Environmental Technology & Innovation 24 (2021) 101848

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Environmental Technology & Innovation

journal homepage: www.elsevier.com/locate/eti

Identification and characterization of marine seaweeds for

biocompounds production
Yang Yang a , Min Zhang a , Adel I. Alalawy b , Fahad M. Almutairi b ,
∗
Mohammed A. Al-Duais b , Junling Wang c , El-Sayed Salama a ,
a
Department of Occupational and Environmental Health, School of Public Health, Lanzhou University, Lanzhou 730000, Gansu
Province, China
b
Department of Biochemistry, Faculty of Science, University of Tabuk, Kingdom of Saudi Arabia
c
Institute of Toxicology, School of Public Health, Lanzhou University, 730000, Gansu Province, China

article info a b s t r a c t

Article history: Seaweeds from three different phyla including Sargassum thunbergii GEEL-15 (Phaeo-
Received 30 June 2021 phyta), Mastocarpus stellatus GEEL-16 (Rhodophyta), and Ulva sp. GEEL-17 (Chlorophyta)
Received in revised form 1 August 2021 were screened, identified, and evaluated for their potential applications. Characteriza-
Accepted 3 August 2021
tions including analysis of the bio-components (carbohydrates, proteins, and lipids),
Available online 5 August 2021
proximate and ultimate analyses were performed along with FTIR and TGA/DTG analyses.
Keywords: High volatile solid (>71%) and carbon (>32%) content in seaweeds suggested the pros-
Seaweeds perous presence of organics such as polysaccharides, phenols, aldehydes, ketones, and
Sargassum thunbergii esters. The content of carbohydrates ranged from 35.08–55.40%, with GEEL-17 having
Mastocarpus stellatus the highest. While the second biocompound was proteins (4.24–9.14%) followed by
Ulva sp lipids (4.63–7.88%). The major functional groups were confirmed by FTIR peaks i.e.
Biocompounds carboxyl, amino, hydroxyl, amide, ester, and sulfate groups. Properties of these seaweeds
showed that they can produce value-added compounds (such as alginate, fucoidan,
and glycoproteins). This study demonstrated that obtained seaweeds are the potential
candidate for biocompound production.
© 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction

Seaweeds, are primary producers in the aquatic food chain, and a major component of ecosystems as they are
considered natural dietary bio-resources for various organisms (Dietz et al., 2021). Seaweeds are multicellular algae with a
wide geographical distribution. Based on pigmentation type and morphological characteristics, seaweeds (macroalgae) are
mainly divided into three categories, which include Phaeophyta (brown algae), Rhodophyta (red algae), and Chlorophyta
(green algae) (Cermeño et al., 2020). A few characteristics of seaweeds such as the growth ability and adaptation to
extreme conditions make them a potential candidate for various applications such as human food, value added products,
pharmaceutical purpose and as energy feedstock (Abomohra et al., 2016). A few years back, seaweeds were only used to
promote the water cycle and their biomass was used for composting.
Recently, seaweeds are used for many purposes such as biofuels feedstock, health benefits, and extraction of bioactive
compounds (Tabassum et al., 2017). Seaweeds comprises of highly valuable nutritional components (polysaccharides,

∗ Corresponding author at: Department of Occupational and Environmental Health, School of Public Health, Lanzhou University, Lanzhou 730000,
Gansu Province, China.
E-mail addresses: salama@lzu.edu.cn, sayed14@hanyang.ac.kr (E.-S. Salama).

https://doi.org/10.1016/j.eti.2021.101848
2352-1864/© 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.
org/licenses/by-nc-nd/4.0/).
Y. Yang, M. Zhang, A.I. Alalawy et al. Environmental Technology & Innovation 24 (2021) 101848

Table 1
The main physicochemical characteristics of the seaweed habitat site at Jinshatan Sea.
Parameters Values
pHa 6.30
Salinity (g L−1 ) 30.81 ± 2.75
Conductivity (ms cm−1 ) 57.01 ± 4.69
Total dissolved solids (g L−1 ) 40.53 ± 3.41
Dissolved oxygen (mg L−1 ) 1.48 ± 0.88
Total organic carbon (g L−1 ) 10.42 ± 2.33
a
No unit.

proteins, polyphenols, vitamins, and >20 amino acids), which are beneficial to improve human health. It has been reported
that bioactive compounds extracted from seaweeds have the potential for anti-coagulation, anti-oxidation, anti-tumor,
anti-viral, and anti-bacterial. Thus, seaweeds can be considered for developing new drugs and functional foods, with few
or no side effects (Ponthier et al., 2020). Based on the factors mentioned above, seaweeds have obtained great attention
in the pharmaceutical and biomedical fields. However, different genera and species of seaweed have a potential variation
in bioactive compound types and functions. A few previous studies reported that, fucosterol and hexadec-4-enoic acid
extracted from Sargassum longifolium, have inhibitory effects on Vibrio parahaemolyticus, Pseudomonas fluorescens, and V.
vulnificus, while focusterol in Eklonia stolonifera showed anti-fat activity in 3T3-L1 adipocytes (Jung et al., 2014; Rajendran
et al., 2013). Therefore, species specific identification is essential before studying the activity of bioactive compounds of
seaweeds.
Morphological identification of seaweeds is a preliminary technique, while molecular identification technology is more
systematic and reliable. Therefore, implementation of molecular biology approaches are essential to disclose potential
species, which is conducive to the subsequent research on the bioactive ingredients of seaweeds. The presence of
biochemical content in seaweeds majorly depends on type of species, harvest season, and geographical area (Abomohra
and Almutairi, 2020). In a recent study, it is reported that the carbohydrate content of Sargassum vulgare was 28% in the
rainy season and 39% in the dry season (de Melo et al., 2021). Therefore, it is necessary to comprehensively characterize
seaweeds, which might help in understanding their potential in the production of value-added compounds. In this
study, seaweeds species from brown, red, and green algae were selected based on physiology, geographical location,
and edibility. The selected seaweeds were identified based on morphological characterization and molecular remark.
Moreover, the characterizations including bio-components analysis (carbohydrates, proteins, and lipids), proximate and
ultimate analyses were performed along with FTIR and TGA/DTG analyses to evaluate their potential applications.

2. Materials and methods

2.1. Collection and identification of seaweed species

Seaweeds having different morphology along with the water from their respective sites were collected in March 2021
from the Jinshatan Sea, Huangdao District, Qingdao City, Shandong Province, China (40◦ N, 120◦ E) (Fig. 1). Jinshatan Sea
is located at the southern tip of shandong peninsula, adjacent to the Yellow Sea. The physicochemical properties of
the seawater were analyzed (Table 1). The pH was measured by PHS-3C+ pH meter. Conductivity, TDS, and salinity
were measured using Multi-Parameter PCSTestr 35 (EUTECH Instruments, Singapore). The dissolved oxygen (DO) was
determined by a portable DO meter (DO300, Alalis instruments, Shanghai, China). Total organic carbon (TOC) was
determined by TOC-1500 (Metash instruments, Shanghai, China). After collection, the biomass was transferred to the
laboratory. Solid impurities, epiphytic plants and animals were removed by repeated washing under tap water. After that,
the clean biomass was air-dried, it was transferred to polypropylene plastic bags and stored at room temperature for
subsequent analysis.
The seaweeds were first recognized by studying their morphological features using botanical approaches (Arif et al.,
2021). The collected seaweeds were then identified using molecular approaches. In brief, the DNA of seaweeds was
extracted by the method that was proposed by Rice and Bird (2019). The eukaryotic primers 1143-510-2-F (AATTGACG-
GAAKGGCA) and 1637-510-2-R (CGACGGGCGGTGTGTA) were used to amplify the ITS regions of 18S rRNA genes to identify
the species. The obtained 18S rRNA gene sequences were downloaded to the database and the genes searched using BLAST
(Arif et al., 2021). The sequences of gene fragments were analyzed and corrected with Genedoc. The phylogenetic tree
was constructed using the neighbor-joining (NJ) Kimura’s two-parameter algorithm, as implemented within the MEGA-5
software. The phylogenetic tree was constructed from molecular data (Arif et al., 2021).

2.2. Analyses of biocomponents

The carbohydrate content in dried biomass was determined by the phenol-sulfuric acid method (Nielsen, 2010). The
extraction of crude lipids was carried out according to the previous method by Bligh and Dyer, with few modifications
(Manirakiza et al., 2001). Protein content in seaweeds was measured by the Lowry method. A standard curve was obtained
using bovine serum protein (BSA) and the absorbance of the sample was measured at 660nm .
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Y. Yang, M. Zhang, A.I. Alalawy et al. Environmental Technology & Innovation 24 (2021) 101848

Fig. 1. The map showing the geographical location of the sample collection site of seaweeds in Jinshatan Sea, Qingdao City, Shandong Province,
China. The red dot inside the map indicating the sampling site.

2.3. Characterization of seaweeds biomass

2.3.1. Proximate and ultimate analyses

The moisture content and the total solids in the samples were determined by heating the samples in an air-drying oven
at 105 ◦ C until a constant weight was reached. The volatile content was determined by weight loss of the samples when
the samples were incinerated at 550 ◦ C using a furnace (LECO TGA 701, America) and the residual mass was measured
as ash. The percentages of carbon, nitrogen, hydrogen, and sulfur elements in dried biomass were calculated by CHNS
element analyzer (Vario EL Cube, Germany). All the analyses were carried out in triplicate and the data is presented
as their average and standard deviation. The higher heating value (HHV) was calculated using an Eq. (1) proposed by
Dulong-Berthelot, based on elemental (Arif et al., 2021).
O
HHV (MJ kg−1 ) = 0.3383 C + 1.422(H − ) (1)
8
where HHV is the higher heating value (MJ kg−1 ), C, H, N, O, and S are the elemental content (%) of each seaweed.

2.3.2. Fourier transform infrared spectroscopy (FTIR)

The FTIR spectra of Freeze-dried seaweed biomass were recorded using a MAGNA 550 Nicolet (Madison, USA), equipped
with a mercury cadmium telluride detector. The spectra were scanned 30 times in the transmission mode over the
wavenumber range of 4000–400 cm−1 , with a resolution of 4 cm−1 . Average spectra were obtained from samples in
triplicate (Arif et al., 2021). The background spectrum was obtained by scanning the air with the same instrument before
measuring the samples.

2.3.3. Thermogravimetric analysis (TGA)

TGA of seaweed biomass was carried out using a thermal analyzer (Linseis Messgerate GmbH, Germany), which,
in combination with a high-resolution microbalance, enables the analyzer to accurately measure mass changes in the
samples under a fixed gas environment and at different heating and cooling rates. The 5 mg sample were transferred
from the alumina crucible to the thermal analyzer in case of the influence of heat and mass transfer on the experiment.
To fully investigate the gradual distribution process of biochemical components (carbohydrates, proteins and lipids) of
seaweeds biomass, the samples were pyrolyzed from 25 ◦ C to 800 ◦ C at a temperature rate of 10 ◦ C min−1 under a
constant atmosphere of nitrogen (100 mL min−1 ). The diagram for thermal degradation behavior, volatile solids, and ash
analyses of seaweed biomass was described. Differential thermogravimetric analysis (DTG) is the curve obtained by the
first derivative of TG concerning time, which represents the relationship between weight loss rate and time (Arif et al.,
2021).
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3. Results and discussion

3.1. Identification and characteristics of seaweed species

The obtained seaweeds are mainly distributed in Liaoning (Fig. 1), Shandong, Jiangsu, Zhejiang, Fujian, and Guangdong
provinces of China. The genomic sequences of the identified seaweed species including S. thunbergii GEEL-15 (Phaeophyta),
M. stellatus GEEL-16 (Rhodophyta), and Ulva sp GEEL-17 (Chlorophyta) were published in the NCBI database under
accession numbers of #MZ219644, #MZ219091, and #MZ447783, respectively (Fig. 2). The growth in full bloom is from
March to July. The typical characteristics are dark brown color, short trunk, distribution of several lateral branches, short
leaves, and filamentous, mostly serrated edge. M. stellatus is a small red alga with forked branches that are dark red-
brown to purple. It has a thicker margin and a distinct groove at the base, mainly distributed in Ireland, Europe, West
Africa, the United States, and the Mediterranean Sea. Ulva sp. are mainly found in the East China Sea, and the South
China Sea, however, a few are found in the Yellow Sea and the Bohai Sea. The leaves are bright green, almost oval, and
often slightly wavy around the edges. According to the external morphology, growing season and geographical location
of seaweeds mentioned above, the three seaweed species in this study have been identified as S. thunbergii, M. stellatus,
and Ulva sp. (Fig. 3). The morphological identification of seaweed species mainly depends on the internal structure, and
physiological characteristics. However, seaweeds under the same phylum have more or fewer similarities in external
morphology and ecological habits. Besides, under the influence of environmental factors, the morphological characteristics
and salient features of seaweeds gradually disappear, which makes it difficult to accurately identify by morphological
analysis. Molecular identification is one of the most accurate and popular way to recognize species. Internal transcribed
spacer (ITS) are most commonly used genetic markers for molecular identification and molecular ecology. ITS sequences
are relatively consistent within species, with obvious interspecific differences, small in length, and more information can
be obtained from shorter sequences (Baldwin et al., 1995). Therefore, the analysis of ITS sequence effectively identified
the species and genus relationships of seaweeds at the sequence level.

3.2. Bio-composition of seaweeds for potential bioactive compounds

The content of biocomponents contained in seaweeds is helpful to determine the potential of each species in extracting
bioactive compounds. Therefore, carbohydrates, proteins, and lipids of all seaweed species in this study were determined
(Fig. 4). The carbohydrates content in the three seaweed species ranged from 35.08% W W−1 – 55.40% W W−1 . Ulva sp.
GEEL-17 (Chlorophyta) exhibited the highest carbohydrates content (55.40% W W−1 ), compared to S. thunbergii GEEL-15
(37.00% W W−1 ) and M. stellatus GEEL-16 (35.08% W W−1 ) (Fig. 4a, b, and c). The content of polysaccharides is related to
the life cycle of seaweed. April to mid-June is the growth period of S. thunbergii, which means the accumulation period
of polysaccharides (Jin et al., 2017). However, the collected seaweeds (S. thunbergii GEEL-15) can consider as potential
candidate to produce higher polysaccharides. The carbohydrate content of S. thunbergii GEEL-15, M. stellatus GEEL-16,
and Ulva sp. GEEL-17 was higher in comparison to the previous studies reported (Table 2). The polysaccharides and
related compounds extracted from seaweeds have been widely used in food additive, cosmetics, and pharmaceutical
engineering, because of their unique anticoagulant, antiviral, anti-tumor, and immunomodulatory effects (Wijesekara
et al., 2011). A glycoprotein with a molecular weight of about 10 kDa isolated from Saccharina Japonica showed antioxidant
activity against a variety of oxidation-induced DNA cleavage (Kim et al., 2012). Studies also provide an evidence that the
polysaccharides extracted from S. thunbergii have anti-complement activity, and the active components of polysaccharides
are sulfated fucan or sulfated galactofucan (Jin et al., 2017). In the similar study it was reported that in the life cycle of
S. thunbergii, the content of sulfate was relatively stable (7.66%–15.17%), which was not affected by the harvest season,
thus ensuring the stability of anti-complement activity of polysaccharides (Jin et al., 2017). The sulphated-galactans of
M. stellatus was associated with a significant increase of caecum reducing capacity and plasma anticoagulant capacity
(Gomez-Ordonez et al., 2012). Previously it is reported that Ulva contains a high content of sulfates up to 17% of its dry
matter, which is almost 14% chemically bound in a soluble polymer (ulvan), a sulphated glu-curonorhamnoglycan (Durand
et al., 1997). The sulfate content is proportional to the antioxidant activity of ulvan (Tziveleka et al., 2019). In addition,
some polysaccharides are considered as dietary fiber and cannot be digested in the gastrointestinal tract due to the lack of
polysaccharides degrading enzymes in the body. Another major advantage of these seaweeds includes formation of drugs
from these polysaccharides can be absorbed orally and also beneficial for the patients suffering from intravenous infusion.
However, previous studies on the biological activity of polysaccharides from S. thunbergii mainly focused on antioxidant
and anti-tumor activities, while those from M. stellatus and Ulva sp. mainly focused on antioxidant activities. Antioxidant
activity was mainly studied in vitro, and the mechanism of antioxidant activity was not clear (Jimenez-Escrig et al., 2012).
In addition, there are only a limited number of cancer types that have been reported to be inhibited by polysaccharides
from S. thunbergii.
The proteins content of S. thunbergii GEEL-15, M. stellatus GEEL-16 and Ulva sp. GEEL-17 were 7.14% W W−1 , 9.14% W
W−1 , and 4.24% W W−1 , respectively (Fig. 4a, b, and c). Compared with previous studies, the proteins content of all
seaweeds used in this study is at a medium or lower than other algal species (Table 2). This may be related to the
low salinity (30.81 g L−1 ) of seawater, which has negative effects on photosynthesis and growth of seaweeds, and the
high content of non-protein nitrogen (Amano and Noda, 1992). Lower protein content reduces the amount of proteolytic
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Fig. 2. The phylogenetic tree of the seaweed species was established by using molecular data and showing the relationships among the sequences
of GEEL-15, GEEL-16, GEEL-17, and the most similar sequences retrieved from the NCBI nucleotide database.

Table 2
The comparison of major biochemical composition (% dry weight of sample) among the collected seaweeds used in this study and previously reported
studies.
Phylum Seaweeds Carbohydrates Proteins Lipids References
Sargassum ilicifolium 32.90 ± 1.08 8.90 ± 0.94 2.00 ± 0.20
Rohani-Ghadikolaei et al. (2012)
Colpomenia sinuosa 32.10 ± 1.75 9.20 ± 1.78 1.50 ± 0.29
Phaeophyta (Brown algae)
Sargassum vulgare 39.07 ± 1.34 10.32 ± 0.04 4.02 ± 0.19 de Melo et al. (2021)
S. thunbergii GEEL-15 37.00 ± 1.64 7.14 ± 0.41 7.88 ± 0.88 This study
Gracilaria corticata 43.00 ± 5.58 19.30 ± 2.19 1.80 ± 0.46 Rohani-Ghadikolaei et al. (2012)
Gracilaria edulis 4.71 ± 0.60 25.29 ± 0.67 4.76 ± 0.73 Rosemary et al. (2019)
Rhodophyta (Red algae)
Hypnea valentiae 30.10 ± 0.51 4.56 ± 0.81 0.73 ± 0.14 Dixit et al. (2017)
M. stellatus GEEL-16 35.08 ± 0.24 9.14 ± 0.41 4.63 ± 0.38 This study
Enteromorpha intestinalis 35.50 ± 2.83 10.50 ± 1.02 2.90 ± 0.25 Rohani-Ghadikolaei et al. (2012)
Ulva lactuca 18.50 ± 0.12 19.34 ± 0.20 3.46 ± 0.10 El-Din (2019)
Chlorophyta (Green algae)
Corallina mediterra 26.60 ± 0.20 17.05 ± 0.20 2.07 ± 0.05 Verma et al. (2017)
Ulva sp. GEEL-17 55.40 ± 5.03 4.24 ± 0.68 6.67 ± 0.62 This study

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Fig. 3. Morphological characteristics, thermal degradation (pyrolysis), and proximate analyses of Sargassum thunbergii GEEL-15 (Phaeophyta),
Mastocarpus stellatus GEEL-16 (Rhodophyta), and Ulva sp. GEEL-17 (Chlorophyta).

enzyme during extraction and purity of polysaccharides. These results indicated that the seaweeds in this study have
more advantages in the extraction of polysaccharides and sulfated polysaccharides. However, they are still a potential
source of polypeptides. Additionally, the proteins content of S. thunbergia, M. stellatus, and Ulva sp. was different from
other studies, possibly due to different growth environments (Blanco-Pascual et al., 2014; Hong-bo et al., 2009). Although
different seaweed species contain higher and lower levels of proteins, the types of amino acids are the same (Biancarosa
et al., 2017). The glycoprotein, a special protein that exists in seaweeds, has attracted the interest of experts in the field
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Fig. 4. The percentages of biocomposition (left) and elements (right) in seaweeds S. thunbergii GEEL-15 (a and d), M. stellatus GEEL-16 (b and e),
and Ulva sp. GEEL-17 (c and f).
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of medicine. For instance, it can induce apoptosis in AGS (gastric adenocarcinoma cell line) human gastric cancer cells
(Han et al., 2011). However, the high content of phenols in seaweeds limits the application of the proteins in the human
body. Therefore, when applying the proteins of a particular species of seaweed to humans, it is necessary to determine
its phenolic content.
The lipids content of S. thunbergii GEEL-15 was higher 7.88% W W−1 than M. stellatus GEEL-16 and Ulva sp. GEEL-
17, which were, 4.63% W W−1 , and 6.67% W W−1 , respectively (Fig. 4a, b, c). This reflected the difference in lipids
accumulation ability among seaweed species. The lipids content of all seaweeds in this study was significantly higher
than that in previous studies (Table 2). The seasonal variation of lipids content in brown algae, red algae, and green algae
was previously studied, and it was found that the highest lipids content occurred in spring (El Maghraby and Fakhry,
2015). Research on the nutritional value of seaweeds mainly focuses on polysaccharides and proteins. However, the lipid
content of seaweeds was reported to be rich in polyunsaturated fatty acids and several essential fatty acids, such as linoleic
acid, linolenic acid, and arachidonic acid (Sánchez-Machado et al., 2003). Therefore, from the perspective of promoting
human health, it can be considered to increase the proportion of seaweeds lipids intake in daily life.

3.3. Physiochemical analysis of seaweeds biomass

The moisture content of all the seaweeds ranged from 9.23–11.28 wt% dry, among which the moisture of Ulva sp.
GEEL-17 was highest (11.28 wt% dry). S. thunbergii GEEL-15 and M. stellatus GEEL-16 were 9.89 wt% dry and 9.23 wt% dry,
respectively, while the relative proportions of total solids were 90.11 wt%, 90.77 wt%, and 88.73 wt%. The main volatile
compounds in Phaeophyta, Rhodophyta, and Chlorophyta are hydrocarbons, alcohols, phenols, ketones, aldehydes, esters,
sulfides, and fatty acids (Gressler et al., 2009). In this study, the volatile solid content of all the obtained seaweeds was
71.84–79.16 wt% dry, which was higher than the previous study (Hu et al., 2019). Therefore, these results suggested
that all seaweeds were rich in nutrients and conducive to the extraction of bioactive compounds. The ash contents in
S. thunbergii GEEL-15, M. stellatus GEEL-16, and Ulva sp. GEEL-17 was 20.84 wt% dry, 28.17 wt% dry, and 21.30 wt% dry,
respectively. The ash mostly represents the sulfate group, uronic acid, and minerals of polysaccharides (Saravana et al.,
2018).
The percentages of carbon, nitrogen, hydrogen, and sulfur in all seaweeds were 32.38%–35.20%, 3.20%–5.49%, 5.46%–
6.63%, and 1.08%–5.41%, respectively (Fig. 4). There is a large amount of total nitrogen in seaweeds, which includes
non-protein nitrogen and protein nitrogen. In a study, it is reported that seaweeds account for 7.2%–59.5% of non-protein
nitrogen from the total nitrogen, while protein nitrogen accounts for 3.1%–17.4% of the total nitrogen (Amano and Noda,
1992). According to the relationship between nitrogen content and protein (1 gram of nitrogen equals 6.25 grams of
protein), the protein content in this study was low, which may be caused by a large amount of non-protein nitrogen in
the seaweeds. The values of the C/N ratio in seaweeds were 6.57, 6.07, and 11.00 for GEEL-15, GEEL-16, and GEEL-17,
respectively. The high N content and low C/N ratio of M. stellatus GEEL-16 refers to the presence of comparatively higher
proteins content. The low N element and highest C/N ratio of Ulva sp. GEEL-17 (almost twice of GEEL-15 and GEEL-16)
was consistent with its high carbohydrates and low proteins content. The values of C/H ratio in seaweeds were 5.93, 5.39,
and 5.31, respectively. The higher heating value (HHV) in seaweeds biomass were 8.74 MJ kg−1 , 11.27 MJ kg−1 and 12.13
MJ kg−1 , respectively. Due to the high ash content, the values of HHV in this study were lower than that of lignocellulose
(19.07–20.72 MJ kg−1 ) (Yanik et al., 2013).

3.4. Spectroscopic analyses

3.4.1. FTIR spectra of seaweeds biomass

The FTIR has become a well-accepted technique for the identification of bioactive compounds in plants, as a qualitative
or semi-quantitative method that can identify functional groups and obtain other structural information (Song et al., 2020).
The FTIR spectra recorded in this experiment were compared with the spectra of other studies as a reference and the
following results were obtained (Fig. 5a, b, c). The broad bands centered at 3550–3200 cm−1 assigned to hydrogen-bonded
O–H and N–H stretching vibrations, which correspond to polysaccharides and amino acids. The peak near 2930 cm−1 was
caused by the C–H asymmetrical stretching on the saturated carbon atom, proving the existence of aliphatic. The weak
peaks between 2140 and 2100 cm−1 were attributed to the stretching of the C≡C. The strong C==O stretching from 1657
to 1648 cm−1 proved the presence of COOH. The N–H stretching at 1547–1525 cm−1 again predicted the presence of
amino acids. Amino acids are associated with amide bonds, and the absorption of amide I and II bands results in stretching
vibration of C==O bond and bending vibration of N==H bond (Thomas et al., 2017). The peaks at 1424–1414 cm−1 indicating
hydroxyl deformation, carbon–oxygen stretching of phenolic hydroxyl groups, and carbon–hydrogen deformation of CH2 .
Fucoidan (main water-soluble sulfated polysaccharide) showed bands in the FTIR spectra at 1257–1254 cm−1 , assigned
to the sulfate group (S==O) (Maciel et al., 2007). The peaks between 1030 and 1058 cm−1 were due to the C–O bond
stretching of the polysaccharides (Yip et al., 2014). The bands between 840 and 790 cm−1 corresponded to a stretch of
C==C.
The key functional groups obtained from FTIR further predicted the possible presence of biocompounds in seaweeds.
By identifying amide group, amino, hydroxyl, and ester groups in the infrared spectrum, it can be proved that there are
essential biological components in seaweeds (such as proteins, amino acids, polysaccharides, and lipids) (Gangidi et al.,
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Fig. 5. Fourier transform infrared spectroscopy (FTIR) of seaweeds (left) identifying the functional groups present in essential compounds.
Thermogravimetric analysis (TGA) and differential thermogravimetric (DTG) indicating thermal degradation of biocompounds in seaweeds (right).

Table 3
The essential biocompounds of seaweeds based on the FTIR peak values and biological activities reported in previous studies Tanna et al. (2019)
and Kumar et al. (2019).
Peak values (cm−1 ) Functional groups Acid groups Associated biocompounds Application
3550–3200 O–H CH2 OH Polysaccharides and glycoproteins Antioxidant, antiviral,
3550–3200, 1547–1525 N–H CNH3 + Amino acids and glycoproteins antitumor,
2930 C–H – Aliphatic, polysaccharides, proteins, and glycoproteins antibacterial,
2140–2100 C ≡C R–C≡C–H Alkynes antifungal,
1657–1648 C==O COOH Proteins, glycoproteins, and alginate anticoagulant,
1257–1254 S==O COSO3 H Fucoidan immunomodulatory,
1058–1030 C–O CH2 OH Polysaccharides and glycoproteins and functional food.

2002). The main function of polysaccharides, glycoproteins, fucoidan, alginate, aliphatic (mainly unsaturated fatty acids),
proteins, and amino acids is to provide nutrients to seaweeds and help them to resist in the adverse environments as
the primary or secondary metabolites. When applied to humans, they can influence metabolic processes and enhance
immunity for the betterment of human health (Srivastava et al., 2021). In vitro and in vivo studies have shown that they
have antioxidant, antiviral, anti-tumor, anti-bacterial, and immunomodulatory functions (Table 3), which provides a new
way of thinking for future research and development of safe and efficient new drugs.
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3.4.2. TGA spectra of seaweeds biomass

The pyrolysis behavior of three species of seaweed was studied by thermogravimetric analysis (TGA) and differential
thermogravimetric analysis (DTG). The seaweeds showed a gradual degradation process corresponding to their biochem-
ical composition (Fig. 5d, e, f). The sequence of degradation and relative proportion of carbohydrates and proteins in
seaweed can be seen from the TGA curve. The thermal decomposition of seaweeds biomass mainly had three stages.
The first stage was from ambient temperature to 170 ◦ C, and the weight loss of seaweeds was mainly caused by the
evaporation of moisture, first free water, and then the combined water in the polysaccharides. The evaporation of bound
water in Ulva sp. GEEL-17 might be related to the presence of ulvan, because ulvan contain hydroxyl groups and loses
huge water during pyrolysis. The second stage pyrolytic decomposition occurred at 170 ◦ C, and no weight loss occurred
when it reached about 550 ◦ C. At this time, the TGA and DTG curve tended to be stable. All seaweeds exhibited the biggest
weight loss between 170 ◦ C and 280 ◦ C, related to the high carbohydrates content of the three species.
The peak temperature of seaweeds biomass degradation in the second stage was lower (237 ◦ C), which indicated that
the alginic acid and fucoidan content were higher. The weight loss of seaweeds from 280 ◦ C – 450 ◦ C represented the
decomposition of proteins. As it can be seen from the figure, the weight loss of the samples caused by the decomposition
of carbohydrates, suggesting that carbohydrates play a dominant role in the biochemical composition of seaweeds. At the
last stage (550–800 ◦ C), the weight of the seaweeds barely changed, and the remaining weight represented as ash content,
which may contain the minerals of polysaccharides, like sulfates, phosphates, and carbonates (Saravana et al., 2018). The
TGA curve showed that the degradation of seaweed biomass was 1.72%–2.65% in the first stage, 64.52%–71.70% in the
second stage, and 1.57%–2.21% in the last stage. The DTG curve represented the degradation rate of seaweed biomass,
and the peak value appeared between 230 ◦ C and 240 ◦ C, indicating the fastest decomposition of biomass (Fig. 5d, e, f).
The peak of seaweed biomass degradation was different at different temperature bands, which was similar to earlier study
(Shekhar et al., 2012). However, the difference is that the seaweeds in this study had higher weight loss in the second
stage.

4. Conclusions

Seaweeds identification and systematic characterization of its composition are the prerequisites for the exploration
of associated biocompounds. The high carbohydrates, volatile solids, and carbon contents of the selected seaweeds make
them potential candidates for the production of value-added compounds. The low proteins content of seaweeds in this
study makes obtained seaweed more advantageous in polysaccharide extraction. The lipids in the seaweeds might be
suitable for studying essential fatty acids. This study demonstrated that these seaweeds are a promising source for
essential bio compound and further use in human (dietary material) and environmental applications (biofuels, potential
biomass and drugs).

CRediT authorship contribution statement

Yang Yang: Methodology, Visualization, Investigation, Data curation, Formal analysis, Writing, Original draft. Min
Zhang: Investigation, Data curation, Formal analysis. Adel I. Alalawy: Visualization, Formal analysis, Writing – review
& editing. Fahad M. Almutairi: Visualization, Formal analysis, Writing – review & editing. Mohammed A. Al-Duais:
Visualization, Formal analysis, Writing – review & editing. Junling Wang: Visualization, Formal analysis, Writing – review
& editing. El-Sayed Salama: Conceptualization, Supervision, Resources, Data curation, Validation, Visualization, Formal
analysis, Writing – review & editing, Funding acquisition, Project administration.

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.

Acknowledgments

This work was supported by the startup fund for the construction of the double first-class project, China (No.
561119201), Lanzhou University, China; and the ‘‘Deanship of Scientific Research, University of Tabuk, KSA’’, under the
Research Group (S-1441-0044).

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