Ijesrt: International Journal of Engineering Sciences & Research Technology

ISSN: 2277-9655
[Pujari* et al., 5(10): October, 2016] Impact Factor: 4.116

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IJESRT
INTERNATIONAL JOURNAL OF ENGINEERING SCIENCES & RESEARCH
TECHNOLOGY
REVIEW ON SYNTHESIS, CHARACTERISATION AND BIOACTIVITY OF
CHITOSAN
Nisha Pujari , S. L. Pandharipande
*
Chemical Engineering Department, Laxminarayan Institute of Technology, Nagpur, India
DOI: 10.5281/zenodo.160867
ABSTRACT
One of the most widespread biopolymer in nature, after cellulose, is chitin. It can be extracted from sources like
crustaceans, microorganisms and insects. However, the main commercial sources of chitin are shells of
crustaceans such as prawns, crabs, lobsters and krill that are supplied in large quantities by the shellfish processing
industries. Extraction of chitin can be done by two processes, chemical and biological. Chemical method requires
acids and bases like sodium hydroxide, hydrochloric acid and acetic acid for deproteination, demineralisation and
decolourisation processes respectively and microbial method includes the use of various micro-organisms for the
similar steps as chemical process. The chitin obtained is further processed by deacetylation method using sodium
hydroxide to convert into chitosan, another product having high industrial significance which possesses valuable
properties like biocompatibility, biodegradability, antibacterial nature, film forming and fibre forming ability
promoting its use in a variety of interesting applications. Applications of chitosan are found in industries such as
textiles, medicine, food, agriculture, paper, cosmetics and wastewater treatment. The paper reviews the methods
of synthesis, characterisation of chitosan with analytical methods like FTIR, SEM, NMR etc. and bioactivity
determination of Chitosan as anti-microbial, anti-blood coagulant etc. properties.
KEYWORDS: Chitin, Chitosan, Characterisation
INTRODUCTION
Chitin is widely found in insects, fungi, and yeast and marine invertebrates. However, in higher plants and animals,
chitin is not present. Generally, the shell of selected crustacean consists of 30-50% calcium carbonate and calcium
phosphate, 30-40% protein and 20-30% chitin. The principal source of chitin is shellfish waste such as shrimps,
crabs, and crawfish. Although chitin itself is insoluble in water, on deacetylation it yields chitosan, a product
having a wide range of viable uses. Chitosan, which is very much similar to cellulose, is a non-toxic, biodegradable
polymer of high molecular weight. It is a co-polymer of glucosamine and N-acetylglucosamine and has generated
interest due to its biocompatibility, high charge density, non-toxicity and mucoadhesion. The biological properties
like biodegradability, adaptability, hemostatic activity and wound healing properties of chitin and chitosan
attracted much attention to their biomedical applications. They are also used in water engineering, in the food and
nutrients industry, film forming and coagulating ability and many more applications [1].
Until now, chemical, microbiological and enzymatic methods have been used for preparing chitosan from prawn
shell powders. The chemical method involves demineralization, deproteinisation, and deacetylation steps using
strong acids and/or alkali. However, the use of such chemicals seriously pollute the ecological environment and
harm human health, produce abundant waste and can hydrolyse the polymer. With increased demands on
environment-friendly culture, more eco-friendly processes have been worked upon by researchers using
microbiological and enzymatic methods for producing chitin and chitosan. The enzymatic method consists the use
of trypsase, papain, and pepsase. However, the high cost of enzymes and the low extraction are some of the pitfalls
of these methods [2].
This paper reviews the methods of synthesis, characterisation and bioactivity determination of Chitosan from
prawn shells.
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Seventeen papers reported in the literature have been studied and the review is presented in three parts: synthesis,
characterisation methods and bioactivity determination.
LITERATURE REVIEW
Synthesis
Chemical method
Abhrajyoti Tarafdar [3] & Gargi Biswas [3] performed the extraction of chitosan from prawn shells and its use in
successfully carrying out various applications in laboratory. They used inedible parts including head, body shells
and tails of P. monodon and P. indica for extraction of chitosan. Experiment were performed using two different
methods. In the first, 10 grams of prawn shell waste was used and washed. Demineralization process was carried
out by adding 1.5N HCl at room temperature for 1hour followed by de-proteinisation with 0.5% NaOH at 100⁰C
for 30 minutes. This method helped to weaken the protein tertiary structure of the shells. The process was repeated
for decolourisation with 3% NaOH at 100⁰C for 30 minutes to obtain chitin slurry. Deacetylation of chitin yielded
chitosan which was prepared by treating with 42% aqueous NaOH at 95°C for 1.5 hour and washed and then
dried. In second method, biomass of shrimp waste collected was 5grams. It was then de-proteinised in 4%
aqueous NaOH at room temperature (25⁰C) for 21 hours. The de-proteinised shell was demineralized by 4%
HCl at room temperature for 12 hours. The chitin was dried at ambient temperature. The Chitosan was obtained
by treating chitin with 50% aqueous NaOH at 40⁰C for 3 days. Finally, the chitosan was dried at ambient
temperature. It was observed that chitosan produced employing process-II was more readily soluble in 1%
acetic acid solution than that produced through process-I. It was confirmed that the chitosan obtained from P.
monodon shells had better activity and quality than that obtained from P. indica shells.
Musarrat H. Mohammed, Peter A. Williams et.al. [4] performed the extraction of chitin and chitosan from prawn
shells. Frozen prawn shells were initially washed with boiling water and then dried in an oven at 60oC. These
were crushed and powdered prawn shells were treated with 5% NaOH and refluxed at 60 oC for 2 h followed by
treatment with acetone to remove pigments at room temperature for 24 h. These were further treated with a 0.5 or
1% HCl solution for 24 h at 25oC to dissolve the calcium carbonate. The prawn shells were then washed several
times with water to obtain. Deacetylation process was done by treating of extracted chitin with sodium hydroxide
(NaOH) solution at elevated temperature and concentration. Effects of various parameters such as temperature,
NaOH concentration and reaction times on the deacetylation process were investigated. Further, the chitosan
produced was washed several times with distilled water, checked for pH neutrality and dried at 60oC in a vacuum
oven.
F. Nessa, Shah Md. Masum et.al. [5] evaluated the influence of deacetylation process in chitosan production on
the physiochemical and functional properties of prawn shell chitosan. The prawn shells obtained were sun dried
and ground into course particles through a centrifugal grinding mill. The dried prawn shell (1kg) was
demineralized with 10% HCl acid at ambient temperature for 22 hrs. The demineralized shells were deproteinised
with 10% sodium hydroxide solution for 24 hrs at 70º C. Samples were decolorized with acetone and dried under
vacuum for 2-3 hrs until the powder was crispy. The resultant product was chitin. Deacetylation of chitin was
achieved by using 60% NaOH solution. Four samples of prawn shell chitosan were prepared. The duration of
deacetylation process was 45 hrs, 55 hrs, 65 hrs and 72 hrs. The resulting chitosan were rinsed to neutrality with
distilled water, and dried at 65º C for 40 hrs in the oven. The yield of chitin was 20% and chitosan ranged from
16.4-19.6%. The prawn shell chitosan samples had a moisture content ranging from 0.3% to 0.4%. The nitrogen
content of the prawn shell chitosan samples varied between 7.91% and 8.33%. The ash content of prawn shell
chitosan range from 0.19% to 0.24%. The degree of deacetylation of prawn shell chitosan samples ranged from
45% to 75%. Three prawn shell chitosan samples B, C, D were found to have excellent solubility ranging from
96.01 to 97.2% with no significant difference but sample A which showed comparatively lower solubility (44.3%),
may be due to lower degree of deacetylation (DD). WBC of the samples B, C & D varied from 738.3 % to 748.4%
and FBC differed among chitosan products, ranging from 335.5% to 589.6%.
V. Mohanasrinivasan, Mudit Mishra et.al. [6] carried out the demineralization of shrimp shells, obtained from the
local market of Vellore, where it was first suspended in 4 % HCl at room temperature and then washed with water
to remove acid and calcium chloride. Then deproteinisation of shells was done further by treating it with 5 %
NaOH at 90C. After the incubation time the shells were sun dried. The product obtained was chitin. Chitosan was
prepared by deacetylating the obtained chitin with NaOH solution at room temperature for 72 h. The residue
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obtained was washed with running tap water, filtered, sun dried and was finely ground. The resultant whitish
flakes obtained after grinding is chitosan. The prepared chitosan had a percentage yield of 17 % and an ash content
of 2.28 % which when compared to commercial chitosan had 2 %. The solubilty of chitosan was found to be 1 %
acetic acid solution and partially soluble in water and moisture content of chitosan was measured to be 1.25 %. In
the present study, DD of the prepared chitosan was found to be 74.82 %. Chitosan prepared from shrimp shells
has WBC and FBC of 1,136 and 772 %.
Divya K, Sharrel Rebello and Jisha M S [7] collected five grams of shrimp shell waste and treated it with 4%
NaOH at room temperature for 24 hrs. The alkali was drained from the shells and the shells were then washed
with distilled water followed by treatment with 4% HCl at room temperature for 12hours to yield chitin. Chitin
was then filtered, washed with distilled water and dried at ambient conditions. Same process was repeated with
2% NaOH and 1% HCl. A slight pink colour was observed in chitin thus obtained. Chitin was soaked in 1%
potassium permanganate for 30 mins and then in 1% oxalic acid for 30 mins to 2hours for decolourisation
to take place. The decolourised chitin was then treated with 65% NaOH for 3 days at room temperature to form
chitosan. This alkali was then drained off and washed repeatedly with distilled water till pH was neutral.
The Chitosan obtained was further dried at room temperature and stored. The chitosan yield was found to be 46%.
The degree of deacetylation was found to be 85% and the concentration of chitosan in acetic acid is 7.7g/L.
Anshar Patria [8] used the randomized design group of 3 x 4 with two factors in this study. Shrimp shells
were procured from market of Banda Aceh city, Indonesia, washed, dried in sun light for 24 h and further dried
in a furnace at a temperature of 80˚C for 24 h. 20 g samples of shrimp shell powders were left for 1 hour at 90˚C
in 3.5 % NaOH. The solution was then filtered and the residue was washed and re-dried in a furnace at a
temperature of 60˚C for 4 hours. Thus, chitin powder was formed. This is then added with 2-N-hydrochloric
acid and allowed to stand for 1 hour at 90˚C. It is washed and dried in a furnace at 60˚C for 4 hours. The
extracted chitin is then treated with acetone for 4 hours in soxhlet and bleached with 0.32 % sodium
hypochlorite for 5 min at room temperature. A total of 5 g of chitin were reacted with 50 ml of 50 % sodium
hydroxide at various temperatures, then filtered and thus chitosan is formed. Chitosan yield were ranged from
50.39 to 88.25 % with average of 67.42 %, while the chitin yield was 40 %.
K. Kamala, P. Sivaperumal and R. Rajaram [9] collected P. stylifera shrimp shell wastes from the Versova landing
centre, Mumbai. Shells were thoroughly washed with running tap water and then placed in hot air oven at
600C for 24 hours. 100 grams of shrimp shell powder was left in 1000 ml of 7% (w/w) HCl at room
temperature for 24 h. Acid was washed with distilled water to neutral. Deproteination was carried out by
immersing the residue in 1000 ml of 10% (w/w) NaOH at 60°C for 24 h for. To wash the residue to neutral,
distilled water was used. To remove ethanol-soluble substances from the obtained chitin, 250 ml of 95% and
absolute ethanol were used. Chitin was dried overnight in an air oven at 50°C. Chitosan was prepared by putting
chitin in 50% NaOH for 8h at 60°C to prepare crude chitosan. After filtration, washings were given with hot
distilled water at 60°C for three times. The crude chitosan was dried in an air oven at 50°C overnight. 1 gram of
obtained chitosan was added in 20 ml of 2% (w/w) acetic acid. After reaction, to adjust the solution to neutrality,
10% NaOH was used. After filteration, two times the volume of ethanol were added to the filtrate. After
incubation at ambient condition overnight, the crystal of water-soluble chitosan was liberated and dried in an air
oven at 50°C. The yield of water soluble chitosan and crude chitosan was 87.8% and 54.3 %.
Sneha Paul, Aiswarya Jayanthe et. al. [10] removed the exoskeletons of the prawns separately, washed with tap
water and distilled water and dried at 55°C for about 24 hrs in a hot air oven. The sample obtained was immersed
in boiling 4% NaOH for 1 hr and then allowed to cool at ambient conditions for 30 minutes. Demineralisation
was carried out using 1% hydrogen chloride in ratio of 1:4 for 24 hrs to remove minerals. Further deproteination
was carried out by treating the samples with 50 ml of 2% NaOH for 1 hr. The remains of the sample were washed
with deionized water. Deacetylation process further carried out by adding 50% NaOH at 100°C to the sample for
2 hrs. The sample was then cooled at ambient conditions for 30 minutes and then washed with 50% NaOH. The
sample obtained is filtered, left uncovered, and oven-dried for 6 hrs at 110°C. The purification process of obtained
chitosan was carried out in three steps - removal of insoluble with filtration, reprecipitation of chitosan with 1 N
NaOH, demetallization of retrieved chitosan. The degree of deacetylation of the prepared chitosan was found to
be 87% and yield upto 67%.
Arafat A., Sabrin A Samad et. al.[11] dried shrimp shells in sun for 2 days. After sun drying, the shrimp were
grounded into powder. This powdered shrimp shell was demineralized with HCl at room temperature with 5%
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HCl for 24 hours with a ratio of 1:6. To remove acid and calcium chloride, these shells were rinsed with water
and dried in an oven to 60°C. Demineralized shells were deproteinized with 5% NaOH solution for 48 hours at
60-70°C at a ratio of 1:10 (w/v). The residue was washed with distilled water to neutral pH and dried for 2 days.
The product found is called chitin. Deacetylation process was carried out by treating samples with 60% NaOH
solution and heated for 2 hours. After rinsing and drying, the deacetylated chitin i.e. chitosan was ready. The
degree of deacetylation and yield of prepared chitosan was found to be 87% and 19% respectively.
Md. Monarul Islam, Shah Md. Masum et. al. [12] collected shrimp shell waste materials from Khulna,
Bangladesh. Shrimp shells were scraped free of loose tissue, washed with cold water and sundried for 2
days. The shells were then suspended in 4% HCl at room temperature in the ratio of 1:14(w/v). After 36
hours, to remove acid and calcium chloride, the shells rinsed with water. Further deproteination was carried
out by treating samples with 5% NaOH at 90°C for 24 hours with a ratio of 12:1(v/w). The washed to neutrality
in running tap water. This obtained chitin was dried in sun. Deacetylation of chitin was achieved by using 70%
NaOH solution with a ratio of 1:14 (w/v) at room temperature for 72 hours. The resulting chitosan was rinsed
neutrality in running tap water and distilled water and dried in sun. The degree of deacetylation was 75% and
yield was upto 15%.
Yateendra Shanmukha Puvvada et. al. [13] collected the shrimps exoskeletons from Suryalanka coast. To dissolve
the proteins and sugars, crushed shrimps exoskeletons were soaked in boiling NaOH (2 and 4% w/v) for 1
hour. After this, the samples were allowed to cool for 30 minutes at room temperature and the exoskeletons were
crushed to pieces of 0.5-5.0 mm. The grounded exoskeleton was demineralized using 1% HCl with four times
its quantity for 24 h to remove the minerals. To decompose the albumen into water soluble amino-acids, the
demineralized shrimp shell samples were subjected to deproteination process for 1 hour with 50 ml of 2% NaOH
solution. This chitin is washed with deionized water, which is then drained off. The deacetylation process is
carried out by adding 50% NaOH and then boiled at 100°C for 2 h. Afterwards the samples were washed
continuously with the 50% NaOH and filtered in order to retain the solid matter, which is the chitosan.
The samples were then oven dried at 110°C for 6 h. The obtained chitosan was purified in three steps, namely,
removal of insoluble with filteration, reprecipitation of chitiosan with 1 N NaOH, demetallisation of retrieved
chitosan. The chitosan yield was found to be 35.49%.
Microbial method
M. Khorrami, G. D. Najafpour et.al. [1] purchased Lactobacillus plantarum 1058 from Persian Type Culture
Collection (PTCC). The MRS medium was used. The medium was autoclaved at 101 kPa for 20 min at 121 °C.
The inoculated culture was cultivated in an incubator shaker at 30 °C and agitation rate of 180 rpm for 24 hours.
The 100 mL shrimp shell powder broth was sterilized, cooled and inoculated with 5% of seed culture. In the
incubator shaker, batch fermentation was carried out at 30°C and 180 rpm. This medium was incubated for 6 days.
The raw chitin obtained, after fermentation was treated with 0.5 mol L–1 HCl for 2 hours at ambient conditions
and then washed. This was further treated with 0.5 mol L–1 NaOH for 2 hours at room temperature and washed.
The purified chitin was put into a flask with 55 % NaOH solution for 4 hours in a water bath at 95 °C, followed
by washing and drying. The final product, chitosan, had a degree of deacetylation of about 83 %.
Hongcai Zhang, Yafang Jin et. al. [2] to extract chitin, fermented shrimp shell powders by successive two-step
fermentation of Lactobacillus plantarum ATCC 8014 and Serratia marcescens B742 was used. The one step
fermentation, identified optimal fermentation conditions were 2% SSP, 4 d of culture time, 2 h of sonication time,
10% incubation level and while that of using L. plantarum ATCC 8014 fermentation conditions were 2% SSP, 2
d of culture time, 10% incubation level and 15% glucose. Successive two-step fermentation resulted in chitin yield
of 18.9% with the final deproteinization rate of 94.5% and demineralization rate of 93.0%. Results showed that
the chitin prepared by the later method exhibited similar structural and physicochemical properties to those of the
commercial one, while using less chemical reagents.
Jag Pal, Hari Om Verma et.al. [14] took lactobacillus cell and transferred it in 5 ml MRS broth and incubated it
at 30oC for 24 hrs. 2 ml of starter culture was taken and transfer in 100 ml sterile MRS broth and incubated at 30
°C for a further 24 hrs, thus making them ready for fermentation. Shell fish waste was ground properly and
10% of any carbon source followed by 10% culture inoculums was added to it. This was then incubated for 180hrs
followed by filtering and the solid cake obtained was dried in hot air oven. In addition, deproteinisation can be
made by adding exo-proteases or by proteolytic bacteria. Lactic acid producing bacteria carries out the calcium
carbonate separation through the conversion of an added carbon source. Thus, chitin was extracted. The advantage

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of this technique is production of liquid fraction which is rich in protein and can be used for animal feed and
humans.
Sumathi, S., Hamsa, D., et.al. [15] focused on the isolation of the prawn shell waste and its conversion to chitosan
using microbial method. Latex milk of Euphorbia neriifolia, which has potent medicinal properties, was attempted
to incorporate with chitosan and check its antimicrobial activity. The prawn fish shell waste was collected from
the Ukkadam fish market in Coimbatore. The prawn shell waste were further washed thoroughly with tap water
to remove the soil particles, adherent protein, soluble organics, and other impurities and then dried for 24 hours
in a forced air oven at 60°C. Chitin was extracted from the prawn shell by deproteinisation and demineralisation.
Chitosan was further prepared from chitin through deacetylation process. Its results proved the antimicrobial
activity exhibited by the samples on test organisms.
Islem Younes, Olfa Ghorbel-Bellaaj et. al.[16] obtained the shrimp shells in fresh condition from a shrimp
processing plant located in Sfax, Tunisia. Shell waste were washed thoroughly with tap water, and cooked
with distilled water at a ratio of 1:2 (w/v) for 20 min at 90 oC. The cooked sample was drained and kept
at −20oC until further use. Two commercial enzymes, bromelain and alcalase were chosen as control for
deproteinization experiments. Shrimp waste homogenate was mixed with distilled water in ratio of 1:3. The
pH and temperature of the mixture, for each enzyme, were adjusted to the optimum conditions. To inactivate
enzymes, the reaction was stopped by heating the solution at 90 oC for 20 min. Demineralization was carried
out in a dilute HCl solution of 1.5 M HCl in 1:10 (w/v) ratio for 6 h at 50◦C under constant stirring.
The chitin product was filtered and washed to neutrality with deionized water and then dried for 1 h at
60oC. To deacetylate the purified chitin, it was treated with 12.5 M NaOH in 1:10 (w/v) ratio at 140 oC for
4 h. This residue was filtered and washed with deionized water, and the crude chitosan was obtained by
drying in a dry heat incubator at 50 oC overnight. Results showed that chitosan dissolved at 50 mg/ml.
Dilyana Zvezdova [17] used different local resources like shrimp shell and crab to extract chitosan.
Demineralization was carried out in all species by treating it with 7% HCl solution at room temperature with a
ratio of 10 (v/w). Distilled water was used to wash the residue till neutral pH. Then the demineralized samples
were dried and weighed. The demineralised shells were subjected to deproteinization using 10% NaOH at 60 °C.
This was repeated several times. Then the resulting solution was washed with water till pH was neutral. The
purified chitin was dried at 50 °C to constant weight. Deacetylation was prepared but putting the chitin obtained
into 50% NaOH for 8 h at 60°C to prepare crude chitosan. After filtration, the residue was washed with hot
distilled water repeatedly. The chitosan was obtained by drying at 50°C in an air oven for overnight. This chitosan
gives higher degree of morphological arrangement than that of standard chitosan.
Characterisation
The chitosan synthesised is analysed for its quality using analytical methods like FTIR, XRD, SEM etc. for
determination of functional groups, crystallinity, Surface morphology respectively. The details have been
presented in Table 1 to 9.
Table 1. Details of characterisaton method of chitosan

SR. NO. TITLE AUTHOR REF. NO.
1. Production of Chitin and Chitosan from Shrimp M. Khorrami, G.D. Najafpour,H. 1
Shell in Batch Culture of Lactobacillus plantarum.
Younesi and M. N. Hosseinpour
CHARACTERISATION METHOD
A). FTIR: OH stretching: 3490-3489; C–H symmetrical stretching: 2920-2925; C=O in amide groups: 1644-
1644; NH2 bending vibration: 1548-1548; C–O group in amide group: 1030-1027.


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2. Production of chitin from shrimp shell powders Hongcai Zhang, Yafang Jin, Yun 2
using Serratia marcescens B742 and Lactobacillus Deng, Danfeng Wang, Yanyun
plantarum ATCC 8014 successive two-step Zhao.
fermentation.
A). SEM: The smashed prawn shell powder displayed smooth microfibrillar crystalline structure, while the SEM
images after demineralisation showed more slightly fracture than that of prawn shell powder after
deproteinsiation with sonication treatment.
B). FTIR: Vibrations of –NH group: 1577 cm-1; vibrations of -C-O group: 1654 cm-1; vibrations of -CO-CH3
group: 2932 cm-1; polysaccharide structures: 890 and 1156 cm-1.
C). The wide-angle X-ray diffraction (WAXD): Purified chitin had WAXD pattern and showed two crystalline
peaks at 2θ=9.3° and 19.1°.

3. Extraction of chitin from prawn shells and Musarrat H. Mohammed, Peter A. 4
conversion to low molecular mass chitosan. Williams, Olga Tverezovskaya
A). FTIR: Splitting of the amide I band: 1658 cm-1; Intramolecular hydrogen bond: 1628 cm-1; NH stretching:
3262 cm-1 and 3114 cm-1; Mineral CaCO3: 1798, 1420-1430 and 876 cm-1; Protein has been removed: absence
of peak at 1540 cm-1; Primary amine groups: 1606-1566 cm-1; NH2 deformation of primary amines: 1632 cm-
1; NH2stretching: 3398 cm-1.
B). NMR Spectroscopy: DD was calculated by using integrals at 3.11 ppm and at 1.99 ppm.
C).Gel Permeation Chromatography: The polymer molecules eluting at peak 1 for 2 h have a Molecular
weight of 1.3*106 while those corresponding to peak 2 have a Molecular weight of 3.5*10 3. At the longer reaction
times of 5 h and 10 h, peak 1 elutes at higher elution times, corresponding to Mw values of 9.6*105 and 3.1*105
respectively. At a reaction time of 10 h average Mw values ranging from 0.6-2.1*105.

4. Studies on heavy metal removal efficiency and antibacterial V. Mohanasrinivasan, 6
activity of chitosan prepared from shrimp shell waste. Mudit Mishra et.al.
A). XRD: The XRD pattern illustrates two characteristics broad diffraction peaks at 2θ = 10 o and 20o.
B). FTIR: OH stretching vibrations: 3,450.65 cm-1; Asymmetric stretching of CH3 and CH2: 2,924.09 and
2,852.72; Bending vibration of NH2: 1,629.85 cm-1.
C). SEM: The micrographs showed non-homogenous and non-smooth surface.

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5. A Simple and Effective Method for Extraction of High Divya K, Sharrel 7
Purity Chitosan from Shrimp Shell Waste. Rebello and Jisha M S.
A). XRD: The XRD pattern showed characteristic peaks at 2θ= 9.280 and 20.180.
B). SEM: Chitosan had a long thin crystal structure on a smooth surface. Non-smooth and Non-homogenous
surface structure of chitosan was also reported.
C). FTIR: Deformation to CH3group: 1375 cm-1 corresponds to symmetrical. N-H deformation of amide II: 1552
cm-1; Vibration of amide I band: 1618 cm-1; Amide I stretching of C=O: 1654 cm-1; Free amino group at C2
position of glucosamine:1029 cm-1; C-O starching of primary alcoholic group: 1375 cm-1.

6. Extraction and Characterization of Water Soluble K. Kamala, P. 9
Chitosan from Parapeneopsis Stylifera Shrimp Shell Sivaperumal, R. Rajaram
Waste and Its Antibacterial Activity.
A). FTIR: Polysaccharide structure: 1155, 1078, 1032, and 899 cm−1; amide I and III bands: 3425, 1651, and
1321 cm−1; vibration of OH and CH: 1418 cm−1; OH group: 3411 cm-1; C–H stretching vibration: 2919 cm-1;
Bound water: 1610 cm-1; the sugar units in polysaccharide: 842 and 877 cm-1

7. Isolation and Characterization of Chitin From Sumathi S., Hamsa D., et.al. 15
Prawn Shell Waste and Incorporation into
Medical Textiles.
A). FTIR: The IR spectrum of the standard chitosan contained 15 major peaks whereas the IR spectrum of the
sample from prawn waste also recorded 13 peaks.

8. Chitin and chitosan preparation from shrimp shells using Islem Younes, Olfa 16
optimized enzymatic deproteinization Ghorbel-Bellaaja et. al.
A). CP/MAS-NM: C1-C6 carbons of N-acetyl glucosamine monomeric: 50 and 110 ppm. The carbonyl group:
173 ppm; methyl group: 23 ppm; CH3: 23 ppm; C-O: 173 ppm

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9. Synthesis and characterization of chitosan from marine Dilyana Zvezdova 17
sources in Black Sea
A). FTIR: N-H in NH2: 3425-2881 cm-1; CH3 group: 2921-2879 cm-1; NH2groups: The band at 1597 cm-1; Amide
II: 1559 cm-1; N–H bending vibrations: 1600–1400 cm−1; N–H bending mode of –NH2: 1600.9 cm−1; N–H bending
mode of amide 1 band: 1647.19 cm−1
Comparative studies of FTIR spectra show that the –OH and symmetrical NH stretching vibrations are indicated
by the wavenumber of around 3454 cm-1 which may be indicative of OH group in chitosan. CH3 and CH2
stretching’s indicated by the peaks of approximately 2924 and 2852cm-1 showed the extent of conversion of chitin
to chitosan. Bending vibrations of NH2 was observed at 1629 cm-1. The mineral matter remains were indicated
by the peaks 1798, 1420-1430, and 876 cm-1. Complete removal of protein was indicated by absence of peak at
1540 cm-1. Sugar units in polysaccharides were shown by the peaks at 842-877 cm-1. XRD analysis showed
characteristic peaks for chitosan at approximately 2θ= 10 o and 20o. Comparative study SEM method on chitosan
showed its thin crystalline, non - homogenous and non-smooth surfaces. Gel permeation chromatography showed
molecular weights ranging from 0.6*105 to 1.3*106 depending on the reaction times.
Bioactivity Measurements
The antimicrobial and shelf life properties of chitosan have been reviewed and summarised. The details are
presented in Table 2.
Table 10. Details of Antimicrobial and Shelf-life studies

SR. NO. TITLE AUTHOR REF NO.
1. Extraction of Chitosan from Prawn Shell Wastes and Abhrajyoti Tarafdar and 3
Examination of its Viable Commercial Applications Gargi Biswas
AREA OF APPLICATION REMARKS
A). Anti-Bacterial Activity: The degree of deacetylation plays an important role in The experiments carried
determining the antibacterial activity of chitosan as well. It was also observed that out by the authors showed
chitosan has profound antagonistic activity against gram negative bacteria than gram chitosan to be an anti-
positive. bacterial and anti-
b). Anti-Fungal Activity: Inhibited the growth of fungi and that the anti-fungal microbial product. It also
activity of Chitosan enhanced with the degree of deacetylation. can be used as a food
c). Chitosan As Food Preservative And Shelf-Life Enhancer: The experimental preservative upto a limited
sets, which were coated by chitosan showed no signs of spoilage after 1 week of number of days and
storage and looked as fresh. The experimental cucumbers showed no microbial or increases the scavenging
fungal attack, though they had stated to ripen on the twelfth. The experimental set activity and also acts as an
of tomato which were coated by chitosan were unchanged & looked as fresh without anticlotting agent for
any sign of spoilage even after seven days. blood.

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d). Blood Anti-Coagulant Activity: For set-I, even after 1 hour 30 minutes, test-
tube containing 7 ml chitosan showed no signs of clotting. For set-II, test tubes
containing 5ml & 7ml chitosan-II solution, even after 1 hour showed no signs of
blood clot formation.
E). DPPH Radical Scavenging Activity: The scavenging activity increased with
increase in chitosan concentrations but the degree of deacetylation of chitosan
affected the rate of radical scavenging.

2. Studies on heavy metals removal efficiency in V. Mohanasrinivasan, Mudit 6

industrial effluents Mishra et.al.
A). Heavy metals removal efficiency in industrial effluents: The results indicated Experimental study of the
that the ability to adsorb the metal ions present in industrial effluents was possessed authors proved chitosan to
by the prepared chitosan. Out of all the metal ions Cu (II) was best absorbed showing remove metal ions
removal of 98.97%. efficiently from industrial
B). Inhibitory activity of chitosan: The chitosan in the liquid medium inhibited the effluents and to possess
growth of Xanthomonas species and very less turbidity in the test flask was inhibitory activity against
observed. Xanthomonas species.

3. Isolation And Characterization Of Chitin From Prawn Sumathi, S., Hamsa, D. 15
Shell Waste And Incorporation Into Medical Textiles et.al.

A). Antibacterial Activity: Among the three, higher concentration of chitosan These authors showed
showed maximum zone of inhibition followed by higher concentration of latex milk chitosan to be
and the chitosan + latex milk. antibacterial and
B). Antifungal Activity: The percentage of spore germination was drastically reduced antifungal in nature.
in Aspergillus flavus, Aspergillus fumigatus and Mucor.

4. Extraction And Purification of Chitosan from Sneha Paul, Aiswarya Jayan, 10
Chitin Isolated from Sea Prawn (Fenneropenaeus Changam Sheela Sasikumar et.
Indicus) al.[]

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ISSN: 2277-9655
A). Antimicrobial activity: Chitosan showed an inhibition towards This study showed
Staphylococcus aureus and Candida albicans types of tests microorganism. More chitosan to be
prominent zone was seen in higher concentration of chitosan. antimicrobial in nature.

5. Extraction and Characterization of Water Soluble K. Kamala, P. 9
Chitosan from Parapeneopsis Stylifera Shrimp Shell Sivaperumal, R. Rajaram
Waste and Its Antibacterial Activity

A). Antibacterial activity: The water soluble chitosan showed higher zone of This study showed chitosan
inhibition range as compared to crude chitosan. It was indicated that both chitosans to be antibacterial in
might have the antibacterial inhibition mechanism. nature.

6. Chitin and chitosan preparation from shrimp shells Islem Younes, Olfa Ghorbel- 16
[]
using optimized enzymatic deproteinization Bellaaja, Rim Nasria et. al.
A). Antimicrobial activity: Chitosan obtained by deproteinisation with B. This study showed chitosan
mojavensis proteases having low degree of acetylation showed higher inhibition to be antimicrobial in
activity when compared with the commercial one with higher degree of acetylation. nature.
CONCLUSION
Chitin and chitosan, both are natural aminopolysaccharides having unique structures and with properties such as
biocompatibility, biodegradability, non-toxicity, with a wide range of applications. The raw material sources for
their production included crustaceans’ shells. An effort has been made in this paper to take a review of recently
published papers on synthesis of chitosan from variety of natural resources of prawns and shrimps. Two types of
synthesis methods have been reviewed that include chemical and microbial methods. The details of the process
parameters employed by various researchers which include quantity and concentration of HCl and NaOH in
Demineralisation, Deproteinisation and Deacetylation step with respective temperature and time conditions. In
microbial methods details about the medium, temperature, quantities and microbial species with respective time
duration have been reviewed and summarised. The paper also reviews the various characterisation methods
employed by researchers in evaluation of quality of chitosan synthesised. These analytical methods include FTIR
for determination of functional groups, SEM and XRD for surface morphology and surface crystallinity and NMR
for magnetic properties of certain atomic nuclei. The paper also covers the findings of various researchers that
studied the antimicrobial property of chitosan. Condensed form of summary of characterisation methods and
antimicrobial properties along with interpretation has been given in tabular form that would help the prospective
researchers.
REFERENCES

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[1] M. Khorrami, G. D. Najafpour, H. Younesi and M. N. Hosseinpour, “Production of Chitin and Chitosan
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[344]

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[Pujari* et al., 5(10): October, 2016] Impact Factor: 4.116

KEYWORDS: Chitin, Chitosan, Characterisation

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Table 1. Details of characterisaton method of chitosan

Table 2. Details of characterisaton method of chitosan

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Table 3. Details of characterisaton method of chitosan

Table 4. Details of characterisaton method of chitosan

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Table 6. Details of characterisaton method of chitosan

Table 7. Details of characterisaton method of chitosan

Table 8. Details of characterisaton method of chitosan

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Table 9. Details of characterisaton method of chitosan

Table 10. Details of Antimicrobial and Shelf-life studies

AREA OF APPLICATION REMARKS

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Table 11. Details of Antimicrobial and Shelf-life studies

2. Studies on heavy metals removal efficiency in V. Mohanasrinivasan, Mudit 6

AREA OF APPLICATION REMARKS

Table 12. Details of Antimicrobial and Shelf-life studies

AREA OF APPLICATION REMARKS

Table 13. Details of Antimicrobial and Shelf-life studies

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Table 14. Details of Antimicrobial and Shelf-life studies

AREA OF APPLICATION REMARKS

Table 15. Details of Antimicrobial and Shelf-life studies

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