Proximate Composition and Nutritional Value of
Three Macroalgae: Ascophyllum nodosum,
Fucus vesiculosus and Bifurcaria bifurcata
JoséM. Lorenzo 1, *ID , Rubén Agregán1, Paulo E. S. Munekata 2, Daniel Franco 1ID ,
Javier Carballo 3, Selin ¸Sahin 4, Ramón Lacomba 5and Francisco J. Barba 6, *ID
1Centro Tecnológico de la Carne de Galicia, Adva. Galicia n◦4, Parque Tecnológico de Galicia,
San Cibrao das Viñas
, 32900 Ourense, Spain; email@example.com (R.A.); firstname.lastname@example.org (D.F.)
Department of Food Engineering, Faculty of Animal Science and Food Engineering, University of São Paulo,
225 Duque de Caxias Norte Ave, Jardim Elite, Pirassununga, São Paulo 13.635-900, Brazil;
Area de Tecnologia de los Alimentos, Facultad de Ciencias de Ourense, Universidad de Vigo, 32004 Ourense,
4Department of Chemical Engineering, Engineering Faculty, Istanbul University, Avcilar, 34320 Istanbul,
5Grupo Alimentario Citrus (GAC), Avda. dels Gremis, Parcela 28 Pol. Ind. Sector 13 del Túria,
Riba-roja de Túria, 46394 València, Spain; email@example.com
6Nutrition and Food Science Area, Preventive Medicine and Public Health, Food Science,
Toxicology and Forensic Medicine Department, Universitat de València, Avda. Vicent Andrés Estellés, s/n,
Burjassot, 46100 València, Spain
*Correspondence: firstname.lastname@example.org (J.M.L.); email@example.com (F.J.B.); Tel.: +34-988548277 (J.M.L.);
+34-963544972 (F.J.B.); Fax: +34-988548276 (J.M.L.); +34-963544954 (F.J.B.)
Received: 8 August 2017; Accepted: 8 November 2017; Published: 15 November 2017
Proximate composition (moisture, protein, lipid and ash content) and nutritional value (fatty
acid, amino acid and mineral proﬁle) of three macroalgae (Ascophyllum nodosum,Fucus vesiculosus and
Bifurcaria bifurcate) were studied. Chemical composition was signiﬁcantly (p< 0.001) different among
the three seaweeds. In this regard, the B. bifurcata presented the highest fat content (6.54% of dry
matter); whereas, F. vesiculosus showed the highest protein level (12.99% dry matter). Regarding fatty
acid content, the polyunsaturated fatty acids (PUFAs) were the most abundant followed by saturated
fatty acids (SFAs) and monounsaturated fatty acids (MUFAs). On the other hand, the three seaweeds
are a rich source of K (from 3781.35 to 9316.28 mg/100 g), Mn (from 8.28 to 1.96 mg/100 g), Na (from
1836.82 to 4575.71 mg/100 g) and Ca (from 984.73 to 1160.27 mg/100 g). Finally, the most abundant
amino acid was glutamic acid (1874.47–1504.53 mg/100 dry matter), followed by aspartic acid
(1677.01–800.84 mg/100 g dry matter) and alanine (985.40–655.73 mg/100 g dry matter).
Ascophyllum nodosum;Fucus vesiculosus;Bifurcaria bifurcate; seaweeds; fatty acid proﬁle;
amino acid content; minerals; chemical composition
Seaweeds have been traditionally consumed as food in many cultures, and have been used as
condiments, fertilizers, and as source of phycocolloids such as alginate, agar, and carrageenan for
industrial applications [
]. Seaweeds are traditionally divided into three main groups corresponding
to the phylum: green (Chlorophyta), red (Rhodophyta) and brown (Phaeophyta), depending on their
chemical composition and nutritional value .
Mar. Drugs 2017,15, 360; doi:10.3390/md15110360 www.mdpi.com/journal/marinedrugs
Mar. Drugs 2017,15, 360 2 of 11
Seaweeds are consumed in Asia as part of the daily diet. Nowadays, brown algae (66.5%) are the
most consumed species, followed by red (33%) and green (5%) algae. Today, Japan, China and South
Korea, are the countries with the greatest seaweed consumption [
]. Seaweeds are an excellent nutrient
source, containing high amounts of macro- and micronutrients [
], as well as bioactive compounds
(e.g., catechins such as gallocatechin, epicatechin and catechingallate, ﬂavonols, and ﬂavonol
glycosides) with high antioxidant and health beneﬁcial properties [
]. Food, pharmaceutical,
and cosmetic industries have shown interest in the recovery of antioxidant compounds (isolated
compounds and/or complex extract mixtures) assisted by conventional (solid-liquid or liquid-liquid
extraction, Soxhlet, etc.) and innovative processing technologies (high-pressure, supercritical-CO
), electrotechnologies, microwave- and ultrasound-assisted extraction, among others) [
Several authors [
] have reported that the chemical composition of seaweeds varies according
to maturity, habitats, environmental conditions, and species. A comprehensive study of nutritional
(protein and amino acids, fat and fatty acids, carbohydrates, minerals, and vitamins) and bioactive
compounds such as polyphenols, carotenoids, etc., from each seaweed, which can exert some beneﬁcial
properties on health, is necessary. There are many brown types of seaweeds found in the Spanish
coast and the details of their chemical and nutritional composition are needed in order to fulﬁl the
growing demand for Spanish seaweeds and their derived products. Thus, the aim of the present study
was to evaluate the chemical and nutritional properties of three different brown seaweeds A. nodosum,
F. vesiculosus and B. bifurcata from the Galician coast.
2. Results and Discussion
2.1. Chemical Composition of Seaweeds
The proximate composition of the three seaweeds is summarized in Table 1. As can be seen the
moisture content showed signiﬁcant (p< 0.001) differences among the three macroalgae, since the
lowest value was observed in B. bifurcata (7.95%).
Proximate composition of the three seaweeds studied (mean
standard deviation values)
A. nodosum F. vesiculosus B. bifurcata
Moisture (g/100 g algae) 11.08 ±0.53 a11.23 ±0.08 a7.95 ±0.06 b
Protein (g/100 g DW) 8.70 ±0.07 a12.99 ±0.04 b8.92 ±0.09 c
Lipid (g/100 g DW) 3.62 ±0.17 a3.75 ±0.20 a6.54 ±0.27 b
Ash (g/100 g DW) 30.89 ±0.06 a20.71 ±0.04 b31.68 ±0.41 c
DW: dry weight of seaweed.
Means in the same row not followed by a common superscript letter are signiﬁcantly
different (p< 0.05; Duncan test).
This ﬁnding is in close agreement with the data reported by Rodrigues et al. [
], who also noticed
that the moisture content of different edible seaweeds species ranged from 8.0 g/100 g of dry weight
(DW) in dried Gracilaria gracilis to 11.8 g/100 g of DW in dried Osmundea pinnatiﬁda. In addition,
Gómez-Ordoñez et al. [
] also reported similar moisture contents (between 6.64% and 9.86%) in
several edible seaweeds from the northwestern Spanish coast. However, Chan & Matanjun [
lower moisture content (5.32%) in freeze-dried Gracilaria changii seaweed.
F. vesiculosus specie presented the highest protein content (12.99 g/100 DW), followed by
B. bifurcata (8.92 g/100 DW) and the A. nodosum (8.70 g/100 DW). These results are in agreement with
the data reported by Fleurence [
], who also noticed low protein content (<15 g/100 DW) in most of
the brown seaweeds industrially exploited (F. vesiculosus,A. nodosum,Laminaria digitata and Himanthalia
elongata). Similar values were found by Gómez-Ordoñez et al. [
] and Alves et al. [
] in B. bifurcata
(10.92 g/100 DW and 8.57 g/100 g DW, respectively) and by Chan & Matanjun [
] in G. changii
Mar. Drugs 2017,15, 360 3 of 11
(12.57 g/100 DW). However, these values were lower than those obtained by
Rodrigues et al. 
for brown (14.4–16.9 g/100 DW), red (20.2–23.8 g/100 DW) and green (18.8 g/100 DW) seaweed
species. In addition, our values were lower than those observed by Fleurence [
] in other seaweed
species such as Porphyra tenera (47 g/100 DW) and Palmaria palmata (35 g/100 DW). On the contrary,
Sánchez-Machado, López-Cervantes, et al. [
] obtained lower protein content (5.46 g/100 DW) in
H. elongata dried seaweed. According to Denis et al. [
], the protein content of seaweed changes
during the year, having the maximum content during winter and the beginning of spring, and the
minimum content during summer and early autumn periods. In addition, the protein level varied
among different algal species, geographic areas, seasons, or environmental conditions .
In general, seaweeds exhibit low fat content (bellow 4%) [
], which varies signiﬁcantly through
the year [
]. Extractable lipid showed signiﬁcant (p< 0.001) differences among seaweeds, since the
highest levels were observed in B. bifurcata (6.54% DW). Our values were similar to those reported
by Peinado et al. [
], who found the contents ranging from 3.95 to 4.64% DW in F. vesiculosus and by
Gómez-Ordoñez et al. [
] and Alves et al. [
], who observed fat levels of 5.67% DW and 5.81% DW
in B. bifurcata, respectively. On the other hand, ash contents were high and ranged from 20.71% DW
to 31.68% DW for F. vesiculosus and B. bifurcata, respectively. These ﬁndings are in agreement with
the data reported by Alves et al. [
] and Gómez-Ordoñez et al. [
] in B. bifurcata (34.31% DW and
30.15% DW, respectively) and by Peinado et al. [
] in F. vesiculosus (21–19% DW). The high ash levels
constitute an important characteristic of seaweeds, and are higher than those observed in terrestrial
vegetables . It is known that high amounts of ash are linked with high levels of minerals.
2.2. Mineral Content of Seaweeds
The mineral content of the three macroalgae is given in Table 2. Among the macrominerals,
K (3781.35–9316.28 mg/100 g DW) was the most abundant element in the three seaweeds studied,
followed by Na (1836.82–4575.71 mg/100 g DW) and Ca (984.73–1160.27 mg/100 g DW). A similar
trend was reported by other authors [
], who found that K was the main mineral element
followed by Na. On the other hand, B. bifurcata presented a Na/K ratio lower than that observed in the
other seaweeds (0.19 vs. 0.58 vs. 1.21, for the B. bifurcata,F. vesiculosus and A. nodosum, respectively),
which is really interesting from the nutritional viewpoint, because high Na/K ratio diets and the
hypertension incidence are closely linked [
]. Thus, B. bifurcata could be useful for the regulation of
the Na/K ratio of diets. In addition, Rodrigues et al. [
] suggested that seaweeds with low ratios of
Na/K are useful as salt replacers.
The values of manganese in the three macroalgae ranged from 528.04 mg/100 g DW to
867.82 mg/100 g DW, for B. bifurcata and A. nodosum, respectively, differing signiﬁcantly (p< 0.001)
among species. These values were higher than those reported by Chan et al. [
] in G. changii
(436.13 mg/100 g DW) and lower than the data previously found by Rodrigues et al. [
] in Sargassum
muticum (1504 mg/100 g DW) and Codium tomentosum (1046 mg/100 g DW).
On the other hand, Ca contents also showed signiﬁcant (p< 0.001) differences among seaweeds,
showing the highest Ca level in F. vesiculosus (1160.27 mg/100 DW). In this regard, Moreiras et al. [
noticed that Wakame and Sea Spaghetti seaweed species contained approximately eight times more Ca
than milk and they could be an excellent source of Ca for the prevention and treatment of osteoporosis,
for growing children, and for pre- and post-menopausal women. Phosphorous, the least abundant
macromineral, was also detected in F. vesiculosus and B. bifurcata, ranging from 169.54 mg/100 g DW to
193.57 mg/100 g DW for F. vesiculosus and B. bifurcata, respectively.
A. nodosum and F. vesiculosus also contained iron (ranged from 13.34 mg/100 g DW to
18.99 mg/100 g DW) and Mg (from 1.96 mg/100 DW to 8.28 mg/100 g DW). Our Fe values were
higher than those obtained by Rupérez [
] for Porphyra tenera (10.3 mg/100 g DW), but less than those
found by Rao et al. [
] for Porphyra vietnamensis (33 mg/100 g DW). In this regard, F. vesiculosus can
be a useful to provide the daily intake of iron and to prevent the anemia caused by iron deﬁciency [
Mar. Drugs 2017,15, 360 4 of 11
Table 2. Mineral proﬁle of the three seaweeds studied (mean ±standard deviation values) (n= 5).
Minerals (mg/100 g DW) Seaweed
A. nodosum F. vesiculosus B. bifurcata
Ca 984.73 ±47.26 a1160.27 ±23.10 b996.42 ±12.83 a
Fe 13.34 ±0.90 a18.99 ±0.32 bn.q.
K 3781.35 ±13.40 a3745.05 ±36.01 a9316.28 ±101.94 b
Mg 867.82 ±12.01 a732.37 ±5.35 b528.04 ±8.25 c
Mn 1.96 ±0.69 a8.28 ±1.07 bn.q.
Na 4575.71 ±50.05 a2187.51 ±36.90 b1836.82 ±52.12 c
P n.q. 193.57 ±1.13 a169.54 ±1.41 b
Zn n.q. n.q. n.q.
Cu n.q. n.q. n.q.
Total 10,224.91 ±64.32 a8045.96 ±94.44 b12,848.97 ±142.01 c
n.q. = not quantiﬁed. DW: dry weight of seaweed.
Means in the same row not followed by a common superscript
letter are signiﬁcantly different (p< 0.05; Duncan test).
2.3. Amino Acid Content of Seaweeds
The amino acid (AA) composition of the three seaweeds evaluated is summarized in Table 3.
The total AA contents were 7.48, 11.90 and 7.32 g/100 g DW (p< 0.001), for A. nodosum,F. vesiculosus,
and B. bifurcata, respectively; and these values were comparable to corresponding crude protein levels
(Table 1), thus showing that the amount of non-protein nitrogenous materials in these seaweeds
Amino acid proﬁle of the three seaweeds studied (mean
standard deviation values) (n= 5).
Amino Acids (mg/100 g DW) Seaweed
A. nodosum F. vesiculosus B. bifurcata
Essential amino acids
Threonine 363.22 ±17.12 a613.08 ±33.62 b360.27 ±38.25 a
Valine 353.89 ±32.95 a582.70 ±36.73 b372.82 ±49.05 a
Methionine 147.59 ±18.71 a218.21 ±20.20 b178.41 ±18.08 a
Isoleucine 295.26 ±25.73 a507.82 ±32.42 b299.73 ±37.74 a
Leucine 537.37 ±38.87 a862.14 ±57.02 b524.59 ±61.38 a
Phenylalanine 340.13 ±17.74 a541.53 ±25.72 b330.05 ±32.32 a
Lysine 431.72 ±38.40 a800.28 ±74.20 b393.06 ±56.57 a
Histidine 126.46 ±10.65 a194.59 ±8.73 b138.76 ±12.70 a
Arginine 316.79 ±14.05 a557.87 ±38.44 b330.11 ±42.41 a
Total EAA 2912.42 ±204.93 a4878.22 ±304.12 b2927.79 ±346.84 a
Non-essential amino acids
Tyrosine 162.85 ±24.50 a327.01 ±30.59 b175.00 ±30.90 a
Asparagine 846.64 ±38.87 a1677.01 ±156.39 b800.84 ±105.55 a
Serine 378.62 ±13.57 ab 630.54 ±47.00 a357.10 ±36.87 b
Glutamic acid 1714.55 ±133.17 a1974.47 ±150.67 b1504.53 ±178.74 a
Glycine 417.70 ±12.89 a651.24 ±30.84 b390.14 ±29.42 a
Alanine 655.73 ±34.75 a985.40 ±69.50 b846.65 ±82.87 c
Proline 399.24 ±11.70 a575.19 ±39.15 b318.40 ±40.96 c
Cysteine 0.00 ±0.00 a205.23 ±25.43 b0.00 ±0.00 a
Total NEAA 4575.33 ±198.91 a7026.10 ±512.60 b4392.67 ±502.38 a
Total AA 7487.76 ±400.31 a11,904.32 ±816.67 b7320.46 ±848.14 a
Relative Amount EAA (%) 38.87 ±0.71 a40.99 ±0.26 b39.99 ±0.31 c
DW: dry weight of seaweed.
Means in the same row not followed by a common superscript letter are signiﬁcantly
different (p< 0.05; Duncan test). EAA: Essential Amino acids.
Mar. Drugs 2017,15, 360 5 of 11
The three seaweeds studied contained all the essential amino acids (EAAs) (excluding cysteine
in the A. nodosum and B. bifurcata). The EAAs content ranged from 3075.28 mg/100 g DW
5205.23 mg/100 g
DW for the A. nodosum and F. vesiculosus, respectively, showing signiﬁcant
differences among species. The EAA/total AA ratio suggests that more than 40% of the AAs were
EAAs. This ratio was lower than the data reported by Chan et al. [
], who observed the ratios (above
55%) in G. changii, but comparable to Porphyra umbilicalis (36.87%), Undaria pinnatiﬁda (42.72%) and
H. elongata (40.82%) reported by Cofrades et al. [
]. In the essential fraction, leucine was the most
abundant, ranging from 524.59 mg/100 g DW to 862.14 mg/100 g DW for B. bifurcata and F. vesiculosus,
respectively, followed by lysine (393.06–800.28 mg/100 g DW), threonine (360.27–613.08 mg/100 g
DW) and valine (353.89–582.70 mg/100 g DW). These ﬁndings were not in agreement with those
reported by Chan et al. [
], who observed that arginine was found to be the highest EAA in G. changii,
representing 18.69% of the total AAs. On the other hand, glutamic and aspartic acids were the major
amino acids found in the non-essential fraction and theses two AAs accounted between 30.67% and
34.20% of the total AAs, for the FV and AN species, respectively. The sum of aspartic and glutamic
acids was higher than data reported by other authors [
] who found values below 25% in different
seaweed species. According to Saini et al. [
], the special ﬂavor and taste of seaweeds in linked to
the glutamic and aspartic acids contents. The next highest NEEA were alanine > glycine > serine >
proline. Finally, the protein quality of FV seaweed is better than those the other ones, because cysteine
is lacking in the AN and BB species.
The nutritional quality of the three seaweeds studied is shown in Table 4. The chemical score
(CS) for each of the essential amino acids with respect to the pattern protein, as proposed by Food and
Agriculture Organization of the United Nations (FAO)/World Health Organization (WHO)/United
Nations (UNU)  for humans (children > 1-year old and adults) was calculated.
Table 4. Nutritional quality of protein for the three seaweeds studied.
Amino Acid IOM/FNB
A. nodosum (CS) F. vesiculosus (CS) B. bifurcata (CS)
Histidine 1.8 1.5 96.8 99.8 103.7
Isoleucine 2.5 3.0 113.1 130.3 112.0
Leucine 5.5 5.9 104.7 112.5 99.7
Lysine 5.1 4.5 110.3 136.9 97.9
Met + Cys 2.5 1.6 105.9 203.8 125.0
Phe + Tyr 4.7 3.8 152.1 176.0 149.0
Threonine 2.7 2.3 181.5 125.0 175.6
Valine 3.2 3.9 104.3 115.0 107.2
IAEE 118.4 133.9 118.8
Seaweeds: A. nodosum =Ascophyllum nodosum;F. vesiculosus =Fucus vesiculosus; and B. bifurcate =Bifurcaria bifurcata.
Pattern proteins are expressed in (g/100 g protein). Values of CS and IEAA (Index Essential Amino Acids) are
referred only respect to FAO/WHO/UNU (2007) protein pattern.
The proﬁle of the Institute of Medicine, Food and Nutrition (FNB) [
] is also shown for
comparative purposes. The analysis of the CS allows the order of the restrictive amino acids to
be determined. Concentration of all the essential amino acids were above the FAO/WHO/UNU 
except for histidine in the A. nodosum and F. vesiculosus and leucine and lysine in B. bifurcata. Thus,
histidine was the most limiting AA found in A. nodosum and F. vesiculosus and lysine seemed to be the
limiting AA in B. bifurcata. This is in agreement with the data found by Cofrades et al. [
], who found
that the most limiting AA in the brown seaweeds was lysine. However, Chan et al. [
] observed that
methionine was the most limiting AA found in G. changii.
2.4. Fatty Acid Proﬁle of Seaweeds
Table 5shows the fatty acid proﬁle of the three seaweeds studied. The polyunsaturated
fatty acids (PUFAs) were the most abundant, ranging from 43.47% to 48.19% for the A. nodosum
Mar. Drugs 2017,15, 360 6 of 11
and F. vesiculosus, respectively. This result is in agreement with the data previously reported by
, who found that PUFAs were the main fatty acids in seaweeds. However,
Pen et al. 
Maehre et al. 
observed higher saturated fatty acid (SFA) content in different
In the present study, the percentage of fatty acid differed signiﬁcantly (p< 0.001) among seaweeds.
In this regard, the highest oleic acid (C18:1n-9) content (27.83–19.94%) was found in A. nodosum and
F. vesiculosus, whereas B. bifurcata presented the highest arachidonic acid (C20:4n-6) level (15.24%).
A similar trend was reported by Peinado et al. [
] and Ortiz et al. [
], who observed that oleic acid was
the main fatty acid in seaweed samples. On the contrary, Chan et al. [
Alves et al. 
that docosahexaenoic acid (C22:6n-3; DHA) and palmitic acid (C16:0) were the most abundant fatty
acids in G. changgi and B. bifurcata, respectively. These differences on the fatty acid proﬁle could be due
to differences among species, as well as other abiotic factors such as light, salinity, and nutrients .
Table 5. Fatty acid proﬁle of the three seaweeds studied (mean ±standard deviation values) (n= 5).
Fatty Acids Seaweed
A. nodosum F. vesiculosus B. bifurcata
C14:0 9.40 ±0.11 a11.38 ±0.11 b4.52 ±0.46 c
C14:1n-5 0.28 ±0.00 a0.10 ±0.00 b0.00 ±0.00 c
C15:0 0.30 ±0.00 a0.37 ±0.00 b0.17 ±0.01 c
C16:0 13.42 ±0.46 a14.66 ±0.36 b17.35 ±0.43 c
C16:1n-7 2.24 ±0.01 a1.18 ±0.02 b2.51 ±0.16 c
C17:0 0.41 ±0.14 a0.82 ±0.15 b0.54 ±0.02 a
C17:1n-7 0.29 ±0.00 a0.20 ±0.00 b1.87 ±0.07 c
C18:0 0.76 ±0.01 a1.06 ±0.08 b1.75 ±0.13 c
C18:1n-11 trans 0.00 ±0.00 a0.00 ±0.00 a3.57 ±0.13 b
C18:1n-9 cis 27.83 ±0.26 a19.94 ±0.31 b12.61 ±0.35 c
C18:1n-7 cis 0.45 ±0.05 a0.39 ±0.04 a0.52 ±0.03 b
C18:2n-6 trans 0.11 ±0.00 a0.06 ±0.00 a5.68 ±0.21 b
C18:2n-6 cis 7.47 ±0.12 a6.43 ±0.08 b1.92 ±0.06 c
C20:0 0.22 ±0.01 a0.39 ±0.01 b1.89 ±0.18 c
C18:3n-6 0.54 ±0.01 a0.56 ±0.01 a0.42 ±0.05 b
C20:1n-9 0.07 ±0.01 a0.53 ±0.01 b4.18 ±0.12 c
C18:3n-3 4.45 ±0.03 a7.59 ±0.11 b3.97 ±0.09 c
C18:2n-7 (CLA) 0.00 ±0.00 a0.00 ±0.00 a0.87 ±0.10 b
C21:0 0.00 ±0.00 a0.00 ±0.00 a0.71 ±0.07 b
C20:2n-6 5.05 ±0.02 a6.46 ±0.09 b1.44 ±0.01 c
C22:0 0.22 ±0.00 a0.22 ±0.00 a0.34 ±0.02 b
C20:3n-6 0.74 ±0.04 a0.69 ±0.02 b0.42 ±0.04 c
C22:1n-9 0.00 ±0.00 a0.00 ±0.00 a0.73 ±0.04 b
C20:3n-3 0.33 ±0.01 a0.21 ±0.00 b0.00 ±0.00 c
C20:4n-6 17.25 ±0.26 a15.86 ±0.24 b15.24 ±0.37 c
C22:2n-6 0.29 ±0.01 a0.39 ±0.01 b1.76 ±0.09 c
C20:5n-3 7.24 ±0.08 a9.94 ±0.14 b4.09 ±0.08 c
C24:0 0.41 ±0.00 a0.36 ±0.01 b0.34 ±0.03 b
C24:1n-9 0.00 ±0.00 a0.00 ±0.00 a0.53 ±0.06 b
C22:6n-3 0.00 ±0.00 a0.00 ±0.00 a11.10 ±1.13 b
SFA 25.14 ±0.49 a29.26 ±0.34 b27.62 ±0.77 c
MUFA 31.15 ±0.23 a22.33 ±0.33 b26.51 ±0.48 c
PUFA 43.47 ±0.54 a48.19 ±0.62 b46.91 ±1.37 b
n-3 12.02 ±0.11 a17.74 ±0.25 b19.16 ±1.03 c
n-6 31.45 ±0.42 a30.44 ±0.38 b26.87 ±0.48 c
n-6/n-3 2.62 ±0.01 a1.72 ±0.01 b1.41 ±0.07 c
Results expressed as percentage of total fatty acid analyzed.
means in the same row not followed by a common
superscript letter are signiﬁcantly different (p< 0.05; Duncan test). Saturated fatty acids: SFA. Monounsaturated
fatty acids: MUFA. Polyunsaturated fatty acids: PUFA.
Mar. Drugs 2017,15, 360 7 of 11
Eicosapentaenoic acid (EPA) (C20:5n-3) represented from 4.09 to 9.94% of the total fatty acids,
whereas docosahexaenoic acid (DHA) was only detected in B. bifurcata (11.10% of the total fatty
acids). Other studies reported similar EPA percentages in brown algae [
]. In another work,
Maehre et al. 
found that none of the algae contained DHA, whereas the EPA content varied
considerably among species.
On the other hand, Western country diets are deﬁcient in n-3 fatty acids due to the low seafood
consumption versus the high intake of n-6 fatty acid from vegetable oil. In this regard, the World Health
Organization (WHO) [
] recommended a n-6/n-3 ratio below 10. In our study, we observed n-6/n-3
ratio ranging from 2.62 to 1.41, placing the three macroalgae studied according WHO recommendations.
This outcome is in agreement with those reported by other authors [
] who found n-6/n-3 ratios
between 4.1 and 0.02.
3. Material and Methods
3.1. Algal Material
The brown seaweeds, A. nodosum,F. vesiculosus and B. bifurcata used in the present study, were
kindly supplied by Portomuiños Company (A Coruña, Spain). They were collected from August to
September 2015, in the Atlantic Ocean, in the area of Camariñas (A Coruña, Spain). The samples were
grinded to obtain powder with a particle size lower than 0.8 mm, using a conventional mincer. Then,
the seaweeds were passed through a 0.8 mm mesh sieve and stored under vacuum in plastics bags at
−20 ◦C until analysis.
3.2. Chemical Composition
Moisture, protein, and ash were determined following the ISO recommendations (ISO
], ISO 937:1978 [
], and ISO 936:1998 [
], respectively). Moisture content was determined
by measuring sample (3 g) weight loss at 105
C in an oven (Memmert UFP 600, Schwabach,
Germany), until constant weight. Kjeldahl total nitrogen method was used to determine protein
percentage (total nitrogen content was multiplied
6.25). Five hundred milligrams of seaweed were
subjected to reaction with H
O was employed as a catalyst) in a digester (Gerhardt
Kjeldatherm KB, Bonn, Germany), then the organic nitrogen was transformed into (NH
distilled in alkali condition (Gerhardt Vapodest 50 carroused, Bonn, Germany). Ash content was
assessed by determining seaweed (3 g) weight loss in a mufﬂe furnace (Carbolite RWF 1200, Hope
Valley, UK) at 600
C until constant weight. Lipids were determined using the method proposed by
Ortiz et al. 
with some modiﬁcations. Lipids from each seaweed (20 g) were extracted with 300 mL
O (1:2:0.8), overnight under dark condition. Then, 79 mL of chloroform and
79 mL of water were added to each sample, obtaining a ﬁnal solvent ratio of CHCl
of 1:1:0.9 by volume. NaCl (5%) was added and then, samples were centrifuged at 4000 rpm during
10 min. Chloroform phase was concentrated under vacuum condition in order to recover the lipids,
which were gravimetrically measured.
3.3. Amino Acid Content
Amino acids were extracted following the method proposed by Lorenzo et al. [
]. Amino acids
were derived using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (Waters AccQ-Fluor reagent
kit) and determined by RP-HPLC (Waters 2695 Separations Module + Waters 2475 Multi Fluorescence
Detector + Waters AccQ-Tag amino acids analysis column). The amino acids content was expressed in
mg/100 g of dry matter.
Mar. Drugs 2017,15, 360 8 of 11
3.4. Protein Quality: Chemical Score of Amino Acids
The chemical score (CS) of the essential amino acids was determined using a protein pattern
recommended by FAO/WHO/UNU [
] as reference protein applying the next equation (Equation (1)):
g EAA in tested protein
g EAA in pattern protein ×100 (1)
The essential amino acids index (EAA) value was also assessed according to the Equation (2) [
EAA =100 ×n
×. . . . . . j
a,b,c, . . . , j= content of Phe, Tyr, Val, Met, Thr, Lys, His, Ile and Leu in seaweeds.
ap,bp,cp, . . . , jp= content of Phe, Tyr, Val, Met, Thr, Lys, His, Ile and Leu in protein standard .
n= number of amino acids used.
3.5. Fatty Acid Proﬁle
The lipids extracted (50 mg) were used to determine fatty acid proﬁle. Total fatty acids
were transesteriﬁed using the method previously by Domínguez et al. [
]. A GC equipment
(GC-Agilent 6890 N; Agilent Technologies Spain, S.L., Madrid, Spain) with a ﬂame ionization
detector was used for the separation and quantiﬁcation of the fatty acids methyl esters (FAMEs)
using the chromatographic conditions proposed by Domínguez et al. [
]. Individual FAMEs were
identiﬁed by comparing their retention times with those of authentic standards (Supelco 37 component
FAME Mix, Sigma-Aldrich, Barcelona, Spain). C18:1n-7 cis (Supelco cis-11-Vaccenic methyl ester),
C18:1n-11 trans (trans-11-vaccenic methyl ester) and C18:2n-7 (CLA) (Matreya LLC Methyl 9(z),
11 (E)-octadecadienoate) were not included in the commercial mix. In addition, nonadecanoic acid
(C19:0) was used as internal standard, which was added to the samples prior to methylation. Data
were expressed in g/100 g of FAME.
3.6. Mineral Proﬁle
The ash samples obtained by ISO recommended standard method [
] were dissolved in 10 mL
of 1M HNO
. Mineral (Ca, Fe, K, Mg, Mn, Na, P, Zn and Cu) was determined by inductively coupled
plasma-optical emission spectroscopy (ICP-OES), using a Thermo-Fisher ICAP 6000 plasma emission
spectrometer (Thermo-Fisher, Cambridge, UK), following the method proposed by Lorenzo et al. [
All determinations were made in triplicate.
3.7. Statistical Analysis
The differences in proximate composition, amino acid, fatty acid and mineral proﬁles among the
three seaweeds studied were examined using an ANOVA test. Least-squares means were compared
among seaweeds using the Duncan’s post hoc test (signiﬁcance level p< 0.05). The values were given
in terms of mean values
standard deviations. All statistical analysis were performed using IBM
SPSS Statistics®21 software (IBM Corporation, Armonk, NY, USA).
Among the three seaweeds studied (A. nodosum,F. vesiculosus, and B. bifurcata),B. bifurcata
had the highest level of lipid and ash. It should also be noted that although B. bifurcata had the
highest total mineral and K contents, F. vesiculosus presented the highest Ca, Fe, Mn, and P contents,
while A. nodosum presented the highest Mg, and Na contents. This fact is of a great importance,
Mar. Drugs 2017,15, 360 9 of 11
especially when seaweeds are used to extract targeted minerals to be used in diets. F. vesiculosus
had the highest protein content. The three seaweeds studied contained all the essential amino acids
(excluding Cys in the A. nodosum and B. bifurcata). Glu and Asp acids were the predominant amino
acids found in the non-essential fraction and theses two amino acids accounted between 30.67% and
34.20% of the total amino acids, for the F. vesiculosus and A. nodosum, respectively. Concentration of
all the essential amino acids were above the chemical score established by FAO/WHO/UNU except
for His in the A. nodosum and F. vesiculosus seaweeds and Leu and Lys in the B. bifurcata. Regarding
fatty acids, polyunsaturated fatty acid (PUFA) were the predominant fatty acids in the three seaweeds
evaluated, ranging from 43.47% to 48.19% for A. nodosum and F. vesiculosus, respectively. The highest
oleic acid content (27.83–19.94%) was found in A. nodosum and F. vesiculosus, whereas B. bifurcata
presented the highest arachidonic acid level (15.24%). Moreover, the n-6/n-3 ratio ranged from 2.62 to
1.41, placing the three macroalgae studied according to WHO recommendations (n-6/n-3 ratio < 10).
The authors thank INIA (Instituto Nacional de Investigaciones Agrarias y Alimentarias,
Spain) for granting Ruben Agregán with a predoctoral scholarship (CPR2014-0128).
JoséM. Lorenzo, Rubén Agregán, Paulo E. S. Munekata, Daniel Franco and Javier Carballo
conceived, designed and performed the experiments; Selin ¸Sahin, Ramón Lacomba and Francisco J. Barba
supervised the study, wrote and reviewed the manuscript. All authors have read and approved the
Conﬂicts of Interest:
The authors declare no conﬂict of interest, and the founding sponsors had no role in the
design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in
the decision to publish the results.
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