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A new method for fast chitin extraction from

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A new method for fast chitin extraction

from shells of crab, crayfish and shrimp
ab bc bc
Murat Kaya , Talat Baran & Muhsin Karaarslan
a
Department of Biotechnology and Molecular Biology, Faculty of
Science and Letters Aksaray University, 68100 Aksaray, Turkey
b
Science and Technology Application and Research Center,
Aksaray University, 68100 Aksaray, Turkey
c
Department of Chemistry, Faculty of Science and Letters,
Aksaray University, 68100 Aksaray, Turkey
Published online: 02 Apr 2015.
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To cite this article: Murat Kaya, Talat Baran & Muhsin Karaarslan (2015): A new method for fast
chitin extraction from shells of crab, crayfish and shrimp, Natural Product Research: Formerly
Natural Product Letters, DOI: 10.1080/14786419.2015.1026341

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 Natural Product Research, 2015
http://dx.doi.org/10.1080/14786419.2015.1026341

SHORT COMMUNICATION
A new method for fast chitin extraction from shells of crab, crayfish
and shrimp
Murat Kayaab*, Talat Baranbc and Muhsin Karaarslanbc
a
Department of Biotechnology and Molecular Biology, Faculty of Science and Letters Aksaray University,
68100 Aksaray, Turkey; bScience and Technology Application and Research Center, Aksaray University,
68100 Aksaray, Turkey; cDepartment of Chemistry, Faculty of Science and Letters, Aksaray University,
68100 Aksaray, Turkey
(Received 27 December 2014; final version received 28 February 2015)
Downloaded by [Aksaray University] at 05:52 16 April 2015

A new method for quick chitin isolation from the shells of crab, crayfish and shrimp is
described. The main difference between the new method and the conventional method
is two sodium hypochlorite (NaClO) treatments for 10 min each before the processes of
demineralisation and deproteinisation. After the NaClO treatment, only 15 min is
adequate for the demineralisation and 20 min for the deproteinisation processes. Newly
extracted chitin from crab, crayfish and shrimp shells and commercial chitin were
characterised using FT-IR, TGA, X-ray diffractometry and elemental analysis. From
the results, it was observed that the chitins isolated with the new method and the
commercial chitin had almost the same physicochemical properties. The advantage of
the new method compared to traditional methods is the relatively rapid chitin
extraction. When compared to the traditional chitin extraction method, the proposed
method appears to be promising regarding its time and energy saving nature.
Keywords: new method; chitin; isolation; characterisation; sodium hypochlorite

1. Introduction
Chitin isolation using the classical chemical method involves demineralisation (1 4 h),
deproteinisation (16 24 h) and decolourisation (1 2 h) (Arbia et al. 2013). In total, the
procedure takes longer than 1 day. In recent years, new methods have been developed for chitin
extraction, such as bacterial fermentation (Ghorbel-Bellaaj et al. 2012) and enzymatic
deproteinisation (Manni et al. 2010; Jellouli et al. 2011; Younes et al. 2012). However, these
methods for the isolation of chitin from crustacean shells require a longer time and high energy
consumption. It seems that these methods could be modified to make the chitin extraction more
feasible. Therefore, any study capable of reducing the cost of chitin isolation is of importance.
Chitin is a valuable biomaterial because of its biocompatibility, non-toxicity, biodegrad-
ability and antioxidant and antimicrobial properties (Benhabiles et al. 2012). It is commonly
used in food, textile, medicine, pharmacy and water treatment (Felse & Panda 1999; Rinaudo
2006; Aranaz et al. 2009; Muzzarelli 2011; Laribi-Habchi et al. 2015). Although chitin is the
second most abundant biopolymer on the earth, the difficulty of its isolation restricts its mass

*Corresponding author. Email: muratkaya3806@yahoo.com

q 2015 Taylor & Francis

 2 M. Kaya et al.

production. However, chitin and its derivative have gained much interest worldwide, and the
demand for new chitin sources and new production methods is on the rise (Park & Kim 2010).
Although some scientists have carried out studies to find new chitin sources from insects, sponge
and fungi (Ehrlich et al. 2007; Sajomsang & Gonil 2010; Ifuku et al. 2011; Bo et al. 2012;
Ehrlich et al. 2013), there is no available extraction method that saves energy and time.
The aim of this study is to develop a quick isolation process for chitin from common
crustacean (crab, crayfish and shrimp) shells using chemical methods. In addition, it aims to
characterise the obtained chitin samples using Fourier transform infrared spectroscopy (FT-IR),
thermogravimetric analysis (TGA), X-ray diffractometry (XRD) and elemental analysis, and to
compare the chitins isolated with the new method to commercial chitin.

2. Results and discussion

2.1. Advantage of the new method compared to the traditional method
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The main difference between the proposed method and the conventional method is the two
treatments with NaClO (10 min each) before the processes of demineralisation and
deproteinisation. The purpose of the NaClO treatments was to remove any pigments and to
ensure complete disintegration. This treatment substantially reduces the time required for
demineralisation and deproteinisation. The introduction of NaClO into the extraction method
enabled us to accomplish the demineralisation and deproteinisation of the chitin samples in
relatively short times. In the traditional chitin isolation method, NaClO is used for bleaching
after the processes of demineralisation and deproteinisation. In our method, NaClO is used
before the demineralisation and deproteinisation processes, and thereby the chitin extraction was
achieved relatively quickly. The percentage chitin yields of the crab, crayfish and shrimp shells
following extraction with the new method was recorded as 13.4%, 15.3% and 14.8%. In previous
studies, the chitin contents of crab, crayfish and shrimp shells were observed to be between 10%
and 20% (Synowiecki & Al-Khateeb 2003). As can be seen, the chitin yields of the shells
extracted by the new method were found to be very similar to the previously reported yields.
In addition, the colour of the isolated chitins was white, which is the same as that of the
commercial chitin.

2.2. FT-IR
The FT-IR spectra of the commercial chitin and chitins isolated using the new methods are given
in Figure S1. There are three major bands that are used to identify a-chitin. These bands are
around 1660 cm21 (CvO secondary amide stretch, Amide I), around 1620 cm21 (CvO
secondary amide stretch) and around 1550 cm21 (NZH bend, CZN stretch, Amide II) (Kurita
et al. 1993; Lavall et al. 2007). In our study, we observed these bands at 1656, 1620 and 1550
cm21 for crab; 1655, 1550 and 1620 cm21 for crayfish; 1655, 1621 and 1551 cm21 for shrimp
and 1654, 1620 and 1551 cm21 for commercial chitin. As can be seen, all the bands of the chitins
isolated with the new method and the commercial chitin were quite similar (Figure S1).

2.3. TGA
The TGA results for the chitins extracted with the new method and the commercial chitin are
shown in Figure S2. Mass losses were observed in two steps for all the chitin samples. The
observed mass losses in the first step (between 25 and 1508C) were due to the evaporation of
water from the structure. In the second step, the observed mass losses between 150 and 6008C
can be attributed to the deterioration of the chitin structures. The mass losses in the first step were
recorded as 5.7% for crab chitin, 5.8% for crayfish chitin, 6% for shrimp chitin and 5.9% for
 Natural Product Research 3

commercial chitin. The mass losses in the second step were observed as 73.9% for crab chitin,
71.3% for crayfish chitin, 70.3% for shrimp chitin and 76.6% for commercial chitin. The
maximum degradation temperature (DTGmax) values were observed as 388.58C for crab chitin,
380.88C for crayfish chitin, 376.48C for shrimp chitin and 391.28C for commercial chitin.
When we looked at the TGA results for chitin samples in earlier studies, the weight losses
were observed in two steps, indicating a similarity to the analysis results observed here (Jang
et al. 2004; Paulino et al. 2006; Sajomsang & Gonil 2010). In addition, the percentage mass
loss rates of the chitins extracted using the new method were found to be very similar to the
results of analyses on commercial chitins (Jang et al. 2004; Paulino et al. 2006; Sajomsang &
Gonil 2010).

2.4. Elemental analysis

The elemental analysis results for the chitin from crab, crayfish, shrimp and the commercial
chitin are given in Supplementary Table S1. All the N, C and H percentages and C/N values of
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the chitin samples were found to be very similar (Supplementary Table S1). Only the H
percentage value of the crab chitin was found to be a bit lower than the others.
In the literature, it is known that the N percentage value of completely acetylated chitin is
6.89 and the DA (degree of acetylation) value is 100% (Sajomsang & Gonil 2010). In this study,
the N percentage values of the chitins (including commercial chitin) were observed to be lower
than the theoretical figure. The DA values of the chitins were found to be higher than 100%.
These N percentage and DA value results are similar to previously published studies (Sajomsang
& Gonil 2010; Liu et al. 2012).
In earlier studies, the N percentage and DA values of chitins isolated from crab, crayfish and
shrimp were quite similar to these results (Sajomsang & Gonil 2010; Liu et al. 2012). It has also
been reported that chitins isolated with the traditional method have very low N percentages and
very high DA values (Ifuku et al. 2011).

2.5. XRD
The XRD peaks and CrI (crystallinity index) values of the crab, crayfish and shrimp shell chitins
obtained by the new method as well as the commercial chitin are given in Supplementary
Table 2S. The XRD peaks of the chitins isolated with the new method and the commercial chitin
were quite similar (Figure S3). According to Jang et al. (2004), there are two sharp (around 98
and 198) and four weak peaks (138, 218, 238 and 268) in a-chitin. In our study, we observed the
same peaks as Jang et al. (2004) from commercial chitin. In addition, the CrI values of the chitin
obtained with the new method and the commercial chitin were very similar (Supplementary
Table S2).

3. Conclusion
Data obtained from TGA, FT-IR, XRD and elemental analysis indicated that the chitin isolated
by the new method, the commercial chitin and reports in the literature have high similarity. The
advantages of the new chitin extraction method over traditional methods are its shorter
processing time and economic feasibility. Based on the findings of this study, it can be said that
the new method can be used commonly in future chitin extraction studies.

Supplementary material
Experimental part of this article is available online, alongside Tables S1 and S2 and
Figures S1 S3.
 4 M. Kaya et al.

Disclosure statement
No potential conflict of interest was reported by the authors.

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