Antioxidant activities in tropical marine macroalgae - CINVESTAV ...

J Appl Phycol (2007) 19:449–458
DOI 10.1007/s10811-006-9152-5
Antioxidant activities in tropical marine macroalgae
from the Yucatan Peninsula, Mexico
Mayalen Zubia & Daniel Robledo &
Yolanda Freile-Pelegrin
Received: 19 July 2006 /Accepted: 16 November 2006 / Published online: 27 February 2007
# Springer Science + Business Media B.V. 2007
Abstract Extracts from 48 marine macroalgae species (17
Chlorophyta, 8 Phaeophyta and 23 Rhodophyta) from the
coasts of Yucatan and Quintana Roo (Mexico) were
evaluated for antioxidant activity. The antioxidant activity
was measured with the DPPH (2,2-diphenyl-1-picrylhydrasyl)
method, and the phenolic content of each extract were
also evaluated. All species exhibited a DPPH radical
scavenging activity, and three species (Avrainvillea longicaulis,
Chondria baileyana and Lobophora variegata)
demonstrated great antioxidant potential with very low
oxidation index EC 50 (1.44±0.01, 2.84±0.07 and 0.32±
0.01 mg mL −1 , respectively), significantly equivalent to
EC50 of some commercial antioxidants such as α-tocopherol,
ascorbic acid, BHA and BHT. Moreover, extracts of
the most active species exhibited reducing activities,
superoxide anion radical scavenging and inhibition of lipid
peroxidation. These results suggest that some macroalgae
from the Yucatan peninsula have a great antioxidant
potential which could be considered for future applications
in medicine, food production or cosmetic industry.
Key words antioxidant activity . macroalgae
Introduction
In the past decade, the search for natural antioxidant
compounds has gained considerable attention and the
number of publications on antioxidants and oxidative stress
has nearly quadrupled (Huang et al. 2005). Antioxidant
M. Zubia : D. Robledo : Y. Freile-Pelegrin (*)
Departamento de Recursos del Mar, CINVESTAV Unidad Mérida,
Km 6 Carretera Antigua a Progreso, A.P. 73 Cordemex,
97310 Mérida, Yucatan, México
e-mail: freile@mda.cinvestav.mx
compounds play an important role against various diseases
(e.g., chronic inflammation, atherosclerosis, cancer and
cardiovascular disorders) and ageing processes (Kohen and
Nyska 2002), which explains their considerable commercial
potential in medicine, food production and the cosmetic
industry. Moreover, interest in employing antioxidants from
natural sources is considerably enhanced by consumer
preference for natural products and concern about the
potential toxic effects of synthetic antioxidants (Safer and
al-Nughamish 1999).
Marine algae, like other photosynthesizing plants, are
exposed to a combination of light and oxygen that leads to
the formation of free radicals and other strong oxidizing
agents. However, the absence of oxidative damage in the
structural components of macroalgae (i.e., polyunsaturated
fatty acids) and their stability to oxidation during storage
suggest that their cells have protective antioxidative defense
systems (Fujimoto 1990; Matsukawa et al. 1997). In fact,
algae have protective enzymes (superoxide dismutase,
peroxidase, glutathione reductase, catalase) and antioxidative
molecules (phlorotannins, ascorbic acid, tocopherols,
carotenoids, phospholipids, chlorophyll related compounds,
bromophenols, catechins, mycosporine-like amino acids,
polysaccharides, etc.) which are similar to those of vascular
plants (Fujimoto 1990; Le Tutour et al. 1998; Rupérez et al.
2002; Yuan et al. 2005).
Antioxidative properties of seaweed extracts have been
studied in several geographic regions, but only a few
studies have been performed on tropical seaweed species
(Anggadiredja et al. 1997; Lim et al. 2002; Fallarero et al.
2003; Santoso et al. 2004). Lack of information about the
antioxidant activity of tropical macroalgae is surprising
since these species are expected to develop a very effective
antioxidant defence system due to the strong UV radiation
in the tropical environment. In fact, previous studies have

450 J Appl Phycol (2007) 19:449–458
demonstrated that UV radiation induces the promotion of
antioxidant defense in macroalgae (Aguilera et al. 2002;
Bischof et al. 2002).IntheGulfofMexicoandCaribbean
coast of Yucatan and Quintana Roo, there are numerous species
ofmacroalgaeexposedtohighsolar irradiation and some of
them have never been studied for their antioxidant activities.
The present study evaluated the antioxidative potential
of 48 species of macroalgae from the coasts of Yucatan and
Quintana Roo (Mexico) by measuring the 2,2-diphenyl-1picrylhydrasyl
(DPPH) radical scavenging activity and total
content of phenolic compounds in alcoholic extracts.
Antioxidant activities have been attributed to various
reactions and mechanisms: prevention of chain initiation,
binding of transition metal ion catalysts, reductive capacity,
radical scavenging, etc. (Frankel and Meyer 2000; Huang
et al. 2005). Therefore, in order to better understand the
antioxidant processes involved, the specific antioxidative
activities of the most active extracts were also characterized,
using different biochemical methods: reducing activity,
superoxide anion scavenging activity and inhibition of
lipid peroxidation.
Materials and methods
Collection and preparation of algal extracts
Forty-eight species of macroalgae were collected from the
Gulf of Mexico and Caribbean coast of Yucatan and
Quintana Roo (Fig. 1) between October 2005 and February
2006. Once harvested, macroalgae were stored in plastic
Fig. 1 Map of Yucatan peninsula indicating the collecting sites in
Yucatan (1 Telchac; 2 Dzilam de Bravo) and Quintana Roo (3 Cancún;
4 Playa del Carmen; 5 Tulum) coasts
bags and placed on ice for transport to the laboratory.
Voucher specimens of all species were pressed and stored in
4% formol for identification according to Wynne (2005).
Samples were washed thoroughly with fresh water to
remove salts, sand and epiphytes, and stored at −20°C.
Entire plants of each macroalgae were lyophilized and
milled into powder before extraction. Lyophilized samples
(5 g) were extracted twice with 75 mL of dichloromethanol:
methanol (2:1) during 20 h. Extracts were combined,
filtered and concentrated under reduced pressure to 10 mL,
and stored at −20°C.
Determination of phenolic content
Total content of phenolic compounds of algal extracts was
determined spectrophotometrically using Folin-Ciocalteu
reagent according to the method described in Lim et al.
(2002). First, extracts (0.3 mL) were diluted with methanol
(2.7 mL). Aliquots of the diluted extracts (0.1 mL) were
transferred into the test tubes; 2.9 mL of distilled water
and 0.5 mL of Folin-Ciocalteu reagent were added. After
10 min, 1.5 mL of 20% sodium carbonate solution was
added and the mixture was mixed thoroughly and allowed
to stand at room temperature in the dark for 1 h. Absorbance
was measured at 725 nm and total content of
phenolic compounds was calculated based on a standard
curve of phloroglucinol and expressed in % of dry
weight.
DPPH radical scavenging activity
DPPH (2,2-diphenyl-1-picrylhydrasyl) radical scavenging
activity was determined according to the method of Brand-
Williams et al. (1995). Briefly, 100 μL of each extract at
various dilutions (pure, 0.5, 0.25, etc., depending on the
activity of each extract) were mixed with 3.9 mL of a DPPH
solution (25 mg L −1 ) prepared daily. Due to color intensity
of the extracts, it was necessary to prepare a blank of 100 μL
of each extract at various dilutions added to 3.9 mL of
methanol. The reaction was complete after 2 h in the dark at
room temperature, and the absorbance was read at 515 nm.
DPPH versus absorbance was calculated by linear regression
(n=8; r=0.99): [DPPH]=36.76 (Abs+0.0044). These
values were plotted against percentage of DPPH (% DPPH)
to estimate 50% reduction of its initial value (EC50;
efficient concentration, also called oxidation index). Every
sample was done in triplicate and mean values were
obtained to calculated the EC50. Positive controls such as
ascorbic acid, α-tocopherol, BHT (butylated hydroxytoluene)
and BHA (butylated hydroxyanisole) were also
measured, and the means of the EC50 of each control were
calculated from five measurements.

J Appl Phycol (2007) 19:449–458 451
Reducing activity
Reducing activity of the extracts was evaluated according
to the method of Oyaizu (1986) in Yen and Chen (1995).
This assay measures the total antioxidant capacity of an
extract evaluating the redox potentials of the compounds.
Extract samples (0.5 mL) at two different concentrations
(0.1 and 0.5 mg mL −1 ) were mixed with phosphate buffer
(1.25 mL, 0.2 M, pH 6.6) and potassium ferricyanide (K3Fe
(CN)6; 1.25 mL, 1%) and incubated at 50°C for 20 min,
cooled and mixed with 1.25 mL of trichloroacetic acid
(10%), and 1.25 mL of this mixture was transferred to other
test tubes in which distilled water (1.25 mL) and
FeCl 3.6H 2O (0.25 mL, 0.1%) were added. The mixture
was centrifuged and kept at room temperature for 10 min
before reading the absorbance at 700 nm. Every sample and
positive controls (ascorbic acid, α-tocopherol, BHT and
BHA) were done in triplicate.
Superoxide anion radical scavenging activity
Superoxide anion is one of the most effective free radical,
implicated in cell damage as precursors of mainly reactive
oxygen species, contributing to the pathological process of
many diseases. The superoxide anion scavenging activity of
the extracts was determined using a non enzymatic system
described in Lim et al. (2002). All reagents were prepared
with Tris-HCl buffer (16 mM, pH 8.0). In the test tubes,
1 mL of 234 μM NADH, 1 mL of 150 μM nitroblue
tetrazolium (NBT) and 0.2 mL of the seaweed extracts at
three concentrations (0.1, 0.5 and 1 mg mL −1 ) were mixed
together with 0.8 mL of 37.5 μM phenazine methosulfate
(PMS); after 5 min the absorbance was measured at
560 nm. The same mixture without sample extract was
used as control, and blanks for each extract were prepared
without PMS. Every sample and positive controls (acid
ascorbic, α-tocopherol and BHT) were done in triplicate.
Inhibition of lipid oxidation
Oxidation of unsaturated fatty acids in biological membranes
leads to the formation of lipid radicals and destruction of
lipid membranes. Antioxidants can disrupt the free-radical
chain reaction by donating their hydrogen to fatty acids
radicals to terminate chain reaction. The ferric thiocyanate
(FTC) method of Larrauri et al. (1996) in Sanchez-Moreno
et al. (1999) used a linoleic acid model to evaluate inhibition
of lipid oxidation. Sample extracts (0.5 mL, 0.5 mg mL −1 )
were mixed with an emulsion of 2.51% linoleic acid in
absolute ethanol (0.5 mL), 0.05 M phosphate buffer pH 7
(1 mL) and distilled water (0.5 mL) in a screw capped tube,
shaken and incubated in an oven at 40°C in the dark. The
same mixture without sample extract was used as control. At
different time intervals (0–10 days), 0.1 mL of each tube was
transferred to the other tube where 9.7 mL of 75% ethanol
and 0.1 mL of 30% ammonium thiocyanate together with
0.1 mL of 0.02 M ferrous chloride in 3.5% hydrochloric acid
were added; after 3 min, the absorbance was measured at
500 nm. Every sample and positive controls (α-tocopherol,
BHT and BHA) were done in triplicate.
Statistical analysis
All statistical analyses were performed with Statistica 6
software. Data were tested for normality (Shapiro–Wilk’s
test) and subjected to Bartlett’s test to verify the homogeneity
of variances groups. One-way analysis of variance
(ANOVA) was used to compare antioxidant activity
between extracts after transformation of data if necessary
and post-hoc test (Tukey HSD) was performed when data
showed significant differences (p

452 J Appl Phycol (2007) 19:449–458
Table 1 Phenolic content of the species collected in Quintana Roo 1 and the Yucatan 2 coasts of Mexico
Species Locality Phenolic content (% dry wt)
Chlorophyta
Acetabularia schenckii K. Möbius Cancun 1
Avrainvillea longicaulis (Kützing) G. Murray & Boodle Puerto Morelos 1
Caulerpa ashmeadii Harvey Dzilam de Bravo 2
Caulerpa cupressoides (H. West in Vahl) C. Agardh Dzilam de Bravo 2
Caulerpa paspaloides (Bory de Saint-Vincent) Greville Dzilam de Bravo 2
Caulerpa prolifera (Forsskål) J.V. Lamouroux Dzilam de Bravo 2
Caulerpa sertularioides (S.G. Gmelin) M. Howe Tulum 1
Caulerpa taxifolia (H. West in Vahl) C. Agardh Tulum 1
Cladophora prolifera (Roth) Kützing Playa de Carmen 1
Cladophora vagabunda (Linnaeus) Hoek Cancun 1
Codium decorticatum (Woodward) M. Howe Dzilam de Bravo 2
Enteromorpha intestinalis (Linnaeus) Nees Cancun 1
Halimeda monile (J. Ellis & Solander) J.V. Lamouroux Telchac 2
Halimeda tuna (J. Ellis & Solander) J.V. Lamouroux Telchac 2
Penicillus dumetosus (J.V. Lamouroux) Blainville Telchac 2
Penicillus pyriformis A. Gepp & E.S. Gepp Telchac 2
Udotea conglutinata (J. Ellis & Solander) J.V. Lamouroux Dzilam de Bravo 2
Phaeophyta
Dictyota cervicornis Kützing Dzilam de Bravo 2
Dictyota ciliolata Sonder ex Kützing Tulum 1
Dictyota crenulata J. Agardh Dzilam de Bravo 2
Lobophora variegata (J.V. Lamouroux) Womersley ex E.C. Oliveira Telchac 2
Padina gymnospora (Kützing) Sonder Playa de Carmen 1
Sargassum pteropleuron Grunow Telchac 2
Sargassum ramifolium Kützing Telchac 2
Turbinaria tricostata E.S. Barton Tulum 1
Rhodophyta
Acanthophora spicifera (M. Vahl) Børgesen Telchac 2
Bryothamnion triquetrum (S.G. Gmelin) M. Howe Tulum 1
Ceramium nitens (C. Agardh) J. Agardh Tulum 1
Champia salicornioides Harvey Cancun 1
Chondria atropurpurea Harvey Tulum 1
Chondria baileyana (Montagne) Harvey Cancun 1
Chondrophycus papillosus (C. Agardh) Garbary & J.T. Harper Tulum 1
Chondrophycus poiteaui (J.V. Lamouroux) K.W. Nam Puerto Morelos 1
Digenea simplex (Wulfen) C. Agardh Telchac 2
Eucheuma isiforme (C. Agardh) J. Agardh Telchac 2
Gracilaria bursa-pastoris (S.G. Gmelin) P.C. Silva Telchac 2
Gracilaria caudata J. Agardh Telchac 2
Gracilaria cornea J. Agardh Telchac 2
Gracilaria cylindrica Børgesen Telchac 2
Gracilaria tikvahiae McLachlan Telchac 2
Gracilariopsis tenuifrons (C.J. Bird & E.C. Oliveira) Fredericq & Hommersand Playa de Carmen 1
Halymenia floresii (Clemente & Rubio) C. Agardh Telchac 2
Heterosiphonia gibbesii (Harvey) Falkenberg Playa de Carmen 1
Hypnea spinella (C. Agardh) Kützing Playa de Carmen 1
Laurencia intricata J.V. Lamouroux Dzilam de Bravo 2
Laurencia obtusa (Hudson) J.V. Lamouroux Tulum 1
Liagora ceranoides J.V. Lamouroux Tulum 1
Nemalion helminthoides (Velley) Batters Playa de Carmen 1
0.76±0.05
3.36±0.05
1.42±0.11
4.36±0.31
4.26±0.21
8.13±0.52
6.48±0.17
6.60±0.23
1.95±0.12
1.02±0.08
0.54±0.04
1.42±0.11
0.47±0.00
1.70±0.02
2.24±0.06
1.35±0.16
2.04±0.21
5.55±0.23
5.53±0.09
1.99±0.06
29.18±0.32
5.58±0.30
0.76±0.04
0.95±0.10
1.05±0.08
1.19±0.09
1.55±0.21
1.92±0.07
2.64±0.33
3.21±0.40
7.30±0.29
1.34±0.17
1.77±0.10
0.64±0.08
0.36±0.08
0.38±0.01
0.34±0.03
0.50±0.12
0.41±0.03
0.36±0.02
2.04±0.13
1.03±0.08
3.45±0.36
0.67±0.06
2.51±0.10
4.17±0.12
0.90±0.10
1.87±0.03

J Appl Phycol (2007) 19:449–458 453
Fig. 2 DPPH radical scavenging
activity expressed in oxidation
index EC 50 given in mg
mL −1 (mean ± SD; n=3) for
Chlorophyta extracts and controls
(ascorbic acid, BHA, BHT
and α-tocopherol). Bars represent
standard deviations. Significant
differences are indicated
by different letters as determined
by Tukey HSD test
(p

454 J Appl Phycol (2007) 19:449–458
Fig. 4 DPPH radical scavenging
activity expressed in oxidation
index EC 50 given in mg
mL −1 (mean ± SD; n=3) for
Rhodophyta extracts and controls
(ascorbic acid, BHA, BHT
and α-tocopherol). Bars represent
standard deviations. Significant
differences are indicated
by different letters as determined
by Tukey HSD test
(p

J Appl Phycol (2007) 19:449–458 455
The most active species of Phaeophyta in our study
(Lobophora variegata, Padina gymnospora and Dictyota
cervicornis) belong to the same family, the Dictyotaceae,
which has been extensively studied for its wide variety of
bioactive compounds (e.g., terpenoids and acetogenins) that
conferred a very effective antiherbivore defense system and
various biological activities (Ballantine et al. 1987; Duran
et al. 1997; Amsler and Fairhead 2006). In previous studies,
low antioxidant activities have been measured in other
species of Dictyotaceae (Yan et al. 1998; Santoso et al.
2004; Cavas and Yurdakoc 2005), except in Spatoglossum
pacificum (Matsukawa et al. 1997).
The genus Sargassum also exhibited relatively high DPPH
radical scavenging activities with an oxidation index EC 50
rangingfrom6.64to7.14mgmL −1 . The genus Sargassum has
been studied extensively showing high antioxidant potential
in vitro (Anggadiredja et al. 1997; Matsukawa et al. 1997;
Yan et al. 1998; Lim et al. 2002; Santoso et al. 2004; Heo
et al. 2005; Kimetal.2005; Parketal.2005; Connan et al.
2006) and in vivo (Mori et al. 2003; Wei et al. 2003).
Rhodophyta
All species of Rhodophyta showed antioxidant activities
with EC 50 from 2.84 to 77.71 mg mL −1 (Fig. 4). Chondria
baileyana had the highest antioxidant activity with the
lowest EC50 (2.84±0.07 mg mL −1 ) significantly equivalent
to EC 50 of all commercial antioxidants tested as well as the
highest phenolic content (7.30±0.29% dry wt) (Table 1).
Chondria atropurpurea also exhibited a relatively high
antioxidant activity (10.43±0.11 mg mL −1 ). In fact, the
genus Chondria is a source of various bioactive compounds
(e.g., terpenoids, novel amino-acids, cyclic polysulfides and
indoles) that have shown anthelmintic (Davyt et al. 1998)
and antibiotic activities (Ballantine et al. 1987). Chondria
crassicaulis and C. tenuissima have not shown a high
antioxidant activity (Huang and Wang 2004) when compared
to our results, but they were collected in temperate
environments.
Various species from the order Ceramiales also exhibited
relatively high DPPH radical scavenging activities: Heterosiphonia
gibbesii (8.15±0.10 mg mL −1 ), Acanthophora
spicifera (12.50±0.24 mg mL −1 ), Bryothamnion triquetrum
(12.89±0.51 mg mL −1 ) and Ceramium nitens (13.89±
0.07 mg mL −1 ). Other studies have shown antioxidant
activity in Polysiphonia urceolata and P. morrowii (Fujimoto
1990), Bryothamnion triquetrum (Fallarero et al. 2003),
and Rhodomela confervoides (Huang and Wang 2004). On
the other hand, although the genus Laurencia and
Chondrophycus represent a prolific source of secondary
metabolites (Blunt et al. 2005), their antioxidant activity
was not comparable to other species from the order
Ceramiales. This observation is in accordance with
previous studies where Laurencia species have not shown
high antioxidant activity (Anggadiredja et al. 1997; Yan et al.
1998; Takamatsu et al. 2003; Kim et al. 2005). Gracilaria
species exhibited very low antioxidant activities (28.94 to
72.51 mg mL −1 ), although Yan et al. (1998) reportedaDPPH
radical scavenging activity around 40% for G. verrucosa
methanolic extract, though results are not comparable
because of the different methods used.
No discernable pattern of antioxidant activity could be
detected within a single algal order or division. It is difficult
to compare the antioxidant activity between all the above
species since they were collected at different periods and
localities. In fact, the production of antioxidant compounds,
like phenolics, is influenced by several factors: extrinsic
(herbivory pressure, irradiance, depth, salinity, nutrients,
etc.), and intrinsic (type, age and reproductive stage) (see
review in Connan et al. 2006). Therefore, antioxidant
activity of seaweeds could be subject to great intraspecific
variation, even at very small scales (Connan et al. 2006),
with intra-thallus variations in antioxidant activity having
been shown for Ascophyllum nodosum. Moreover, it is very
difficult to compare macroalgae antioxidant activities with
other studies because each author used different extraction
protocols, assay methods and units. This is why the use of
the oxidation index EC 50 represents a valuable tool for
comparisons.
Antioxidative activities of the most active species
Our study showed a very strong antioxidant potential in three
species of macroalgae: Lobophora variegata (0.32±0.01 mg
mL −1 ), Avrainvillea longicaulis (1.44±0.01 mg mL −1 )and
Chondria baileyana (2.84±0.07 mg mL −1 ). In fact, they are
equivalent to the activities of some commercial antioxidants.
Moreover, reducing activities of the extracts at two concentrations
(0.1 and 0.5 mg mL −1 ) confirmed the results of the
DPPH method: L. variegata had the greatest reducing
activity, and its activity increased as extract concentration
increased (Fig. 5a). The highest levels of reducing activity
were measured for BHA, ascorbic acid and BHT, but the one
from L. variegata is significantly higher than those of
commercial antioxidant α-tocopherol. Most of the published
studies with macroalgae have demonstrated reducing activity
(Karawita et al. 2005; Kudaetal.2005; Yuanetal.2005;
Senevirathne et al. 2006).
Superoxide anion radical O 2 scavenging activity of
the extracts are summarized in Fig. 5b. Highest values were
for ascorbic acid and L. variegata (71.84% and 70.46% for
1 mg mL −1 , respectively) which confirm the strong
antioxidant activity of L. variegata as was the case with
various species of Phaeophyta (Siriwardhana et al. 2003;
Karawita et al. 2005; Kim et al. 2005; Kuda et al. 2005;
Senevirathne et al. 2006), but only Ecklonia cava compares

456 J Appl Phycol (2007) 19:449–458
Fig. 5 Antioxidant activity of
A. longicaulis, C. baileyana and
L. variegata extracts compared
to commercial antioxidants
(ascorbic acid, BHA, BHT and
α-tocopherol). a Reducing activity
(absorbance at 700 nm); b
superoxide anion radical scavenging
activity (%). Bars represent
standard deviations.
Significant differences are indicated
by different letters as
determined by Tukey HSD test
(p

J Appl Phycol (2007) 19:449–458 457
the addition of algal extracts or commercial antioxidants
was accompanied by a rapid increased in absorbance that
reached 1.540 in 10 days (control). The inhibitory effect of
C. baileyana at 0.5 mg mL −1 is equivalent to BHT and
significantly higher than the effects of commercial antioxidants
tested. A. longicaulis also showed inhibitory effect of
lipid peroxidation, significantly higher than BHA and αtocopherol.
In this assay, the antioxidant activity of L.
variegata showed the lowest antioxidant activity; nevertheless,
its inhibitory effect is equivalent to that of α-tocopherol.
These data are in accordance with previous studies that have
demonstrated the inhibition of lipid peroxidation by extracts
from Rhodophyta (Athukorala et al. 2003; Yuanetal.2005),
Phaeophyta (Lim et al. 2002; Siriwardhana et al. 2003;
Karawita et al. 2005; Senevirathne et al. 2006) and
Chlorophyta (Cavas and Yurdakoc 2005). Moreover, these
results suggest that antioxidants from C. baileyana and A.
longicaulis are more effective as chain breaking molecules
rather than reductors, whereas those of L. variegata have
very good reductive capacity, but low chain breaking
capacity. In the future, identification of these molecules will
be helpful to understand the different antioxidant mechanisms
observed in this study.
Conclusion
This work represents the largest screening of antioxidant
activity in tropical macroalgae to date. All species collected
from the coasts of Quintana Roo and Yucatan showed
antioxidant activities, suggesting that tropical macroalgae
develop an effective antioxidant defense system which may
reflect an adaptation to high solar radiation. This screening
emphasized the great antioxidant potential (free-radical,
superoxide anion radical scavenging, reducing activity, and
inhibition of lipid peroxidation) of three species: Lobophora
variegata, Avrainvillea longicaulis and Chondria baileyana.
Identification of the antioxidant compounds of these extracts
will lead to their evaluation in medicine, food production
and cosmetic industry.
Acknowledgments This research was financed by SAGARPA-
CONACYT (Contract 2002-C01-1057). The authors thank J.L. Godinez
for identification of the macroalgae species and C. Chávez and M.L.
Zaldivar for technical assistance.
References
Aguilera J, Bischof K, Karsten U, Hanelt D, Wiencke C (2002)
Seasonal variation in ecophysiological patterns in macroalgae
from an Arctic fjord. II. Pigment accumulation and biochemical
defense systems against light stress. Mar Biol 140:1087–1095
Amsler CD, Fairhead VA (2006) Defensive and sensory chemical
ecology of brown algae. Adv Bot Res 43:1–91
Anggadiredja J, Andyani R, Hayati, Muawanah (1997) Antioxidant
activity of Sargassum polycystum (Phaeophyta) and Laurencia
obtusa (Rhodophyta) from Seribu Islands. J Appl Phycol 9:477–479
Athukorala Y, Lee KW, Song C, Ahn CB, Shin TS, Cha YJ, Shahidi F,
Jeon YJ (2003) Potential antioxidant activity of marine red alga
Grateloupia filicina extracts. J Food Lipids 10:251–265
Ballantine DL, Gerwick WH, Velez SM, Alexander E, Guevara P
(1987) Antibiotic activity of lipid-soluble extracts from Caribbean
marina algae. Hydrobiologia 151/152:463–469
Bischof K, Kräbs G, Wiencke C, Hanelt D (2002) Solar ultraviolet
radiation affects the activity of ribulose-1,5-bisphosphate carboxylase-oxygenase
and the composition of photosynthetic and
xanthophylls cycle pigments in the intertidal green alga Ulva
lactuca L. Planta 215:502–509
Blunt JW, Copp BR, Munro MHG, Northcote PT, Prinsep MR (2005)
Marine natural products. Nat Prod Rep 22:15–61
Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free
radical method to evaluate antioxidant activity. Lebensm-Wiss
U-Technol 28:25–30
Cavas L, Yurdakoc K (2005) A comparative study: assessment of the
antioxidant system in the invasive green alga Caulerpa racemosa
and some macrophytes from the Mediterranean. J Exp Mar Biol
Ecol 321:35–41
Choo KS, Snoeijs P, Pedersen M (2004) Oxidative stress tolerance in
the filamentous green algae Cladophora glomerata and Enteromorpha
ahlneriana. J Exp Mar Biol Ecol 298:111–123
Connan S, Delisle F, Deslandes E, Ar Gall E (2006) Intra-thallus
phlorotannin content and antioxidant activity in Phaeophyceae of
temperate waters. Bot Mar 49:34–46
Davyt D, Entz W, Fernandez R, Mariezcurrena R, Mombru AW,
Saldaña J, Dominguez L, Coll J, Manta E (1998) A new indole
derivative from the red alga Chondria atropurpurea. Isolation,
Structure determination, and anthelmintic activity. J Nat Prod
61:1560–1563
Duran R, Zubia E, Ortega MJ, Salva J (1997) New diterpenoids from
the alga Dictyota dichotoma. Tetrahedron 53:8675–8688
Fallarero A, Loikkanen JJ, Mansito PT, Castañeda O, Vidal A (2003)
Effects of aqueous extracts of Halimeda incrassata (Ellis)
Lamouroux and Bryothamnion triquetrum (S.G. Gmelin) Howe
on hydrogen peroxide and methyl mercury-induced oxidative
stress in GT1-7 mouse hypothalamic immortalized cells. Phytomedicine
10:39–47
Fenical W, Paul VJ (1984) Antimicrobial and cytotoxic terpenoids
from tropical green algae of the family Udoteaceae. Hydrobiologia
116/117:135–140
Frankel EN, Meyer AS (2000) The problems of using one-dimensional
methods to evaluate multifunctional food and biological
antioxidants. J Sci Food Agric 80:1925–1941
Fujimoto K (1990) Antioxidant activity of algal extracts. In: Akatsuka
I (ed) Introduction to applied phycology. SPB Academic
Publishing, The Hague, pp 199–208
Hay ME, Duffy JE, Paul VJ, Renaud PE, Fenical W (1990) Specialist
herbivores reduce their susceptibility to predation by feeding on
the chemically defended seaweed Avrainvillea longicaulis.
Limnol Oceanogr 35:1734–1743
Heo SJ, Park EJ, Lee KW, Jeon YJ (2005) Antioxidant activities of
enzymatic extracts from brown seaweeds. Biores Technol
96:1613–1623
Huang HL, Wang BG (2004) Antioxidant capacity and lipophilic
content of seaweeds collected from the Qingdao coastline. J Agric
Food Chem 52:4993–4997
Huang D, Ou B, Prior L (2005) The chemistry behind antioxidant
capacity assays. J Agric Food Chem 53:1841–1856
Karawita R, Siriwardhana N, Lee KW, Heo MS, Yeo IK, Lee YD,
Jeon YJ (2005) Reactive oxygen species scavenging, metal
chelation, reducing power and lipid peroxidation inhibition

458 J Appl Phycol (2007) 19:449–458
properties of different solvent fractions from Hizikia fusiformis.
Food Res Technol 220:363–371
Kim SJ, Woo S, Yun H, Yum S, Choi E, Do JR, Jo JH, Kim D, Lee S,
Lee TK (2005) Total phenolic contents and biological activities of
Korean seaweed extracts. Food Sci Biotechnol 14:798–802
Kohen R, Nyska A (2002) Oxidation of biological systems: oxidative
stress phenomena, antioxidants, redox reactions, and method for
their quantification. Toxicol Pathol 30:620–650
Kubanek J, Jensen PR, Keifer PA, Sullards MC, Collins DO, Fenical
W (2003) Seaweed resistance to microbial attack: a targeted
chemical defense against marine fungi. Proc Natl Acad Sci USA
100(12):6916–6921
Kuda T, Tsunekawa M, Hishi T, Araki Y (2005) Antioxidant
properties of dried ‘kayamo-nori’, a brown alga Scytosiphon
lomentaria (Scytosiphonales, Phaeophyceae). Food Chem
89:617–622
Le Tutour B, Benslimane F, Gouleau MP, Gouygou JP, Saadan B,
Quemeneur F (1998) Antioxidant and pro-oxidant activities of the
brown algae, Laminaria digitata, Himanthalia elongata, Fucus
vesiculosus, Fucus serratus and Ascophyllum nodosum. J Appl
Phycol 10:121–129
Lim SN, Cheung PCK, Ooi VEC, Ang PO (2002) Evaluation of
antioxidative activity of extracts from a brown seaweed,
Sargassum siliquastrum. J Agric Food Chem 50:3862–3866
Matsukawa R, Dubinsky Z, Kishimoto E, Masaki K, Masuda Y,
Takeuchi T, Chihara M, Yamamoto Y, Niki E, Karube I (1997) A
comparison of screening methods for antioxidant activity in
seaweeds. J Appl Phycol 9:29–35
Mori J, Matsunaga T, Takahashi S, Hasegawa C, Saito H (2003)
Inhibitory activity on lipid peroxidation of extracts from marine
brown alga. Phytother Res 17:549–551
Nakamura T, Nagayama K, Uchida K, Tanaka R (1996) Antioxidant
activity of phlorotannins isolated from the brown alga Eisenia
bicyclis. Fish Sci 62:923–926
Oyaizu M (1986) Studies on products of browning reaction prepared
fromglucoseamine. Jpn J Nutr 44:307–314
Park PJ, Heo SJ, Park EJ, Kim SK, Byun HG, Jeon BT, Jeon YJ
(2005) Reactive oxygen effect of enzymatic extracts from
Sargassum thunbergii. J Agric Food Chem 53:6666–6672
Pavia H, Cervin G, Lindgren A, Åberg P (1997) Effects of UV-B
radiation and simulated herbivory on phlorotannins in the brown
alga Ascophyllum nodosum. Mar Ecol Prog Ser 157:139–146
Ragan MA, Glombitza KW (1986) Phlorotannins, brown algal
polyphenols. In: Round FE, Chapman DJ (eds) Progress in
phycological research. Biopress, Bristol, pp 129–241
Rice-Evans CA, Miller NJ, Paganga G (1997) Antioxidant properties
of phenolic compounds. Trends Plant Sci 2:152–158
Robak J, Gryglewski RJ (1988) Flavonoids are scavengers of
superoxide anions. Biochem Pharmacol 37:837–841
Rupérez P, Ahrazem O, Leal JA (2002) Potential antioxidant capacity
of sulphated polysaccharides from the edible marine brown
seaweed Fucus vesiculosus. J Agric Food Chem 50:840–845
Safer AM, al-Nughamish AJ (1999) Hepatotoxicity induced by the antioxidant
food additive, butylated hydroxytoluene (BHT), in rats: an
electron microscopical study. Histol Histopathol 14:391–406
Sanchez-Moreno C, Larrauri JA, Saura-Calixto F (1999) Free radical
scavenging capacity and inhibition of lipid oxidation of wines,
grape juices and related polyphenolic constituents. Food Res Int
32:407–412
Santoso J, Yoshie-Stark Y, Suzuki T (2004) Anti-oxidant activity of
methanol extracts from Indonesian seaweeds in an oil emulsion
model. Fish Sci 70:183–188
Senevirathne M, Kim SK, Siriwardhana N, Ha JH, Lee KW, Jeon YJ
(2006) Antioxidant potential of Ecklonia cava on reactive
oxygen species scavenging, metal chelating, reducing power
and lipid peroxidation inhibition. Food Sci Tech Int 12:27–38
Siriwardhana N, Lee KW, Kim SH, Ha JH, Jeon YJ (2003)
Antioxidant activity of Hizikia fusiformis on reactive oxygen
species scavenging and lipid peroxidation inhibition. Food Sci
Tech Int 9:339–346
Sun HH, Paul VJ, Fenical W (1983) Avrainvilleol, a brominated diphenylmethane
derivative with feeding deterrent properties from the tropical
green alga Avrainvillea longicaulis. Phytochemistry 22:743–745
Takamatsu S, Hodges TW, Rajbhandari I, Gerwick WH, Hamann MT,
Nagle DG (2003) Marine natural products as novel antioxidant
prototypes. J Nat Prod 66:605–608
Targett NM, Boettcher AA, Targett TE, Vrolijk NH (1995) Tropical marine
herbivore assimilation of phenolic-rich plants. Oecologia 103:170–179
Wei Y, Li Z, Hu Y, Xu Z (2003) Inhibition of mouse liver lipid
peroxidation by high molecular weight phlorotannins from
Sargassum kjellmanianum. J Appl Phycol 15:507–511
Wynne MJ (2005) A checklist of benthic marine algae of the tropical
andsubtropical western Atlantic: second revision. Nova Hewigia
Beih 129:1–152
Yan XJ, Li XC, Zhou CX, Fan X (1996) Prevention of fish oil
rancidity by phlorotannins from Sargassum kjellmanianum. JAppl
Phycol 8:201–203
Yan XJ, Nagata T, Fan X (1998) Antioxidative activities in some
seaweeds. Plant Foods Hum Nutr 52:253–262
Yen GC, Chen HY (1995) Antioxidant activity of various tea
extracts in relation to their antimutagenicity. J Agric Food
Chem 43:27–32
Yuan YV, Bone DE, Carrington MF (2005) Antioxidant activity of
dulse (Palmaria palmata) extract evaluated in vitro. Food Chem
91:485–494
Zhang P, Omaye ST (2001) Antioxidant and prooxidant roles for βcarotene,
α-tocopherol and ascorbic acid in human lung cells.
Toxicol In Vitro 15:13–24