ALGINATE EXTRACTION

By :
Name : Angelin Marhavyna Cristy
Student ID : B1B015030
Entourage : IV
Group :1
Asisstant : Lukman Adi Nugroho

PHYCOLOGY LABORATORY REPORT

MINISTRY OF RESEARCH, TECHNOLOGY, AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY OF BIOLOGY
PURWOKERTO
2018
 I. INTRODUCTION

A. Introduction

Shrinking development industries developed countries have production of

various products that are required to meet the needs of a modern society, good for
food sources, medicines, cosmetic material, textile and so on. One of the ingredients
that are required is generally phyllocoid products, produced from the seaweed. One
of the ingredients phyllocoid which is needed in the industry is the alginat
compound. Compound of alginat needs for the textile industry in the Asia Pacific
region nowdays reach 8.000-10,000 kg,while compounds of alginat needs in
developed countries around 15,000 kg that most imported process (Sugiarto, 1978).
Potential alginate source is found in brown sea macroalgae. The main species
include: Ascophyllum, Ecklonia, Durvillaea, Laminaria, Lessonia, Macrocystis,
Sargassum and Turbinaria. In the cold climates the most important species are
Laminaria, Macrocystis, and Ascophyllum. While the tropical regions of the
Sargassum, Turbinarian, and Hormophysa are the main species of alginate producers.
The spread of brown algae in Indonesia grows to occupy almost along the coast of
the islands. The common macroalgae found abundantly is the Sargophysa clan found
in relatively small numbers. The utilization of brown microalgae is still in limited
circles, while the benefits on its downstream products have spread like beverage,
cosmetics, textile, paper, food and medicines. Alginate is an organic polymer
composed of 2 units of L-acid monomer Guluronat and D-Mannuronic acid.
Polymers are colloidal, gelled, hydrophilic, causing alginate compounds to be used
as emulsifying agents, thickening agents and stabilizing agents (Yulianto, 2009).
According to Deepika (2017) states algin is extracted from genera of Phaeophyta
is a linear polymer based on two monomeric units, β- D mannuronic acid and α- L
guluronic acid. Alginate is one of the clusters of polysaccharides formed in brown
algal cell walls, with levels reaching 40% of total dry weight and plays an important
role in maintaining the structure of algal tissues. Alginate was first synthesized by
Stanford in 1880. Alginate consists of three kinds of structures,
homopolysaccharides, which are alternate forms of α-1,4-L-guluronic acid and β-1,4-
D-mannuronic acid (Sugiarto, 1978) .
 Alginate is widely used in the cosmetics industry to make soaps, creams, lotions,
and shampoos. In the pharmaceutical industry requires for the manufacture of
suspensions, emulsifiers, stabilizers, tablets, ointments, capsules and plasters (Halim
et al., 2011). Alginate is utilized in various industries such as food, beverage,
medicine, cosmetics, textile, paint and other usage industries. Utilization of alginate
as emulsifying agent, disintegrating agent, moisturizing agent, utilization is based on
physical and chemical properties of alginate (Rasid & Rocmaniar, 2004).
The source of alginate derived from seaweed Sargassum sp. green which has
been obtained only from Sargassum chocolate (Sukma et al., 2014). According to
(Oliveira et al., 2014), Sargassum sp. belongs to Phaeophyta group (brown
seaweed),whose cellular walls are constituted of a fibrillar skeleton and an
amorphous matrix. The amorphous matrix is linked to the fibrillar skeleton via
hydrogen bonds. The most common fibrillar skeleton is cellulose (2–10% of dry
mass). The amorphous matrix is mainly composed of alginic acid (about 40% of dry
mass) or alginate (deprotonated form) besides small amounts of fucoidan (5–20% of
dry mass). Fucoidan contains 1-fucose- 4-sulfate monomers as major component
with sulfate ester ramifications. Despite alginate and fucoidan are also present in
fibrillar skeleton, cellulose remains as the main structural component.
Alginate can be determined through optimization of the extraction process which
is very important. Especially hydrolysis acid process because if the extraction is done
on the acidic atmosphere and the temperature is too high cause alginate will be easily
hydrolyzed so it will decrease the yield and quality of alginate flour obtained. If the
alginate can be extracted well then it can produce added value on the alginate-
producing seaweed (Winarno, 1996).
The extraction process is an application of mass transfer process, temperature.
The extraction process is one of the factors that influence mass transfer rate. The
amount of solvent affects the area of the solid contact with the solvent, the more the
greater the contact solvent will be, the greater the distribution of the solvent to the
solid. The uniform distribution of the solvent to the solid agar will increase the
resulting yield, the amount of solvent will reduce the saturation level of the solvent,
so that the alginate component in the seaweed will be extracted perfectly (Jayanudin
et al., 2014).
B. Objective

The objective of this laboratory activity is to know the yield of alginates and the
extraction process of alginate from seaweed Sargassum sp.
 II. MATERIAL AND METHOD

A. Material

The tools that used in this laboratory activity are pan, tray, beaker glass,
camera,filter cloth, strring rod (spatula), ph paper, and stove.
The matrials that used in this laboratory activity are 50grSargassum sp, Na2CO37
%, KOH 0,7 %, NaOCl 13 %, HCL 5 %, and NaOH 2 %.

B. Method

The method that used in this practicum, that are :

Sargassum sp are soaked in KOH 0,7 %

for 30 minute

Seaweed are cleaned with running water

Sargassum sp are soaked in HCL 5 % for 1

hour

Seaweed are cleaned with running water and weighed until 50 gr

350 ml of Na2CO37 % are added and boiled for 15 minute

150 ml of NaOCl13 %are added and boiled for 15 minute

350 ml of HCL5%are added and boiled for 15 minute

NaOH sufficiently are added until the Ph 7

The result is poured into tray and dried under the

sunlight. After that, calculated the yield content
 Calculation the water content of agar.
( B ) ( gram)
Yield content ( )= x 100
( A ) (gram)
Detail: A: Raw material
B: End product
 III. RESULT AND DISCUSSION

A. Result

Table 3.1 Calculation Results of Alginate

Extraction of Entourage IV
No Group Weight product End product
1 1 1.20 0,30
2 2 1.42 0,52
3 3 1.23 0,33
4 4 1.42 0,52

The calculationof water content of group 1 entourage IV

Initial weight of seaweed = 50 gram
Final weight of extract agar = 0,30
( B)
Yield content ( )= x 100
(A)
0,30
¿ x 100
50
¿ 0,6

Figure 3.1. Sargassum sp. is Figure 3.2. 350 ml of Na2CO3

put into pan to be 7% is added to the
boiled solution
Figure 3.3. 150 ml of NaOCl Figure 3.4. 350 ml of HCl 5% is
13% is added to added to the solution
the solution

Figure 3.5. NaOH is added to Figure 3.6. pH meter showed 7

the solution until the
pH turned 7
Figure 3.7. Alginate solution Figure 3.8. Alginate solution
are filtered using are poured into tray
filter cloth

Figure 3.9. Alginate solution Figure 3.10. Alginate solution

are dried under already dried
sunlight

Figure 3.11. Alginate yield

B. Discussion

The result of level of alginate yield from Sargassum sp. that obtained from our
group 1 in entourage IV is 0,6 %. It is a calculation between the final weight of
seaweed which is divided with the initial weight of seaweed. The weight of seaweed
after extracted and dried has reached 0,30 gram, while the initially weight of seaweed
before being treated is 50 gr. These results indicate that the alginate level of
Sargassum sp. seaweed extraction that obtained is low. It is not accordance with
Anggadireja et al. (1993), alginate from normal sargassum is 8 to 32% depending on
the type, season and condition of the waters where it grows. Environmental
conditions such as species, seasons and conditions of waters where the growth of
Sargassum sp. affect the rate of photosynthesis of seaweed so that it affects the
growth of seaweed which in turn also affects the alginate produced. The water
content of the alginate salt shows the amount of water still trapped in the alginate
molecule and this is proportional to the viscosity of alginate which also indicates its
binding strength. The long immersion will also cause the softening of seaweed cell
walls. The softening of this cell wall causes more alginate ingredients out of the
seaweed tissue during extraction, including minerals in seaweed tissue. Some water
is trapped in the dried alginate matrix so that the moisture content is still relatively
high (Zailanie et al., 2001).
Alginate is a prominent water-soluble polysaccharide found in brown
seaweed consisting of (1-4)-linked β-D-mannuronic acid (M) and α-L-gluronic acid
(G) units. It has very high water absorption as can absorb 200-300 times its own
weight in water. Alginate is able to be invented into a variety forms such as film,
microspheres and fibers because of their reversible solubility. However, the
application of alginate is limited owing to several weaknesses that high water
solubility and low mechanical strength. Its hydrophilicity can be the major problem
in DMFC performance because over water uptake in polymer electrolyte membrane
will leads to the high swelling ratio thus burst the membrane during working process.
Other than that sodium alginate membrane easy to dissolve in wet condition thus
lowered its mechanical stability (Shaari & Kamarudin, 2017).
Extraction procedure of alginate according to Rasyid (2010), seaweed is
weighing 5 grams was soaked in a 0.5% HCl solution for 30 min, at a ratio of 1:15
b/v (seaweed: HCl solution) in order to increase alginate levels and salt free
minerals. Subsequent submersion was performed using 0.5% NaOH solution at a
ratio of 1:15 b/v (seaweed: 0.5% NaOH) for 30 min, in order to remove the protein
compound. After immersion, seaweed was extracted by incubating the seaweed in
7% Na2CO3 solution at 60oC for 60 minutes. Next, the filtrate was filtered with a
filter paper, and the resulting filtrate was added 13% NaOCl solution by 2% of the
amount of filtrate produced, stirred until the color changed to yellow. Then, the
obtained filtrate was adjusted to a pH of 1-2 by adding 5% HCl solution, allowed to
stand for 30 minutes, and filtered using a 40 mesh filter. The gel obtained was then
dissolved in 10% Na 2 CO 3 solution, stirred until homogeneous, and the pH
adjusted to neutral. Further, the solution is dissolved in an isopropyl alcohol solution
while stirring. Finally, the obtained fiber is then dried, and is an alginate rendement.
Alginate is believed to be in the form of Na-alginate salt extracted using Na 2CO3 salt
(Winarno, 1990).
The solution used in alginate extraction by rashid method is KOH 2%, 0.5%
NaOH, 0.5% HCl and 15%, Na 2CO3 7% and H2O2. The solutions employed include
KOH for removing the gel from seaweed, NaOH to alginate algae acidic acid
(Sodium), HCl for demineralization or reducing the mineral salts still attached to
seaweed, H2O2 for brighten alginate, and Na2CO3 to remove alginate compounds.
According to Basmal et al. (2001), the process of extraction of brown seaweed was
done in an alkaline atmosphere aimed at separating cellulose and alginate. Na 2CO3
serves to extract the alginate content contained in the talus of brown seaweed. The
rate of alginate extraction present in the talus is highly dependent on the
concentration of Na2CO3, the temperature and duration of the given extraction.
NaOH which is one group of alkaline compounds in the process of seaweed
extraction function to form sodium alginate from alginic acid.
The process of extraction of brown seaweed is done in an alkaline atmosphere
aimed to separate cellulose and alginate. Extracting materials that can be used are
Na2CO3 and NaOH. Na2CO3 serves to extract the alginate content contained in the
talus of brown seaweed. The rate of alginate extraction present in the talus is highly
dependent on the concentration of Na2CO3, the temperature and time of extraction
given. NaOH which is one group of alkaline compounds in the process of seaweed
extraction serves to form sodium alginate from alginic acid (Sukma et ai., 2017). The
alginate extraction process also uses a functioning HCl in demineralization.
Subsequent immersion using 0.5% HCl. The atmosphere is too alkaline can cause
hydrolysis some alginate in seaweed so that when reacted with the acid (HCl)
amount of alginic acid obtained slightly. The color after this immersion is fixed.
Acidity causes the solution to be foaming, the color is blackish brown, and somewhat
thick(Susanto et al., 2001). According to Glicksman (1998) the use of HCl in
alginate, will break down the cell wall so as to facilitate the extraction, because HCl
is a strong acid and will be perfectly ionized.
Widyastuti (2009), also states that the high rendement alginate depends on the
type, condition of the growing place and climate. Added by Susanto et al. (2001),
alginate rendement is also influenced by habitat (light intensity, large or small waves
and aquatic nutrients). According to Sukma et al., (2017), alginate extraction is
influenced by several factors such as temperature and duration of heating extraction,
extortion, drying, and smoothing. the physical factors affecting the properties of the
alginate solution are the temperature, concentration and size of the polymer. Physical
characteristic of alginate salt is in the form of flour or fiber, white to yellowish,
almost odorless, and tasteless. While the chemical factors that influence the pH and
the presence of metal binders, as well as monovalent and polyvalent cations (Bahar
et al., 2012)
According to Wenten et al., (2009), alginate can be obtained by extracting brown
seaweed with an alkaline solution (generally Sodium Carbonate) to obtain a thick
paste of Sodium Alginate. Good quality alginates are cellulose-free alginates and are
pure white. High quality alginates are used primarily in the pharmaceutical and food
industries as thickening agents and emulsifiers, while the textile, paint and dye, paper
and paperboard industries can use low quality alginates. The complete standard of
alginate quality can be seen in table 3.2 below
Table 3.2 Quality standards of alginate from several grade.

Alginate that has the quality of food grade, must be free of cellulose and color
already bleached so bright or white, pharmaceutical grade, usually also free of
cellulose. Besides the grade, there is another so-called industrial grade usually still
allow some part of cellulose, with the color of brown until white. Alginate pH also
varies from 3,5 - 10, with viscosity 10 - 5000 cps, water content 5 - 20% and particle
size 10 – 200 mesh (Winarno, 1990). Standart quality can Seen in Table 1.
Table. Quality standards of alginic acid and alginate salts

Characteristics Alginic acid Alginate salt

Purity (% dry weight) 91-104% 90.8-106%

Rendemen > 20% > 18%

CO 2 levels <23% <21%

Levels of As <3 ppm <3 ppm

Pb content <0.004% <0.004%

Ash Content <4% 18-27%

Drying losses <15% <15%

Recent research about alginate who researched by Gao et al. (2017), study
plasticized alginate films with different biobased polyols (glycerol and sorbitol) and
their mixtures weresuccessfully prepared by thermo-mechanical mixing instead of
the usual casting-evaporation procedure.The microstructure and properties of the
different plasticized alginate formulations were investigatedby SEM, FTIR, XRD,
DMTA and uniaxial tensile tests. SEM and XRD results showed that native
alginateparticles were largely destructured with the plasticizers (polyols and water),
under a thermo-mechanicalinput. With increasing amount of plasticizers, the samples
showed enhanced homogeneity while theirthermal and mechanical properties
decreased. Compared to sorbitol, glycerol resulted in alginate filmswith a higher
flexibility due to its better plasticization efficiency resulting from its smaller size
andhigher hydrophilic character. Glycerol and sorbitol mixtures seemed to be an
optimum to obtain the bestproperties. This work showed that thermo-mechanical
mixing is a promising method to produce, at largescale, plasticized alginate-based
films with improved properties.
 IV. CONCLUSION & SUGGESTION

A. Conclusion

Based on result above, can be conlcude :

1. The result of alginate yield that obtained from group 1 entourage IV is 0,6 %.
2. Alginate yield is obtained from the extraction of Sargassum sp., whereas these
seaweeds were firstly soaked in KOH 0,7% for 30 minutes and then washed in
water flow. After that seaweed was soaked in a 0.5% HCl solution for 30
minutes. in order to remove the protein compound. After immersion, seaweed
was extracted by incubating the seaweed in 7% Na2CO3 solution at 60oC for 60
minutes. Next, the filtrate was filtered with a filter paper, and the resulting filtrate
was added 13% NaOCl solution by 2% of the amount of filtrate produced, stirred
until the color changed to yellow. Then, the obtained filtrate was adjusted to a pH
of 1-2 by adding 5% HCl solution, allowed to stand for 30 minutes, and filtered
using a 40 mesh filter. The gel obtained was then dissolved in 10% Na 2CO3
solution, stirred until homogeneous, and the pH adjusted to neutral.

B. Suggestion

The suggestion for this laboratory activity is when the boiling process or cook
needed optimum heat so as to soften the cell walls of seaweed is optimal.
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