Algal Research 57 (2021) 102349

Contents lists available at ScienceDirect

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

Short communication

Sustainable exploitation of macroalgae species from Chilean coast:

Characterization and food applications
Eduardo Caballero *, Agustín Flores, Araceli Olivares
Centro Regional de Estudios en Alimentos Saludables – CREAS, Av. Universidad #330, Placilla, Sector Curauma, Valparaíso, Chile

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

Keywords: The use of macroalgae has been always linked to direct consumption, however, current trends have increased the
Macroalgae applications of algae in the formulation of foods to replace animal proteins. Hence, we must ensure the safety of
Extraction the increasing food products based on macroalgae. In this regard, and due to the worldwide variability of its
Alginate
composition, we determined the nutritional characterization of Durvillaea Antarctica, Macrocystis pyrifera, and
Viscosity
Lessonia nigrescens to evaluate their potential at nutritional level. Also, as they are accumulating organisms, we
Heavy metals
characterized their minerals and heavy metals content, to determine its potential applications and consumption
risks. The results indicated that the content total As is into the safe range to be consumed (16.92 and 60.75 mg/
kg dw), but Cd (between 3.97 and 6.15 mg/kg dw) is higher than 0.5 mg/kg of final product (regarding Chilean
Normative), and in this case, we have two possibilities: To add no more than 12.5% of M. pyrifera and no more
than 8% of D. antarctica or L. nigrescens flour in the final product; or avoid the heavy metals by the extraction of
alginates to enhance applications in food such as increasing the viscosity or improving dispersion stability. From
the last point of view, we have compared the alginates extracted from D. antarctica, M. pyrifera, and L. nigrescens
through the use of two methodologies to obtain calcium alginate (10.55–16.75% extraction yield) and sodium
alginate (12.45–20.8% extraction yield), where the methodology to extract sodium alginate showed the best
perform for the three macroalgae. The results showed that viscosities at 20 ◦ C and alginate concentration of 40 g/
L were: 1201.4 cP, 544.0 cP and 722.4 cP for D. antarctica, M. pyrifera, and L. nigrescens respectively. With these
results we can conclude that the macroalgae D. antarctica is the one with both the highest extraction yield and
technological potential as it generates a higher viscosity.

1. Introduction
Chile is one of the leading fishing countries, with a vast coastline of
more than 6000 km, and like the rest of the world, it must face the
overexploitation of a large part of its fisheries due to an excess of catch.
Due to this situation, artisanal fishermen have had to change their ac­
tivities and focus on sustainable exploitation of various marine resources
like seaweeds or macroalgae. That is the case in Laguna Verde area
(Valparaíso Region, Chile), where the extraction of macroalgae of Dur­
villaea antarctica, Macrocystis pyrifera and Lessonia nigrescens, is been
studied and characterized by the first time in terms of macronutrients,
minerals, and heavy metals to evaluate the strategy to be sustainable
exploited in the best commercial and innocuous way. Regarding the FAO
[1], 28.4 million tons of macroalgae were obtained as food or as in­
gredients for food processing in 2014. Macroalgae also has been used in
pharmaceutical and cosmetic industries, due to its high content of
nutrients and functional compounds, reason why macroalgae are good
alternatives for innovations in functional foods [2]. Despite the nutri­
tional profile, macroalgae can contain some contaminants and other
unwanted compounds (e.g., cadmium (Cd) and arsenic (As)). Some
studies have reported that the accumulation rate of As, is not only
related to its availability but also to biological, chemical, and environ­
mental variables. Also the bioavailability and bioaccumulation of As in
macroalgae can be linked with biotic and abiotic factors [2,3,4,5]. This
is a fact that gives importance to characterize each macroalgae species in
different conditions and geographic zones like Laguna Verde. That is
also the reason why macroalgae are considered as an effective envi­
ronmental indicator [6]. Hence, within the study of Ownsworth et al.
[6], investigated the trace of heavy metals of macroalgaes in Scotland.
They found that the overall trend in minerals and heavy metals (Os ≪
Re < Ag < U < Cd < Co < Ni < Pb < Cu < As < Zn ≪ I), and identified
the variations along the estuary. The importance of the study of several

* Corresponding author.
E-mail address: ecaballero@creas.cl (E. Caballero).

https://doi.org/10.1016/j.algal.2021.102349
Received 20 October 2020; Received in revised form 21 April 2021; Accepted 2 May 2021
Available online 26 May 2021
2211-9264/© 2021 Elsevier B.V. All rights reserved.
E. Caballero et al.
species is due to, depending on macroalgae, some elements increase
towards the sea and others decrease or have a peak in the middle of the
estuary. These changes are controlled by different factors, such as
salinity, mixing and macroalgae species [6]. This is another very sig­
nificant fact, and gives heavy metal characterization a great value when
we are looking for some food application approaches of macroalgae for
human consumption. For example, a pretreatment of feedstock by
soaking and thermal treatment, decreases the inorganic arsenic con­
centration [6].
In terms of regulations, the Commission of the European Commu­
nities (2003/40/EC) [7] determined the limit of total arsenic in water
(10 mg/L), while the values for the specific As sources (organic or
inorganic) have not defined limits [2]. Furthermore, for food products
including seaweeds on its formulation, there is no defined threshold [2].
However, since the inorganic As methodologies are available for some
types of food products, a concentration of 0.2 mg of inorganic As/kg
white rice was defined by the EU, and 0.1 mg of inorganic As/kg rice-
based food [2,8].
Despite the above mentioned, if the macroalgae have a heavy metal
profile that avoids using them directly for consumption, we can consider
that macroalgae have a good profile of functional and bioactive com­
pounds. The nutritional properties of macroalgae consumption have
been reported in several researches, such as the interesting compositions
of dietary fiber [9,10], proteins, polyunsaturated fatty acids and anti­
oxidant [11], and low percentage in fat (specifically saturated fats).
Those nutritional advantages are complemented with its functional
compounds, such as antibacterial, antiviral and antifungal bioactive
compounds [12]. Among those compounds, one of the opportunities of
processing macroalgae, is to take advantage of macronutrients that ex­
press technological functions. That is the case of the hydrocolloids,
where the growth of the algal hydrocolloids market has shown a slow
but continue rate of 2–3% per year, reaching a growth of 20,535 t, which
represents 93,035 t of hydrocolloids in 2015 (equivalent to USD 414
million).
[13]. The main countries producing macroalgal hydrocolloids are
China, Indonesia, Korea, Philippines, Japan, and other Asian countries,
with estimated amounts in 2015 of 10,000 t, 31,844 t and 27,200 t for
agar, alginate and carrageenan, respectively, which represents more
than 50% of the total market of hydrocolloids [13,14].
As it was mentioned, alginate is the most demanded and produced
hydrocolloid in the world. Alginates are structural compounds of cell
walls in macroalgae (Phaeophyceae), such as Durvillaea Antarctica,
Macrocystis pyrifera and Lessonia nigrescens, and make the tissue flexible
and strong. Polysaccharides content in macroalgae are in the range from
4 to 76% d.b., which depends on the season, and the section and species
of macroalgaes [13,15]. Alginate, agar, and carrageenan have demon­
strated biological and physicochemical functionalities of interest for the
food industry [16]. Thereby increasing the inherent ability of these
hydrocolloids to form hydrogels and be used as thickening, gelling or
film-forming ingredients [13,17,18,19], to form coatings for pharma­
ceutical products, to be included in packagings, or to increase the shelf-
life of food [13,20,21,22,23,24,25]. In addition, macroalgae carbohy­
drates are relevant from the technological point of view, allowing pro­
cesses related with emulsification, texturization, fat replacers, and water
absorption in food product formulation. Garcia-Vaquero et al [26] re­
ported a strong relationship between consumer acceptance of food
products including polysaccharides from macroalgaes, and its taste and
texture. In the case of alginate, the viscosity depends on the length of the
polymer. Besides, hydrocolloids are biocompatible, biodegradable and
nontoxic natural polysaccharide [27].
On the other hand, the uses of hydrocolloids in food formulations
could be increased, but the intensive extraction process at industrial
level [13,28,29,30,31] or adaptations used in this work [19,32], must be
optimized and new clean technologies must be studied and scaled up.
The current methodologies to extract hydrocolloids use large amounts of
solvents and reagents, which increase the processing cost [33,34,35].
2
Algal Research 57 (2021) 102349
Therefore, the extraction of functional compounds from macroalgaes
using new clean technologies are currently reported, however, not all
those technologies are available at industrial level [13,36].
Taking into account all the information, the future application as a
food ingredient and the potential of three brown macroalgae species
were evaluated. The nutritional, functional, and heavy metal charac­
terization are valuable tools to define if a specific species from a specific
location is feasible to be exploited to direct human consumption or to
generate alginate extracts. In the case of alginate extraction, we studied
the effect of two extraction methodologies, and evaluated their extrac­
tions yield and alginate viscosity as a measurement of its potential as a
thickener.
2. Materials and methods
2.1. Macroalgae source
Three macroalgae (Fig. 1): Durvillaea antarctica, Macrocystis pyrifera
and Lessonia nigrescens, were sustainably collected on July 2019 (winter
season in South Hemisphere) by local fisherman from Laguna Verde
area, Valparaíso, Chile (33◦ 06′ 16′′ S and 71◦ 40′ 03′′ W).
2.2. Pretreatment
Macroalgae were first washed with abundant fresh water. Then,
samples were grounded using a Robot Cook grinder (Robot Coupe ®)
and dehydrated by infrared equipment at 50 ◦ C by 24 h, where macro­
algae showed right colour and flavour (no toasted flavour) or in an oven
at 105 ◦ C by 15 min or until constant weight. The resulting dried macro
algae were processed in a centrifugal mill (Retsch ZM 200) with a mesh
of 0.1 mm to obtain a powder used in food formulation. A portion of
three fresh macro algae were frozen at − 80 ◦ C and then lyophilized at
− 53 ◦ C and 50mTorr (Ilshin freeze-dryer equipment). These samples
were used to carry out the proximal analysis. In all cases, samples were
maintained at 20 ◦ C into polyethylene bags.
2.3. Proximate analysis of macroalgae
Chemical composition of macroalgae was determined by Official
Methods (AOAC, 1995) [37]. Proteins were determined by distillation at
Kjeldahl analysis and total ash by calcination of macroalgae overnight in
a furnace at 550 ◦ C. Lipids were determined in a Soxhlet extractor (Büchi
®). Total carbohydrates were estimated by rounding up. Analysis were
performed in triplicate.
2.4. Antioxidant capacity of macroalgae
The antioxidant capacity of macroalgae was assayed by the ORAC
method, according to Soto-Maldonado et al. [38]. Shortly, a macroalgae
sample, a Trolox standard and AAPH (2,2′ -azobis(2-methyl­
propionamidine)dihydrochloride), were prepared with phosphate buffer
at pH 7.4. 1 g of dry macroalgae was dissolved in a 17 mL phosphate
buffer. Then, 20 μL of macroalgae sample or phosphate buffer used as
blank, and fluorescein (200 μL, 1.5 mM) were added to each well in a 96-
well opaque plate. Then, the plate was incubated at 37 ◦ C for 10 min.
After that, AAPH (75 μL, 79.7 mM) was added to each well. Fluorescence
was measured and recorded at 37 ◦ C every 1 min (wavelengths of
excitation 485 nm, and emission 538 nm). The final antioxidant capacity
of macroalgae was calculated as Trolox equivalents and the calibration
curves were calculated using Trolox as the standard (20–90 μM) [38].
The analysis was carried out in triplicate.
2.5. Mineral and heavy metals analysis of macroalgae
Minerals content and heavy metals were determined using methods
TR-ICP MS (Inductively Coupled Plasma Mass Spectrometry (ICP-MS))
E. Caballero et al.
Fig. 1. Macroalgae: Durvillaea antarctica (left), Macrocystis pyrifera (center) and Lessonia nigrescens (right).
[39]. The analysis was carried out in triplicate and the blank for the most
of minerals was less than 0.0001 mg/kg.
2.6. Alginic acid extraction methods
Alginic acid extraction from three macroalgae was carried out using
a powder of each macroalgae. Two extraction methods were used
(Fig. 2).
A. Method (GREEN) 20 g of dried and milled macroalgae was mixed
with 100 mL of a 2% CaCl2 solution and then was mixed 1/23 with
water (pH 3.4) at 83 ◦ C for 4 h with constant agitation according to
Yu and Chao [32]. The mix was centrifuged at 3000 ×g for 20 min
(HERMLE LaborTechnik GmbH, German, mod Z446K). The super­
natant was mixed with ethanol (96%v/v) at 4 ◦ C, for 4 times and
centrifuged again at the same conditions mentioned above. Six vol­
umes of ethanol were added to supernatant to precipitate the algi­
nate as calcium alginate and finally alginate was filtered and dried to
a constant weight in an oven at 50 ◦ C. The extractions were carried
out in triplicate.
B. Method (ORANGE) 25 g of dried and milled macroalgae was mixed
with 2% (v/v) formaldehyde solution (ratio 1/32) for 24 h to remove
phenolic compounds and pigments according to Khajouei et al. [19]
and insoluble fraction was rinsed with MilliQ water (3 times). After
that, 800 mL of hydrochloric acid solution HCl (0.2 M) was added
and incubated at 60 ◦ C for 3 h in an orbital shaker (250 rpm) for acid
treatment. Then the suspension was centrifuged at 3000 ×g for 20
min, and pellets were washed with MilliQ water (3 times). Pellets
were steeped with Na2CO3 solution (3% w/v) at 60 ◦ C for 2.5 h, and
the mixture was centrifuged at 3000 ×g for 30 min. Then three
volumes were added to the supernatant to precipitate, and pellets
(including sodium alginate) were dissolved in MilliQ water and
precipitated with ethanol again and repeated twice [19]. Finally, the
precipitate containing sodium alginate was dried. The extractions
were carried out in triplicate.
2.7. Water holding capacity and fat binding capacity
The extracts obtained from both extraction methods were separated
and dehydrated to get an alginate-rich powder as a hydrocolloid. Then,
water holding capacity (WHC) and fat binding capacity (FBC) of alginate
obtained from three macroalgae were determined according to Jeya-
Shakila et al. [40]. 10 mg of sample was taken in a centrifuge tube
3
Algal Research 57 (2021) 102349
(Eppendorf). 0.5 mL of distilled water or 0.1 mL of sunflower oil was
added to measure WHC and FBC, respectively. The solution was mixed
in a vortex every 15 min by 5 s at room temperature for 1 h. Then, the
solution was centrifuged at 3000 ×g for 20 min. The measurements were
performed in triplicate. The upper portion was removed and drained for
30 min on filter paper at a 45◦ angle. WHC and FBC were calculated by
Eq. (1) (E.1):
(
g
)
Wt.of the content of the tube, g (after drained)
WHC o FBC =
100g Wt.of the dried sample, g
(1)
2.8. Viscosity
A solution of 40 g/L of alginate-rich powder was prepared (for three
macroalgae and both methods). Each solution was measured for the
viscosity (cP) in a Digital Viscometer (VISCO™ – 895, Japan) using N◦ 3
spindle from 3 to 250 rpm at room temperature (24 ± 1 ◦ C). A solution of
alginic acid sodium salt from brown algae (SIGMA cod. 71,238) at 10
and 40 g/L was used as control. The measurements were carried out in
triplicated.
2.9. Statistical analysis
The results are expressed as the mean ± standard deviation of trip­
licate experiments. Single-factor ANOVA was used to compare results
and determine statistically significant difference (P < 0.05).
3. Results and discussion
3.1. Proximate and antioxidant composition
Tables 1 and 2 show proximate composition and antioxidant activity
of Durvillaea antarctica, Macrocystis pyrifera, and Lessonia nigrescens,
respectively.
Different works report chemical analysis of dried samples of brown
macroalgae [18,19] and Macrocystis pyrifera is one of main commer­
cialized macroalgae as an alginate source [18,41]. Gao et al. [18] report
similar contain of proteins and ash in M. pyrifera, meanwhile fat re­
ported is higher than our value for M. pyrifera. In the case of carbohy­
drates Gao et al. [18] report 43.3% for M. pyrifera. Among the factors
that impact in the nutritional composition of macroalgae and alginic
acid containing, we can find the seasonal factor, seawater temperature,
E. Caballero et al.
Fig. 2. Schematic representation of extraction methods.
nutrient availability, salinity and/or light exposure. For example,
Mansilla and Ávila [42] reported the seasonal analysis of M. pyrifera
flour; showing that spring is the best time to collect macroalgaes to
obtain the highest protein and ash contents, meanwhile, if the lipids are
the target, the best season is summer and for carbohydrates is winter,
coinciding with our values obtained in July (winter season in South
Hemisphere). Ortiz et al. [43] report identical values for proteins from
D. antarctica (10.4 and 11.6% in leaves and stem, respectively) respect to
our value (10.79%) and for ashes our value is similar to obtained from
stem.
D. antarctica shows greater antioxidant activity, being 63% greater
4
Algal Research 57 (2021) 102349
than M. pyrifera and 172% greater than L. nigrescens. However, in terms
of antioxidant activity ORAC, the values are not interesting, because the
vegetables in general have a range between 6000 and 60,000 μmol ET/
100 g dry weight.1 Regarding the statistical analysis (ANOVA), the only
significant statistically differences in ORAC values are between D.
Antarctica and L. nigrescens (p value <0.05).
1
(http://www.portalantioxidantes.com/base-de-datos-de-actividad-antio
xidante-orac-y-de-contenido-de-polifenoles-totales-pft-en-hortalizas/.)
E. Caballero et al. Algal Research 57 (2021) 102349

Table 1 Table 4
Proximate composition (g/100 g macroalgae). Heavy metals content of macroalgae.
Composition Durvillaea Macrocystis Lessonia Heavy metals Concentration ± SD (mg/kg DW)
antarctica DA pyrifera MP nigrescens LN
Durvillaea Macrocystis Lessonia
Energy (kcal) 265.31 251.97 235.03 antarctica pyrifera nigrescens
Fresh moisture 83.86 ± 0.11 82.23 ± 0.64 76.95 ± 0.2
Fe (Iron) <0.01 <0.01 <0.01
Moisturea 8.15 ± 0.014 6.8 ± 0.027 8.5 ± 0.13
Cu (Copper) 4.37 ± 0.35 5.33 ± 0.39 5.4 ± 0.43
Proteina 10.79 ± 0.08 9.81 ± 0.04 9.88 ± 0.1
Zn (Zinc) 37.66 ± 2.98 21.65 ± 1.65 15.87 ± 1.26
Lipida 0.43 ± 0.022 0.21 ± 0.02 0.23 ± 0.03
As (Arsenic) 47.75 ± 0.8a 60.75 ± 0.86b 16.92 ± 0.21c
Asha 26.06 ± 0.031 30.47 ± 0.096 33.03 ± 0.068
Se (Selenium) <0.0002 <0.0002 <0.0002
Carbohydratea, 54.57 52.71 48.36
b Sn (Tin) <0.0001 <0.0001 <0.0001
Hg (Mercury) 0.012 ± 0.00 0.026 ± 0.00 0.021 ± 0.00
n = 3. Pb (Lead) 7.33 ± 0.01 1.45 ± 0.00 2.73 ± 0.01
a
Lyophilized samples. Cd 6.15 ± 0.02d 3.97 ± 0.01e 6.01 ± 0.03f
b
Included TDF (Total Dietary fiber). (Cadmium)

Different letters (a, b, c) and (d, e, f), represents statistically significant differ­
ences.
Table 2 n = 3.
Antioxidant activity (ORAC) of macroalgae.
Antioxidant activity Durvillaea Macrocystis Lessonia carcinogenic effect in humans [49]. Hence, the economic relevance of
antarctica DA pyrifera MP nigrescens LN macroalgae consumption in several reports, is focused at the beginning,
ORAC (μmol ET/100 g 2506 ± 13.0 1534 ± 361.5 920 ± 108.6 by As concentration [5,50,51,52,53,54]. Arsenic in macroalgaes can be
dry weight) in its organic form (arsenosugars) [55], which has lower toxicity than its
n = 3. inorganic form [56], been thiodimethylarsinic acid (one of its metabolic
products) cytotoxic in culture human lung cells [57]. Regarding As, the
3.2. Mineral and heavy metals analysis Table 4 considers total As. In that case, the macroalgae tested are in the
range between 16.9 and 60.8 mg/kg dry weight, being Lessonia nigres­
Minerals content and heavy metals are shown in Tables 3 and 4, cens the species with the lowest content. When we analyze one of the
respectively. most extracted and produced species in Chile as D. antarctica (total As
Macroalgae are a relevant mineral source [44]. The main mineral 47.75 mg/kg dry weight), we found some studies carried out by Díaz
content in macroalgaes, related with human health, are Na, Ca, K, Mg et al. [5], where 13 samples of D. antarctica were analyzed for total and
and P, also considering micronutrients as I, Fe, Zn, Cu, Mn and Se inorganic arsenic concentration (mg/kg) 49.0 ± 6 and 0.31 ± 0.08,
[44,45,46]. In terms of concentrations, Brito et al., [47] have been respectively, concluding that the species are not health risky for human
measured K, Ca, and Mg for several varieties of macroalgae, among consumption. In addition, seaweed processing such as washing, soaking
them, the brown algae presented concentrations ranging between 780 and cooking may reduce the total arsenic concentration by as much as
and 56,390 mg/Kg for Ca, 2830–115,790 mg/kg for K, and 60% [58], which could be a good strategy in the case of the three
1030–33,000 mg/kg for Mg. According to these results, and in com­ macroalgae tested in the present study due to the results shown in
parison with the results shown in Table 3, the three macroalgae tested Table 4.
are into the range for Ca, K and Mg. Other heavy metal present in macroalgae worldwide is Cd, which
Regarding Na, if we want to use the macroalgae flour in food for­ could fluctuate among the species and classifications (green, brown, and
mulations, we have to take into account that the current legislation in red macroalgae) [48]. Regarding the amounts of cadmium in D.
Chile, consider limits for critical nutrients and salt (Na). In that case, we antarctica, Almela et al. [59], reported values around 2.46 mg/kg dry
cannot formulate food products with over 400 mg Na/100 g product. weight, which exceeds the norm by around five times. This is a property
When we develop food products we have to consider no more than 19% considering the D. Antarctica as a bioadsorbent of cadmium in contam­
of macroalgae flour in the final product. Hence, it could increase the inated ground [60]. Cd concentrations are higher than 0.5 mg/kg of
potential of L. nigrescens as macroalgae with lower Na content. final product (regarding Chilean Normative), and in this case, we can
Cd in most samples exceed limits, and the total and inorganic As in add no more than 12.5% of M.P and no more than 8% of D. antarctica or
Hizikia fusiforme (Sargassum fusiforme), reported by Besada et al. [48], L. nigrescens flour in the final product to accomplish the normative.
was 103–147 mg total As/kg dry weight; 32–70 mg inorganic As/kg dry However, for human consumption and taking into account all the results
weight, which would inhibit the human consumption [2,48] due to its shown including the Table 4, this has further implications if the mac­
roalgae is used, for example, to cook soups (e.g., Dashi), as the leached
elements become a significant component of the soup, which is an
Table 3 important point for food applications and food processing [6]. This is an
Mineral content of macroalgae. important reason to generate and develop pretreatments or focus on the
main compounds of interest, for example, bioactive substances antiox­
Mineral Concentration ± SD (mg/kg dw)
idants have been widely studied due to the positive effects against
Durvillaea Macrocystis Lessonia serious disorders [61] or as in the present study, the extraction of
antarctica pyrifera nigrescens
compounds with technological properties like alginates.
Na (Sodium) 20,364.4 ± 28,869.9 ± 15,556 ± 1102.7c
1425.5a 1987.2b
K (Potassium) 10,447.1 ± 815.8 71,048.9 ± 5583.9 37,795.7 ±
3.3. Alginic acid extraction methods
3056.1
Ca (Calcium) 14,144.1 ± 747.2 22,520.2 ± 1096 14,897.2 ± 756.6
Mg 14,403.7 ± 457.1 11,108 ± 363.2 2702.7 ± 86.9 The extraction yield of alginate, depending on extraction methods,
(Magnesium) was calculated as % w/w, on the basis of the initial dry weight of
Y (Iodine) 0.102 ± 0.003 0.082 ± 0.02 0.103 ± 0.003 macroalgae.
Different letters (a, b, c), represents statistically significant differences. Alginate extraction yield obtained by A and B methods, are presented
n = 3. in Table 5.

5
E. Caballero et al. Algal Research 57 (2021) 102349

Table 5 shows WHC and FBC expressed as g/100 g (E.1) for both methods.
Alginate extraction yield. The functional properties, like hydration of alginates obtained, are
Extraction Extraction yield ± SD (% w/w) described by measuring the water holding capacity (WHC). WHC is
method taken as a measurement of how different types of flours are able to hold
Durvillaea Macrocystis Lessonia
antarctica pyrifera nigrescens different amounts of water. This is useful to identify ingredients for food
applications. For example, crackers and cookies require flour with low
A Method 10.97 ± 0.62a,e 16.75 ± 0.81b,g 10.55 ± 0.35a,h
B Method 20.8 ± 1.69c,f 18.0 ± 1.27c,g 12.45 ± 0.93d,h
WHC to make the product crispy, conversely in buns it is positive with
flour with a high WHC to soften the product [63]. In this case, the WHC
a, b, c, d represents the differences statistically significant among species in of sodium alginate (B method) using M. pyrifera, expresses the lowest
method A or method B.
WHC, which could be a good choice to formulate crispy snacks.
e, f, g, h represents differences statistically significant between methods for each
WHC values obtained with A method are higher than obtained with B
macroalgae.
method for three macroalgae. They depend on the extraction methods;
different types of alginates were obtained. Then, calcium alginate and
The values presented in Table 5, are in the range previously reported.
sodium alginate, for A and B methods, were obtained, respectively.
In the case of M. pyrifera, [18] reported yields between 16.31% to
Values for WHC are not relevant to soluble polysaccharides, how in the
26.32% of sodium alginate obtained from different pre-treatment. Yu
case of sodium alginate, that is more soluble than calcium alginate,
and Chao [32] obtained an extraction yield of 1.26% of kelp poly­
explaining the minor values of WHC and the variability of the results
saccharides employing a similar methodology to extraction method A.
expressed in the standard deviation. Due to calcium alginate can form
Khajouei et al. [19] reported a yield of 24% of sodium alginate from
bigger cross-linked polymers than sodium alginate, the first one is more
Nizimuddinia zanardini. Regarding the process, A method require less
insoluble in water at room temperature, which is demonstrated in the
reaction volumes and solvent than B method, however, B method
results of Table 6 for WHC. The comparison of different WHC values is
showed higher extraction yields and better alginate performance (Figs. 3
complicated, because it depends on the type of macroalgae, alginate
and 4) than A method.
extraction conditions, and methods of measurement, even the season
Some innovative processes such as Enzyme-assisted Extraction
that was extracted the macroalgae (mentioned previously). Another
(EAE), Ultrasound-assisted Extraction (UAE), Supercritical Fluid
aspect to take into account is that WHC is determined by the composi­
Extraction (SFE), Microwave-assisted Extraction (MAE), Pressurized
tion of the sample and its characteristics such as total dietary fiber and
Liquid Extraction (PLE), Ultra-High Pressure Extraction (UPE), Pres­
β-glucan content, proteins, starch and particle size [64]. Considering
surized Fluid Extraction (PFE) and auto-hydrolysis processes [13,62] are
that all the dried macroalgae were milled under the same conditions, the
the most promising processes currently explored to increase the
differences could be determined by the differences in terms of the
extraction performance of hydrocolloid from macroalgae. Nevertheless,
proximal characterization shown in Table 1. Hence, D. antarctica has the
each technology has its limitations that must be studied and optimized,
highest content of protein and carbohydrates among the macroalgae
depending on both the biological source to be processed and the appli­
tested and if a food formulation needs more WHC it would be better to
cations of the final product [30].
use the whole macroalgae flour to retain humidity in the final food
On the other hand, taking into account the information in Table 1,
product. Despite WHC can give information about the applications, it is
the importance to give an added value to a by-product of both meth­
not a good parameter to compare extraction methods for two different
odologies is discussed. When alginate is removed from macroalgae, the
salts of alginate from different macroalgae sources.
main compounds that remains in the solid residues are represented by
On the other hand, fat binding capacity (FBC) is another important
minerals, protein and another carbohydrate, which could be used as an
property for the ingredients used in the formulation and stabilization of
organic fertilizer in agricultural applications. However, further studies
food with high percentages of fat and emulsion. Our results show that
must be carried out to establish the heavy metal content in the solid
FBC is higher when A method was used. Calcium alginate is less soluble
residues.
than sodium alginate, and the triplicated demonstrated less variability
(see standard deviations for FBC in Table 6). The highest value was
obtained for L. nigrescens. These results are interesting from the tech­
3.4. Water holding capacity and fat binding capacity
nological point of view, allowing us to recognize the calcium alginate
from L. nigrescens, like a good ingredient to be applied in food formu­
WHC and FBC were determined for alginate of three macroalgae:
lations that need to retain oil or fat. For example, to reach the texture
Durvillaea antarctica, Macrocystis pyrifera and Lessonia nigrescens ob­
and palatability of vegetable based burgers, and others vegetarian meat
tained with both extraction methods (A: green and B: orange). Table 6

Fig. 3. Viscosity of alginate of macroalgae obtained by two extraction methods (A and B) at different rpm. MP (M. pyrifera), LN (L. nigrescens), DA (D. Antarctica). The
standard deviation of each point is ranging between 1.3 and 8.3% of the mean value.

6
E. Caballero et al. Algal Research 57 (2021) 102349

Fig. 4. Viscosity of alginate of macroalgae obtained by two extraction methods (A and B) at 50 rpm (Control at 40 g/L was 6878 cP). MP (M. pyrifera), LN (L.
nigrescens), DA (D. Antarctica). Different letters mean a significant difference (P < 0.05) among viscosities of alginate obtained into each method.

mayonnaise.
Table 6
Regarding the alginate extraction method, we can mention that
WHC and FHC (g/100 g).
different solvent used could also extract another constituent from the
Durvillaea Macrocystis Lessonia wall cells like β-glucan or even proteins or starch. The presence of
antarctica pyrifera nigrescens
β-glucan increases the viscosity of a sample [65].
WHC (Method 291.00 ± 132.28 269.24 ± 19.73 337.67 ± 67.69
A) 4. Conclusion
WHC (Method 216.95 ± 0.00 156.43 ± 105.30 230.62 ± 69.29
B)
FBC (Method A) 267.49 ± 10.48 373.10 ± 34.44 462.47 ± 16.17 The characterization of Durvillaea Antarctica, Macrocystis pyrifera,
FBC (Method B) 268.46 ± 10.01 287.48 ± 12.58 232.13 ± 0.24 and Lessonia nigrescens to evaluate their potential at nutritional level,
and their minerals and heavy metals content, revealed that the content
total As is into the safe range to be consumed (16.92 and 60.75 mg/kg
substitutes and emulsions.
dw), but Cd (between 3.97 and 6.15 mg/kg dw) is higher than 0.5 mg/kg
of final product (regarding Chilean Normative). To avoid the restrictions
3.5. Viscosity about the heavy metals, the extraction of alginates allows to enhance
applications in food products as an ingredient to increase the viscosity or
Fig. 3 shows the viscosity of alginate as a function of speed equip­ improve dispersion stability. The comparison among the alginates
ment (rpm) for three macroalgae and two extraction methods (A and B). extracted from D. antarctica, M. pyrifera, and L. nigrescens through the
B method showed higher viscosity for three macroalgae (M. pyrifera < L. use of two methodologies to obtain calcium alginate (10.55–16.75%
nigrescens < D. antarctica). extraction yield) and sodium alginate (12.45–20.8% extraction yield),
Jeva-Shakila et al. [40] determined viscosity in a digital viscometer allows us to identify sodium alginate extraction as the best methodol­
at 60 rpm in different gelatine solutions (66.7 g/L) obtained values ogy. Into this methodology, we evaluate the viscosity from the three
between 13.8 and 18.5 cP. Meanwhile, Gao et al. [18] determined vis­ sodium alginate sources (D. antarctica, M. pyrifera, and L. nigrescens),
cosity of alginate, obtained from M. pyrifera using different pre- concluding that sodium alginate from D. antarctica shows both the
treatment extraction methods, in a digital viscometer at 50 rpm, highest extraction yield (20.8%) and viscosity (1201.4 cP) at 20 ◦ C for a
where values between 63.1 and 381.9 cP were reported. 40 g/L alginate solution.
Our values at 50 rpm (solution at 40 g/L) were 67.2 cP, 54.6 cP and
69.8 cP for D. antarctica, M. pyrifera, and L. nigrescens, respectively for A CRediT authorship contribution statement
extraction method; and 1201.4 cP, 544.0 cP and 722.4 cP for B extrac­
tion method (Fig. 4). Corresponding Author(Eduardo Caballero):
It's not only the starch, or in this case the alginate, that expressed an -The corresponding author is responsible for conceptualization and
effect on viscosity. The viscosity is also influenced for proteins, soluble design of the study. Responsible for the research activity.
fiber, and lipids present in the sample. In addition, the complications to -Designthe methodology.
explain the increasing or decreasing of viscosity values, is given by the -Performing the experiments.
specific characteristic of proteins [64]. Particle size is another variable -Formal analysis, graphics, application of statistical.
that impact on viscosity, hence, it takes more time for get fully wet by a -Preparation, creation, writing and editing the original draft
liquid in the case of bigger particles [64]. manuscripts.
In Fig. 4, we can see the difference between two methods tested with -Supervision.
the same macroalgae flours. In all cases, the conventional B method, -Acquisition of the financial support for the project leading to this
demonstrated the highest viscosity at room temperature and 40 g/L in publication.
distilled water. The comparison is validated for the results shown in Co-Author (Agustín Flores):
Fig. 3, which allow ensuring that under the conditions tested, the vis­ -Performing the experiments and data analysis.
cosity was already stabilized. Taking into account the B method, we can -Observation of draft manuscripts.
identify D. antarctica like the sodium alginate source with more potential Co-Author (Araceli Olivares):
to be applied in food formulation like thickener and increase the vis­ -Conceptualization and design of the study.
cosity, for example in vegetable soups or to give texture to low calories

7
E. Caballero et al.
-Formal analysis, graphics, application of statistical.
-Preparation, creation, writing and editing the original draft
manuscripts.
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.
Acknowledgements
This work was supported by CORFO project 19SN-112378 and co-
financed by “Empresa Portuaria de Valparaíso” (EPV). Macroalgae
samples were provided by “Sindicato de Trabajadores Independientes
Pescadores Artesanales y de Rivera de caleta Sud Americana”.
Statement of informed consent
No conflicts, informed consent, or human or animal rights are
applicable to this study.
References
[1] FAO, The State of World Fisheries and Aquaculture 2016. Contributing to Food
Security and Nutrition for All. Rome, 200 pp, http://www.fao.org/3/a-i5555e.pdf,
2016.
[2] V.A.T. Reis, A.C. Duarte, Analytical methodologies for arsenic speciation in
macroalgae: a critical review, Trends Anal. Chem. 102 (2018) 170–184, https://
doi.org/10.1016/j.trac.2018.02.003.
[3] B. Radke, L. Jewell, J. Namiesnik, Analysis of arsenic species in environmental
samples, Crit. Rev. Anal. Chem. 42 (2012) 162e183, https://doi.org/10.1080/
10408347.2011.634637.
[4] T. Llorente-Mirandes, M.J. Ruiz-Chancho, M. Barbero, R. Rubio, J.F. Lopez-
Sanchez, Measurement of arsenic compounds in littoral zone algae from the
Western Mediterranean Sea. Occurrence of arsenobetaine, Chemosphere 81 (2010),
867e875, https://doi.org/10.1016/j.chemosphere.2010.08.007.
[5] O. Díaz, Y. Tapia, O. Mu~noz, R. Montoro, D. Velez, C. Almela, Total and inorganic
arsenic concentrations in different species of economically important algae
harvested from coastal zones of Chile, Food Chem. Toxicol. 50 (2012) 744e749,
https://doi.org/10.1016/j.fct.2011.11.024.
[6] E. Ownsworth, D. Selby, C.J. Ottley, E. Unsworth, A. Raab, J. Feldmann, A.
D. Sproson, J. Kuroda, C. Faidutti, P. Bücker, Tracing the natural and
anthropogenic influence on the trace elemental chemistry of estuarine macroalgae
and the implications for human consumption, Sci. Total Environ. 685 (2019)
259–272, https://doi.org/10.1016/j.scitotenv.2019.05.263.
[7] Commission Directive, 2003/40/EC of 16 May, Off. J. Eur. Union (2003). http://
eur-lex.europa.eu/legal-content/en/ALL/?uri1/4CELEX%3A32003L0040.
[8] Commission Regulation (EU), 2015/1006 of 25 June 2015 amending regulation
(EC) no 1881/2006 as regards maximum levels of inorganic arsenic in foodstuffs,
Off. J. Eur. Union (2015). L 161/14–L 161/16, https://www.fsai.ie/uploadedFil
es/Reg2015_1006.pdf.
[9] D.I. Sánchez-Machado, J. López-Cervantes, J. López-Hernández, P. Paseiro-Losada,
J. Simal-Lozano, Determination of the uronic acid composition of seaweed dietary
fibre by HPLC, Biomed. Chromatogr. 18 (2004) 90–97, https://doi.org/10.1002/
bmc.297.
[10] H. Yaich, H. Garna, B. Bchir, S. Besbes, M. Paquot, A. Richel, C. Blecker, H. Attia,
Chemical composition and functional properties of dietary fibre extracted by
Englyst and Prosky methods from the alga Ulva lactuca collected in Tunisia, Algal
Res. 9 (2015) 65–73, https://doi.org/10.1016/j.algal.2015.02.017.
[11] D.I. Sánchez-Machado, J. López-Cervantes, J. López-Hernández, P. Paseiro-Losada,
Fatty acids, total lipid, protein and ash contents of processed edible seaweeds, Food
Chem. 85 (2004) 439–444, https://doi.org/10.1016/j.foodchem.2003.08.001.
[12] C.S. Kumar, P. Ganesan, P.V. Suresh, N. Bhaskar, Seaweeds as a source of
nutritionally beneficial compounds-a review, J. Food Sci. Technol. 45 (2008) 1–13.
[13] L.P. Gómez, C. Alvarez, M. Zhao, U. Tiwari, J. Curtin, M. Garcia-Vaquero, B.
K. Tiwari, Innovative processing strategies and technologies to obtain
hydrocolloids from macroalgae for food applications, Carbohydr. Polym. 248
(2020), 116784, https://doi.org/10.1016/j.carbpol.2020.116784.
[14] T. Chopin, A. Buschmann, The 22nd international seaweed symposium: academia
meets industry, J. Appl. Phycol. 29 (2017) 2155–2158, https://doi.org/10.1007/
s10811-017-1243-y.
[15] S.L. Holdt, S. Kraan, Bioactive compounds in seaweed: functional food applications
and legislation, J. Appl. Phycol. 23 (3) (2011) 543–597, https://doi.org/10.1007/
s10811-010-9632-5.
[16] A. Synytsya, J. Čopíková, W.J. Kim, Y.I. Park. Cell wall polysaccharides of marine
algae, in: Kim, S.-K. Springer Handbook of Marine Biotechnology. pp. 543–590. htt
ps://hdl.handle.net/10.1007/978-3-642-53971-8_22.
8
Algal Research 57 (2021) 102349
[17] V. Stiger-Pouvreau, N. Bourgougnon, E. Deslandes, Chapter 8: carbohydrates from
seaweeds, in: Seaweed in Health and Disease Prevention, 2016, pp. 223–274,
https://doi.org/10.1016/B978-0-12-802772-1.00008-7.
[18] F. Gao, X. Liu, W. Chen, W. Guo, L. Chen, D. Li, Hydroxyl radical pretreatment for
low-viscosity sodium alginate production from brown seaweed, Algal Res. 34
(2018) 191–197, https://doi.org/10.1016/j.algal.2018.07.017.
[19] R.A. Khajouei, J. Keramat, N. Hamdami, A. Ursu, C. Delattre, C. Laroche,
C. Gardarin, D. Lecerf, J. Desbrières, G. Djelveh, P. Michaud, Extraction and
characterization of an alginate from the Iranian brown seaweed Nizimuddinia
zanardini, Int. J. Biol. Macromol. 118 (2018) 1073–1081, https://doi.org/10.1016/
j.ijbiomac.2018.06.154.
[20] S. Roohinejad, M. Koubaa, F.J. Barba, S. Saljoughian, M. Amid, R. Greiner,
Application of seaweeds to develop new food products with enhanced shelf-life,
quality and health-related beneficial properties, Food Res. Int. 99 (2017)
1066–1083, https://doi.org/10.1016/j.foodres.2016.08.016.
[21] S. Kraan, Algal polysaccharides, novel applications and outlook, in: Carbohydrates
Comprehensive Studies on Glycobiology and Glycotechnology, 2012, https://doi.
org/10.5772/51572.
[22] D.S. Lakshmi, N. Trivedi, C. Reddy, Synthesis and characterization of seaweed
cellulose derived carboxymethyl cellulose, Carbohydr. Polym. 157 (2017)
1604–1610, https://doi.org/10.1016/j.carbpol.2016.11.042.
[23] Y. Qin, Applications of bioactive seaweed substances in functional food products,
in: Bioactive Seaweeds for Food Applications, 2018, pp. 111–134, https://doi.org/
10.1016/B978-0-12-813312-5.00006-6.
[24] F. Sedlmeyer, Xylan as by-product of biorefineries: characteristics and potential use
for food applications, Food Hydrocoll. 25 (8) (2011) 1891–1898, https://doi.org/
10.1016/j.foodhyd.2011.04.005.
[25] B. Wolf, Polysaccharide functionality through extrusion processing, Curr. Opin.
Colloid Interface Sci. 15 (1–2) (2010) 50–54, https://doi.org/10.1016/j.
cocis.2009.11.011.
[26] M. Garcia-Vaquero, M. Lopez-Alonso, M. Hayes, Assessment of the functional
properties of protein extracted from the brown seaweed Himanthalia elongata
(Linnaeus) SF Gray, Food Res. Int. 99 (2017) 971–978, https://doi.org/10.1016/j.
foodres.2016.06.023.
[27] Z. Wu, J. Wu, R. Zhang, S. Yuan, Q. Lu, Y. Yu, Colloid properties of hydrophobic
modified alginate: surface tension, ζ-potential, viscosity and emulsification,
Carbohydr. Polym. 181 (2018) 56–62, https://doi.org/10.1016/j.
carbpol.2017.10.052.
[28] H.P. Calumpong, A.P. Maypa, M. Magbanua, Population and alginate yield and
quality assessment of four Sargassum species in Negros Island, central Philippines,
Hydrobiologia. 398/399 (1999) 211–215.
[29] M. Fertah, A. Belfkira, E. Dahmane, M. Taourirte, F. Brouillette, Extraction and
characterization of sodium alginate from Moroccan Laminaria digitata brown
seaweed, Arab. J. Chem. 10 (2017) S3707–S3714, https://doi.org/10.1016/j.
arabjc.2014.05.003.
[30] M.L. Torres, J.M. Fernandez, F.G. Dellatorre, A.M. Cortizo, T.G. Oberti, Purification
of alginate improves its biocompatibility and eliminates cytotoxicity in matrix for
bone tissue engineering, Algal Res. 40 (2019), 101499, https://doi.org/10.1016/j.
algal.2019.101499.
[31] M. Torres, A. Sousa, E. Silva, D. Melo, J. Feitosa, R. De Paula, M. Lima, Extraction
and physicochemical characterization of Sargassum vulgare alginate from Brazil,
Carbohydr. Res. 342 (14) (2007) 2067–2074, https://doi.org/10.1016/j.
carres.2007.05.022.
[32] P. Yu, X. Chao, Statistics-based optimization of the extraction process of kelp
polysaccharide and its activities, Carbohydr. Polym. 91 (2013) 356–362, https://
doi.org/10.1016/j.carbpol.2012.08.043.
[33] M. Garcia-Vaquero, G. Rajauria, B. Tiwari, Conventional extraction techniques:
Solvent extraction, in: M.-D. Torres, S. Kraan, H. Dominguez (Eds.), Sustainable
Seaweed Technologies. Cultivation, Biorefinery and Applications, Elsevier, 2020.
[34] M. Garcia-Vaquero, V. Ummat, B. Tiwari, G. Rajauria, Exploring ultrasound,
microwave and ultrasound–microwave assisted extraction technologies to increase
the extraction of bioactive compounds and antioxidants from brown macroalgae,
Mar. Drugs 18 (3) (2020) 172, https://doi.org/10.3390/md18030172.
[35] H. Khalil, T. Lai, Y. Tye, S. Rizal, E. Chong, S. Yap, M. Paridah, A review of
extractions of seaweed hydrocolloids: properties and applications, Express Polym
Lett 12 (4) (2018), https://doi.org/10.3144/expresspolymlett.2018.27.
[36] M. Garcia-Vaquero, J.V. O’Doherty, B.K. Tiwari, T. Sweeney, G. Rajauria,
Enhancing the extraction of polysaccharides and antioxidants from macroalgae
using sequential hydrothermal-assisted extraction followed by ultrasound and
thermal technologies, Mar. Drugs 17 (8) (2019) 457, https://doi.org/10.3390/
md17080457.
[37] AOAC, Official Methods of Analysis, Association of Official Analytical Chemists,
Washington DC, USA, 1995.
[38] C. Soto-Maldonado, M. Vergara-Castro, J. Jara-Quezada, E. Caballero-Valdés,
A. Müller-Pavez, M.E. Zúñiga-Hansen, C. Altamirano, Polyphenolic extracts of
walnut (Juglans regia) green husk containing juglone inhibit the growth of HL-60
cells and induce apoptosis, Electron. J. Biotechnol. 39 (2019) 1–7, https://doi.org/
10.1016/j.ejbt.2019.02.001.
[39] B. Racionero-Gómez, A.D. Sproson, D. Selby, A. Gannoun, D.R. Gröcke, H.
C. Greenwell, K.W. Burton, Osmium uptake, distribution, and 187Os/188Os and
187
Re/188Os compositions in Phaeophyceae macroalgae, Fucus vesiculosus:
implications for determining the 187Os/188Os composition of seawater, Geochim.
Cosmochim. Acta 199 (2017) 48–57, https://doi.org/10.1016/j.gca.2016.11.033.
[40] R. Jeya-Shakila, E. Jeevithan, A. Varatharajakumar, G. Jeyasekaran, D. Sukumar,
Functional characterization of gelatin extracted from bones of red snapper and
E. Caballero et al.
grouper in comparison with mammalian gelatin, LWT Food Sci. Technol. 48 (2012)
30–36, https://doi.org/10.1016/j.lwt.2012.03.007.
[41] G. Hernández-Carmona, R. Reyes-Tisnado, Y.E. Redríguez-Montesinos, J.I. Murillo-
Álvarez, D.L. Arvizu-Higuera, M. Muñoz-Ochoa, Technological advance for
alginate production in Mexico, Ingeniería Investigación y Tecnología. XIII (2)
(2012) 155–168.
[42] A. Mansilla, M. Ávila, Using Macrocystis pyrifera (L.) C. Agardh from southern Chile
as a source of applied biological compounds, Braz. J. Pharmacogn. 21 (2) (2011)
262–267, https://doi.org/10.1590/S0102-695X2011005000072.
[43] J. Ortiz, N. Romero, P. Robert, J. Araya, J. Lopez- Hernández, C. Bozzo,
E. Navarrete, A. Osorio, A. Rios, Dietary fiber, amino acid, fatty acid and
tocopherol contents of the edible seaweeds Ulva lactuca and Durvillaea antarctica,
Food Chem. 99 (1) (2006) 98–104, https://doi.org/10.1016/j.
foodchem.2005.07.027.
[44] P. Matanjun, S. Mohamed, N.M. Mustapha, K. Muhammad, Nutrient content of
tropical edible seaweeds, Eucheuma cottonii, Caulerpa lentillifera and Sargassum
polycystum, J. Appl. Phycol. (2009) 75–80, https://doi.org/10.1007/s10811-008-
9326-4.
[45] C. Reilly, Metal Contamination of Food: Its Significance for Food Quality and
Human Health, third ed., Blackwell Science Ltd, 2002.
[46] R.M. Welch, The impact of mineral nutrients in food crops on global human health,
Plant Soil 247 (2002) 83–90, https://doi.org/10.1023/A:1021140122921.
[47] G.B. Brito, L.S.G. Teixeira, M.G.A. Korn, Direct analysis of marine macroalgae for
determination of macro minerals by energy dispersive X-ray fluorescence,
Microchem. J. 134 (2017) 35–40, https://doi.org/10.1016/j.microc.2017.05.001.
[48] V. Besada, J.M. Andrade, F. Schultze, J.J. González, Heavy metals in edible
seaweeds commercialized for human consumption, J. Mar. Syst. 75 (1–2) (2009)
305–313, https://doi.org/10.1016/j.jmarsys.2008.10.010.
[49] IARC, Arsenic and Arsenic Compounds, International Agency for Research on
Cancer, 2012.
[50] S. Ichikawa, S. Nozawa, K. Hanaoka, T. Kaise, Ingestion and excretion of arsenic
compounds present in edible brown algae, Hijikia fusiforme, by mice, Food Chem.
Toxicol. 48 (2) (2010) 465–469, https://doi.org/10.1016/j.fct.2009.09.037.
[51] N. Khan, K.Y. Ryu, J.Y. Choi, E.Y. Nho, G. Habte, H. Choi, M.H. Kim, K.S. Park, K.
S. Kim, Determination of toxic heavy metals and speciation of arsenic in seaweeds
from South Korea, Food Chem. 169 (2015) 464–470, https://doi.org/10.1016/j.
foodchem.2014.08.020.
[52] J.M. Ronan, D.B. Stengel, A. Raab, J. Feldmann, L. O'Hea, E. Bralatei, E. McGovern,
High proportions of inorganic arsenic in Laminaria digitata but not in Ascophyllum
nodosum samples from Ireland, Chemosphere 186 (2017) 17–23, https://doi.org/
10.1016/j.chemosphere.2017.07.076.
Algal Research 57 (2021) 102349
[53] M. Rose, J. Lewis, N. Langford, M. Baxter, S. Origgi, M. Barber, H. MacBain,
K. Thomas, Arsenic in seaweed–forms, concentration and dietary exposure, Food
Chem. Toxicol. 45 (2007) 1263–1267, https://doi.org/10.1016/j.fct.2007.01.007.
[54] K. Yokoi, A. Konomi, Toxicity of the so-called edible hijiki seaweed (Sargassum
fusiforme) containing inorganic arsenic, Regul. Toxicol. Pharmacol. 63 (2012)
291–297, https://doi.org/10.1016/j.yrtph.2012.04.006.
[55] V.F. Taylor, Z. Li, V. Sayarath, T. Palys, K. Morse, R. Scholz-Bright, M. Karagas,
Distinct arsenic metabolites following seaweed consumption in humans, Sci. Rep. 7
(2017) 3920, https://doi.org/10.1038/s41598-017-03883-7.
[56] C. Almela, S. Algora, V. Benito, J. Clemente, V. Devesa, M.A. Súñer, D. Vélez,
R. Montoro, Heavy metal, total arsenic, and inorganic arsenic contents of algae
food products, J. Agric. Food Chem. 50 (2002) 918–923, https://doi.org/10.1021/
jf0110250.
[57] M. Bartel, F. Ebert, L. Leffers, U. Karst, T. Schwerdtle, Toxicological
characterization of the inorganic and organic arsenic metabolite Thio-DMA in
cultured human lung cells, J. Toxicol. (2011) (2011) 373141, https://doi.org/
10.1155/2011/373141.
[58] K. Hanaoka, K. Yosida, M. Tamano, T. Kuroiwa, T. Kaise, S. Maeda, Arsenic in the
prepared edible brown alga hijiki, Hizikia fusiforme, Appl. Organomet. Chem. 15 (6)
(2001) 561–565, https://doi.org/10.1002/aoc.195.
[59] C. Almela, M. Clemente, D. Vélez, R. Montoro, Total arsenic, inorganic, lead and
cadmium contents in edible seaweed sold in Spain, Food Chem. Toxicol. 44 (2006)
1901–1908, https://doi.org/10.1016/j.fct.2006.06.011.
[60] C. Gutiérrez, H.K. Hansen, P. Hernández, C. Pinilla, Biosorption of cadmium with
brown macroalgae, Chemosphere. 138 (2015) 164–169, https://doi.org/10.1016/
j.chemosphere.2015.06.002.
[61] Y.V. Yuan, Antioxidants from edible seaweeds, in: Antioxidant Measurement and
Applications. ACS Symposium Series, 2017, pp. 268–301, https://doi.org/
10.1021/bk-2007-0956.ch019. Chapter 19pp.
[62] S. Kadam, C. O’Donnell, D. Rai, M. Hossain, C. Burgess, D. Walsh, B. Tiwari,
Laminarin from Irish brown seaweeds Ascophyllum nodosum and Laminaria
hyperborea: ultrasound assisted extraction, characterization and bioactivity, Mar.
Drugs 13 (7) (2015) 4270–4280, https://doi.org/10.3390/md13074270.
[63] N. Haas, Optimizing Wheat Blends for Customer Value Creation: A Special Case of
Solvent Retention Capacity, Thesis, Kansas State University, Manhattan, 2006, htt
p://krex.k-state.edu/dspace/bitstream/handle/2097/8387/NikolasHaas2011.pdf.
[64] G. Crosbie, A. Ross, The RVA Handbook. 1, AACC International, Minnesota, 2007,
https://doi.org/10.1002/star.200790062.
[65] S. Berggren, Water Holding Capacity and Viscosity of Ingredients From Oats,
Bachelor thesis project in chemistry, Linnaeus University, Sweden, 2017.