Undergraduate Research Proposal

Department of Fisheries Science

Proposed on:- 7th June 2017

By:- Miss. Thapeetha Yoganathan

Supervisor:- Prof. S. Kuganathan

Proposed Title

Quantitative Study on the Seasonality and Sexuality of Chitin, in Selected

Commercial Crabs available in Jaffna.

Introduction

Every year, some 6 million to 8 million tonnes of waste crab, shrimp and lobster

shells are produced globally, about 1.5 million tonnes in South Asia alone. Whereas

75% of the weight of a tuna fish can be extracted as fillets, meat accounts for only

around 40% of a crab’s mass (Yan and Chen, 2015). These shells are regarded as

waste and dumped into the landfill or to the sea. Yet shells harbor useful chemicals-

protein, calcium carbonate and chitin. This chitin can be extracted simply using

chemical and biological methods.

Potential and usual applications of chitin and its derivatives mainly chitosan, are

estimated high and diverse (Brzeski,1987). Researchers have found that chitin and

chitosan as biocompatible, biodegradable and non toxic which have made wide

applicability in different fields, eg:- in cosmetics, agriculture, food, pharmacy,

biomedical, paper industry, and also as absorbent materials for waste water treatment

as well.

Yan and Chen, (2015) stated that the crab shell content was divided as calcium

carbonate (20-50%), chitin (15-40%), protein (20-40%). The degree of acetylation of

the extracted chitin, derived from the X-ray diffraction intensity values of chitin

characteristic peaks, revealed that the extracted chitin is purer than the commercially

acquired chitin.

As crab shell contain large amount of chitin, which can be effectively used in

industries and in-addition, thus helps in reduction of waste. Extraction of chitin is in

trend nowadays. Therefore, specific attention should be paid towards both biological

and chemical methods.

The paper titled on Extraction and characterization of chitin; a functional biopolymer

obtained from scales of common carp fish; reports about proposed chitin synthesis

process using fish scales as a raw material. Abdulkarim et al., (2013) carried out a

study on extraction and characterization of chitin and chitosan from mussel shell. Paul

et al., (2014) extracted and purified chitosan from chitin isolated from shrimp and

analyzed it for its physiochemical parameters, antibacterial, and antifungal activity.

Rasti et al., (2017) carried out a study on chitin extraction from mollusc chiton.
Biological extraction methods of chitin were conducted by Hajji et al., (2015) by

using Bacillus bacteria. Hamdi et al., (2015) carried out chitin extraction from blue

crab (Portunus segnis) and shrimp (Penaeus kerathurus) shells using digestive

alkaline proteases from P. segnis viscera.

Biological method of chitin extraction from crustacean shells is an advanced and new

technique which is eco friendly and low cost. Eventhough, it is limited in laboratory

condition as because demineralization and deproteinization have not yet reached upto

the yield as compared to chemical method (Pal et al., 2014).

Pandharipande and Prakash, (2016) investigated the extraction of chitin from crab to

produce nanostructured film. Sakthivel et al., (2016) extracted chitin from mangrove

crab Sesarma plicatum and characterized it and checked the antibacterial activity and

antifungal activity of it. A study was carried on Philippine Blue swimming crab by

Fernando et al., (2016) found that the extracted chitin has purity better than that of

commercially acquired chitin. Koli et al., (2015), isolated and characterized chitin

derivative chitosan from crab (Scylla serrata) shell waste.

The proposed work intends on the extraction of chitin from different species of crabs

and study seasonal variations, through which investigate the efficiency of chitin.

Objectives

 Assess the variability of chitin present in crab shells with regard to species.

 Determination of seasonal influence in crab shell chitin content.

 Study the chitin content difference with sex and crab shell types (soft and

hard).
 Time frame:-

September

November

December

January
October
August
June
May

July
Activities

Proposal
writing and
submission

Literature
review

Sample
collection

Laboratory
experiment
s
Analysis of
results

Writing
final report

Submission
of final
report

Research methodology:-

Sample collection

Chitin extraction will be conducted by using crab shell waste from the crab industries.

Neptunus pelagicus, Scylla serrata and Thalamita crenata the most abundant species

in Jaffna peninsula and where the availability is more constant year around.

Therefore, the study will be intented to concentrate on above species. Crab shells will

be cleaned by flushing in sea water, then dried and crushed. At the processing centre,

crushed samples will be considered for the further studies in laboratory.

Market purchased fresh crabs’ hard shells will be gathered after deshelling. Then,

after being boiled, soft shells will be gathered. The crab shells will be cleaned, dried

and crushed. Further study will be continued in laboratory using crushed samples.

Chemical method of chitin extraction

Cleaned crab shells

Grinding into powder

Weighed 25g
Demineralization 100ml of 1.0% HCl added
Allowed to soak for 24 hours

Filtration

Deproteinization 50ml 2% NaOH, 1hr

Filtration

Washed with distilled water

Wet cake

Drying (in oven)

Bleaching Washed with

Hydrogen peroxide

Drying

Analysis using IR
White chitin
spectroscopy
Expected outcome

 The research will assist the determination of chitin content in different species

of crabs.

 The results will provide knowledge on effect of seasonal variation on chitin.

 The results will provide knowledge about the influence of factors; such as sex,

and shell type on chitin content of the crab shell.

 Thus the project will be useful to establish chitin extraction industry in Jaffna

from waste crab shells to focus on high yield and will resolve the problem of

excess crab shell offs.

References

1) Abdulkarim, A., Isa, M. T., Abdulsalam, S., Muhammad, A. J. and Ameh,

A.O.(2013). Extraction and Characterization of Chitin And Chitosan from

Mussel Shell. Civil and Environmental Research. 3(2).

2) Arbia, W., Arbia, L., Adour, L. and Amrane, A. (2013). Chitin Extraction

from Crustacean Shells Using Biological Methods- A Review. Food

Technology, Biotechnology. 51(1): 12-25.

3) Burrows, F., Louime, C., Abazinge, M. and Onokpise, O. (2007). Extraction

and Evaluation of Chitosan from Crab Exoskeleton as a Seed Fungicide and

Plant Growth Enhancer. American- Eurasian journal of Agriculture and

Environmental Sciences. 2(2): 103-111.

4) Fernando, L. A. T., Poblete, M. R. S., Ongkiko A. G. M. and Diaz, L. J. L.

(2015). Chitin Extraction and Synthesis of chitin based Polimer films from
 Philippine Blue Swimming Crab (Portunus pelagicus) shells. Procedia

Chemistry. 19: 462-468.

5) Gaikwad, B. V., Koli, J. M. and Desai, A.S. (2015). Isolation and

Characterization of Chitosan from crab (Scylla serrata) ahell waste.

International Journal of Sciences And Applied Research. 2(8): 78-84.

6) Hajji, S., Bellaaj, O. H., Younes, I., Jellouli, K. and Nasri, M. (2015). Chitin

Extraction from crab shells by Bacillus bacteria. Biological activities of

fermented crab supernatants. International Journal of biological

macromolecules.

7) Hamdi, M., Hammami,A., Hajji, S., Jridi, M., Nasri, M. and Nasri, R. (2017).

Chitin extraction from blue crab (Portunus segnis) and shrimp (Penaeus

kerathurus) shells using digestive alkaline proteases from P. segnis viscera.

Internatinal Journal of Biological Macromolecules.

8) Kaur, K., Dattajirao, V., Shrivastava, V. and Bhardwaj, U. (2012). Isolation

and Characterization of Chitosan-Producing Bacteria from Beaches of

Chennai, India. Enzyme Research. 2012.

9) Maruthiah, T. and Palavesam, A. (2017). Characterization of haloalkalophilic

organic solvent tolerant protease for chitin extraction from shrimp shell waste.

International Journal of Biological Macromolecules. 97: 552-560.

10) Pal, J., Verma, H. O., Munka, V. K., Maurya S. K., Roy, D. and Kumar J.

(2014). Biological Method of Chitin Extraction from Shrimp Waste an Eco-

friendly low cost Technology and its Advanced Application. International

Journal of Fisheries and Aquatic Studies. 1(6): 104-107.

11) Pandharipande, S. L. and Bhagat, P. H. (2016). Synthesis of Chitin from Crab

Shells and its Utilization in Preparation of Nanostructured film. International

Journal of Science, Engineering and Technology Research. 5(5): 1378-1383.

12) Paul, S., Jayan, J., Sasikumar, C.S. and Cherian, S.M. (2014). Extraction and

purification of Chitosan from Chitin isolated from sea prawn (Fenneropenaeus

indicus). Asian journal Of Pharmaceutical and clinical Research. 7(4).

13) Rasti, H., Parivar, K., Baharara, J., Iranshahi, M. and Namvar, F. (2017).

Chitin from the Mollusc Chiton: Extraction, Characterization and Chitosan

Preparation. Iranian Journal Of Pharmaceutical Research. 16(1): 366-379.

14) Sakthivel, D.(2015). Extraction of Chitin and Chitosan from Mangrove crab

Sesarma plicatum from Thengaithittu Estuary Pondicherry Southeast coast of

india. International journal of Pharmacy and Pharmaceutical Research. 4(1):

12-24.

15) Yan, N. and Chen, X. (2015). Sustainability: Don’t waste seafood waste.

Nature 524: 155-157.