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Preparation, optimization and characterization of chitosan polymer from

shrimp shells

Article in Journal of Food Measurement and Characterization · September 2017

DOI: 10.1007/s11694-017-9490-9

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Food Measure
DOI 10.1007/s11694-017-9490-9

ORIGINAL PAPER

Preparation, optimization and characterization of chitosan

polymer from shrimp shells
Bachir Ben Seghir1 · M. H. Benhamza1

Received: 5 September 2016 / Accepted: 6 February 2017

© Springer Science+Business Media New York 2017

Preparation, optimization and characterization of chi- Introduction

tosan polymer from shrimp shellsBachir Ben Seghir, M.
H. BenhamzaAbstract In this study, chitosan is extracted Chitin is found in several organisms such as an important
from shrimp Parapenaeus longirostris shells by a chemi- exoskeleton constituent of crustaceans, insects, mollusks
cal process involving demineralization, deproteinization, and shrimp. It is also found as the main polymer of cell
decolorization and deacetylation. The effect of chitosan walls in some fungi and certain algae [1, 2]. In general, a
degree of deacetylation DD% is investigated using a model dry shrimp waste contains (30–40%) of protein, (30–50%)
of response surface methodology (RSM). The using of of calcium carbonate and (20–30%) of chitin [3]. The latter
ANOVA regression equation gives a determination coeffi- can generate about 80% of chitosan, and the term chitosan
cient R2 = 0.9853, indicating the precision of the predicted usually refers to a family of polymers obtained after chi-
model. Chitosan derived from chitin treated under opti- tin deacetylation to varying degrees. Chitosan is a natural,
mized conditions of 28.6% of NaOH concentration, a tem- non-toxic, biodegradable and high molecular weight poly-
perature of 81.15 °C and a reaction time of 9.55 h, showed cationic polymer [4, 5]. It is described as ‘nature’s most
the highest value of DD% exceeding 98%. The obtained versatile biomaterial’ and has been reported to have numer-
Chitosan characterization results using a spectral analysis ous applications especially in pharmaceutical, biomedical,
infrared (FTIR), Differential Scanning Calorimetry (DSC), biological, environment and food technology [6].
X-ray spectroscopy (DRX) and Scanning electron micros- Chitin is a naturally abundant polymer consisting of
copy (SEM) confirmed the purity of chitosan. 2-acetamido-2-deoxy-D-glucopyrnose [7, 8], and chi-
tosan is constituted primarily of glucosamine or 2-amino-
Keywords Chitosan · Shrimp · Deacetylation · Response 2-deoxy-d-glucose linked together by β(1–4) glycosidic
Surface · Characterization bonds [9–11]. Several studies have clearly demonstrated
that the chitin extraction steps can be achieved mainly
by two methods, either chemically or biologically. The
chemical process of chitin preparation has been devel-
oped by many researchers, it consist firstly by a deminer-
alization to remove carbonate and phosphate calcium [12,
13], and then deproteinization is done to eliminate pro-
* Bachir Ben Seghir tein, after that decolorization is made to remove pigments
[email protected] and finally chitin is converted into chitosan by alkaline
M. H. Benhamza deacetylation [14, 15]. Chitosan can be characterized in
[email protected] terms of its quality intrinsic properties such, molecular
1
weight, viscosity and degree of deacetylation (DD%).
Laboratory of Industrial Analysis and Materials Engineering,
Department of Process Engineering, Faculty of Sciences
The most widely used methods for the latter are acid-
and Technology, University 8 May 1945 Guelma, BP 401, base titration [16], X-ray spectroscopy (DRX) [17, 18],
Guelma 24000, Algeria infrared (IR) [19–21], UV-Vis-spectroscopy [22], nuclear

13
Vol.:(0123456789)
 Bachir Ben Seghir, M. H. Benhamza

magnetic resonance spectroscopy (NMR) [23–25], gas Experimental and methods

chromatography, pH-metric titration, mass spectrometry
(MS) and chromatography HPLC [26]. Materials
The purity of chitosan is established by its characteri-
zation using spectrometry IR, DRX, differential scan- Shrimps of Parapenaeus longirostris variety (Fig. 1),
ning calorimetry (DSC) and scanning electron micros- obtained from local fish market at Guelma city (Algeria)
copy (SEM) [10, 12]. Many research works on optimized are used in this study. Chemicals used in the process are
conditions by considering various significant variables Hydrochloric Acid HCl and Sodium Hydroxide NaOH
have been reported, and one of the most important is the (purchased from Sigma-Aldrich, USA) and Hydrogen
degree deacetylation (DD%) in a range of conditions [16, peroxide ­H2O2 of 30% concentration (purchased from
27]. Merck KGaA, Darmstadt, Germany).
Several factors can influence chitin deacetylation pro-
cess and different quality of chitosan can be obtained.
Reports indicate that concentrated NaOH in the process Preparation of chitin and chitosan
at different conditions is one of the important factors. The
quality of chitosan is also depending upon shrimp species The chemical process of chitin and the extraction of chi-
and seasonal variations. In order to differentiate chitin from tosan method are described below:
chitosan, the average degree of deacetylation (DD%) is
determined [7, 28]. Therefore, there is a need for process
optimization. Demineralization
Several factors cause difficulty in understanding and
controlling products’ results; thus statistical response sur- Demineralization is typically achieved by acid treatments
face methodology (RSM) based on the Box-Behnken design which dissolve the calcium carbonate and calcium chlo-
(BBD) is used to investigate chitosan production by opti- ride found in shrimp shells. The acid used is hydrochloric
mizing the chitin deacetylation process [29]. acid of 4% concentration. Duration of the acid treatment
The chitin deacetylation process optimization depends is 24 h at a room temperature of 25 °C and with a ratio of
on several parameter such as number of successive solid to solvent v/w equals to 15/1(ml/g). During the pro-
baths used, reaction time, temperature, alkali reagent cess of demineralization, the presence of carbon dioxide
(NaOH–KOH), atmospheric conditions (Nitrogen–Air), use gas ­(CO2) emission is an indication to the extent of min-
of sodium borohydride (with–without) and finally alkali eral matter present in the process [6, 7, 9].
concentration [30]. The temperature is found to be the
major factor that control chitosan acetylation degree. The
lowest degrees deacetylation is obtained when using NaOH
at high concentration instead of KOH, and increasing alkali
concentration toward saturation has little effect on deacety-
lation and viscosity [31–33].
The objective of the present research work is to inves-
tigate chitosan production by optimizing the chitin deacet-
ylation process in order to increase its purity and that by
using chemical methods. The influence of the following
variables: NaOH concentration (C), temperature (T), reac-
tion time (t) and solvent to solid ratio (r) have been investi-
gated by means of Response Surface Methodology (RSM).
Then the extracted chitin and chitosan is characterized
using FTIR, XRD, DSC and SEM techniques.
The novelty in the present research work is the jointly
use of the most important controlled parameters (temper-
ature T, NaOH concentration C, reaction time t and solid
to solvent ratio r) and that in wide and extended interval
range. In addition information on chitosan extracted from
Parapenaeus longirostris variety is not available. Further-
more, RSM statistical method and FTIR technique are both
employed to optimize the chitin deacetylation process. Fig. 1  Shrimp of Parapenaeus longirostris (a) and shells (b)

13
Preparation, optimization and characterization of chitosan polymer from shrimp shells

Deproteinization thoroughly mixed with KBr, the dried mixture is then

pressed to form a homogeneous sample/KBr disc. The
After demineralization the dried shrimp powder is placed absorption bands at 1655 and 3450 cm−1 are used to cal-
in 10% sodium hydroxide (NaOH) solution during 2 h at culate the DD% according to the following Eq. 1 [34, 35]:
70 °C and at a ratio of solvent to solid (v/w) of 15/1 ml/g.
(1)
[ ( ) ]
DD% = 100 − 100 × A1655 ∕A3450 ∕1.33
This is done in order to remove the remaining proteins
as well as other organic materials. After that, it is filtered where: DD% is deacetylation degree, A1655 and A3450 ­cm−1
under vacuum then washed with tap water for 30 min until are absolute heights of absorption bands of amide and
the solution reaches a neutral pH (pH = 7). Thus the prod- hydroxyl groups respectively. The factor 1.33 denotes the
uct obtained is a chitin [8, 10]. value of the ratio ­A1655/A3450 for fully N-acetylated chi-
tosan [3, 34, 35].
Decolorization

The crustacean shells pigment which still found in the chi- Differential scanning calorimetry (DSC)
tin should be removed by reagents such as ethanol, ether,
sodium hypochlorite NaOCl or by hydrogen peroxide Differential scanning calorimetry measurements of chi-
­H2O2. During bleaching process, the reagent used must not tosan are performed using a DSC Q200 type (TA Instru-
have an effect on the chitin physicochemical properties. ments, NJ, USA). The mass of chitosan sample is of
Therefore, in this work the hydrogen peroxide H
­ 2O2 is used 5 mg and the empty cup is used as reference. Samples are
[8]. analyzed under a continuous flow of dry nitrogen gas at
10 ml/min and at a heating rate of 10 °C/min from 50 to
Deacetylation 450 °C [27, 36].

Deacetylation of chitin is performed to remove acetyl

group. It is generally achieved by concentrated sodium X‑ray diffraction (XRD)
solution (40–50%) treatment, Fig. 2. This process is per-
formed at temperature of 100 °C, at a ratio of solvent to Structural and crystalline characterization is made using
solid (v/w) of 10/1(ml/g) and the duration reaction time is X-Ray diffraction meter (D500 Siemens diffractometer
of 12 h. After deacetylation chitosan is obtained, it is then mode, Germany). The measurement conditions are as fol-
washed and rinsed with hot distilled water at 90 °C, after lows: Cu-Ka radiation λ = 1.54 Å, 2θ angle varying from
that it is filtered and oven dried at 50 °C for 16 h [12, 14, 5° to 70° at a scanning rate of 4.8 min−1, the voltage is of
15]. 40 kV and the intensity is of 30 mÅ [10, 12].

Characterization of chitosan
Scanning electron microscopy (SEM)
Fourier transforms infrared spectroscopy (FTIR)
The morphology of the prepared chitosan is illustrated by
The spectra of chitosan are obtained using an infrared a SEM micrograph. The scanning electron microscopy
spectroscopy of a type Perkin Elmer FTIR 1600 (Per- (SEM) is conducted using a JEOL JSM−630 J Scanning
kin Elmer Spectroscopy, USA), with spectral frequency Electron Microscope type (JEOL Ltd., Tokyo, Japan)
ranging from 4000 to 400 cm−1. The chitosan sample is operating at 5 and 20 kV [10, 12].

Fig. 2  Conversion of chitin to

chitosan by deacetylation

13
 Bachir Ben Seghir, M. H. Benhamza

Statistical analysis using a response surfaces design consists of twenty-four factorial points each one with three
method (RSM) levels coded −1, 0, and +1 and three axial points to form
a central point, (Table 1). Derived result is in the form of
A statistical analysis using a response surfaces design second-order polynomial Eq. 2:
method (RSM) is performed to find an optimum DD%
in order to obtain a chitosan of high purity. The chosen Y = a0 + a1 x1 + a2 x2 + a3 x3 + a4 x4 + a12 x1 x2 + a13 x1 x3
(RSM) method applies a Box–Behnken Design (BBD) + a23 x2 x3 + a14 x14 + a24 x24 + a34 x34 + a11 x21
and the mathematical model used is of the second degree + a22 x22 + a33 x23 + a44 x24 … .. + e
with two-way interactions [13]. The experimental design (2)
where Y is the predicted response; ­a0 is a model constant;
­a1, ­a2, ­a3 and a­ 4 are the linear coefficients; ­a12, ­a13, ­a14, ­a23,
Table 1  Experiment design levels for various parameters ­a24 and a­ 34 are the interaction coefficients; a­ 11, ­a22, ­a33 and
Parameters Codes Level
­a44 are the quadratic coefficient; ­x1, ­x2, ­x3 and ­x4 are the
independent variables; ­x1x2, ­x1x3, ­x2x3 are interaction fac-
−1.0 0.0 1.0 tors terms and x­12, ­x22, ­x32 and x­42 are quadratic terms
Concentration NaOH (%) x1 25 50 75 (Table 2) [14, 15]. The RMS procedure is performed using
Temperature (°C) x2 40 70 100 Design Expert 9 statistics software.
Time (h) x3 6 15 24
Ratio v/w (ml/g) x4 5/1 10/1 15/1

Table 2  Experimental runs Run x1 x2 x3 x4 DD%

and responses of degree of
deacetylation (DD%) Experimenta* Predicted

1 −1 1 0 0 80.00 79.45
2 0 0 0 0 71.00 72.01
3 0 0 1 −1 65.62 65.20
4 1 1 0 0 54.65 56.80
5 0 −1 −1 0 40.00 43.45
6 0 0 −1 −1 90.00 90.48
7 0 0 1 1 64.00 61.37
8 0 0 0 0 72.05 72.01
9 1 0 −1 0 61.11 60.38
10 0 −1 1 0 45.00 47.73
11 1 0 1 0 61.00 61.89
12 0 1 0 −1 81.24 81.99
13 1 −1 0 0 61.24 59.64
14 −1 0 0 −1 83.00 83.79
15 0 −1 0 −1 69.00 67.86
16 1 0 0 1 75.00 74.75
17 −1 0 0 1 52.68 53.70
18 0 1 −1 0 89.79 87.60
19 −1 0 −1 0 73.96 74.66
20 −1 −1 0 0 34.00 29.70
21 −1 0 1 0 38.00 40.32
22 1 0 0 −1 70.53 70.05
23 0 0 0 0 73.00 72.01
24 0 1 1 0 53.40 50.49
25 0 −1 0 1 45.00 47.73
26 0 1 0 1 75.90 78.63
27 0 0 −1 1 70.65 68.92

*Degree Deacetylation (DD %) obtained using FTIR spectroscopy

13
Preparation, optimization and characterization of chitosan polymer from shrimp shells

Results and discussion f. Interaction between time and temperature show that at
lower values of the latter, time has a little effect on DD%,
Fitting of model Fig. 3d.
On the other hand, a different phenomenon effect is
The effect of the four variables on chitosan degree of dea- observed for v/w ratio ­(x4) on temperature (­ x2). At higher
cetylation (DD %) is simultaneously examined. The data temperature (>70 °C), ­(x4) has a slight effect on DD%, in
obtained in the Box-Behnken Design (BBD) are gener- contrast at lower values a greater influence is observed
ated into a response function model expressed by: and DD% is extremely reduced Fig. 3e.
Therefore, response surface plots reveal an extremely
Y = 72.01 + 1.82x1 + 11.79x2 − 8.34x3 − 6.34x4 − 3.15x1 x2
significant influence of NaOH concentration ­(x1), temper-
+ 8.96x1 x3 − 10.34x2 x3 + 8.70x1 x4 + 4.66x2 x4 ature ­(x2), reaction time (­ x3) and the v/w ratio (­ x4) on the
+ 4.43x3 x4 − 6.81x21 − 8.80x22 − 5.89x23 + 5.73x24 deacetylation efficiency. The High Degrees of Deacetyla-
(3) tion (DD% >90%) are found for lower NaOH concentra-
where: Y is the predicted Degree of Deacetylation DD%, tions (<50%), higher temperatures above 70 °C, reduced
­x1, ­x2, ­x3 and ­x4 are the coded values of NaOH concentra- time (<11 h) and the v/w ratio around less 7/1 ml/g.
tion, reaction temperature, time, and solvent / solid ratio
respectively. Optimization using the desirability functions
Table 2 shows observed and predicted DD% at dif-
ferent combination of the independent variables. When To optimize and validate the predicted mathemati-
comparing the predicted values with the experimental cal model, a supplementary Expert Design runs have
ones, the results indicate that these data are in practically been carried out under the same experimental condi-
close agreement. tions. Higher desirability function results in an improved
Analysis of variance for response surface methodol-
ogy (ANOVA) of the quadratic model is shown in Table 3.
As revealed in this table the model is significant with a P Table 3  Analysis of variance ANOVA results for experimental
design
value < 0.0001. The confidence level for linear terms ­x1
(P = 0.0391), ­x2, ­x3 and x­ 4 (P < 0.0001) have all a signifi- Source SS DF MS F value P value
cant effect. Every square terms and interactions are also
Model 6045.04 14 431.79 57.47 <0.0001a
found with significant P values < 0.05. The Lack of Fit
x1 40.30 1 40.30 5.36 0.0391*
P value relative to the pure error for the model is 0.1062
x2 1650.65 1 1650.65 219.70 <0.0001*
which is not significant. Moreover, R2 and adjusted R2
x3 808.36 1 808.36 107.59 <0.0001*
values 0.9853 and 0.9682 respectively are closed to 1,
x4 484.63 1 484.63 64.50 <0.0001*
suggesting a high correlation between experimental and
x1x2 691.43 1 691.43 92.03 <0.0001*
fitted values. Therefore the model is found to be adequate
x1x3 321.31 1 321.31 42.77 <0.0001*
for prediction within the range of variables used.
x1x4 300.85 1 300.85 40.04 <0.0001*
x2x3 428.28 1 428.28 57.00 <0.0001*
x2x4 87.05 1 87.05 11.59 0.0052*
Response analysis
x3x4 78.59 1 78.59 10.46 0.0072*
x12 246.79 1 246.79 32.85 <0.0001*
Response surface showed as contour plots in Fig. 3, pro-
x22 413.84 1 413.84 55.08 <0.0001*
vide a method to predict DD% for different values of test
x32 185.34 1 185.34 24.67 0.0003*
variables and to identify their interaction.
x42 154.30 1 154.30 20.54 0.0007*
Interactive effects observed between the four inde-
Residual 90.16 12 7.51 – –
pendent variables ­ (x1x2, ­x1x3, and ­ x1x4) exert signifi-
Lack of fit 88.16 10 8.82 8.81 0.1062b
cant influence on DD%. The interaction between ­x1 and
Pure error 2.00 2 1.00 – –
­x2 reveals that the degree of deacetylation increased by
Total 6135.20 26 – – –
increasing temperature, however at concentration higher
R2 0.9853 – – – –
than 50%, DD% reduced; Fig. 3a. Interaction related
Adj ­R2 0.9682 – – – –
to reaction time reveals that DD% is diminishing when
increasing both concentration and time, Fig. 3b. Mean- SS sum of square, DF degree of freedom, MS mean square
while, the effect of v/w ratio (­x4) is perceived to have a
Significant under 95% level of confidence
little effects on DD% for lower NaOH concentration and b
Not significant
time with practically no effect for higher values, Fig. 3c, *Significant

13
 Bachir Ben Seghir, M. H. Benhamza

Fig. 3  Contour plots for the

effects of different variables on
degree of deacetylation DD%

accurateness model, thus with a desirability value of 0.99 Chitosan characterization

optimum parameters correspond to a DD%=98.86%,
Fig. 4. Using these conditions, the maximum reached Chitosan sample prepared under optimum conditions are
DD% corresponds to NaOH concentration of 28.6%, characterized by FTIR, DSC, DRX and SEM techniques.
temperature of 81.15 °C, reaction time of 9.55 h and v/w
ratio equals to 6/1 ml/g. With these conditions a sample FTIR spectroscopy
of chitosan is prepared and a characterization technique
is carried out. Chitosan spectra are obtained by Fourier Transform
Infrared Spectroscopy and the main strips are shown in
Fig. 5. Analyzes of this spectrum show a broader band

13
Preparation, optimization and characterization of chitosan polymer from shrimp shells

Fig. 4  Desirability ramp for

optimization

Fig. 5  FTIR of prepared chitosan

Fig. 6  DSC curves for chitin and chitosan

at 3100–3500 cm−1, this corresponds to –OH stretching

vibrations of water and hydroxyls as well as –NH stretch- DSC analysis
ing vibrations of free amino groups. The wave length of
2860 cm−1 is corresponding to C ­ H2 and ­CH3 as a result of DSC scans of chitosan and chitin samples are obtained in
stretching vibration attributed to the ramification of C–H order to compare their thermal stability (Fig. 6). The first
[16]. The band that appears at 1652 cm−1 is an amide I endothermic peaks obtained of both chitin and chitosan
resulting from hydrogen and hydroxyl groups interactions, are wide centered between 100 and 125 °C, and this can
this is due to removal of acetyl group (deacetylated chitin). be ascribed to the loss of water. The second chitin peak
The band found at 1587 cm−1 corresponds to an amide II appears at 300 °C, which is corresponding to the residual
(–NH2 bending). The peak at 1429 cm−1 corresponds to the of proteins and chitin pigments [28]. The chitosan second
C–H stretching, whereas the peak appeared at 1373 cm−1 peak is located between 295 and 325 °C with maximum
is an amide III resulting from C–N stretching of the value at 310 °C evaluating at gas flow of 8–50 ml ­min− 1
N-acetyl–glucosamine. The absorption peak at 1148 cm−1 and heating rate of 10 °C ­min−1. This indicates a decom-
shows a symmetric glycosidic linkage (C–O–C), in addi- position of amine groups and showing the conversion of
tion the presence of absorption band is found at 1027 cm−1 acetylated chitin to chitosan [36, 38].
showing a stretching vibration of C–O ring. The absorption
peak around 880 cm−1 is referred to the glycosidic linkage
of β–anomer (1–4) (C–O–C) [37].

13
 Bachir Ben Seghir, M. H. Benhamza

Fig. 7  X-ray of chitosan prepared

X‑ray diffraction analysis

X-ray analysis is used to detect the chitosan polymer Crys-

tallinity Index (CrI). The Crystallinity Index (CrI) is deter-
mined by the following Eq. [16, 36]:

(4)
( )
CrI = I110 − Iam ∕I110
where ­I110 is the maximum intensity (arbitrary unit) of the
diffraction peak at 2θ = 19.51° and ­Iam is the intensity of Fig. 8  SEM image of pure chitosan prepared: a 5 Kv and b 25 Kv
amorphous diffraction at 2θ = 12.30°.
X-ray diffraction spectrum of chitosan is shown in
Fig. 7. At ­I110 the spectrum exhibited crystalline reflections Results and comparison
at 2θ ≈ 20°, however at ­Iam which is a typical fingerprints of
semi-crystalline chitosan as revealed at 2θ ≈ 10°. These two From this study, all process variable parameters i.e. NaOH
peaks are approximately identical of those found by [10, concentration, temperature, time reaction and v/w ratio
12, 16]. Though, these results show that the chitosan sam- contributed towards the experiment optimization. The
ples have a lower crystallinity because of the intra-molecu- obtained DD% of 98.86% in the present investigation is a
lar hydrogen bonds resulting after the chitin deacetylation. great finding compared to those of previous studies, even
These findings are similar to those given in the literature though using different material sources. In the most previ-
[34, 36, 37]. ous result studies, optimized DD% values are not exceed-
The degree of crystllinity (CrI) is calculated in order to ing 96%. Comparisons with various research works show
evaluate the purity of chitosan and the obtained value is that the majority of studies utilizing only three variables
equal to 75%. This decreased value is due to intermolecular [32, 33, 40–42], less than that of our work where we use
hydrogen bonds after the deacetylation, and this indicates the four mainly significant variables. Moreover, the NaOH
that the product molecular structure is in an amorphous concentration variable taken in the present research work
state [12]. is wide-ranging than that of other studies; (C%=from 25%
to 75%). Furthermore, temperature, time reaction and v/w
ratio variables are as much as widespread too, and that in
SEM analysis order to explore and to optimize the degree of deacetyla-
tion. A detailed and a comprehensive comparison are
The morphology of the chitosan sample is examined under shown in Table 4.
a scanning electron microscope (SEM). Images with dif- In conclusion, results from the present work provide an
ferent magnifications 100 and 500 µm, (Fig. 8a, b) respec- important alternative towards the production of high purity
tively show that the polymer surface became very downy; chitosan with a greater degree of deacetylation (DD%).
these are the same as those found by [34, 36, 37]. This is due to the operating conditions and the analyzing
techniques used, therefore the introduction of the most

13
Preparation, optimization and characterization of chitosan polymer from shrimp shells

Table 4  Comparison of results

Raw material sources Controlled Statistical method Measurement technique DD (%) References
variables

Chitin C: 32%–58% RSM NMR 67.3–95.7% [32]

T: 40–120 °C
t: 2–34 h
r: 3/1–9/1 (ml/g)
Metapenaeus monoceros C: 50% Factorial NMR 94.25% [33]
T: 90–140 °C
t: 3–6 h
r: 20/1 (ml/g)
NSB, SB, AC
Squid Pens C: 20–60% RSM NMR 90% [39]
T: 40–100 °C
t: 15–120 min
r: 10/1–20/1 (ml/g)
Streptocephalus sirindhornae C: 60% RSM Titration 86.13% [40]
T: 110 °C
t: 180 min
r: 1/0.08 (ml/g)
Penaeus indicus C: 30–50% RSM Titration 89.34% [41]
T: 60–100%
t: 90–300%
Farfantepenaeus brasiliensis T: 120–130 °C Factorial Potentiometric 90% [42]
t: 90−150 min
Parapenaeus longirostris C: 25–75% RSM FTIR 98.86% Present study
T: 40–100 °C
t: 6–24 h
r: 5/1–15/1 (ml/g)

C NaOH concentration, T temperature, t reaction time, r v/w ratio, NSB number of successive baths, AC atmospheric conditions, NSB use of
sodium boro-hydride, AC alkali concentration

important controlled parameters (T, C, t, and r) as well as followed by the NaOH concentration then the reaction time.
increasing factors interval range; give better results show- Thus, deacetylation degree is highly affected by tempera-
ing the usefulness of the present investigation regarding to ture, NaOH concentration and reaction time; but slightly
other research studies. On the other hand, FTIR spectros- affected by solid to solvent ratio. Finally, the obtained
copy technique employed in this study shows its reliabil- highest degree of deacetylation of the prepared chitosan is
ity. Consequently, using the RSM method optimization tool of 98.86% and the chitosan purity is confirmed by FTIR,
together with better performance parameters give improved DSC, XRD and SEM characterization techniques.
results compared to previous studies.
Acknowledgements This work is supported by the research labora-
tory LAIGM of Guelma University, Algeria.

Conclusion

The present research work shows that higher degree of

deacetylation chitosan can be obtained from Parapenaeus
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