AGAR EXTRACTION

By:
Name
NIM
Group
Cluster
Assistance

: Gibran Muhammad Tri R

: B1K014025
:1
: IV
: Dewi Masfuah

PRACTICUM REPORT OF PHYCOLOGY

MINISTRY OF RESEARCH TECHNOLOGY AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
BIOLOGI FACULTY
PURWOKERTO
2016

I.

INTRODUCTION

A. Background
Chemically, agar is a polymer made up of subunits of the sugar galactose, and
is a component of the cell walls of several species of red algae that are usually
harvested in eastern Asia and California. Dissolved in boiling water and cooled,
laboratory agar looks gelatinous. Although agar's chief use is as a culture medium for
various microorganisms, particularly for bacteria, its other less well-known uses
include serving as a thickening for soups and sauces, in jellies and ice cream, in
cosmetics, for clarifying beverages, and for sizing fabrics (Aslan, 1991).
Agar is one fast consumable food from seaweed. When dissolved in hot water
and cooled, agar-agar is like gelatin: soft solids with many pores in it so textured
'springy'. This trait appealing sensory so many food preparations involve gelatin:
thickening

soups,

pudding

(jelly),

and

mixture

of

ice

cream.

Gelatin is widely known in Asia Tropika as a healthy food because it contains fiber
(fiber) high soft and low calories. Soft high fiber content help to expedite the
disposal of the remains of food in the intestine (laxatives). Besides being used as
food, gelatin is also widely used in the laboratory as a compactor kemikalia in the
experiment, growing medium for plant tissue culture and microbial cultures, as well
as stationary phase in gel electrophoresis. In the laboratory, jelly (usually packaged
in powder form) is known as agar or agarose alone (Cirik, 2010).
Gracilaria is a genus of red algae (Rhodophyta) notable for its economic
importance as an agarophyte, as well as its use as a food for humans and various
species of shellfish. Various species within the genus are cultivated among Asia,
South America. Species of Gracilaria is a seaweed cultivated in estuaries or in ponds,
although originally derived from the marine habitat. This happens because gracilaria
are include in euryhaline plant that can tolerate wide span salinity). Gracilaria usually
are used as raw material for making agar. In Indonesia, gracilaria is used as raw
material suppliers for agar factory (Afrianto, 1989).

B. Objective
To understand the process of extraction chemical content from seaweed such
as agar and the amount of it yield.

C. Literature Review
Agar or agarose is a gel substance that is usually prepared from seaweed
or algae. Some types of seaweed from the group Phaeophycophyta (Gracilaria and
Gelidium) can also be used as a source of agar. Gelatin exported from Melaka since
1871. Gelatin actually is a high molecular weight carbohydrate that filled with cell
walls of seaweed. It classified as pectin group and a polymer composed of monomer
galactose.

Agar

can

be

formed

as

powder,

solid

or

liquid.

Gel form because when heated in water, molecules of gelatin and water to move
freely. When cooled, the molecules of agar start closer together, condense and form
the lattice confined water molecules, forming a solid-liquid colloidal system. The
grille is used in agarose gel electrophoresis to hinder the movement of molecular
objects due to the voltage difference between the two poles. The density of agar gel
is also strong enough to hold small plants so it is often used as a medium in tissue
culture (Gessner, 1972).
Agar is an unbranched polysaccharide extracted from the cell walls of some
species of red algae or seaweed and having major economic importance. Chemically,
agar is a polymer made up of subunits of the sugar galactose, a monosaccharide.
Agar polysaccharides serve as the primary structural support for the algae's cell
walls. The seaweeds are known to contain other valuable materials in addition to the
fermentable carbohydrates, thereby providing carbohydrates as byproducts after
extraction of economically important products such as agaragar and phycocolloids.
The utilization of no-value byproduct for ethanol production will economize the
bioprocess and eventually will meet the concept of developing biorefinery with zerowaste. Through human creativity, it also serves a variety of purposes in human
culture and science. Dissolved in hot water and cooled, agar becomes gelatinous. Its
chief use is as a culture medium for microbiological work. Other uses are as a
laxative; a thickener for soups; in jellies, ice cream and Japanese desserts such as
anmitsu; as a clarifying agent in brewing; for paper-sizing fabrics; and as a
vegetarian gelatin substitute (Savindra, 2013).

Agar is a phycocolloid extracted from a group of red-purple marine algae

(Class Rhodophyceae) including Gelidium, Pterocladia and Gracilaria. Gelidium is
the preferred source for agars. Impurities, debris, minerals and pigment are reduced
to specified levels during manufacture. Agar is a gel at room temperature, remaining
firm at temperature as high as 65C. Agar melts at approximately 85C, a different
temperature from that at which it solidifies, 32-40C. This property is known as
hysteresis. Agar is generally resistant to shear forces; however, different agars may
have different gel strengths or degrees of stiffness. Agar is typically used in a final
concentration of 1-2% for solidifying culture media. Smaller quantities (0.05-0.5%)
are used in media for motility studies (0.5% w/v) and for growth of anaerobes (0.1%)
and microaerophiles. Specifications for bacteriological grade agar include good
clarity, controlled gelation temperature, controlled melting temperature, good
diffusion characteristics, absence of toxic bacterial inhibitors and relative absence of
metabolically useful minerals and compounds (Luning, 1990).
Kingdom : Eukaryota
Phylum

: Rhodophyta

Class

: Florideophyceae

Order

: Gracilariales

Family

: Gracilariaceae

Genus

: Gracilaria

Spesies

: Gracilaria verucosa

Seaweed Gracilaria verrucosa is seaweed that is included in the class of red

algae (Rhodophyta) this seaweed has characteristic such as: Thallus shape is
flattened or cylindrical, forming clumps with a type of irregular branching, narrowed
at the base thallus branching Gracilaria thallus nature of the substance as cartilage
(cartilagenous), the ends of the thallus generally tapered, smooth surface or berbintilnodule, Thallus middle line ranges from 0.5 to 4.0 mm, the length of Gracilaria can
reach 30 cm or more, the special feature of morphologically have thorns that grow
lined circling thallus with varying intervals so as to form segments in the circle of
thorns thallus. Gracilaria verrucosa usually used as raw material for agar, and in
Chinese it usually used as medicine to reduce fever, heartburn, laxative urine,
sputum, laxatives, taking care of the stomach, anti-dysentery, softeners and antitumor (Atmajaya, 1996).

II.

MATERIALS AND METHOD

A. Materials
The materials used in process of agar extraction are aquades 1000ml, KOH
100ml, KCl 5% 100ml, H2O2 6% 100ml, Gracilaria verrucosa 100gr.
The tools used in process of agar extraction are pan, analytical scale, mixer,
stove, tray, blender, filter cloth, and 100 ml measuring glass.
B. Methods
The process of making agar extraction are as:
1. All materials and tool are prepared.
2. Seaweed is crushed using blender.
3. Seaweed cooked and add aquades 500ml for 15 minute
4. KOH 10% 100ml and KCl 5% 100ml added and cook again for 15 minute
5. The mixture filtered and add aquades 500ml
6. Mixture cooked again and add H2O2 6% 100ml for 10 minute
7. Mixture filtered
8. Mixture dried for 4 days.
9. Yield Mixture measured

III.

RESULT AND DISCUSSION

A. Result
Formula to measure agar yield:
??? ???????(?)

Yield (%) = ??? ? ???????(?) X 100%

Yield measurement cluster VI:
?,?

Group 1 = ??? X 100 = 5,5%

?,?

Group 2 = ??? X 100 = 4,5%

?

Group 3 = ??? X 100 = 5%

?,?

Group 4 = ??? X 100 = 3,5%

?,?

Group 5 = ??? X 100 = 9,5%

Agar making process

B. Dicussion
Based on Sulistijo (1985) there are some characteristic of agar:
1. SOLUBILITY
Agar-agar is insoluble in cold water, but it swells considerably, absorbing as
much as twenty times its own weight of water. It dissolves readily in boiling water
and sets to a firm gel at concentrations as low as 0.50%. Powdered dry agar-agar is
soluble in water and other solvents at temperatures between 95 and 100 C.
2. GELLING
The gelling portion of agar-agar has a double helical structure. Double helices
aggregate to form a three-dimensional structure framework which holds the water
molecules within the interstices of the framework. Regarding its gelling power, agaragar is outstanding among other hydrocolloids. Agar-agar gels can be formed in very
dilute solutions, containing a fraction of 0.5% to 1.0% of agar-agar. These gels are
rigid, brittle, have well defined shapes, as well as sharp melting and gelling points.
Gelling occurs at temperatures far below the gel melting temperature. A 1.5%
solution of agar-agar forms a gel on cooling to about 32 to 45 C that does not melt
below 85 C.. The pH noticeably affects the strength of the agar gel; as the pH
decreases, the gel strength weakens. Sugar content has also a considerable effect over
agar gel. Increasing levels of sugar make gels with harder but less cohesive texture.
3. VISCOSITY
The viscosity of an agar solution at temperatures above its gelling point is
relatively constant at pHs 4.5 to 9.0, and is not greatly affected by age or ionic
strength within the pH range 6.0 to 8.0. However, once gelling starts viscosity at
constant temperature increases with time.
4. STABILITY
An agar-agar solution is slightly negatively charged. Its stability depends
upon two factors: hydration and the electric charge. The removal of both factors
result in flocculation of the agar-agar. Prolonged exposure to high temperatures can
degrade solutions of agar-agar, resulting in a lower gel strength after temperature
decrease and gel formation. The effect is accelerated by decreasing pH . Agar-agar
solutions and gels are fertile media for bacteria and/or molds and appropriate
precautions should be taken to avoid the growth of microorganisms.

Manufacture of gelatin paper by Suminarsih (1999) begins with the process:

1. Washing
By using 20x seaweed weigh, Seaweed washed with fresh water until
clean. Dirt that attched such as sand, coral, mud and other kinds
eliminated.
2. Bleaching
Bleaching is done by soaking the seaweed in 0,25% chlorine for one hour.
This will help to make seaweed clean.
3. Smoothing and coloring
To further facilitate extraction, the cell walls need to be broken by adding
KOH 1%. Seaweed soaked in KOH 1% for 15 minutes. Also give H2O2 to
brighther the agar color.
4. Cooking
Seaweed is cooked in water as much as 40 times the weight of the
seaweed. After boiling (90-1000 C), add lemon extract to obtain a pH of 67. If> 7, the pH is lowered by the addition of lemon extract, and if <6,
added NaOH. Cooking is done approximately 45 minutes but can also 2-4
hours depending on how the stirring.
5. Pressing and printing
The results of cooking and then filtered with calico cloth and pressed. The
discharge is collected in a vessel and neutralized by the addition of soda
water so that the pH becomes 7-7.5. if the pH has been reached, the liquid
is then cooked back while stirring. After boiling, the result is poured into
the mold. Approximately 6 hours of agar was cold and freezing.
6. Cooling
That had frozen liquid cooled in the refrigerator at a temperature of -200C
for 4-5 days. Cooling is done so that compaction is really happening
perfectly.
7. Drying
Gelatin is removed from the mold. The results obtained are gelatin sticks. If
desired gelatin sheet-shaped, gelatinous cut 0.5 cm thick. Can be used as a
cutting tool cutting tool, can be used fine wire of steel. Gelatin sticks or
slabs are then dried in the sun.

The function of materials and tool that used in agar making process according
to Rachmad (2002) are:
1. Materials
a. Gracilarium verrucosa: as raw materials for agar
b. KOH 1% 100ml: to breakdown cell wall of the seaweed and increase the
hydrogen potential
c. KCl 5% 100ml: to increase the agar mixture viscocity
d. H2O2 100ml: as a coloring agent to brighther the agar color
e. Aquades: to solvent the agar mixture
2. Tools
a. Pan: are used as container while cooking
b. Analytical scale: to measure the seaweed weight
c. Stirer: to stir the seaweed with other materials so it can blend perfectly
d. Stove: to cook the seaweed and blend it with other materials
e.

Tray: to accommodate the agar mixture after filtered

f.

Blender: to crush the seaweed so it can be easily cooked

g. filter cloth: to filter the cooked seaweed so it will become pure agar
mixture
Based on group 1 cluster IV result, the yield that extracted from 100gr
seaweed are 5,5% from 5,5gr dry weight seaweed. According to Zatnika (2008) the
yield amount in agar extraction is affected by temperature and drying method. When
the temperature is too high it can easily break the agar structure, while when the
temperature too it will slow down the drying process of agar extraction.

IV.

CONCLUSION AND SUGGESTION

A. Conclusion
Based on result and discussion we can conclude that:
1. Agar extraction need at least 4 days to get the yield
2. In agar extraction process the materials such as KOH 1%, KCl 5%, H2O2,
and environment factor while drying is important to yield weight in final
product.

B. Suggestion
When dried the agar yield in rainy season should use oven, so it will dry
faster and not much time to waste.

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Laut Indonesia. Puslitbang Oseanologi LIPI, Jakarta.
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Papenfuss and Determination of Chemical Composition. Turkish Journal of
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Laboratorium Biologi Struktur dan Fungsi Tumbuhan Jurusan Biologi
FMIPA UNDIP : 32-38