International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:14 No:01 29

Biochemical Composition of Some Seaweed From

Hurghada Coastal Along Red Sea Coastal, Egypt
Shimaa M. EL-Shafay
Botany Department, Faculty of science, Tanta University, Egypt
1-Corresponding Author: Dr. Shimaa Mohamed El-Shafay
Current address: Botany Department, Faculty of science, Tanta University, Egypt
Contacts: Phone: 00201009570818,
E-mail: [email protected]

Abstract-- Variations in protein, carbohydrate, lipid, ash, The amino a c i d s composition in free or bound
moisture, fatty acids, amino acids, β-carotene and total phenolic form has been studied in several species of marine algae
compounds content of the seaweeds Ceramium rubrum, (Qasim 1991), whereas pronounced differences were
Sargassum vulgare, Sargassum fusiforme and Padina pavonia observed in protein and amino acids between different algal
were studied. C. rubrum characterized by the highest protein groups.
content, while the highest lipid and carbohydrates contain was Carotenoids are an important group of natural
observed in S. vulgare. The highest concentration of essential pigment with specific applications as colourants, feed
amino acid and β-carotene were observed in P. pavonia. S.
supplements, nutraceuticals, and medicine, cosmetic and
fusiforme characterized by high concentration of total phenolic
compound and total unsaturated fatty acids. biotechnological purposes. Despite the avail ability of a
variety of natural and synthetic carotenoids, only a few have
been exploited commercially, including b -carotene, lycopene,
Index Term-- Seaweeds, Protein, Carbohydrates, Lipid, Fatty
astaxanthin, canthaxanthin, lutein and capxanthin (Bhosale
acid, β-carotene, Total phenol
2004). The β- carotene accumulation and the rate of synthesis
depend on certain environmental parameters (Shariati and
1.0 INTRODUCTION Hadi 2011). Antioxidants in particular carotenoids, help to
Seaweeds have been used since ancient times food, prevent the free radicals damage associated with the aging
fodder and fertilizer and as source of medicinal drugs. process itself. The review by the National research council
Seaweeds are one of the commercially important marine living (NRC 1982) concluded “the epidemiological evidences
and renewable resources. Variability in chemical sufficient to suggest that food rich in carotenes or vitamin A
components and growth of algae may be inter specific, are associated with a reduced risk of cancer”. Thus foods rich
intra-annual or inter-annual. Certain seaweeds contain in carotenoids in this case β-carotene may not only be able to
significant quantities of protein, lipids, minerals and prevent but also reverse cancers.
vitamins, w h i l e although nutrient contents vary with Phenolic compound are one of the most abundant
species, geographical location, season and temperature class of phytochemicals present in algae. Among natural
(Haroon 2000). antioxidants, phenolic antioxidants are commonly found in
Protein, carbohydrates and Lipids, are the most plants, including seaweeds. Presence of phenolic compounds
important biochemical components of algae. A few studies in any medicinal preparation may attribute to various
were done on fatty acids of microalgae and seaweeds. medicinal properties, earlier many algae have been reported to
Macroalgal biomass can store large amounts of oil which can possess a variety of phenolic compounds with antioxidant and
be exploited for the production of biodiesel John and Anisha hepatoprotective properties (Song el al. 2003). Phenolic
(2011). compounds have been reported to have several biological
Seaweeds belonging to the Rhodophyta possess high effects including antioxidant,antiapoptosis, anti-aging, anti
levels of proteins (10–30% DW) (Darcy-Vrillon 1993). In carcinogen and have been highly considered for their
some red seaweed, such as Palmaria and Porphyra tenera, important dietary roles as antioxidant and chemoprotective
the protein contents are 35 and 47% DW, respectively. agents.
These levels are even comparable to that of the soybeans The aim of this work is to investigate the biochemical
(35% DW). composition (in protein, carbohydrate, lipid, ash, moisture,
Some evidences suggest that fatty acids and sterol fatty acids, amino acids, β-carotene and total phenolic
composition may be useful for taxonomic purposes compounds) in Ceramium rubrum, Sargassum vulgare,
(Herbreteau et al. 1997). Rhodophyta are characterized by Sargassum fusiforme and Padina pavonia
high content of C 20 polyunsaturated fatty acids, mainly
arachidonic and eicosapentaenoic acids. Other abundant
fatty acids in this class are palmitic and oleic acids.

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 International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:14 No:01 30
2. MATERIAL AND METHODS adipate (PEGA). The column oven temperature was
programmed at 8 C/min from 70 oC to 190 oC, then
o
2.1. Sampling
Four marine algal species were collected during isothermally at 190 oC for 25 min with nitrogen at 30
March 2013 these collected species including the one species ml/min. Identification of fatty acid methyl esters was
from Rhodophyta (Ceramium rubrum) and three species from accomplished by comparing the retention times of
Phyophyta (Sargassum vulgare, Sargassum fusiforme and experimental samples to those of known standards. The fatty
Padina pavonia). The algae were collected from Hurghada acids analysis was performed in the National Institute of
coastal along the Red sea coastal, Egypt. All samples were Oceanography and Fisheries, Alexandria, Egypt.
brought to laboratory in plastic bags containing sea water to
prevent evaporation. Epiphytic and extraneous matter were 2.8. Ash content
removed by washing first with sea water and then washed Ash content was estimated by ashing the ground
with distilled water to separate potential contaminants. The dried samples overnight in muffle furnace at 525 ºC.
samples were dried in the air at the room temperature and
kept in plastic bags for biochemical analysis. 2.9. Estimation of amino acids
Algal sample of 3 g was prepared for hydrolysis
2.2. Protein estimation according to Walker (1996) before determination of amino
The protein fraction (% of DW) was calculated from acids. Amino acid analyses were carried out using amino
the elemental N determinations using the nitrogen-protein acid analyzer LC 3000 eppendorf / Biotronik using column
conversion factor of 6.25 according to AOAC (1995). type H 125 x in the National Institute of Oceanography and
Fisheries, Alexandria, Egypt.
2.3. Carbohydrate estimation
The total carbohydrate was estimated by following 3. STATISTICAL ANALYSIS
the Phenol-sulphuric acid method of Dubois et al. (1956) The given data were expressed in terms of the mean
using glucose as standard. of three replicates ± SD (standard deviation). In addition, data
concerning the environmental parameters, content of
2.4 Lipid estimation proteins, carbohydrates, lipids, ash and moisture for the
The extraction of lipid was done by the chloroform- algal species were analyzed by Duncan’s multiple range test
methanol mixture (2:1 v/v). The lipids in chloroform were at p < 0.05 by using SPSS Inc. program version15 (2006).
dried over anhydrous sodium sulfate, and then the solvent
4. RESULTS AND DISCUSSION
was removed by heating at 80 ºC under vacuum (AOAC The biochemical contents of the four algal species
2000). are documented in Table I. The Ash contents were
considerably low, a nd Sargassum vulgare have the lowest
2.5. Estimation of β-Carotene: ash content when compared to the tree other species, however,
β-carotene was determined in diethyl ether at 440 nm as Ceramium rubrum have the highest ash content. As shown in
mentioned in the method of Neeld and Pearson (1963), and table I there were significant relation between the all studied
expressed as mg/100g dry wt. algae. The high element contents in seaweed are shown by
their ash contents, which are in the range of 8–40% DW.
2.6. Total phenols: Sanchez-Machado et al. (2002) f o u n d ash of 24.21% in
Total content of phenolic compounds of algal extracts was Caulerpa lentillifera against 17.58% in U. reticulate, while
determined spectrophotometrically at 725 nm using Folin- comparable amounts were measured in
Ciocalteu reagent according to the method described in Lim et Himanthalia elongata (26.78%), Laminaria ochroleuca
al. (2002). The total content of phenolic compounds was (29.47%) and Porphyra sp. (19.07%). The differences in
calculated based on a standard curve of phloroglucinol and ash contents depend on seaweed species, physiological
expressed in % of dry weight. factors, environmental changes, methods of
mineralization and type of processing.
2.7. Fatty acids methyl ester Results showed that, the protein content in species
The lipids in chloroform were dried over anhydrous C. rubrum and S. fusiforme was almost h ig her than in
sodium sulfate, and then the solvent was removed by heating S.vulgare and P.pavonia (table I). S. vulgare demonstrated the
at 60oC under vacuum. Fatty acid methyl esters were minimum protein contain as compared to the other algal
prepared according to Vogel (1975). The analysis was species. The statistic analysis showed that the relationship
performed in a gas liquid chromatography (Pye Unicam between the different studied species of algae was significant
Series 304 Gas Chromatograph) equipped with dual flam (one-way ANOVA, p < 0.05) Variation in protein content of
ionization detector and duel channel recorder. The separation seaweeds can be due to different species, seasonal period
of fatty acid methyl ester was performed through a coiled and geographic area (Haroon 2000, Ratana-arporn and
glass column (1.5 X 4 mm) packed with Diatomite (100 – Chirapart 2006).
120 mesh) and coated with 10% polyethylene glycol

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 Table I
The mean (±SD) contents of ash, moisture, lipid, protein, carbohydrate, β-carotene and total phenolic compound.

Parameters Ash Moisture Protein Carbohydrates (%) Lipid β-carotene Total phenol
Algae (%) (%) (%) (%) (IU\100mg) (ppm)

C.rubrum 0.53±0.003a 0.0285±0.00025a 9.27±0.03a 88.76 ±1.17acd 0.0082±0.0003a 2565.01 ± 3.9a 43.83 ± 0.52a
S.vulgare 0.19± 0.003b 0.0894±0.00025b 5.85±0.03b 93.34±1.01b 0.0403±0.0005b 1679.69 ±3.45b 0.38 ± 0.001b
S.fusiforme 0.33± 0.004c 0.0686±0.00026c 8.85± 0.03c 90.71±1.06a 0.0204±0.0005c 201.47 ± 0.94c 74.97 ± 0.25c
P.pavonia 0.51± 0.003d 0.0374±0.00026d 8.35 ±0.02d 90.50 ±1.25a 0.006± 0.0003d 4260.14 ± 2.17d 54.33 ± 0.88d

F –value 7434.01 35687.33 8912.63 8.44 4582.53 99999.99 10668.69

P-value 0.0001 0.0001 0.0001 0.007 0.0001 0.0001 0.0001

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Each value is the mean of three replicates ±SD
Values with the same letters in the same column showed insignificant differences (p≤0.05)
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 International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:14 No:01 32
In this study , the protein contents of the studied species al. 2002). Carbohydrate is the most important component
were higher than that of Sargassum polycystum, Gelidium for metabolism as it supplies the energy needed for
domingensis and Gelidium birdiae . Similar values recorded respiration and other metabolic processes. Changes in
for P. capillacea , the red alga Palmaria sp. , notably lower carbohydrate content at the different studied algae were
values in the green alga U. lactuca , and t h e brown algae, observed during the present study (Table I). The results
Himanthalia elongata and Laminaria ochroleuca. Haroon showed that, the four studied species contain a very high
(2000) reported range o f 9.42 ± 4.62 t o 20.60 ± 5.0 % of concentration of carbohydrates ranged from (88.88% in C.
DW for Enteromorpha spp., while protein in U. reticulata rubrum to 93.65% in S. vulgare). According to statistic
was three times o f that contained in U. lactuca and analyses there were significant relation between C. rubrum
slightly lower than that in Porphyra sp. (Sanchez-Machado et and S. vulgare, however the other relations was insignificant.
Table II
Fatty acid composition
Fatty acids C.rubrum S.vulgare S.fusiforme P.pavonia
Saturated
C8:0 ND ND ND 0.031
C10:0 0.057 ND ND 0.112
C11:0 0.105 ND ND 0.201
C12:0 0.491 0.073 0.077
0.144
C13:0 3.384 4.217 5.001 4.344
C14:0 3.912 1.956 3.851
3.415
C15:0 3.238 3.492 4.635 3.603
C16:0 32.401 23.991 26.368
28.923
C17:0 0.297 0.257 0.206
0.248
C18:0 5.545 3.368 2.824 3.893
C20:0 1.024 0.564 0.781
0.539
C21:0 0.751 ND 0.595 0.461
C22:0 0.493 ND ND ND
Sum of saturated 46.845 49.202 39.896 43.928
Unsaturated

C14:1 2.468 2.904 3.587 3.070

C15:1 1.759 1.915 2.046 1.669
C16:1 1.109 1.217 1.611 1.831
C17:1 0.787 1.144 1.196 0.980
C18:1 1.699 1.554 1.177 1.823
C20:1 ND 1.024 ND
0.579
C22:1 0.446 ND ND
1.030
Sum of mono 9.431 7.265 10.641 9.373
Polyunsaturated
C18:2 0.851 1.333 1.547
1.238
C18:3 0.778 0.427 0.404 0.422
C20;5 0.102 ND 0.950 ND
Sum of poly 2.118 1.278 2.687 1.969
Total unsaturated 11.549 8.543 13.328 11.342
ND= not detected

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 International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:14 No:01 33
John and Anisha (2011) reviewed that; c arbohydrates respectively). The palmitic acid as the major fatty acid
obtained from macroalgal biomass can be used for (85.36%) was also recorded in Gellidum micropterum,
fermentative production of bioethanol. Methane and hydrogen 63.19% in Porphyra spp. and Ergrezia menziesii,
can readily be produced from macroalgal biomass through Chondracanthus canaliculatus, and Ulva lobata. C13:0,
biologically mediated degradation. Anantharaman et al. C14:0, C15:0 and C18:0 was also present by relatively high
(2013) found that the carbohydrate content of five studied percentages in the all studied species.
species of seaweeds ranged from 10.63% and 28.58%, the As observed from table II, the unsaturated fatty acids
maximum carbohydrate content was recorded in the green present by nearly low concentration in the four studied
seaweed Enteromorpha intestinalis and the brown seaweed species, while the monounsaturated fatty acids present by
Dictyota dichotoma recorded the minimum value. moderate concentration among the all tested species. S.
The total lipid contents in the studied seaweed fusiforme contain the highest concentration of total
species were relatively low (Table I), the highest value was unsaturated fatty acid; however S. vulgare have the lowest
found in S. vulgare, however the lowest value was recorded in concentration of total unsaturated fatty acid. They were the
P. pavonia. The relationship between all algal species was next most common fatty acids as represented by Shanmugam
significant (one-way ANOVA, p < 0.05). Typically, seaweeds a n d Palpandi ( 2008), and oleic acid demonstrated high
are not considered to be good source of lipid (Ratana-arporn concentration in Porphyra spp. In the present study species S.
and Chirapart 2006), and the total lipid content was always fusiforme was characterized by the highest amount of
found less than 4% (Herbreteau et al. 1997). The present polyunsaturated fatty acids among the studied algae, and it
results a r e supported by the findings of Shanmugam and characterized by the present of EPA (C20:5).
Palpandi (2008) in Sargassum wightii (0.16 - 1.55 %), in As observed in table I the highest concentration of
Gracilaria spp. (0.78 -3.97%), in Enteromorpha intestinalis β-carotene was recorded in species P. pavonia however, the
(1.2 %) and in U. reticulate (1.7%). These differences could lowest concentration was recorded in species S. fusiforme.
b e due to environmental factors affecting the growth of the The main source of β-carotene was red and brown algae
seaweed. The literature has established that in seaweeds in (Sachindra et al. 2007).
general the lipid content is less than 4%. The differences could A lot of studies demonstrated the antioxidant
have been due to factors such as climate and geography of properties of algal carotenoids and the role they play in
development of the seaweed (Herbetreau et al. 1997). preventing many diseases linked to oxidative stress (Banerjee
As observed from table II, saturated fatty acids were et al. 2009). The β-carotene (with provitamin A activity)
the predominant in types present in the all studied species. content ranges from 36 to 4,500 ppm dry weight, with
Species S. vulgare contain the highest percentage of total Porphyra with the highest content and Palmaria palmata with
saturated fatty acids, while, species S. fusiforme contain the the second highest of 456 ppm. However, other studies
lowest content of total saturated fatty acids. C16:0 was the suggest that β-carotene could induce lung cancer in smokers.
predominant unsaturated fatty acid among the all studied Furthermore, experimental studies strongly suggest that β-
species. Species S. vulgare contain the higher percentage of carotene could prevent the onset of cancers, especially lung
C16:0. This results coincides with Shanmugam a n d cancer. However, natural source of β-carotene is highly
Palpandi (2008) who found that the saturated fatty acids recommended as they protect against the development of
c o n s t i t u t e d 70.01% of the total fatty acids, w i t h t h e cancer (NRC 1982).
d o m i n a n c e of C16:0 and C14:0 (50.76% and 11.77%,
Table III
Amino acid composition.
Amino acids C.rubrum (µM) S.vulgare (µM) S.fusiforme (µM) P.pavonia (µM)

Essential
Lysine 9.06 9.94 6.99 17.76
Thereonine 9.85 19.34 28.94 14.08
Valine 9.66 ND 13.42 14.36
Isoleucine 60.5 ND ND ND
Phenylalanine ND 62.59 30.61 46.39
Sum essential 89.07 91.86 79.96 92.59
Non essential
10.92 ND ND ND
Alanine
Cystine ND 8.13 5.05 7.43
Glutamine ND ND 15 ND
Sum non 10.92 8.13 20.05 7.43
ND= not detected

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 International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:14 No:01 34
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