Algal Research 66 (2022) 102798

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Algal Research
journal homepage: www.elsevier.com/locate/algal

Changes in antioxidant activity of fresh marine macroalgae from the Canary

Islands during air-drying process
Marcos Adrián Ruiz-Medina *, Marta Sansón, Águeda María González-Rodríguez
Departamento de Botánica, Ecología y Fisiología Vegetal, Universidad de La Laguna, 38200 Santa Cruz de Tenerife, Canary Islands, Spain

A R T I C L E I N F O A B S T R A C T

Keywords: Marine macroalgae develop structurally distinct molecules that have been long recognized as an important
Bioactive compounds source of natural products, e.g., bioactive compounds and antioxidants, such as carotenoids, amino acids, pro­
DPPH teins, lipids, vitamins and polyphenols, with promising properties as anticoagulant, anti-proliferative and anti­
Ferrous iron-chelating
microbial. In this study we analyzed the antioxidant activity of 24 fresh methanolic extracts from marine
Flavonoids
Phenolic compounds
macroalgae from the Canary Islands. Additionally, the effects of air-drying process in the antioxidant activity
Phytoconstituent were also evaluated. Plant material was collected in three locations on the island of Tenerife. Total antioxidant
activity methods included DPPH-free radical scavenging activity and ferrous iron chelating activity. Phytocon­
stituents evaluated were total carotenoids, proline, phenols, flavonoids and condensed tannins content. Our
results indicated that from fresh material, species Cladophora liebetruthii, Dasycladus vermicularis and Dictyopteris
polypodioides presented the highest scavenging activity, supported by high correlation with phenolics and fla­
vonoids content, however, in air-dry extracts, Anadyomene saldanhae and D. polypodioides showed the highest
antioxidant potential, correlated with a high phenolic compounds content. Asteronema breviarticulatum and
D. polypodioides presented a high content of carotenoids in fresh and air-dry state. The highest ferrous iron
chelating activity (>60 %) was recorded in extracts from fresh material in Grateloupia imbricata and Lophocladia
trichoclados. However, only L. trichoclados was able to maintain this high activity after air-drying supported by a
high proline content. A. breviarticulatum was the species that showed the highest condensed tannins content both
in fresh and dried extracts. The study revealed that fresh extracts of D. vermicularis, D. polypodioides and
L. trichoclados possess promising properties as raw materials to obtain biologically active substances in
alimentary and pharmaceutical industry. Besides, these properties were maintained after the air-drying period in
the last two species, which makes them species of great interest to be used under drying preservation methods.

1. Introduction
Oxidative stress is an unavoidable consequence of life in an oxygen-
rich atmosphere. The human body maintains a constant oxide-reduction
balance, preserving the balance between the production of substances
that induce oxidative stress and the antioxidant defense systems [1]. The
loss in this oxide-reduction balance leads to a state of oxidative stress
that is characterized by an increase in the levels of free radicals (FRs)
and reactive species, which is not compensated by the antioxidant de­
fense systems, causing damage and cell death [2]. Reactive species
include oxygen species (ROS) and there are many pathological processes
reasonably attributable to ROS attack, some of the most significant are:
aging and cell death, cancer, mutations and heart, muscular, pulmonary
and liver diseases, among others [3–5]. Fortunately, the negative effects
of oxidative stress may be mitigated by antioxidants [4]. Marine or­
ganisms, including macroalgae, have attracted the attention of many
researchers as a source of bioactive compounds and antioxidants due to
their properties, the diversity of their molecules, and their new chemical
structures that are complex and difficult to synthesize chemically [6].
Many studies have already demonstrated the promising properties of
marine macroalgae extracts, being an excellent source of bioactive
compounds such as carotenoids, amino acids, proteins, lipids, pigments,
vitamins and polyphenols, among other substances [7,8]. Carotenoids
are largely distributed in lipoproteins and membranes, usually together
with vitamin E [9]. Main antioxidant capabilities of carotenoids include
pro-vitamin A activity, immunological, endocrine and metabolic

Abbreviations: dry wt, dry weight; TPC, total phenolic content; TFC, total flavonoid content; CTC, condensed tannin content; FIC, ferrous iron-chelating.
* Corresponding author.
E-mail addresses: alu0101051642@ull.edu.es (M.A. Ruiz-Medina), msanson@ull.edu.es (M. Sansón), aglerod@ull.edu.es (Á.M. González-Rodríguez).

https://doi.org/10.1016/j.algal.2022.102798
Received 2 February 2022; Received in revised form 16 July 2022; Accepted 17 July 2022
Available online 31 July 2022
2211-9264/© 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
M.A. Ruiz-Medina et al.
activities [10,11]. In addition, the amino acid proline, an essential
osmolyte in the management of water stress in plants, has pharmaco­
logical characteristics that are still unknown [12]. It is currently known
that proline is involved in collagen synthesis, as well as the preservation
of joints and the prevention of arthritis, the improvement of tissue and
mucous membrane healing, the prevention or improvement of diseases
like ulcers, cardiovascular disorders and the maintenance of skin, bone
and muscle tissue [12,13].
Even though in nature there are multiple compounds with antioxi­
dant activity, phenolic compounds are probably the most studied. They
have also been shown to have potential as antidiabetics, anticancer,
anti-inflammatory, antihypertensive, anticoagulant, antimicrobial and
antiallergic, which gives them great value in the alimentary and phar­
maceutical industry [14]. Furthermore, within this large group are fla­
vonoids, which are a particular type of polyphenols present in plants
that are also well known [15]. The growing interest in flavonoids is due
to the appreciation of their broad pharmacological activity. Due to this
fact, protective effects have been described in pathologies such as can­
cer, heart diseases, viral infections, stomach and duodenal ulcer and
inflammations [16]. Flavonoids include condensed tannins, which are
known by their usefulness in the treatment of allergies, cancers, car­
diovascular disorders, and platelet aggregation [17].
Macroalgae are commonly classified into three higher taxa, brown
(Class Phaeophyceae), red (Phylum Rhodophyta) and green (Phylum
Chlorophyta) [18]. According to Guiry & Guiry [19] about 2092 species
of brown algae, 6984 of green algae and 7470 of red algae are worldwide
currently recognized. All these organisms, like other photosynthetic
plants, are exposed to a combination of light and oxygen that leads to the
formation of FRs and other strong oxidizing agents, besides, some of
them live in complex habitats exposed to extreme conditions (high
salinity, low or high temperature, high pressure, low availability of
nutrients, and low or high exposure to sunlight) that further increase
oxidative stress [20]. In addition, the absence of structural and photo­
dynamic induced damage suggests that algae possess mechanisms for a
rapid adaptation, producing secondary metabolites, which protect them
against effects of oxidative stress in unfavorable periods [21,22].
An immense algal biodiversity develops on the Canary Islands,
currently >600 species are recognized [23,24]. Many of these species
have a ubiquitous distribution and therefore have already been studied
to find compounds with biological activity, for example, ethyl acetate
extract of Ulva rigida presented a high concentration of total phenols
while U. lactuca presented also a high total phenolic content [25,26].
Other species that have shown high antioxidant, anti-inflammatory and
antiproliferative activity have been those of the Cystoseira s.l. [27].
However, many other species are distributed regionally and their
morphological and physiological characteristics (small size,
morphology, growth habits) make their collection difficult and, there­
fore, have led to few or incomplete studies on their biological potential.
Fresh algae collected or cultivated from the sea are usually dried
before being used in any nutritional evaluation or industrial processing
as they originally consist of 70–90 % water [28]. This large amount of
moisture irretrievably leads to rapid microbial decomposition and the
destruction of biologically active substances [29]. The best procedure to
preserve the flavor, texture, and nutritional value of macroalgae after
harvesting is freezing because considerably reduces the physical and
biochemical changes involved in molecules degradation, as well as the
development and reproduction of spoilage microbes [30,31]. However,
when large warehouses with freezers are not available, air-drying could
be a preservation method. Drying is the traditional algae preservation
procedure and could be an important factor affecting the nutritional
value of species either through chemical modifications or direct losses of
the nutrients [32]. Exposure to the atmosphere places a stress on marine
macroalgae when their tissues dry out. Previous studies have described
that prolonged period of drying induce oxidative stress through ROS
overproduction, alterations in cell membranes and protein denaturation,
disruption of the life cycle and finally cell death [33,34]. However, there
2
Algal Research 66 (2022) 102798
are many physiological mechanisms that can be activated to attenuate
oxidative stress, for example, increasing the antioxidant response or
accumulating compatible solutes or osmolytes that help avoid osmotic
stress [35,36].
Even though marine macroalgae are known to contain a wide variety
of secondary compounds, comprehensive chemical analysis, and the
identification of bioactive components in many species, as well as the
effects of drying procedure on the antioxidant properties, remain un­
known [37,38]. Therefore, the objectives of this study were (1) to
evaluate the antioxidant activity of methanolic extracts from 24 species
of marine macroalgae from the island of Tenerife (Canary Islands) and
(2) to analyze the effects of air-drying as a processing and storage
method on this activity. Total antioxidant activity methods included
DPPH-free radical scavenging activity and ferrous iron chelating activ­
ity. Phytoconstituents evaluated were total carotenoids, proline, phe­
nols, flavonoids and condensed tannins content.
2. Material and methods
2.1. Study area and macroalgae sampling
In Canary Islands, subtropical waters are characterized by a quasi-
permanent thermocline caused by the strong surface heating
throughout the year. The thermocline is only eroded from January to
March, allowing cold nutrient-rich waters to enter the euphotic zone and
thereby increasing primary productivity during the so-called “late
winter bloom” [39]. Besides, the islands are located at the outer edge of
the intertropical convergence zone, so they get a quantity of Saharan
dust, which can fertilize the water column with phosphorus and iron
(1–3 nM), among other trace metals [40,41]. In Tenerife, sea surface
temperature annually ranges from 17 ◦ C (February) to 24 ◦ C
(September) [42]. In this study, 24 species of macroalgae (3 Phaeo­
phyceae, 7 Chlorophyta and 14 Rhodophyta) belonging to 10 orders
were collected between October–November 2021 from three sites
located on the north of Tenerife (Table 1): 15 species in Punta del Hi­
dalgo (28◦ 34′ 15.57′′ N, 16◦ 19.59′ 83′′ O), 4 in Punta Brava
(28◦ 24′ 45.08′′ N, 16◦ 33.57′ 68′′ O) and 5 in El Penitente (28◦ 25′ 07.30′′
N, 16◦ 32′ 55.08′′ O) (Fig. 1).
2.2. Plant material processing
Samples, 10 g per individual (3 individuals per species) were washed
to remove sand, salts and cleaned off epiphytes. To analyze antioxidant
activity in fresh, half of the cleaned material were immediately frozen
and stored at − 80 ◦ C. The other half was subjected to a drying treatment,
which consisted in air-dry on filter paper during one week in a darkness
chamber with constant temperature (19 ± 2 ◦ C) and air humidity con­
ditions (80 ± 5 %), measured with a thermo-hygrometer (EMS33, EMS,
Brno, CZ). After one week, the samples were frozen and stored at
− 80 ◦ C. The frozen material was lyophilized, ground until obtaining a
fine powder and finally stored at − 20 ◦ C until later assays.
2.3. Preparation of algal extracts for antioxidant activity and phenolic
compounds determination
To execute radical DPPH scavenging activity and ferrous iron
chelating activity assays, in addition to determining the content of total
phenolics, flavonoids and condensed tannins, the same extraction was
performed following Xu and Chang [43] protocol with slight modifica­
tions. 0.015 g of dry powder was suspended in 2 mL of solvent [80:20
methanol/water (80 % methanol)] and vortexed vigorously during 30 s.
Subsequently, it was kept stirring for 1,5 h at 400 rpm and then allowed
to stand over the night. Next day the supernatant was collected in a test
tube that was reserved. The pellet was resuspended again in another 2
mL of solvent and vortexed vigorously during 30 s. Finally, both su­
pernatants were unified, filtered with a syringe through a 0.45 μm
M.A. Ruiz-Medina et al. Algal Research 66 (2022) 102798

Table 1 according to the equation:

Taxonomic classification and collection sites of the 24 marine macroalgal [ ( / )]
species. Percentage of discoloration = 1 − Asample Acontrol × 100

Name of the species and authority Taxonomy Collection The FR scavenging activity of the extracts was expressed as equiva­
site lent to that of Trolox. Results were calculated and expressed as milli­
Anadyomene saldanhae A.B.Joly & E.C. Ch, Cladophorales, PH grams of Trolox equivalents (TE) per gram of dry weight using the
Oliveira Anadyomenaceae Trolox calibration curve. The linearity range of the calibration curve was
Asteronema breviarticulatum (J.Agardh) Ph, Scytothamnales, PE
50 to 350 μg/mL (r = 0.99).
Ouriques & Bouzon Asteronemataceae
Canistrocarpus cervicornis (Kützing) De Ph, Dictyotales, PH
Paula & De Clerck Dictyotaceae 2.5. Ferrous ion-chelating activity assay
Caulerpa webbiana Montagne Ch, Bryopsidales, PH
Caulerpaceae
The ferrous iron-chelating (FIC) activity assay was adapted from a
Cladophora albida (Nees) Kutzing Ch, Cladophorales, PH
Cladophoraceae method described by Gülçin et al. [45] with Chew et al. [46] modifi­
Cladophora liebetruthii Grunow Ch, Cladophorales, PH cations. A reaction mixture was prepared by mixing 0.1 mM ferrous
Cladophoraceae sulphate, 0.25 mM FerroZine and macroalgae extract in 1:1:1 propor­
Codium intertextum Collins & Hervey Ch, Bryopsidales, PH tion. The reaction mixture was allowed to stand for 10 min before the
Codiaceae
absorbance measurements were taken at 562 nm using a UV–visible
Dasycladus vermicularis (Scopoli) Ch, Dasycladales, PH
Krasser Dasycladaceae spectrophotometer (UV-160 A, Shimadzu). The ferrous ion-chelating
Dictyopteris polypodioides (A.P.De Ph, Dictyotales, PH property of extracts was calculated as a percentage using the
Candolle) J.V.Lamouroux Dictyotaceae following formula:
Galaxaura rugosa (J.Ellis & Solander) J. Rh, Nemaliales, PH [( )/ ]
V.Lamouroux Galaxauraceae Chelating activity (%) = Acontrol − Asample Acontrol × 100
Grateloupia imbricata Holmes Rh, Halymeniales, PB
Halymeniaceae
where Acontrol and Asample are the absorbance of the control and extract,
Heterodasya mucronata (Harvey) M.J. Rh, Ceramiales, PH
Wynne Rhodomelaceae respectively. The negative control contained 80 % methanol, ferrous
Hypnea spinella (C.Agardh) Kützing Rh, Gigartinales, PH sulphate and FerroZine in 1:1:1 proportion. EDTA was used as positive
Cystocloniaceae control at same the concentration of extracts.
Hypnea sp. Rh, Gigartinales, PB
Cystocloniaceae
Jania pedunculata var. adhaerens (J.V. Rh, Corallinales, PE 2.6. Determination of total phenolic content
Lamouroux) A.S.Harvey, Woelkerling Corallinaceae
& Reviers The total phenolic content (TPC) was determined by a Folin-
Jania virgata (Zanardini) Montagne Rh, Corallinales, PE Ciocalteu assay [47,48] using gallic acid (GA) as the standard. The
Corallinaceae
Laurencia dendroidea J.Agardh Rh, Ceramiales, PE
mixture of the sample solution (50 μL), distilled water (1.5 mL), 250 μL
Rhodomelaceae of 2 N Folin-Ciocalteu's reagents solution, and 7 % sodium carbonate
Laurencia pyramidalis Bory ex Kützing Rh, Ceramiales, PB (750 μL) was vortexed and incubated for 8 min at room temperature.
Rhodomelaceae Then, a dose of 950 μL of distilled water was added. The mixture was
Lophocladia trichoclados (C.Agardh) F. Rh, Ceramiales, PH
allowed to stand for 2 h at room temperature. Finally, the absorbance
Schmitz Rhodomelaceae
Palisada perforata (Bory) K.W.Nam Rh, Ceramiales, PH was measured immediately against the blank (distilled water) at 765 nm
Rhodomelaceae using a UV–visible spectrophotometer (UV-160 A, Shimadzu) and the
Rytiphlaea tinctoria (Clemente) C. Rh, Ceramiales, PH results were calculated and expressed as milligrams of GA equivalents
Agardh Rhodomelaceae (mg GAE/g dry wt) using the GA calibration curve (10–400 μg/mL; r =
Spyridia filamentosa (Wulfen) Harvey Rh, Ceramiales, PB
Callithamniaceae
0.99).
Valonia utricularis (Roth) C.Agardh Ch, Cladophorales, PE
Valoniaceae 2.7. Determination of total flavonoid content
Wrangelia argus (Montagne) Montagne Rh, Ceramiales, PH
Wrangeliaceae
The total flavonoid content (TFC) was determined by a previously
The taxonomic abbreviations Rh, Ph and Ch represent Phylum Rhodophyta, described colorimetric method with slight modifications [49]. Briefly, to
Class Phaeophyceae and Phylum Chlorophyta, respectively. The locality ab­ 250 μL of extract or standard solution of (+)-catechin was added 1.25
breviations PH, PB and PE represent Punta del Hidalgo, Punta Brava y El Pen­ mL of distilled water in a test tube, followed by the addition of 75 μL of a
itente, respectively.
5 % sodium nitrite solution. After 6 min rest, 150 μL of a 10 % aluminum
chloride solution were added and allowed to stand for another 5 min
Millipore filter, and stored at − 20 ◦ C in the dark until further analysis. before adding 0.5 mL of 1 M sodium hydroxide. The mixture was made
The extractions were performed in triplicate. up to 2.5 mL with distilled water and mixed well. The absorbance was
measured immediately against the blank (the same mixture without the
2.4. DPPH free radical scavenging activity assay sample) at 510 nm using a UV–visible spectrophotometer (UV-160 A,
Shimadzu). Results were calculated and expressed as micrograms of
DPPH-free radical scavenging capacity of leaf extracts was evaluated (+)-catechin equivalents (mg CAE/g dry wt) using the (+)-catechin
according to the method of Chen and Ho [44] with slight modifications. calibration curve. The linearity range of the calibration curve was 10 to
Briefly, 0.2 mL of the extract was added to 3.8 mL of 0.25 mM ethanolic 400 μg/mL (r = 0.99).
DPPH solution. The mixture was vigorously stirred for 1 min and
allowed to stand at room temperature in the dark for 30 min. Next, the 2.8. Determination of condensed tannin content
absorbance of the sample plus DPPH solution (Asample) was measured
using the UV–visible spectrophotometer (UV-160 A, Shimadzu) at 517 Analysis of condensed tannin content (CTC) was carried out ac­
nm against an ethanol blank. A negative control (Acontrol) was taken after cording to the method of Broadhurst and Jones [50]. To 100 μL of the
adding DPPH solution to 0.2 mL of the solvent (80 % methanol). The sample extract, 2 mL of a 4 % vanillin methanol solution and 1 mL of
percentage of discoloration by DPPH of the sample was calculated concentrated hydrochloric acid (12 M) were added. The mixture was

3
M.A. Ruiz-Medina et al.
Fig. 1. Geographical location of the Canary Islands and collection sites in Tenerife Island: Punta Brava, El Penitente and Punta del Hidalgo.
vortexed during 5 s and left to stand for 15 min, and the absorption was
measured using a UV–visible spectrophotometer (UV-160 A, Shimadzu)
at 500 nm against 80 % methanol as a blank. CTC was calculated and
expressed as mg catechin equivalents (mg of CAE/g dry wt) using the
calibration curve of (+)-catechin. Linearity range of the calibration
curve was 10 to 400 μg/mL (r = 0.99).
2.9. Determination of total carotenoids content
For carotenoids determination, 0.01 g of dry powder were extracted
with 2 mL 100 % acetone saturated with calcium carbonate in order to
avoid acid traces that modify the pigment composition. They were
centrifuged for 10 min at 4 ◦ C and syringe-filtered through 0.45 μm
Millipore filter. Total carotenoids were quantified with a double beam
spectrophotometer (UV-160 A, Shimadzu) following the method pro­
posed by Lichtenthaler [51], for which it is necessary to calculate the
concentration of chlorophylls a and b. The equations for calculating total
carotenoids are shown below:
Total carotenoids = [(1000 × A470 ) − (1.9 × Ca ) − (63.14 × Cb ) ]/214
Chlorophyll a (Ca ) = (11.24 × A661.5 ) − (2.04 × A644.5 )
Chlorophyll b (Cb ) = (20.13 × A644.5 ) − (4.19 × A661.5 )
Results are expressed as mg/g dry weight (dry wt). The extraction
was conducted in triplicate.
2.10. Determination of total proline content
The colorimetric analysis of proline was carried out according to
Bates et al. [52] based on proline's reaction with ninhydrin. Dry powder
(0.03 g per sample) was extracted with 2 mL 3 % sulfosalicylic acid and
centrifuged 5 min at 1400 rpm. For determinations, 1:1:1 solution of
supernatant, ninhydrin acid and glacial acetic acid was incubated at
100 ◦ C for 1 h. The content was poured into a tube with 2 mL of toluene
to extract the chromophore, then vortexed 1 min and centrifuged 5 min
at 1000 rpm. Finally, the chromophore was placed in the upper layer
and its absorbance at 520 nm was determined in a UV–visible spectro­
photometer (UV-160 A, Shimadzu). The extraction was conducted in
triplicate. The method was calibrated for each determination with
standard L-proline solutions within the detection range of the method
(1–300 μM; r = 0.99).
2.11. Statistical analysis
Statistical analyses were performed using SPSS 25.0 software pack­
age (SPSS, Inc., Chicago, IL, USA). Normality and homoscedasticity of
4
Algal Research 66 (2022) 102798
samples distribution was assessed using Shapiro Wilk and Levene's test,
respectively. Differences between fresh and dried macroalgae extracts
were analyzed by Student t-test. Duncan's multiple range test was car­
ried out to test any significant differences among species, while Pearson
correlation test was conducted to determine the correlation coefficient
(r) among quantitative variables. Significant levels were defined using p
≤ 0.05.
3. Results
3.1. Total antioxidant activity
Antioxidant activity of marine macroalgae extracts is summarized in
Table 2. In eight species, the radical scavenging activity increased
significantly after air-drying, for example in A. saldanhae, H. spinella and
J. pedunculata var. adhaerens, whose dried samples (23.12 ± 0.64, 16.43
± 0.73 and 15.16 ± 0.49 mg TE/g dry wt, respectively) were up to nine
times higher than the fresh samples (8.12 ± 0.30, 1.86 ± 0.08 and 1.82
± 0.04 mg TE/g dry wt, respectively). However, two species
(C. liebetruthii and D. vermicularis) showed an opposite effect, with a
radical scavenging activity in fresh extract significantly higher (29.15 ±
0.75 and 58.71 ± 0.81 mg TE/g dry wt) than in dry extract (10.37 ±
0.87 and 21.53 ± 0.72 mg TE/g dry wt, respectively).
Significant differences were found among species. In extracts made
from fresh material, D. vermicularis presented the highest activity (58.71
mg TE/g dry wt), which translated into a discoloration percentage of 47
%, significantly higher than the rest of the species. It was followed by
C. liebetruthii and D. polypodioides which presented a discoloration
around 23 %. In the dry extracts, the highest values were recorded in
D. polypodioides (29.92 ± 0.55 mg TE/g dry wt) and A. saldanhae (23.12
± 0.64 mg TE/g dry wt), significantly different from each other but
higher than the rest of the species.
The FIC activity of the analyzed extracts is shown in Fig. 2. FIC ac­
tivity decreased significantly after the drying period in 50 % of the
macroalgae studied. This was revealed in C. cervicornis and G. imbricata
whose activity in fresh (59.22 ± 0.45 and 73.81 ± 0.67 %, respectively)
was significantly higher than in dry (31.96 ± 0.72 and 30.56 ± 0.68 %,
respectively). However, in 5 species this activity was favored after water
loss, for example in W. argus, whose values were 55.11 ± 0.55 % and
29.22 ± 0.45 % in dry and fresh, respectively. After a week of air-drying,
only J. virgata lost all the FIC activity. Significant differences were also
found between species, the highest values compared to the control
(EDTA, 92.70 ± 0.74 %) were recorded in G. imbricata in fresh (73.81 %)
and in L. trichoclados in fresh (67.89 %) and dry (74.48 %). In general, all
species presented high FIC activity values, being in 5 species >50 %
(C. cervicornis, D. vermicularis, A. breviarticulatum, L. trichoclados and
G. imbricata).
M.A. Ruiz-Medina et al. Algal Research 66 (2022) 102798

Table 2 significantly higher than the rest of the species. Finally, A. saldanhae and
DPPH-free radical scavenging activity from 24 marine macroalgae extracts R. tinctoria also presented a high content, about 21 mg GAE/g dry wt.
(fresh and after a week of air-drying). Data are expressed as mean ± standard Additionally, the TFC decreased after the drying period in half of the
error (n = 3). studied species (Table 3), being the only exception R. tinctoria that
Plant name DPPH (mg TE/g dry wt) DPPH (discoloration %) showed a value of 5.42 ± 0.26 mg CAE/g dry wt in fresh condition and
Fresh Dried algae Fresh Dried algae 9.10 ± 0.24 mg CAE/g dry wt in dry condition. The decrease in flavo­
algae algae noid content after the drying period was significant in 54 % of the
Heterodasya mucronata 1.79 ± 2.21 ± 0.00 ± 0.35 ±
species, being in some cases up to 5 times lower, such as in P. perforata,
0.00A 0.15AB 0.00A 0.13AB whose TFC was 4.15 ± 0.17 mg CAE/g dry wt in fresh state and 0.74 ±
Lophocladia 1.82 ± 7.43 ± 0.03 ± 4.70 ± 0.12 mg CAE/g dry wt after drying. Among species, D. vermicularis
trichoclados 0.04A 0.95*C 0.03A 0.79*C (16.23 ± 0.18 mg CAE/g dry wt) exhibited the highest value in the
Jania pedunculata var. 1.82 ± 15.16 ± 0.03 ± 11.14 ±
extracts obtained from fresh material, which was significantly higher
adhaerens 0.04A 0.49*E 0.03A 0.41*E
Wrangelia argus 1.82 ± 3.64 ± 0.03 ± 1.55 ± than the rest of the species. The following were A. saldanhae and
0.04A 0.71AB 0.03A 0.59AB D. polypodioides, with values close to 10 mg CAE/g dry wt. After drying,
Hypnea spinella 1.86 ± 16.43 ± 0.06 ± 12.21 ± the highest values corresponded to A. saldanhae (9.42 ± 0.28 mg CAE/g
0.08A 0.73*E 0.06A 0.61*E dry wt) and R. tinctoria, which were significantly higher than the rest of
Valonia utricularis 1.86 ± 1.90 ± 0.06 ± 0.10 ±
0.08A 0.07AB 0.06A 0.06AB
the species.
Cladophora albida 1.90 ± 1.90 ± 0.10 ± 0.10 ± Within the flavonoids, CTC showed greater sensitivity to drying time
0.12A 0.12AB 0.10A 0.10AB (Table 3), since in most species their concentration decreased signifi­
Codium intertextum 1.90 ± 1.86 ± 0.10 ± 0.06 ± cantly after a week of air drying, even zero values were recorded, as for
0.12A 0.04AB 0.10A 0.03AB
example in S. filamentosa and G. imbricata, which initially had values of
Spyridia filamentosa 1.90 ± 6.89 ± 0.10 ± 4.25 ±
0.12A 0.61*C 0.10A 0.51*C 3.56 and 1.26 mg CAE/g dry wt, respectively. Among species, the
Galaxaura rugosa 1.94 ± 1.86 ± 0.13 ± 0.06 ± highest concentration of condensed tannins was presented by
0.10A 0.04AB 0.09A 0.03AB A. breviarticulatum (20.96 ± 0.71 mg CAE/g dry wt), significantly higher
Laurencia pyramidalis 1.94 ± 3.52 ± 0.13 ± 1.45 ± than the other species. C. cervicornis, D. polypodioides and C. webbiana
0.10A 0.57AB 0.09A 0.48AB
Palisada perforata 1.94 ± 1.82 ± 0.13 ± 0.03 ±
showed the next highest values from fresh material, with a concentra­
0.10A 0.04A 0.09A 0.03A tion around 12 mg CAE/g dry wt. The same trend was observed from
Hypnea sp. 1.98 ± 2.87 ± 0.16 ± 0.90 ± dried material.
0.10A 0.20*AB 0.09A 0.17*AB
Grateloupia imbricata 2.09 ± 3.06 ± 0.26 ± 1.06 ±
3.3. Total carotenoids and proline content
0.17A 0.35AB 0.14A 0.30AB
Canistrocarpus 2.13 ± 5.88 ± 0.29 ± 3.41 ±
cervicornis 0.18A 0.76*C 0.15A 0.63*C Significant differences in the total carotenoids content were found
Caulerpa webbiana 2.13 ± 2.37 ± 0.29 ± 0.48 ± between fresh and dry samples in all species (Fig. 3), being higher in
0.20A 0.13AB 0.17A 0.11AB fresh than in dry material. In some species carotenoids decreased
Laurencia dendroidea 2.52 ± 3.10 ± 0.61 ± 1.10 ±
0.22A 0.27AB 0.18A 0.23AB
significantly, up to five or ten times the content in the extracts from fresh
Jania virgata 2.91 ± 2.02 ± 0.93 ± 0.19 ± macroalgae, which was the case of the Hypnea spp. and S. filamentosa. In
0.41A 0.13AB 0.34A 0.11AB the rest of the species, the total carotenoid values tended to decrease up
Rytiphlaea tinctoria 4.45 ± 18.83 ± 2.22 ± 14.20 ± to 50 % after drying, with respect to the values in fresh. Among the
0.40B 0.85*F 0.33B 0.71*F
species, the highest values were presented in fresh material of
Anadyomene saldanhae 8.12 ± 23.12 ± 5.28 ± 17.78 ±
0.30C 0.64*H 0.25C 0.53*H A. breviarticulatum (1.09 ± 0.00 mg/g dry wt), H. spinella (1.15 ± 0.00
Asteronema 17.59 ± 20.22 ± 13.17 ± 15.36 ± mg/g dry wt) and D. polypodioides (1.19 ± 0.01 mg/g dry wt), which
breviarticulatum 0.67D 0.57*FG 0.56D 0.48*FG differed significantly with the rest of the species. Regarding the dry
Dictyopteris 28.07 ± 29.92 ± 21.90 ± 23.45 ± material, the total content of carotenoids was highly variable among
polypodioides 0.4E 0.55I 0.40E 0.45I
Cladophora liebetruthii 29.15 ± 10.37 ± 22.80 ± 7.15 ±
species. A. breviarticulatum showed the highest value after air-drying
0.75*F 0.87D 0.62*F 0.73D (0.94 mg/g dry wt), in addition to other species that also maintained
Dasycladus vermicularis 58.71 ± 21.53 ± 47.44 ± 16.46 ± good levels such as A. saldanhae and V. utricularis, (0.4 and 0.57 mg/g
0.81*G 0.72G 0.68*G 0.60G dry wt, respectively). Therefore, these values were significantly higher
Asterisks indicate significant differences between fresh and dry extracts in each than other species that presented higher values when fresh but that after
species and different capital letters indicate significant differences among air-drying were more affected, for example, Hypnea spp. and
different species in each state: fresh or air-dried (p ≤ 0.05). H. mucronata.
As observed in Fig. 4, the dried samples presented a higher quantity
3.2. Total phenols and phenolic compounds content of total proline content, with some exceptions such as in L. dendroidea,
whose total content of this amino acid from fresh material was 31.38 ±
The TPC of the fresh and dry algae material is shown in Table 3. After 0.48 μmol/g dry wt, significantly higher than the material after drying
the drying period, no significant differences were found between fresh (8.54 ± 0.14 μmol/g dry wt). In other species, such as R. tinctoria, no
and dried material in the studied species. Only two species of green algae significant differences were observed between the dry and fresh states.
showed a decrease in the TPC after the loss of water, C. liebetruthii and In the comparison among species, the highest values in both fresh and
D. vermicularis, from fresh values of 33.14 ± 1.55 and 41.59 ± 1.75 mg dry material were obtained in L. trichoclados (105.62 ± 0.54 and 127.38
GAE/g dry wt, to 12.83 ± 0.83 and 17.97 ± 0.50 mg GAE/g dry wt after ± 0.29 μmol/g dry wt, respectively) that were significantly higher than
drying, respectively. In fresh samples, the highest significant TPC was the rest of the species (up to 100 times higher in some cases).
reached by D. vermicularis followed by A. breviarticulatum, C. liebetruthii
and D. polypodioides, which showed a similar high content, around 33 3.4. Correlation analyses between phytochemicals and total antioxidant
mg GAE/g dry wt. From dry material, the highest values observed were activity
presented by D. polypodioides (31.43 ± 0.90 mg GAE/g dry wt) and
A. breviarticulatum (29.65 ± 1.31 mg GAE/g dry wt), which were Regardless of the material (fresh or dried), correlation analyses be­
tween phytochemicals content and total antioxidant activities among all

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M.A. Ruiz-Medina et al. Algal Research 66 (2022) 102798

Fig. 2. Ferrous ion-chelating (FIC) activities of methanol extracts from 24 species of algae fresh and after a week of air drying. EDTA (ethylenediaminetetraacetic
acid) was used as positive control. Data are expressed as mean ± standard error (n = 3). Asterisks indicate significant differences between fresh and dry extracts in
each species and different capital letters indicate significant differences among different species in each state: fresh or air-dried (p ≤ 0.05).

Table 3
Total phenolic, flavonoid and condensed tannin content from 24 marine macroalgae extracts (fresh and after a week of air-drying). Data are expressed as mean ±
standard error (n = 3).
Plant name TPC (mg GAE/g dry wt) TFC (mg CAE/g dry wt) CTC (mg CAE/g dry wt)

Fresh algae Dried algae Fresh algae Dried algae Fresh algae Dried algae

Jania pedunculata var. adhaerens 2.54 ± 0.61A 3.17 ± 0.72AB 1.52 ± 0.07A 1.16 ± 0.24A 1.71 ± 0.52ABC 0.22 ± 0.13AB
Valonia utricularis 2.67 ± 0.29A 3.17 ± 0.66AB 4.17 ± 0.31*CD 2.54 ± 0.28BC 3.41 ± 0.52*DEF 0.15 ± 0.15A
Codium intertextum 2.73 ± 0.50A 3.43 ± 0.77ABC 4.70 ± 0.37*DE 2.73 ± 0.13BC 0.44 ± 0.26AB 0.15 ± 0.07A
Spyridia filamentosa 2.73 ± 0.44A 3.49 ± 0.61ABC 2.98 ± 0.28B 2.14 ± 0.15B 3.56 ± 0.51*DEF 0.00 ± 0.00A
Galaxaura rugosa 3.24 ± 0.50AB 3.43 ± 0.83ABC 1.93 ± 0.20*A 1.04 ± 0.15A 2.96 ± 0.33*CDE 0.07 ± 0.07A
Jania virgata 3.43 ± 0.40AB 3.68 ± 0.44ABC 1.54 ± 0.15*A 0.68 ± 0.09A 0.00 ± 0.00A 0.00 ± 0.00A
Grateloupia imbricata 4.19 ± 0.72ABC 2.79 ± 0.35A 1.74 ± 0.11A 1.44 ± 0.11A 1.26 ± 0.39*ABC 0.00 ± 0.00A
Wrangelia argus 4.25 ± 1.00ABC 4.25 ± 0.88ABCD 1.71 ± 0.19A 1.12 ± 0.12A 7.78 ± 0.77*H 1.93 ± 0.71CD
Hypnea sp. 4.32 ± 0.46ABC 5.02 ± 0.39ABCD 2.22 ± 0.10*A 1.25 ± 0.12A 5.04 ± 0.45*FG 0.00 ± 0.00A
Cladophora albida 5.27 ± 0.50ABC 5.90 ± 0.83CDEF 3.05 ± 0.17B 3.04 ± 0.24CD 2.89 ± 0.71CDE 0.96 ± 0.45ABC
Hypnea spinella 5.52 ± 0.40BC 5.59 ± 0.35BCDE 1.97 ± 0.13*A 1.14 ± 0.13A 3.41 ± 0.52*DEF 0.67 ± 0.13AB
Laurencia pyramidalis 5.78 ± 0.61BCD 6.60 ± 0.61DEFG 4.85 ± 0.28*EF 3.56 ± 0.24D 3.18 ± 0.58*DE 0.22 ± 0.13AB
Lophocladia trichoclados 6.29 ± 0.66CDE 8.89 ± 0.72G 3.94 ± 0.36CD 2.58 ± 0.33BC 2.52 ± 0.45CDE 3.33 ± 0.64EF
Heterodasya mucronata 6.41 ± 0.61CDE 4.38 ± 0.88ABCD 1.50 ± 0.14*A 0.78 ± 0.19A 2.15 ± 0.45BCD 1.19 ± 0.32ABC
Canistrocarpus cervicornis 6.48 ± 0.61CDE 5.65 ± 0.78BCDE 6.71 ± 0.20H 5.42 ± 0.44E 12.52 ± 0.71*I 5.48 ± 0.77G
Laurencia dendroidea 8.32 ± 0.78DE 7.05 ± 0.83EFG 6.07 ± 0.13GH 5.86 ± 0.28EF 4.15 ± 0.85*EFG 0.89 ± 0.34ABC
Palisada perforata 8.70 ± 0.92EF 7.81 ± 0.61FG 4.15 ± 0.17*CD 0.74 ± 0.12A 2.37 ± 0.39*CDE 0.37 ± 0.20AB
Caulerpa webbiana 10.92 ± 0.66F 7.68 ± 0.94FG 5.97 ± 0.22G 5.59 ± 0.33EF 11.11 ± 0.77*I 4.07 ± 0.45F
Rytiphlaea tinctoria 18.92 ± 0.72G 20.76 ± 0.99J 5.42 ± 0.26FG 9.10 ± 0.24*H 5.71 ± 0.58G 3.48 ± 0.52EF
Anadyomene saldanhae 21.21 ± 1.27G 20.70 ± 0.99J 9.67 ± 0.38J 9.42 ± 0.28H 2.30 ± 0.32*CDE 0.15 ± 0.15A
Cladophora liebetruthii 33.14 ± 1.55*H 12.83 ± 0.83H 8.47 ± 0.20*I 6.31 ± 0.32FG 3.56 ± 0.51*DEF 1.48 ± 0.27BC
Asteronema breviarticulatum 33.59 ± 1.28H 29.65 ± 1.31K 6.69 ± 0.15H 6.60 ± 0.17G 20.96 ± 0.71*J 15.48 ± 0.61I
Dictyopteris polypodioides 33.97 ± 0.88H 31.43 ± 0.90K 9.67 ± 0.24*J 5.84 ± 0.26EF 11.19 ± 0.96I 7.70 ± 0.77H
Dasycladus vermicularis 41.59 ± 1.75*I 17.97 ± 0.50I 16.23 ± 0.18*K 6.96 ± 0.37G 2.00 ± 0.34BCD 2.74 ± 0.27DE

Asterisks indicate significant differences between fresh and dry extracts in each species and different capital letters indicate significant differences among different
species in each state: fresh or air-dried (p ≤ 0.05).

macroalgae samples were performed (Table 4). Among the analyzes
carried out from fresh material, except for the total proline content and
CTC, the rest of the phytoconstituents were linearly correlated signifi­
cantly (p ≤ 0.01) with the total antioxidant activity measured with the
DPPH assay. By comparing the correlation coefficients (r) between the
variables, it was found that antioxidant activity was mainly enhanced by
TPC (r = 0.88) and TFC (r = 0.85). On the other hand, FIC activity had a
significant correlation (p ≤ 0.01) with total proline content (r = 0.37)
and CTC (r = 0.26), although in this case it was lower. However, when
the material was dried, DPPH radical scavenging activity was strongly
and positively correlated (p ≤ 0.01) with TPC (r = 0.84), TFC (r = 0.62)
and CTC (r = 0.50), while FIC activity in fresh was similar to dry ma­
terial. In general, carotenoids were significantly correlated with TPC and
phenolic compounds. Proline content was not significantly correlated
with any phytoconstituent.

6
M.A. Ruiz-Medina et al. Algal Research 66 (2022) 102798

Fig. 3. Total carotenoids content in methanol extracts from 24 species of algae fresh and after a week of air drying. Data are expressed as mean ± standard error (n
= 3). Asterisks indicate significant differences between fresh and dry extracts in each species and different capital letters indicate significant differences among
different species in each state: fresh or air-dried (p ≤ 0.05).

Fig. 4. Total proline content in methanol extracts from 24 species of algae fresh and after a week of air drying. Data are expressed as mean ± standard error (n = 3).
Asterisks indicate significant differences between fresh and dry extracts in each species and different capital letters indicate significant differences among different
species in each state: fresh or air-dried (p ≤ 0.05).

4. Discussion addition, other studied macroalgae, such as A. breviarticulatum and

An in-depth screening has been performed for determination of the
antioxidant properties of 24 marine macroalgae extracts, emphasizing
the effect of air-drying process (under controlled conditions) as a pro­
cessing and storage method in these properties. It has been found that,
regardless of their phylogenetic group or genus, the studied species
exhibited a variable antioxidant activity. In some species this activity
decreased after the air-drying experiment. Therefore, the role of pro­
cessing and storage must be considered for preservation of the antioxi­
dant activity in macroalgae.
In this study the highest potential scavenging activity was presented
by D. vermicularis, whose percentage of discoloration was above other
species of macroalgae such as Chondrus and Porphyra spp. [21]. In
D. polypodioides, also showed a high antioxidant capacity. Similar data
was obtained by Karaki et al. [53], where D. polypodioides showed high
antioxidant and anticoagulant capacity from isolated polysaccharides.
Moreover, those species possess the ability to maintain the antioxidant
activity after air-drying, which make them candidate species for com­
mercial use in contrast to other species of edible macroalgae which
maintain a lower total antioxidant activity after air-drying, e.g., Petal­
onia binghamiae [54] as well as Fucus sp., which antioxidant activity by
DPPH decreased by 96 % [21].
On the other hand, all methanolic extracts analyzed in this study
presented FIC activity that mean that the transition metal ion Fe2+
possess the ability to move single electrons by virtue of which it can
allow the formation and propagation of many radical reactions [55].

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M.A. Ruiz-Medina et al. Algal Research 66 (2022) 102798

Table 4
Correlations between phytoconstituents and total antioxidant activity of 24 marine macroalgae extracts (fresh and air-dry after one week air-drying).
Correlation coefficients (r) DPPH FIC activity Total carotenoids Total proline TPC TFC CTC

Fresh algae (N = 72)

DPPH – 0.26* 0.32* − 0.14 0.88** 0.85** 0.14
FIC activity – − 0.03 0.37* 0.20 0.21 0.26*
Total carotenoids – − 0.19 0.43* 0.34* 0.60**
Total proline – − 0.10 − 0.05 − 0.10
TPC – 0.85** 0.40*
TFC – 0.23
CTC –

Dry algae (N = 72)

DPPH – 0.19 0.44* 0.01 0.84** 0.62** 0.50**
FIC activity – 0.18 0.58** 0.23 0.18 0.46*
Total carotenoids – − 0.14 0.62** 0.55** 0.72**
Total proline – 0.03 − 0.01 0.08
TPC – 0.73** 0.72**
TFC – 0.44*
CTC –

* Correlation is significant at 0.05 level (2-tailed).

** Correlation is significant at 0.01 level (2-tailed).

Among analyzed species, only J. virgata lost its activity after air-drying,
which did not occur in J. pedunculata var. adhaerens, what it is indicative
that each species has its own bioactive molecules that allow them to live
and adapt to their environment. In our study, a high capacity as a
chelating agent (>60 %) was observed in fresh extracts of G. imbricata
and L. trichoclados (close to the EDTA control). This capacity was
maintained after the air-drying process in L. trichoclados. Today we
know that this chemical substance, as well as its sodium salts, have
numerous applications in industry, cosmetic products, laboratory and in
medicine, where it is used for the treatment of mercury and lead
poisoning through the chelation therapy [56]. Thus, these properties
make L. trichoclados a species of high interest.
Antioxidant activity of phenolic compounds is correlated to their
chemical structures. In general, FRs scavenging and antioxidant activity
of phenolics mainly depends on the number and position of hydrogen-
donating hydroxyl groups on the aromatic ring of the phenolic mole­
cules [57]. In our study we found two species (C. liebetruthii and
D. vermicularis) with a high content of total phenolic compounds, how­
ever, after air-drying this content decreased significantly as happened in
Hormosira banksii and other edible marine macroalgae [21,58]. Excep­
tionally, A. breviarticulatum and D. polypodioides presented a high
phenolic content that was maintained after drying, being even in
D. polypodioides up to 4 times higher than fresh extract from other spe­
cies of the genus, for example D. divaricata [59]. In our study,
D. vermicularis presented the highest concentration of flavonoids in fresh
material, which was positively correlated with the highest antioxidant
activity (DPPH assay). Nevertheless, although other studies have shown
the ability of this species to produce high concentrations of polyphenolic
compounds in response to high radiation and low salinity [60,61], we
have demostrated that the opposite effect occurs after prolonged air-
drying, having a significant decrease in flavonoids content. Otherwise,
tannins like phlorotannins are mostly found in marine algae and they are
phloroglucinol oligomers produced in brown algae and have tannin
activities. Possibly the activity of phlorotannins in macroalgae is related
to the presence of condensed tannins [62], and therefore in this study
the highest concentration of tannins was found in brown algae
A. breviarticulatum, C. cervicornis and D. polypodioides, which is consis­
tent with that found in other species of brown algae such as Sargassum
ringgoldianum [63] and Macrocystis integrifolia [64].
Among the various defense strategies to reduce oxidative stress, ca­
rotenoids are most likely involved in the scavenging of two of the
reactive oxygen species, singlet molecular oxygen (1O2), but mostly
peroxyl radicals [65]. In our study, the range of variation of total ca­
rotenoids from fresh material varied between 0.15 and 1.2 mg/g dry wt,
while in other studies on edible macroalgae did not detect the presence
of carotenoids e.g., Eisenia bicyclis, Sargassum horneri or Cystoseira
hakodatensis [66] nor even in microalgae, e.g., Isochrysis galbana [67],
However, among all the antioxidants studied, carotenoids were the most
affected by air-drying and might be attributed to both down-regulation
of biosynthesis and consumption by scavenging superoxide anion [68].
After a prolonged air-drying period, these molecules lose their activity
because of their poor stability and high sensitivity to oxidation and
degradation, which accordingly limit their applications as health-
beneficial components [69].
Proline is a compatible osmolyte, whose concentration increases in
plants exposed to air-drying stress. The exogenous application of proline
has been seen to significantly improve the content of rosmarinic acid
and endogenous proline, which may be indicative of the stimulation of
the pentose phosphate pathway. The earliest precursor of rosmarinic
acid is erythrose-4-P, which is one of the end products of the pentose
phosphate pathway. In consequence, high proline synthesis induces the
pentose phosphate pathway and the synthesis of shikimate, rosmarinic
acid and other phenolic compounds [70,71]. Therefore, proline content
of edible plants may be accepted as a measure of its antioxidant capacity
[12,72]. Nevertheless, in our study we did not find a correlation be­
tween proline content and phenolic compounds, so further studies may
be required to evaluate the role of proline as an antioxidant in marine
macroalgae. Among red macroalgae, L. dendroidea, R. tinctoria and
L. trichoclados presented the highest proline values, both from fresh and
dry material extracts. To our knowledge, the proline concentration of
L. trichoclados (127 μmol/g dry wt) is one of the highest recorded in
algae, up to 100 times more than in other edible species such as those of
the genus Chlorella (C. salina and C. vulgaris). This suggests a great po­
tential for endogenous proline synthesis in this species that could be
applied in the pharmaceutical industry due to the benefits that this
amino acid offers to human health [12].
High correlation detected among total antioxidant activity and
phenolic compounds was found in our study, what have been also shown
in other studies [73,74]. However, we noted that correlations of the
antioxidant activities from different assay methods were also affected by
the state of the material (fresh or dried). The findings from the corre­
lation analyses indicated that extracts from different material had
different degrees of contributions to the overall antioxidant activities of
macroalgae. Therefore, for antioxidant studies, the state of the algae
material must be carefully selected in each case.
For the first time, a comprehensive study was carried out on phyto­
constituents with antioxidant activity in 24 marine macroalgae located
in the Canary Islands, comparing the fresh state of the material after

8
M.A. Ruiz-Medina et al.
harvesting and its state after a week of air-drying process. It has been
observed that from fresh material, C. liebetruthii, D. vermicularis and
D. polypodioides presented the highest total antioxidant activity by the
DPPH method, supported by high correlation with carotenoids and
phenolics compounds content. After the drying period to which the
macroalgae were exposed, A. saldanhae and D. polypodioides showed the
highest values referred to total antioxidant activity, which makes them
potentially exploitable species due to the advantages offered by the air-
drying method. The high FIC activity recorded in fresh material from
G. imbricata and L. trichoclados could be used for the treatment of mer­
cury and lead poisoning through the chelation therapy, further studies
should be carried out in this line. Only L. trichoclados was able to
maintain this high activity after air-drying supported also by a high
proline content, whose high values make this species an exception
among most marine macroalgae. The applications of algae are unlimited
in the biotechnology sector, and its extracts, source of unexplored nat­
ural resources, are essential to take advantage of and provide a solution
to the growing social demand for food, energy and pharmaceuticals.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgments
We would like to thank PhD student Daniel Álvarez-Canali and Dr.
Nereida Rancel for their help in fieldwork. We thank the Editor-in-Chief
and two anonymous reviewers for their valuable comments that
improved the quality of this article.
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