extracts was evaluated,
using the Folin–Ciocalteu method and the results are presented
in Table 2. The phenolic content varied from 8.2 to 32.0 mg gallic
acid equivalent/g of extract weight (mg GAE/g EW). The highest
phenolic content was found in Nannochloropsis gaditana extract
with 32.0 ± 0.5 mg GAE/g EW, followed by Tetraselmis sp. with
25.5 ± 1.5 mg GAE/g EW. The extracts of Dunaliella sp., Phaoedac-
tilum tricornutum and Navicula sp. had also high total phenolic
contents (>15 mg GAE/g), while lowest phenolic content was found
in Chlorella sp. extract (8.1 ± 0.1 mg GAE/g EW). Phenolic contents
of analyzed extracts were higher than those found by Goiris et al.
(2012) in extracts of Isochrysis sp. (4.6 mg GAE/g DW), Chaeto-
ceros calcitrans (1.8 mg GAE/g DW), Nannochloropsis sp. (1.4 mg
GAE/g DW), P. tricornutum (3.8 mg GAE/g DW), and Tetraselmis sp.
(3.8 mg GAE/g DW). To our knowledge, only few reports evaluated
phenolic content of microalgae (López et al., 2011; Goiris et al.,
2012). Moreover, when comparing the obtained data with other
studies described in literature, should takes in consideration that
antioxidant compounds compositions are strongly influenced by
microalgae growth conditions (nutrients availability, temperature,
stress application . . .). Furthermore, the extracting solvents used
affect significantly phenolic content and the ethanol might dissolve
more radical scavenging active polyphenols than other solvents
(López et al., 2011; Goiris et al., 2012). Moreover, the content of
phenolic substances in microalgae increases upon exposure to UV-
light (Duval et al., 2000; Kovácik et al., 2010) suggesting that they
indeed play a role in the antioxidative response to this type of stress.
 A. Maadane et al. / Journal of Biotechnology 215 (2015) 13–19 17

Table 3
FAME profile of microalgae ethanol extracts, in percentages of total FAME. Values represent mean ± standard deviation of three independent experiments.

Fatty acids Tetraslmis sp. Dunaliella Nannochlorpsis Chlorella sp. Dunaliella sp. Isochrysis sp. Phaeodactylium Navicula sp. Chaeotoceros sp.
salina. gaditana. tricornetum

SFA (%)
C14:0 1.7 3.9 5.5 2.3 1.8 6.8 5.8 5.6 8.9
C15:0 0.5 0.8 0.8 0.4 0.1 1.0 0.9 0.3 2.0
C16:0 18.6 12.9 24.1 22.2 25.9 24.7 16.4 17.9 15.2
C17:0 0.3 0.1 0.4 0.4 – – 0.1 – 0.9
C20:0 – 0.9 – 2.5 1.3 – – – –

MFA (%)
C16:1 10.6 4.5 28.0 4.8 2.1 5.2 22.5 27.7 28.4
C18:1 48.8 1.5 15.3 – 22 33.2 6.5 7.8 9.6
C20:1 2.9 – – – 0.5 – – – –

PUFA (%)
C16:2 – 3.1 1.2 – 1.8 1.7 1.2 – 7.7
C16:3 – 2.2 – 17.9 3.9 – 6.5 – 3.2
C16:4 – 8.6 – – 6.0 – 5.5 – –
C17:3 – – 0.4 – – – – – 2.0
C18:2 36.4 – 3.6 10.2 13.9 6.3 3.1 6.5 0.9
C18:3 3.9 45.3 1.5 40.7 16.0 3.7 – 1.3 1.3
C20:3 – – – – – – – – 1.2
C20:4 2.9 14.3 3.6 – 4.8 – 8.6 0.9 2.6
C20:5 9.7 2.7 14.0 – 1.4 16.3 20.1 26.7 14.5
C21:5 1.8 0.1 3.1 – – – 7.4 – 4.5
C22:6 1.03 – – – – 5.9 – 0.8 1.5

3.4. FAMEs analysis and PUFA profiling Table 4

FAME profile of microalgae ethanol extracts, in percentages of total FAME (2). Values
represent mean ± standard deviation of three independent experiments.
Microalgae are an important source of long-chain fatty acids,   
whose a large part is PUFA (Plaza et al., 2009). These compounds are Microalgae PUFA (%) SFA (%) MUFA (%)
known to be good radical scavengers, and could be partially respon- Strains

sible for the antioxidant activity of studied microalgae. Gabriel et al. Tetraselmis sp. 12.9 ± 0.03 22.9 ± 0.01 63.0 ± 0.03
(2007) provide, on a previous study, the cellular evidence for the Dunaliella salina 76.9 ± 0.10 17.9 ± 0.10 4.3 ± 0.00
Dunaliella sp. 38.2 ± 0.08 22.7 ± 0.09 25.3 ± 0.10
antioxidant potential of polyunsaturated fatty acids, namely those
Navicula sp. 30.6 ± 0.09 26.6 ± 0.04 42.9±0.05
of the omega 3 series. Chlorella sp. 68.6 ± 0.10 27.3 ± 0.10 4.6 ± 0.01
FAMEs analysis by GC–MS, after transesterification of all ethanol Nannochloropsis 25.5 ± 0.01 28.8 ± 0.00 43.4 ± 0.01
extracts, was performed and the percentage in PUFA, MUFA gaditana.
(monounsaturated fatty acids) and SFA (saturated fatty acids) Isochrysis sp. 32.2 ± 0.02 27.8 ± 0.00 40.0 ± 0.01
Phaeodactylum 31.1 ± 0.10 29.7 ± 0.02 39.1 ± 0.09
was determined and shown in Table 3. The studied microalgae tricornutum
strains displayed high PUFA content ranging from 12.9 to 76,9 % Chaetoceros sp. 35.5 ± 0.10 25.1 ± 0.20 38.6 ± 0.05
of the total FAMEs. The highest PUFA content was observed in
Dunaliella salina (76.9 ± 0.1%) followed by Chlorella sp. (68.6 ± 0.2%).
Dunaliella sp., Isochrysis sp., Phaeodactylium. tricornutum, Nav-
icula and Chaetoceros sp. had similar PUFA content (30–38%), could be an important source of polyunsaturated fatty acids
while the lowest percentage was found in Tetraselmis sp. extracts. Table 4.
Eicosapentenoic acids (C20:5, EPA) was detected in all studied
microalgae extracts except Chlorella sp.. Linolenic acid (C18:3, 3.5. Correlation between antioxidant activity and
ALA) was detected in Dunaliella salina, (45.3%) Chlorella sp. (40.7 carotenoids/phenolic content
%) and Dunaliella sp. (13.8%). and docosahexaenoic acid (DHA,
C22:6) was detected at low percentage only in Isochrysis sp. (5.8%), The correlation coefficient (R2 ) between the antioxidant capac-
Chaetoceros sp. (1.5 %), Tetraselmis sp. (1.03%), Navicula sp. (0.8%). ity and phenolic or carotenoids content of the ethanol extracts
These results corroborate with the reports previously published of the 9 microalgae strains was determined (Fig. 3). The correla-
in literature. The SFA and PUFA in Dunaliella salina are domi- tion between the antioxidant capacities and the phenolic contents
nant, and the principal identified fatty acids were palmitic acid (R2 = 0.154) and the carotenoids content (R2 = 0.154) were found
(C16:0), linolenic acid (C18:3) and oleic acid (C18:1) (Herrero to be insignificant. These results indicate that the phenolic and
et al., 2006; Hemaltaha et al., 2013). In chlorella sp., according to carotenoids compounds might not be major contributor to the
Mendes et al. (2008) the main fatty acids are oleic acid, palmitic antioxidant capacities of these microalgae. There are few other
acids and linolenic acids. Additionally, other studies reported that studies on the relationship between these two parameters for
Isochrysis galbana presented a highly PUFA percentage (29.2%) microalgae and results were contradictory. Some previous works
(Custódio et al., 2014), Navicula sp. and Phaeodactylium tricornu- demonstrated an insignificant correlation between antioxidant
tum showed a highly lipid content with an important percentage activity and phenolic or carotenoids content. (Li et al., 2007; Goh
of PUFA (Plaza et al., 2009). The FA profile of Nannochlorpsis sp. et al., 2010). However, other reports showed opposite results.
and Chaeotoceros mulleri consisted predominantly of C16:0, C16:1 Goiris et al., (2012) reported that both carotenoid and phenolic
and C20:5 (Mendiola et al., 2007). Tetraselmis sp. had amounts content significantly contributed to the antioxidant capacity of
of C16:0, C18:1 and C18:3 as main fatty acids (Cakmak et al., microalgae. Hajimahmoodi et al., 2009 reported the positive cor-
2014). These results show that the studied microalgae strains relation between antioxidant capacity and phenolic content only
in FRAP (Ferric Reducing Antioxidant Potential) assay and not in
18 A. Maadane et al. / Journal of Biotechnology 215 (2015) 13–19

A) Acknowledgments

100 y = 0.4744x + 73.084 The authors thank MAScIR (Moroccan Foundation for Advanced
R² = 0.1547
DPPH Radical Scavenging Activuty %

90 Science Innovation and research)for the funding of this project.

80
70
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DPPH–HPLC method. It’s well-known that microalgae could pro-
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phenolic compounds, polysaccharides and long-chain polyunsat-
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contribute to the antioxidant activities of microalgae. Further
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ongoing.
4. Conclusion
The ethanolic extracts of all tested microalgae exhibited higher
antioxidant activities. The higher antioxidants potentials were
obtained in Dunalliela sp., Tetraselmis sp. and Nannochloropsis gadi-
tana. These microalgae displayed high total carotenoids content,
total polyphenol content and PUFA content, indicating these strains
could be a potential new source of natural antioxidants. Further
identification of the phenolic components content in active extracts
is needed to investigate possibility to isolate new compounds.
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