Journal of Hazardous Materials 229–230 (2012) 346–353

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Journal of Hazardous Materials

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Synthesis and characterization of a novel fish scale-immobilized chitosan

adsorbent—Preliminary features of dichlorophenol sorption by solution
calorimetry
Jackeline A. Mota a , Renata A. Chagas b , Eunice F.S. Vieira b , Antonio R. Cestari b,∗
a
Department of Materials Science/CCET, Federal University of Sergipe, CEP 49100-000, São Cristóvão, Sergipe, Brazil
b
Laboratory of Materials and Calorimetry, Department of Chemistry/CCET, Federal University of Sergipe, CEP 49100-000, São Cristóvão, Sergipe, Brazil

h i g h l i g h t s

 A novel fish scale-immobilized chitosan adsorbent was synthesized and characterized.

 It was evaluated the sorption of dichlorophenol on the new adsorbent.
 Simultaneous sorption data of quantity and energy were found by solution calorimetry.
 The interactions of fish scales/dichlorophenol are due to surface reactions.
 Excellent sorption features at adsorbent/dichlorophenol interface were found.

a r t i c l e i n f o a b s t r a c t

Article history: Brazilian Corvina fish scales were cross linked with polyglutaraldehyde and chemically modified with
Received 27 March 2012 chitosan gel. Characterization has pointed that chitosan has good and stable adhesion on the fish scales.
Received in revised form 4 June 2012 The sorption of dichlorophenol-2,6-indophenol (DPI) on the novel material was studied by isothermal
Accepted 7 June 2012
solution calorimetry. The non-symmetric shapes of the calorimetric plots indicate that the DPI sorption
Available online 21 June 2012
sites of the adsorbent are not energetically uniform. The enthalpies of the DPI sorption processes were
highly exothermic (from −536.7 to −50.9 kJ mol−1 ). The analysis of both the characterization of the mate-
Keywords:
rials and the calorimetric results has suggested that the interactions at the fish scales/DPI interface are
Chitosan
Fish scales
due to surface reactions. The present work underlines the excellent features of the new fish scale-based
Adsorption adsorbent for use in phenol sorption applications at solid/solution interfaces.
Dichlorophenols © 2012 Elsevier B.V. All rights reserved.
Solution calorimetry
Sorption mechanisms

1. Introduction performance [3,4]. Fish scales can be an alternative adsorbent, due

Most phenols including phenol, chlorophenols, nitrophenols
and aminophenols are pollutants of high priority because of their
toxicity and possible accumulation in the environment. Phenols
are introduced into surface water from industrial effluents such
as those from the coal tar, gasoline, rubber proofing, pharmaceu-
tical and steel industries and chemical spills [1]. Current methods
for removing phenolics from wastewater are expensive, and may
lead to contamination of ground water [1].
Adsorption has been a promising option for the removal of non-
biodegradable organics from aqueous streams [2]. Many researches
have focused on the use of alternative adsorbents from recy-
clable materials, due to their cost effectiveness and good removal
∗ Corresponding author. Tel.: +55 79 21056656; fax: +55 79 21056684.
E-mail address: arcestari@gmail.com (A.R. Cestari).
to the presence of specific chemical groups of fish scale collagen,
such as hydroxyl, carboxyl, amine and amide [5,6]. The struc-
tural stability of collagens is enhanced by chemical cross linking
the molecule by polyglutaraldehyde (PGA) reaction [7]. However,
the fibrillar nature of fish scale collagen and the presence of PGA
decrease the amount of available sorption sites of fish scale collagen
[8].
The rationale for this work is that fish scale-immobilized chi-
tosan would be a new potential adsorbent for use in sorption
studies at solid/solution interfaces. Chitosan bears amine and alco-
hol functions, and may interact with inorganic/organic pollutants
by covalent, electrostatic and/or hydrophobic interactions [9]. The
specific properties of fish scale collagen and chitosan may be used to
produce materials with unique properties for sorption of important
pollutants in water streams.
A novelty of this work is the use of solution calorimetry to
determine both the quantity and the energy of sorption without

0304-3894/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.jhazmat.2012.06.010
 J.A. Mota et al. / Journal of Hazardous Materials 229–230 (2012) 346–353 347

C
O
CHO CHO
L
L
A
NH2 + OHC CHO
G n
E
N

C C
O H NH2
CHO CHO I
L
L T
A
N=C
H n
CHO + H2N O
G S
E A NH2
N N

C C
O H NH2
R R
L I
L T
N=C C=N O
Fig. 1. Chemical structures of dichlorophenol-2,6-indophenol (above) and chitosan A H n H
(below). G S
E A NH2
N N
C
H NH2
the need for additional analytical investigation [10,11]. The calori-
I
metric results give information on thermodynamics of the process, T
R= C=N O
energetics and analysis, which are all essential for characterizing
the sorption mechanisms [10,11]. However, less attention has S
A NH2
been paid to the direct calorimetric investigations of phenols N
interactions on naturally occurring materials. The calorimetric
data are invaluable to assess many sorption features at the fish Fig. 2. Idealized schematic sequence of the fish scales cross linking with PGA and
scale/dichlorophenol interface. their chemical modification with chitosan.

2.3. Characterization of the materials

2. Materials and methods
2.1. Reagents and solvents
All chemicals/reagents used were of analytical reagent grade.
The scales of Corvina fish (Micropogonias furnieri) were collected
from a Fishermen’s Market located in Aracaju, state of Sergipe,
Brazil. Mature fish scales were washed repeatedly with water to
remove adhering dust and soluble impurities from their surface
and allowed to dry at 35 ◦ C for 12 h. The chitosan powder (CHIT)
was a free gift from C.E. Roeper, Hamburg, Germany. The degree of
deacetylation (82%) was determined by FTIR [12]. Dichlorophenol-
2,6-indophenol (DPI) was from Sigma–Aldrich and used without
purification. The chemical structures of chitosan and DPI are shown
in Fig. 1.
2.2. Preparation of the adsorbent
The fish scale collagen was crosslinked under mild conditions
using a 0.10% (w/v) pH 7.0 PGA aqueous solution as described ear-
lier [13]. A pale brown material was obtained (hereafter described
as PGA-scale for simplicity). Chitosan was dissolved in 1.0 wt%
acetic acid solution. The PGA-scale (50 g) was then added to 100 mL
of the chitosan gel and the suspension was mechanically agitated
for 3 h at room temperature. The suspension was filtered and the
solid residue washed repeatedly with water and dried at 45 ◦ C
for 6 h. A dark brown material was obtained (hereafter described
as ESC-QUIT for simplicity) and cut into small 5 mm × 5 mm
square membranes of 0.5 mm of thickness. A schematic sequence
of chitosan immobilization on the PGA-scale is shown in
Fig. 2.
The determination of the pH of point of zero charge (pHpzc )
of the adsorbent (ESC-QUIT) was carried out using a procedure
similar as described earlier [14]. The morphological characteri-
zation of the materials was carried out with a scanning electron
microscope (SEM, JEOL-JSM 6360-LV). The samples were previously
coated with gold (thickness of about 12 nm). The Raman spectra
were acquired with a Bruker Senterra Raman System equipped with
an Olympus microscope with a 50× objective to focus a Melles
Griot laser beam on the sample. The spectra were excited by the
785 nm line from an air-cooled He–Ne laser. The thermogravi-
metric analyses (TG and DTG) were made using about 10 mg of
material, under nitrogen atmosphere from 25 to 800 ◦ C, in a SDT
2960 thermoanalyzer, from TA Instruments. XRD analysis were per-
formed in a Shimadzu diffractometer, in the 2 range from 5◦ to
60◦ (accumulation rate of 0.02◦ min−1 ), using Cu K␣ radiation. The
solid-state reflectance spectra of the samples were recorded on an
Ocean Optics UV–vis spectrophotometer from 400 to 900 cm−1 at
a resolution of 4.0 cm−1 .
2.4. Solution calorimetry experiments
Microcalorimetric determinations were performed in a C80
microcalorimeter from Setaram, using the membrane breaking
(thin Teflon® ) technique [15]. Briefly, 100 mg of ESC-QUIT and
0.5 mL acetic acid/sodium acetate buffer solutions at pH 3.0 were
put into the lower of the calorimetric vessel. In this work, HAc/NaAc
buffered solutions at pH 4.0 were used in all calorimetric experi-
ments. It was found that the chemical structure of fish scale collagen
can be denatured in both very acidic and very alkaline media [7].
Additionally, 2.5 mL of a DPI aqueous solution were put in the lower
part of the calorimetric vessel. Calorimetric output was of thermal
348 J.A. Mota et al. / Journal of Hazardous Materials 229–230 (2012) 346–353
power (dq/dt; mW) as a function of time (t; s) and consequently
the integral of this data to time t is equal to the interaction energy
(J). The calorimetric experiments were carried out using DPI solu-
tions of 0.01, 0.10 and 1.00 × 10−3 mol L−1 , at 25 ◦ C. The detectable
calorimeter signals (power vs. time) were analyzed using the SET-
SOFT software (SETARAM). Each experiment was performed in
triplicate runs and values are the average of them. After the end
of the calorimetric experiment, the calorimetric cell was opened
and the DPI concentration in the vessel was determined using a
spectrophotometer (700 Plus, Femto, Brazil) at 515 nm wavelength
(detection level less than 1.0 mg L−1 ). The amount of DPI sorbed in
each calorimetric experiment was calculated by using the following
Eq. (1) [10]:
(Ci − Cf )V
nint =
m
where nint is the fixed quantity of DPI per gram of ESC-QUIT after
at sorption equilibrium, in mol g−1 , Ci is the initial concentration of
DPI in mol L−1 , Cf is the concentration of DPI present after sorption
equilibrium, in mol L−1 , V is the volume of the solution in L, and m
is the mass of ESC-QUIT in g.
3. Results and discussion
3.1. Preliminary considerations
Fish scales are composed of an extracellular matrix, mainly type
I collagen, and hydroxyapatites [16]. Cross linking reactions have
been used for efficient insolubilization of collagen structure [7].
Glutaraldehyde (GA) possesses unique characteristics that render
it one of the most effective collagen cross linking reagents. It can
react with several functional groups of collagens, such as amine,
thiol, phenol, and imidazole by several means such as aldol conden-
sation or Michael-type addition [7,17]. However, most of solutions
of GA is usually polymeric and contain ␣,␤-unsaturated aldehydes
(PGA) that are able to form rings by loss of water molecules by aldol
condensation [7].
In this work, one of the important purposes using PGA is pro-
viding aldehyde groups as reacting sites to immobilized chitosan
on fish scale collagen surface. Typically, this reaction involves a
reductive amination in which an aldehyde function of PGA reacts
with the NH2 group of chitosan [18,19]. In addition, the OH
groups of hydroxyproline in collagen are capable of forming hydro-
gen bonds with OH and NH2 groups in chitosan. Moreover, the
end groups COOH and NH2 in collagen may also form hydrogen
bonds with OH and NH2 groups from chitosan. The properties of
surface-immobilized chitosan are mainly controlled by its molec-
ular structure, length and number [20].
3.2. Characterization of the materials
The Raman spectra of the materials are shown in Fig. 3. Using
the Raman technique, spectra can be obtained for surface samples
just a few microns thick and with minimal interference from the
surrounding water. In addition, spectra can be obtained rapidly and
extensive sample preparation is not needed. However, analysis of
Raman spectra of naturally-occurring materials is difficult due to
the presence of broad bands and band overlappings [21]. The most
prominent peaks were found in the range of Raman shift from
400 to 1800 cm−1 and only the strongest absorbing modes were
assigned. For the raw fish scale, the strong Raman bands centered
at 1670, 1460 and 1287 cm−1 have been assigned, respectively, to
amides I, II and III vibrational modes, which involves C O and C N
stretching, C C N bending, and N H in-plane bending groups or
vibration of thioester forms due to the presence of R O S C
groups, all of the peptide groups of the fish scale collagen [21]. A
960
1050 590 430
D
1385 1326
993
1463 975
895
595 512
1420
C
960
887 827 595
1667
B
1460 1287
(1) 1670
1075 595
870 A
1800 1600 1400 1200 1000 800 600 400
Wavenumber (cm-1)
Fig. 3. Raman spectra of the materials, where (A) Raw scale, (B) PGA-scale, (C) ESC-
QUIT, (D) ESC-QUIT + DPI.
band centered around 1075 cm−1 is due to vibrations of C C, which
is characteristic of ␣-helices of collagen. Bands for proline residues
appear around 870 and 975 cm−1 . Bands centered at 595 cm−1 are
attributed to the symmetric O P O bending mode in the apatite
lattice and the band centered at 870 cm−1 corresponds to carbon-
ate anions substituted for phosphate ions in the apatite lattice [21].
It suggests that the fish scale is a composite consisting of type I
collagen and calcium-deficient apatite containing carbonate ions.
Chemical modification with PGA might cause effects on some
amino acids of fish scale collagen. It is suggested by the observa-
tion of absence of the peak at 1075 and 1287 cm−1 , due to reactions
of PGA with amide III of ␣-helices of collagen [21,22]. The bands at
1670 and 975 cm−1 decreased their intensities and were shifted to
lower wavenumbers, due probably to reactions of PGA with amide I
groups. The broad band centered at 1420 cm−1 has been attributed
to the CH2 bending mode of PGA [22]. For ESC-CHIT the absorption
band centered at 985 cm−1 may be relative to the N H stretching,
due to the formation of imines bonds of chitosan-PGA interac-
tions. Two other bands centered at 1385 and 1326 cm−1 have been
assigned to C H stretching and R N H bending modes of chitosan
[20]. The Raman spectra of the raw scale and the PGA-scale, both
with DPI sorbed (not shown) are very similar to these materials
before DPI interaction. This feature suggests that the interaction of
DPI with these materials is very low and not detectable by RAMAN
spectroscopy. On the other hand, the Raman spectrum of ESC-QUIT
with DPI sorbed (Fig. 3D) has shown almost total absence of the
bands in relation to ESC-QUIT spectrum (Fig. 3C). However, the min-
eral phosphate spectral region (900–1200 cm−1 ) seems not to be
affected. So, it is more likely that chemically-immobilized chitosan
seems to be the main DPI sorption sites at the surface of ESC-QUIT.
Molecular associations are also responsible for the involvement
of ionic sites of fish scale collagen and chitosan. If collagen is
placed at a pH about 6.0 (the pH value of the chitosan gel in acetic
acid medium), the carboxylic functions of collagen are almost all
in a COO− form. The collagen/chitosan interaction is reported
to be maximized in this pH value. Evidently, the polymer blend
collagen–chitosan can only give rise to a restricted number of elec-
trostatic interactions between these two polymers. Typically, in
relation to a pure electrostatic interaction, the interacted colla-
gen is mainly identified by the C O amide and carboxylic bands
present in the range 1600–1700 cm−1 . We have observed (Fig. 3C)
the absence of the C O band at about 1670 cm−1 , which leads us
 J.A. Mota et al. / Journal of Hazardous Materials 229–230 (2012) 346–353 349

100
90
80
ESC-CHIT

Mass loss (%)

70
PGA-SCALE
60
PGA-SCALE
50
RAW SCALE
40
30 ESC-CHIT
RAW SCALE
0 200 400 600 800
Temperature (oC)

10 20 30 40 50 60 70 0.00
2
-0.05
Fig. 4. X-ray diffractograms of the materials.
PGA-SCALE

DTG (%/oC)
-0.10

to suppose that the carboxylic groups of fish scale collagen were
completely reacted with chitosan [20].
The X-ray diffractograms of the materials are shown in Fig. 4. The
presence of broad diffraction peaks suggests that the crystals were
small or structurally disordered or both. The main peaks are found
in 2 = 22.5, 32.5, 40.0, 50.0 and 63.0◦ with d spacings from 0.170
to 0.345 nm. These values are comparable with biological apatite
containing structures. The peak at about 2 = 27.5◦ has been asso-
ciated with the presence of collagen [23]. The XRD diffractogram
of PGA-scale is very similar to the raw scale, suggesting that the
reaction with PGA did not change the main structural features of
the fish scale. The diffractogram of ESC-CHIT shows a new peak at
2 = 32.5◦ , due to the presence of chemically immobilized chitosan
[24,25].
Typically, solid-state chitosan usually exhibits an orthorhombic
unit cell with 2-fold helical chains stabilized by hydrogen bonds
and two independent polymer sheets with the same direction form
a repeating unit piled up along the a-direction [24]. The influence
of chitosan chain packing and crystallinity are important parame-
ters in the characterization of the ability of chitosan to sorb many
chemical species in solution. These parameters control the number
of available free amine groups and the accessibility of water. How-
ever, it was found that chemical cross linkings inhibit close packing
of chitosan chains by reducing the degree of freedom in the 3-D
conformation, limiting or preventing the formation of crystalline
regions [24]. The reduction of crystallinity plays a crucial role on
influencing thermal degradability, water absorption and swelling,
and the sorption properties of chitosan-based materials [24].
The TG–DTG plots of the materials are shown in Fig. 5. The
identification of components of biocomposites, such as fish scales
using TG/DTG is not easy due to overlapping processes [25]. Upon
severe heating, fish scales have shown a continuous sequence of
more or less irreversible decomposition reactions. For the TG/DTG
plots of the raw scale, it has shown four main mass loss regions:
25–215, 215–410, 410–500, 500–750 ◦ C. The first event has been
related to the superficial water releasing and the denaturation of
fish scale collagen. The others correspond to the thermal degrada-
tion of the polymeric chains of collagen, possible dehydroxylation
of hydroxyapatite and carbon material elimination. A slightly
change of slope of the TG curve at about 120 ◦ C is due to the water
removal of the first hydration shell of the collagen structure [26].
At low temperatures, collagen-water interactions are stabilized by
hydrogen bonding between water molecules and terminal OH
groups, both of the primary and the secondary hydration layers.
-0.15
-0.20 ESC-CHIT
-0.25
RAW SCALE
-0.30
200 400 600 800
Temperature( oC)
Fig. 5. TG (upper part) and DTG (lower part) curves of the materials.
As the temperature increases, the loosely bound water molecules
become free resulting in a lower residence time of the water
molecules in the hydration layer of collagen [26].
For type I collagen, complete denaturation is observed at about
205 ◦ C, due to the breaking of the hydrogen bonds between ␣-
chains of collagen [27]. When collagen is subjected to high tem-
peratures, its triple helix unfolds to produce random chains of
denaturated collagen that can remain covalently linked to each
other or not depending on the degree of heating. The denaturation
phenomenon -distinct from degradation- implies that the rupture
of interchain hydrogen bonds leads to the formation of an amor-
phous polymer, typically called as gelatin [28].
For the TG/DTG plots of PGA-scale, the peak due to the com-
plete fish scale collagen denaturation is narrower in relation to the
raw scale. In addition, the denaturation temperature of PGA-scale
collagen is shifted toward a higher temperature (about 220 ◦ C). It
allows concluding that the thermal stability of fish scale collagen is
increased due to the presence of PGA. Typically, PGA cross-linking
in collagen restricts water intrusion into the collagen structure due
to the decrease of the hydrophilic groups in the cross linked mate-
rial [29].
An estimated amount of 10 wt% of immobilized chitosan was
found in the TG/DTG plots of ESC-QUIT (details not shown).
Chitosan immobilization results in electrostatic and covalent inter-
actions involving the groups of both collagen and PGA-cross linked
collagen [30]. When chitosan is added, it is observed that the
thermal stability of fish scale collagen was slightly increased to
about 225 ◦ C. Typically, the thermal decomposition of raw solid-
state chitosan occurs in the temperature range 270–337 ◦ C with
peak maxima at 330 ◦ C [30,31]. However, the temperature of ini-
tial degradation of fish scale-immobilized chitosan is about 400 ◦ C
350 J.A. Mota et al. / Journal of Hazardous Materials 229–230 (2012) 346–353
Fig. 6. SEM images of the raw scale (upper part), PGA-scale (middle) and ESC-QUIT
(lower part), all at magnification of 5000×.
and the maximum degradation is about 480 ◦ C; finally a residue of
about 35 wt% was produced at about 750 ◦ C.
Some SEM micrographs of the materials are shown in Fig. 6.
The internal sections of fish scales are rich in inorganic material
containing high proportions of calcium and phosphorus, whereas
their surfaces are rich in collagen [32]. The collagen fibers are orga-
nized into lamellae that are superimposed to produce an orthogonal
0.50
0.50
0.40
0.30
0.40
Absorbance (a.u.)
0.20
0.30
0.10
400 425 450 475 500
0.20
RAW SCALE ESC-QUIT
PGA-SCALE
0.10
400 500 600 700
Wavelenght (nm)
Fig. 7. Diffuse reflectance spectra of the raw scale, PGA-scale and ESC-QUIT.
and/or a double-twisted plywood-type pattern [33]. In the upper
external layer of the raw fish scales, electron microscopic investiga-
tion revealed sheet-like structures composed of vertically oriented
collagen fibers. Needle-like or flaky crystals of apatites are also
observed in the outer layer of the scales. After reaction with PGA,
the fish scale surface exhibited a randomly rougher surface and
micrometer-sized aggregates. Some authors state that chitosan can
also function as “a bridge” to increase the cross-linking efficiency
in collagen-based materials [20]. The roughening of the fibers was
likely due to partial disruption of the ordered fibrillar structure of
the fish scale collagen due to PGA cross linking reaction [32,20]. The
different aspects of the samples, such as modification of color and
rigidity are also indicative of chemical and/or structural modifica-
tions. The crosslinking reaction can induce the collagen fibers to
be combined to form sheets, leading to the fusion of some smaller
pores to generate larger ones [33]. It is observed a relatively com-
pact arrangement on the surface of ESC-QUIT after being immersed
in the chitosan gel. This suggests that chitosan has good adhesion
on PGA-modified fish scale collagen, which would be favorable for
a new and stable sorption material. Some pores are also detectable
along the chitosan coated surface of ESC-QUIT, probably as a con-
sequence of trapped liquids or air-drying [34].
In this work, solid-state diffuse reflectance (DR) is used to com-
plement the morphological studies of the fish scales. However,
band overlapping and poor spectral resolutions have generally pre-
cluded details of the chemical environment of the materials by
DR [35]. The DR spectra are shown in Fig. 7. For the raw scale, it
is observed a broad and small peak centered at 415 nm, due to
the presence of chromophores of collagen [36]. For PGA-scale, this
broad peak is shifted to about 450 nm, due to PGA itself [29]. No
appreciable changes were observed in the spectra after chitosan
immobilization. This suggests that the chemical structure of colla-
gen is preserved after interaction with DPI.
The pH where the net total particle charge is zero is called the
point of zero charge (pzc). If the pH is above its pzc the adsor-
bent surface will have a net negative charge and predominantly
exhibit an ability to exchange, while the adsorbent will mainly
retain anions (electrostatically) if its pH is below its pzc. The raw
Corvina scale, PGA-scale and ESC-QUIT display pHpzc values at 7.20,
6.90 and 6.60, respectively (Fig. 8). The collagen net charge depends
on the number and location of amino acids and the solvent pH [37].
The ␣-carboxyl group of proteins has a pKa of about 2 and the ␣-
amino group has a pKa of around 9.5. Asp and glu have pKa around
4.0, cys, thr and lys have around 10, arg has around 12.5, and his
at around 6.0. At neutral pH, aspartic and glutamic acids carry a
 J.A. Mota et al. / Journal of Hazardous Materials 229–230 (2012) 346–353 351

3.00 Table 1
Calorimetric parameters of DPI sorption on ESC-QUIT.
pHpzc (raw scale) = 7.20 Temp. (◦ C) Ci /10−3 /mol L−1 −Qint /J g−1 a nint /␮mol g−1 −int H/kJ mol−1
2.00
0.01 1.46 2.72 536.7
25 0.10 3.60 33.9 106.2
1.00 1.00 2.30 45.2 50.9
a
Average SD of the calorimetric results were less than 5.0%.
pH

0.00

contact time. The calorimetric parameters of DPI sorption on ESC-

-1.00
pHpzc (ESC-QUIT) = 6.60
-2.00
4.00 5.00 6.00 7.00
pHinitial
Fig. 8. Plots for determination of pHpcz of the materials.
negative charge and arg and lys carry negative charges. When the
pH becomes more alkaline, lys and arg residues lose their posi-
tive charge and become neutral at pH 12. If the pH is made more
acidic, aspartic and glutamic acids shed their negative charge and
become neutral. Collagen possesses amine-containing amino acids
having pKa ∼11, which become more positively charged at low pH
values and the majority of these residues should remain positive
even at high pH [37]. This effect can explain why many adsorbates
irreversibly bound to collagen substrate under neutral and basic
conditions [37].
The value of pHpzc for ESC-QUIT seems to be directly related to
the average pKa value of chitosan of 6.5 [19]. The species involved
are NH3 + and COOH (at pH 1.0–3.0), NH2 and COO− (at pH 7.0–13.0)
and NH3 + and COO− or NH2 and COOH (at pH 4.0–7.0). In the pH
range of 4.0–7.0, the majority of the base and acid groups are as
NH3 + and COO− or NH2 and COOH forms, and ionic interactions of
NH3 + and COO− species or hydrogen bonding between amine and
carboxylic acid may occur.
3.3. Analysis of DPI sorption on ESC-QUIT by solution calorimetry
All chemical reactions are accompanied by the generation or
absorption of heat. Thermodynamic data are the foundation of
mechanistic investigations, but many of the systems of interest at
solid/liquid interfaces present considerable difficulties in analysis.
Fig. 9 shows a typical calorimetric plot of heat flow as a function of
0.50
0.40
Heat flux (mW)
QUIT are presented in Table 1. The integrations under the curves
were made by the “horizontal way drawn from the first point”
methodology. The sorption heat measured from solution calorime-
try is an integral heat since there are different sites in ESC-QUIT for
8.00 9.00 DPI sorption. The non-symmetric shapes of the calorimetric plots
indicate that the DPI sorption sites of ESC-QUIT are not energet-
ically uniform [38]. This calorimetric behavior is typical for most
sorption systems at solid-solution interfaces [39,40]. In general, a
sharp increase in heat flow occurred in the first 2 min of interac-
tion followed by a sharply decreasing with coverage. The sorption
is very fast between t = 0 and 15 min, and only a few calorimetric
points could be measured in this period with the calorimeter set
up. At t > 15 min the DPI sorption takes place at a much slower rate.
Although it seems to reach completion at around 50–65 min, some
long-term calorimetric experiments showed that sorption can con-
tinue for longer times, but very slowly. Indeed, after 65 min, the
evolved heat becomes too low to be accurately separated from the
instrumental noise.
The integral heats of sorption of DPI on ESC-QUIT (Qint ) were
found to be 1.46 ± 0.07, 3.60 ± 0.08 and 2.30 ± 0.03 J g−1 , when
the initial DPI concentrations in solution were 1.00 × 10−5 and
1.00 × 10−4 and 1.00 × 10−3 mol L−1 , respectively. The heats of
sorption increase with surface coverage due to repulsive lateral
interactions between the sorbed DPI molecules [41]. An estimate
of the energy of activation of sorption (Ea ) of DPI on ESC-QUIT can
be obtained by smooth extrapolation of the data to zero coverage
[42]. The estimated value of 1.10 ± 0.12 J g−1 found is higher than
those normally expected for adsorption processes controlled by dif-
fusion in aqueous solutions [43]. So, this value of Ea lies within
ESC-QUIT/DPI interactions not only at surface sites of ESC-QUIT
[44]. Indeed, the value of Ea is likely to be a cumulative effect
of breaking and reforming of the intermolecular bonds and their
realignment (or relocation) in a solid surface, as well as a cumula-
tive contribution from hydration and diffusion phenomena. Other
processes, such as surface diffusion or surface rearrangements of
DPI molecule, could take place [45].
The adsorption enthalpy (int H) can be calculated directly by
the following Eq. (2) [38,40]:
tf
dQint
ti

int H = (2)
0.30 nint

The enthalpies of DPI sorption on ESC-QUIT are highly exother-

0.20 mic (from −536.7 to −50.9 kJ mol−1 ). The magnitude of int H
increased as the DPI sorption is increased. In fact, the increasing
0.10 of int H with sorbent coverage is the most frequent situation. At
higher loading, repulsive lateral interactions between the sorbed
species, which are endothermic, might increase, decreasing the
0.00
exothermic net int H at higher loadings [46].
0 500 1000 1500 2000 2500 3000 3500
The int H value (calorimetrically measured) can also be used as
EXO Sorption time (s) a measure of the interaction forces between adsorbate and adsor-
bent, giving an indication of the bonding strength. Exothermic
Fig. 9. Typical solution calorimetry plot of DPI sorption on ESC-QUIT. Experimental
conditions: Initial DPI concentration in solution: 1.0 × 10−4 mol L−1 , mass of ESC- int H may suggest the dominance of attractive forces between
QUIT: 100 mg, temperature: 25 ◦ C. the sorption sites and the adsorbing species or attractive forces
352 J.A. Mota et al. / Journal of Hazardous Materials 229–230 (2012) 346–353
between sorbed molecules on the sorption sites. Free energy is
defined as G = H − TS, where S is entropy. For a spontaneous
reaction to occur, the free energy (G) associated with the process
should decrease or G should be negative. A spontaneous sorp-
tion process suggests a negative change in G. If H is positive
(endothermic), the product (TS) has to be >H to obtain a nega-
tive G value. If S is negative, the product – S) is a positive term
and, in this case, enthalpy must have a large enough negative value
(exothermic) to give a negative H. However, the situation may
be different in the case of sorption from solution due to the effects
of changes in solvation and other factors [34]. However, we are
unable to determine the numerical values of the free energies and
entropies of the ESC-QUIT/DPI interactions using solely the solution
calorimetry data of this work.
Several possible mechanisms have been proposed for sorption
processes at solid/solution interfaces, including surface precip-
itation, intraparticle diffusion or diffusion into pores, surface
binding heterogeneity and others [34]. Some effects may con-
tribute to the sorption heat, rendering it a complex quantity:
the energy of the surface bond, changes in degrees of freedom
of the atoms/molecules, the energy of interaction between the
sorbed species, surface relaxations or rearrangements. For amor-
phous materials, such as ESC-QUIT the surface sites are typically
heterogeneous, and the number of these sites depends on their dis-
tribution and the structural defects of sorption sites on the surface
[34].
The values of adsorption enthalpies at solid/solution interfaces
are average results of exothermic chemical bonding and endother-
mic diffusional interaction processes [40]. Typically, interactions
that occur with intense sorbate diffusion present small and rel-
atively similar values of int H. In addition, solvent and solute
transport through adsorbent materials is generally described fol-
lowing adsorption–diffusion mechanisms, where molecules first
diffuse from the bulk phase to the adsorbent surface. Next, they
adsorb to the sites on the surface and/or diffuse through the adsor-
bent structure, driven by the chemical potential gradient within
the adsorbent pores. Most covalent bond energies fall in the range
of −100 to −1000 kJ mol−1 and generally decrease in strength as
the bond length increases [34,47]. Some useful information can be
obtained by analyzing the effect of varying concentration of DPI on
the heat realizing of the sorption processes.
The forces involved in sorption processes include non specific
van der Waals forces, ionic or electrostatic forces, and specific forces
involved in the formation of chemical bonds. Sorption processes
that involve only non specific interactions are generally referred to
as “physical adsorption” while those in which stronger interactions
occur are termed “chemisorption”. In this work, the high exother-
mic results observed are in agreement with interaction of DPI to
sorption sites at the ESC-QUIT surface and chemisorption may be
an important mechanism at very low coverage. However, from data
in Table 1 it is clear that the sorption energies and enthalpies do
not vary linearly with the initial concentration of DPI in solution.
Thus, a simple adsorption mechanism in a single step cannot be
postulated and a more complicated sorption mechanism seems to
be operating.
4. Conclusion
In this work, Brazilian Corvina fish scales were cross linked
with polyglutaraldehyde and chemically modified with chitosan
gel. Characterization has pointed out the presence of micrometer-
sized aggregates on the fish scale collagen surface after the cross
linking and the presence of about 10 wt% of fish scale-immobilized
chitosan. The pHpzc of the adsorbent was 6.60, which also lies within
the presence of the immobilized chitosan on the fish scales. The
morphological analysis of the materials suggests that chitosan has
good adhesion on the cross linked fish scales.
The interaction of DPI with ESC-QUIT from aqueous solu-
tions was studied by isothermal solution calorimetry. The
non-symmetric shapes of the calorimetric plots indicate that
the sorption sites of the adsorbent are not energetically uni-
form. The exothermic integral heats of adsorption were found to
be 1.46 ± 0.07, 3.60 ± 0.08 and 2.30 ± 0.03 J g−1 , when the initial
DPI concentrations in solution were 1.00 × 10−5 and 1.00 × 10−4
and 1.00 × 10−3 mol L−1 , respectively. The values of int H of DPI
sorption on ESC-QUIT were highly exothermic (from −536.7 to
−50.9 kJ mol−1 ). The magnitude of int H increased as the sur-
face coverage is increased, suggesting repulsive lateral interactions
between adsorbed species. The analysis of both the characteriza-
tion of the materials and the calorimetric results has suggested that
the main DPI sorption sites are located at the surface of the adsor-
bent and chemisorption is an important interaction mechanism at
the interfaces of DPI/ESC-QUIT.
Additional features of the ESC-QUIT/DPI interactions will be
determined in future using sorption isotherms. The results of the
present work underline the excellent features of the new fish scale-
based adsorbent for use in sorption of dichlorophenol from aqueous
solutions.
Acknowledgements
The authors are indebted to the Brazilian agencies CAPES andC-
NPq for financial support and fellowships.
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