Vol. 2, No. 2, September 2009
J. Ris. Kim.
OPTIMATION STUDY OF CARRAGEENAN EXTRACTION
FROM RED ALGAE (Eucheuma cottonii)
Refilda1*, Edison Munaf1, Rahmiana Zein1, Abdi Dharma1, Indrawati1
Lee Wah Lim2 and Toyohide Takeuchi2
1
Department of Chemistry, Faculty of Mathematics and Natural Sciences Andalas University
2
Department of Chemistry, Faculty of Engineering, Gifu University
*Corresponding author: telp +6281-26-628-806
email address: refilda_59@yahoo.com
ABSTRACT
Carrageenan is a general name for polysacharides, it is made up of sodium, potassium, magnesium,
and calcium sulphat esters of galactose and 3.6-anhydrogalactose units. Research about parameters
influencing extraction process of carrageenan from red algae (Eucheuma cottonii) has been studied.
The parameters were particle size of red algae, pH of NaOH solution, heating time and heating
temperature. About 64.30% of carrageenan was extracted from red algae (Eucheuma cottonii)
under optimum condition, 425 µm particle size of red algae, solution pH 8.5, heating time 18 h, and
heating temperature 95°C. The FTIR spectra of extracted carrageenan showed the same spectra as
the pure carrageenan.
Keywords : carrageenan, red algae (Eucheuma cottonii), and extraction
INTRODUCTION
Seaweed very simple chlorophyll-containing
organisms: some said that they are plants;
other said that they are not, calling them
protists or protoctists. We can say that they are
simple organisms composed of one cell, or
grouped together in colonies, or as organisms
with many cells, sometimes collaborating
together as simple tissues. Seaweed is
harvested throughout the world as a food
source as well as an export commodity for the
production of agar and carrageenan
products[1,2]. Seaweed has been cultured
traditionally for decades and probably for
centuries in several Asian nations such as
China, Korea and Japan. Until about 1980,
most of the seaweed production from other
nations in the region has been from the harvest
of wild stocks although limited culture took
place in nations such as the Philippines and
Indonesia. Species harvested include varieties
of it is a wholly natural ingredient obtained
from certain species of the red seaweed, class
Rhodophyceae.
Popular
sources
for
carrageenan are the Chondrus crispus,
Eucheuma cottonii and Eucheuma spinosum
120
species were originally harvested from natural
stocks growing in Indonesia and the
Philippines[3,4].
In Indonesia, the farmers usually sell
Eucheuma cottonii and Eucheuma spinosum to
middlemen; there are active middlemen and a
few local companies making semi-refined
carrageenan (SRC). The marketing chain for
Eucheuma cottonii in Indonesian, after
harvesting, the seaweed is dried, baled, and
sent to the carrageenan manufacturer and this
is also typical of the situation in other countries,
such as the Philippines. Indonesia still
importing carrageenan, because of still limited
processing industry of red algae, and still less
precisely extraction method of carrageenan. A
large proportion of the seaweed is exported to
Japan and the Republic of Korea. In Zanzibar
Carrageenan extractors actively pursue a
policy of assistance to those interested in
establishing seaweed farms, so future prospects
for the industry are very encouraging[5].
Cottonii seaweed grows well in salinities
ranging from 33 to 35 parts per thousand, and
will tolerate even higher salinities. The species
ISSN : 1978-628X
J. Ris. Kim.
Vol. 2, No. 2, September 2009
is intolerant of lower than normal salinities,
which are generally rare on outer-island coral
reef flats, occurring only for short periods
when heavy rain coincides with a period of low
water movement. Lower than normal salinities
are more common in lagoon environments, and
cottonii survival will be dependent on the
proximity to flushing with oceanic water, and
the duration of low salinity periods.
Commercial growth rates are achieved in
shallow water temperatures ranging from 24°C
to 30.5°C[6]. In Indonesia eucheuma farms
were found in Sulawesi, Bali, Lombok,
Sumbawa, Seribu Island, Ratu Harbour,
Lampung, Aceh and West Sumatera[7].
Carrageenan is a substance with hydrocolloid
properties owing to the presence of sodium,
potassium, magnesium and calcium sulfate
esters of galactose and 3, 6-anhydrogalactose
units. Carrageenans are large, highly flexible
molecules which curl forming helical
structures. This gives them the ability to form a
variety of different gels at room temperature.
They are widely used in the food and other
industries as thickening and stabilizing
agents[8,9]. A particular advantage is that they
are thixotropic—they thin under shear stress
and recover their viscosity once the stress is
removed. This means that they are easy to
pump but stiffen again afterwards[10].
Most carrageenan was extracted from
Kappaphycus alvarezii (Eucheuma cottonii)
and Eucheuma spinosum into water or aqueous
dilute alkali and may be recovered by alcohol
precipitation[11,12]. The original source of
carrageenan was Chondrus crispus, and this is
still used to a limited extent. Betaphycus
gelatinum is used for a particular type of
Kappa Carrageenan
carrageenan. Some South American species
that have previously been used to a limited
extent are now gaining favour with
carrageenan producers as they look for more
diversification in the species available to them
and the types of carrageenan that can be
extracted. Gigartina skottsbergii, Sarcothalia
crispata and Mazzaella laminaroides are
currently the most valuable species, all
collected from natural resources in Chile.
Small quantities of Gigartina canaliculata are
harvested in Mexico. Hypnea musciformis has
been used in Brazil[13].
Carrageenan is used in processed foods for
stabilization, thickening, and gelation. It has
been successfully used by the food industry in
the US since the 1950s, and with increasing
demand driven by the consumers need for
convenience,
appealing
food
textures,
advances in food processing, and new food
products. It is used worldwide to enhance ice
creams, chocolate milk, custards, cheeses,
jellies, confections, meats, and protein drinks.
Carrageenan is an approved food additive.
Carrageenan from red algae is benefit to health,
food industry, beverage, cosmetic, pharmacy,
paint, textile and others. Indonesia still
importing carrageenan, because of still limited
processing industry of red algae, and still less
precisely extraction method of carrageenan[14].
In order to increase production of carrageenan,
research about some parameters that
influencing extraction process of carrageenan
from red algae (Eucheuma cottonii) by alkaline
solution were studied. The parameters were
particle size of red algae, pH of NaOH solution,
heating time and heating temperature.
Iota Carrageenan
Lambda Carrageenan
Fig. 1 Three types of carrageenan[10]
ISSN : 1978-628X
121
Vol. 2, No. 2, September 2009
J. Ris. Kim.
MATERIAL AND METHODS
RESULT AND DISCUSSIONS
Treatment of Red algae (Eucheuma cottonii)
Effect of Particle Size on Extraction Yield
Carrageenan
Eucheuma cottonii, 40 days age, was harvested
from Seurapong village of Pulau Aceh district
at Aceh Besar sub-province of NAD.
Eucheuma cottonii was extensively washed
with distilled water to remove particulate
material from their surface and dried under
sunlight. Dried biomass was cut, ground in a
grinder and then was screened to particle size
of 150-425 μm.
Chemicals and Apparatus
All reagents in this research were pure. The
reagents were NaOH, isopropyl alcohol, KCl,
Standard Buffer Solution pH 4.0 and 7.0, filter
paper (Merck pro analysis) and distilled water.
The apparatus were used in this study were
grinder, octagon 200, Endcots, London,
Inggeris screener, analytical balance (AA-200,
Denver Instrument Company), pHmeter
(Denver Instrument Company), FTIR (Bio-Rad
FTS 60), water bath, vacuum pump, oven,
desiccators, and glasses that usually used in
laboratory.
Procedure
Carrageenan is obtained by extraction of 10 g
dried Eucheuma cottonii at various particle
size into 500 mL of alkaline solution (pH 8)
with steam boiling to maintain temperature
about 90°C and continues stirring for about 3
hours. Filtered and precipitated in isopropyl
alcohol 1:3, than dried in oven at 60°C,
weighted and milled to find powder.
The same procedures were done for various pH
of solutions, heating time and heating
temperature. FTIR Spectroscopy Analysis was
done to see functional groups of carrageenan.
For the IR studies, 5% (wt/wt) of ground, dried
extracted carrageenan and pure carrageenan
were pressed in KBr discs.
The FTIR spectra were recorded in the 1500500 cm-1 spectral range using a Bio-Rad FTS
60 instrument. A total of 128 scans were
averaged for each sample with a resolution of 2
cm-1.
122
The carrageenan extraction of the red algae
with
alkaline
solution
was
studied.
Percentation of carrageenan was influenced by
particle size of read algae and carrageenan, as
shown in Figure 1.
Figure 1 shows that percentage of carrageenan
increases with increasing particle size of
Eucheuma cottonii from 150 μm to 425 μm. At
particle size higher than 425 μm, the
percentage of carrageenan remind constant,
and particle size lower than 425 μm gave lower
percentage of carrageenan because it was
difficult to separated filtrate and residu by
filtering and gave much waste. Although
percentage of carrageenan exracted from
Eucheuma cottonii was optimum at particle
size 425 μm, the result indicated that for other
size still satisfactory. For the further
experiment particle size 425 μm was selected.
Effect of Heating Time on Extraction Yield
of Carrageenan
The effect of heat treatment on the yield of
carrageenan extracted from Eucheuma cottonii
was examined. The result is shown in Figure 2.
It indicated that the highest percentage of
carrageenan was produced after heating 18 h at
90°C. Heating time lower than 18 h gave lower
percentage of carrageenan because not all
carrageenan was extracted into alkaline
solution yet and heating time higher than 18 h
gave high solution concentration and it was
difficult to separated between filtrate and
residue.
Effect of pH on Yield of Extracted
Carrageenan
In Figure 3 the extraction yield of carrageenan
from Eucheuma cottonii depends on pH, the
percentage of carrageenan increased with the
increasing pH from 8.0 to 8.5 and then
decreased in the range 8.5-10. As the pH
increased, the negative charge density on the
solution will increase and increasing the
attraction of metallic ions with positive charge
ISSN : 1978-628X
J. Ris. Kim.
Vol. 2, No. 2, September 2009
on the Eucheuma
carrageenan[15,16].
cottonii
and
formed
Effect of Heating Temperature
Extraction Yield of Carrageenan
on
Caragenan Result (%)
The extraction yield of of carrageenan was
affected by heating temperature of Eucheuma
cottonii. The carrageenan diluted in hot
alkaline solution from 70-100C. In this
investigation heating temperature 95C gave
the higher percentage of carrageenan result. At
heating temperature lower than 95C not all
carrageenan was produced and heating
temperature higher than 95 C the concentrated
solution was produced and it was difficult to
separate filtrate and residue. Extracted
carrageenan from red algae (Eucheuma
cottonii) by alkaline solution in optimal
condition is 64.31%. It is higher than the result
was found by Syamsuar [4]
52
50
48
100
200
300
400
500
Particle Size (um)
Figure 1. Extraction yield of carrageenan from various particle size of Eucheuma cottonii treated with NaOH
solution at pH 8.5
Conditions: weight of material =10 g, heating time =12 h and heating temperature = 90 oC
Carrageenan result (%)
Carrageenan result (%)
60
55
50
45
40
35
30
70
65
60
55
50
45
40
35
30
25
20
0
5
10
15
20
25
30
Heating time (h)
Figure. 2. Extraction yield of carrageenan from
various heating time of Euchemia cottonii
Conditions: weight of material = 10 g, particle size
= 425 µm, solution of pH = 8.5 and heating
temperature = 90C
ISSN : 1978-628X
7.5
8
8.5
9
9.5
10
10.5
pH
Figure. 3. Extraction yield of carrageenan from
several solutions of pH of Eucheuma cottonni
Conditions: weight of material 10 g, particle size =
425 µm, heating time = 18 h, heating temperature =
90C.
123
J. Ris. Kim.
Vol. 2, No. 2, September 2009
FTIR Analysis
The functional groups of carrageenan that
produced by alkaline solution extraction and
pure carrageenan ware determined by FTIR[17].
The spectra are shown in Figure 5. The
functional group of both carrageenan almost
the same. The spectra could explained that the
extracted carrageenan from Eucheuma cottonii
was almost pure.
The study of carrageenans by FTIR
spectroscopy shows the presence of very
strong absorption bands in the 1266 cm-1
region (due to the S=O of sulfate esters) and
1068 cm-1 region (described to the glycosidic
linkage) in all carrageenan types. The other
chemical groups are characteristics of a given
carrageenan type, namely 3,6-anhydro-Dgalactose at 929 cm-1, D-galactose-4-sulfate at
846 cm-1. D-galactose-2-sulfate at 820 – 830
cm-1 and 3,6-anhydro-D-galactose-2-sulfate at
800-805 cm-1. The spectra could explain that
the extracted carrageenan from Eucheuma
cottonii almost pure.
Mainly carrageenans of the k-type has strong
band at approximately (bands at 933 and 847
cm-1) and i-type (bands at 933, 847 and 805
cm-1)[18]. The FTIR spectrumof alkali-modified
polysaccharide resembled k-carrageenan from
red alga Phacelocarpus peperocarpos with
absorption at 930 cm-1 (indicative of AnGal)
and 850 cm-1 (Gal 4-sulfate) and at 820 cm-1
indicating the presence of equatorial sulfate
ester substitution at O-6 of gal residues[19].
Carrageenan result (%)
80
70
60
50
40
30
20
80
85
90
95
100
105
110
Heating temperature (oC)
Figure. 4. Extraction yield of carrageenan from various heating temperature of Eucheuma cottonii.
Conditions: weight of material 10 g, particle size = 425 µm, heating time = 18 h
%T
A
B
4000
450
Cm-1
Figure 5. FT-IR spectra of pure carrageenan (A) and extracted carrageenan from Red algae (Eucheuma
cottonii)
ISSN : 1978-628X
125
J. Ris. Kim.
Vol. 2, No. 2, September 2009
a
b
c
Fig. 6. SEM image of a. red algae (Eucheuma cottonii), b. extracted carrageenan and c. pure carrageenan
Figure 6 shown that surface of extracted
carragenan from red algae is identical with
pure carageenan. Carrageenan has much pore
and big hole compared with red algae.
CONCLUSION
In conclusion, carrageenan could be extracted
from Eucheuma cottonii by alkaline solution
and precipitate by isopropyl Alcohol ander
optimal condition, particle size 425 µm
alkaline solution pH 8.5, heating time 18 h and
heating temperature 95C. Based on FTIR
spectra and SEM image the extracted
carrageenan shown the same functional groups
and particle pore as the pure carrageenan.
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