Synopsis of the proposed Research Project for National Science and Technology (NST)

Fellowship 2022-2023
Title of the project:
Extraction of Agar from the Red Seaweed Gracilaria, Gelidiella and Eucheuma Employing Simplified
Protocols and Testing Physico-Chemical Properties of Extracted Agar.
Title in Bangla:
লাল সামুদ্রিক শৈবাল জেদ্রলদ্রিয়েলা,গ্র্যাদ্রসলাদ্রি়ো এবং ইউযকদ্রম়ো জথযক সিলীকৃত পদ্ধদ্রতযত অ্যাগাি প্রস্তুতকিণ, এবং
প্রাপ্ত অ্যাগাযিি জ ৌত-িাসা়েদ্রিক শবদ্রৈষ্ট্য পিীক্ষা কিা।
Introduction:
Agar is a mixture of polysaccharides that are present in the cell matrix of red algae (Rhodophyta),
particularly in members of the Gelidiaceae and Gracilariaceae families. It has numerous uses in
biotechnology, medicine, and processed foods as a gelling agent. (Marinho-Soriano and Bourret, 2005).
Agar gels can be employed in a wide range of pH conditions, they are more stronger and more durable at
low concentrations (1 to 1.5%) with simply water. They can also survive temperatures exceeding 100 0C.
Agar gels could be repeatedly gelled and melted without losing its property. FAO/ WHO Codex
Aluminates have permitted the use of agar in human food industry in countries such as United Kingdom,
Germany, Russia, France and Poland. Food and Drug Administration (FDA) of United States (US)
assigned agar as a grading of Generally Recognized as Safe (GRAS) (FAO 2003). Agar is used in various
applications in food industries as thickening agents in the preparation of fruit salads, fruit jellies, yogurt,
bakery products, as a preservative in canned foods and meat industry and in liquor industry to increase the
viscosity. Higher concentration of agars is also used in fabricating moulds for sculpture, archaeological
and dental impressions (Mc Hugh 2003). Agar tends to decrease the concentration of blood glucose and
exerts an anti-aggregation effect on red blood cells and effects absorption of ultraviolet rays (Murata and
Nakazoe 2001). Gracillaria, Gelidiella and Eucheuma these three red algae are used for the extraction of
agar and their chemical properties with gel ability are tested. After that it has to be determined which one
is more suitable for preparation of agar in context with our country and which one is posing good quality
index.
Research Objectives:
 To extract agar from marine seaweeds (three species) of bay of Bengal by using modern techniques
 Testing gel criteria of agar for knowing which one is better for agar extraction
 Testing other physico-chemical properties of extracted agar
Materials and methods:
Sample collection and study area:
Seaweed samples will be collected from the Bay of Bangle coast of cox’s bazar and St. Martin Iceland of
Bangladesh. They will be bought in the CVASU laboratory for further analysis.
Bleaching:
The dry field sample will be soaked in tap water or even in seawater if the bleaching is done in the field.
Soaked sample will spread on bamboo mats, allowed to dry and then wet again by sprinkling water. This
process will be repeated until the Gracilaria, Gelidiella, Eucheuma sample is completely bleached. The
bleached seaweed remains brittle and dry.
Bleaching with lime:
We will make a lime suspension and sprinkle this on the drying seaweed. Lime even helps in increasing
the gel strength of the agar. Will repeat the sprinkling of lime water on the seaweed sample until bleached.
Alkaline pre-treatment and Extraction of agar:
Firstly, we will weigh two 1g samples. We will cut the dried seaweed thallus properly. Each sample will
put into each beaker and add 20mL, 5% NaOH. The samples will be heated on a water bath at 95°C for
one and a half hour. This technique will be repeated with acid solutions. This will be boiled on a hot plate
with continuously stirring with a glass rod till the volume reduced to one-fourth of the previous thickness.
Then the sample will be transferred on a muslin cloth and squeeze the cloth to find the agar extract falling
on the petri-dish. The yields of agars will be calculated based on the dry weight of the starting algae.
Determination of physico-chemical properties of extracted agar:
i. Sulphate content
Agar powder was hydrolysed in 6 mL of 0.5 M HCl at 105-110ºC for 12 h. After cooling, the solutions
will mix thoroughly and filter with a Whatman No. 1 filter paper. A portion will transfer to a 10-mL tube
containing 3.8 mL of 3% trichloroacetic acid and 1 mL of BaCl 2-gelatin reagent. The solution was mixed
vigorously and incubated at room temperature for 20 min. The absorbance at 360nm was measured using a
UV spectrophotometer. The sulphate content was calculated and expressed as the percentage.
ii. Determination of gel properties:
Agars will be dissolved in boiling distilled water to obtain a final concentration of 1.5% (w/v). The
solution will stir using a hot plate stirrer until agar will be solubilised completely. The agar solutions will
transfer to a cylindrical mould with 3cm diameter and 2.5 cm height. The solutions will incubate at a
refrigerated temperature (40C) for 12 h. The gels will equilibrate at room temperature (25–300C) for 2 h
before analysis. A commercial agar was used as a reference.
iii. Gel strength:
The gel strength was determined at 25-300C using a texture analyser with a load cell of 5 kg, cross-head
speed of 1 mm/s, equipped with a 1.27 cm diameter flat faced cylindrical Teflon plunger as per the
method of Lee, Namasivayam and Ho (2014). The maximum force (gram), taken when the plunger had
penetrated 5 mm into the agar gels, was recorded.
iv. Proximate composition profile:
Analytical methods Proteins, carbohydrate, lipids, fibre, ash and moisture contents in agar will be
determined by standard AOAC methods.
v. Bacteria Culture trial with extracted agar:
The agar provides a solid growth surface for the bacteria. We will suspend 6g of nutrient agar powder in
25ml of distilled water. Then this mixture will be heated while stirring to fully dissolve all components.
Dissolved mixture will autoclave at 1210C for 15 minutes. Once the nutrient agar will be autoclaved, allow
it to cool but not solidify. Then we will pour nutrient agar into each plate and leave plates on the sterile
surface until the agar will solidify. Then three kinds of microbes samples will inoculated and observe
weather they are growing on that agar surface or not.
Expected timeframe of the research:
2022 2023
Description of work with timeline Jan-Mar Apr-Jun Jul-Sep Oct-Dec Jan-Mar Apr-Jun
Review of Literature
Planning and Designing
Seaweed collection
Laboratory works and testing procedures
Result analysis
Thesis writing
Expected results, future outcome and conclusions:
Agar will be successfully extracted from three seaweed species, Gracilaria,Gelidiella and Eucheuma. It is
predicted that alkaline pre-treatment using NaOH will increase the yield of agar and improved gelling
property. NaOH pre-treatment showed the higher efficacy in improvement of agar properties than KOH as
indicated by higher increase in 3, 6-AG and lower sulphate content of resulting agar (Yarnpakdee, S,
Benjakul, S et al.,2015). We will also able to recommend one agar species among these three that which
one is more appropriate for producing agar commercially in our country. That can generate future agar
industries in our country in a low cost than importing agar from other country.
Financial Plan:
SL. No. Description of the Equipment Tk in amount
1. Research material expenses ( Chemicals, Glassware and others) 50,000
2. Sample collection and analysis 5,000
3. Report writing, printing, binding 10,000
4. Miscellaneous 5,000
Total 70,000

References:
Marinho-Soriano, E., Bourret, E., 2005. Polysaccharides from the red seaweed Gracilaria dura
(Gracilariales, Rhodophyta). Bioresour. Technol. 96, 379–382.
Murata M and Nakazoe J (2001) Production and use of marine algae in Japan. Jarq Japan Agric Res 35(4):
281–290.
FAO (2003) Aguide to the seaweed industry, FAO fisheries technical paper 44, Rome.
Mc Hugh DJ (2003) Aguide to seaweed Industry, FAO Fisheries Technical paper.No.441. Rome, FAO. Pp
105.
Yaphe, W., & Arsenault, G. (1965). Improved resorcinol reagent for the determination of fructose, and of
3, 6-anhydrogalactose in polysaccharides. Analytical Biochemistry, 13(1), 143-148.
Lee, W.K., Namasivayam, P., & Ho, C. L. (2014). Effects of sulfate starvation on agar polysaccharides of
Gracilaria species (Gracilariaceae, Rhodophyta) from Morib, Malaysia. Journal of Applied
Phycology, 26(4), 1791-1799.
Yarnpakdee, S., Benjakul, S., Kingwascharapong, P., Physico-chemical and gel properties of agar from
Gracilaria tenuistipitata from the lake of Songkhla, Thailand, Food Hydrocolloids (2015), doi:
10.1016/j.foodhyd.2015.05.004.

Name and Signature of Supervisor Name and Signature of Researcher

--------------------------------------- ----------------------------------------
Dr. Md. Faisal Fahmida Ali Ria
Associate Professor Roll No: 0122/01
Department of Fishing and Post-Harvest Technology Reg No: 1123
Faculty of Fisheries, Session: January,2022- June,2023
Chattogram Veterinary and Animal Sciences Department of Fishing and Post-Harvest
University, Chattogram-4225, Bangladesh Technology
Email: [email protected] Faculty of Fisheries,
Cell: 01719002289 Chattogram Veterinary and Animal Sciences
University, Chattogram-4225, Bangladesh
Email: [email protected]
Cell: 01729758439