Chemical Engineering Journal 166 (2011) 890895

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Chemical Engineering Journal
j our nal homepage: www. el sevi er . com/ l ocat e/ cej
Adsorption of melanoidins by chitin nanobers
Rujira Dolphen
a
, Paitip Thiravetyan
b,
a
Pilot Plant Development and Training Institute, King Mongkuts University of Technology Thonburi, Bangkok 10150, Thailand
b
School of Bioresources and Technology, King Mongkuts University of Technology Thonburi, Bangkok 10150, Thailand
a r t i c l e i n f o
Article history:
Received 22 September 2010
Received in revised form
17 November 2010
Accepted 17 November 2010
Keywords:
Melanoidins
Chitin nanobers
Adsorption
a b s t r a c t
Melanoidins, the complex bio-polymer of amino-carbonyl compounds are widely distributed in food,
drinks but if it was discharged into a water resource system that affect as environmental pollutants.
Adsorption is a potential method to remove the color from the food or environment. Therefore, this
research studied on the adsorption of synthetic melanoidins by chitin nanobers prepared from shrimp
shell waste. The results showed that the adsorption of melanoidins with chitin nanobers was increased
when increasing temperature, giving values of 131, 331 and 353mg/g at 20

C, 40

C and 60

C, respec-
tively, which were higher than other chitin-derived adsorbents. In addition, the b value of chitin
nanobers also had a high afnity and strength for melanoidins adsorption at low concentration higher
than other chitin-derived adsorbents. The results from Fourier transform infrared (FT-IR) spectroscopy
and elution studies conrmed that the interaction between melanoidins and chitin nanobers involved
both electrostatic and chemical adsorption. For application in the sugar industry, chitin nanobers can
be used for adsorption of melanoidins and other pigments from sugar syrup.
2010 Elsevier B.V. All rights reserved.
1. Introduction
Melanoidins, which are also known as Maillard reaction prod-
ucts, are polymeric and colored products that are formed by
a condensation reaction involving sugars and amino acids [1].
Despite extensive investigations using techniques such as infrared
(IR) spectroscopy, mass spectrometry and advanced multidimen-
sional nuclear magnetic resonance (NMR) spectroscopy analyses
[2], the structures of melanoidins are still not completely under-
stood. Melanoidins occur extensively in food and biological
material [3] andhave signicant effects onthe quality of food; since
colors and avors are important food attributes and key factor in
consumers acceptance [1,4]. Foodanddrink having brown-colored
melanoidins may offer substantial health promoting effects, but
in vitro studies have also revealed some mutagenic, carcinogenic
and cytotoxic properties [5,6].
Melanoidins in wastewaters released from distilleries and fer-
mentation industries may cause aquatic pollution and several
studies of the decolorization and degradation of melanoidins have
been performed using techniques such as occulation treatment
[7], and physicochemical treatment [8]. Activated carbon adsorp-
tion is widely used as an adsorbent such as remove color from
organic compounds from wastewater [9], the molasses wastew-
ater [10], melanoidin [11], and claried juice in sugar reneries

Corresponding author. Tel.: +66 2 470 7535; fax: +66 2 452 3455.
E-mail address: paitip.thi@kmutt.ac.th (P. Thiravetyan).
[12]. Biological decolorization of melanoidins has been success-
fully achieved [1316], but it has limited environmental factors.
In prior reports, chitin nanobers have been successfully prepared
from other materials based on methods described elsewhere for
crab shells [17,18], squid pens [19], Riftia tubes [20] and shrimp
shells [21]. However, there have not been prior reports the use of
chitin nanobers for adsorption of melanoidins. Moreover, a sig-
nicant amount of shrimp waste is produced in Asia, primarily in
Thailand, the main shrimp exporter [22] that large quantities of
shrimp waste constitute a potential source of chitin production.
Thepresent studyreports thealternativeuseof chitinnanobers for
adsorption of water containing melanoidins, the Langmuir adsorp-
tion isotherm and the interaction between melanoidins and chitin
nanobers studies by FTIR and elution study. In addition, the study
of sugar syrup by using chitin nanobers was also investigated.
2. Materials and methods
2.1. Chitin and chitosan
Chitin (48% DD) and chitosan (90% DD) fromshrimp shell waste
was supplied by Seafresh Chitosan (Lab) Co. Ltd., Thailand. The
average particle sizes were 400m1mm.
2.2. Preparation of chitin nanobers
The preparation of chitin nanobers was adapted from the
method reported in Nair and Dufresne [18]. The chitin was
1385-8947/$ see front matter 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.cej.2010.11.063
R. Dolphen, P. Thiravetyan / Chemical Engineering Journal 166 (2011) 890895 891
hydrolyzed with 3M HCl (the ratio of 3M HCl to chitin was 10g/L)
at 8090

C under stirring to produce a protein-free product. After

acid hydrolysis, the suspensions were centrifuged at 36,000rpm
for 15min, and the process was repeated three times. Next, the
suspensions were transferred to a dialysis bag, dialyzed in running
distilled water for 2h and then overnight in distilled water until
a pH 3.5 was reached and then air dried. The size distribution is
150800nm length and 1050nm width.
2.3. Preparation of modied chitin
Chitinwas bleachedwith6%(w/v) sodiumhypochlorite (ratio of
1:30 (w/v)); the mixture was being shaken at 150rpm at 302

C
for 1h. The modied chitin was washed with an excess of distilled
water until the pHwas 7, and then dried. The average particle size
was 400nm1mm.
2.4. Preparation of melanoidins
Melanoidins were prepared using a modication of a previous
method [9] by mixing 4.5g (0.025M) of glucose, 1.88g (0.025M) of
glycine and 0.42g (0.005M) of sodiumbicarbonate (NaHCO
3
) with
100mL of distilled water, then heating overnight at 80

C until dry.
100mL of water was then added and the sample was dialyzed in
1L of distilled water using 10cm dialysis tubing, with a change of
water every 810h. The total dialysis time was 136h. After dialysis,
the samples were ltered through cotton wool and freeze-dried.
2.5. Determination of surface area
The BET surface area, total pore volume and average pore vol-
ume of adsorbents were determined using a surface area analyzer
(Autosorb-1, Quantachrome, USA).
2.6. IR analysis
Functional groups of adsorbents before and after melanoidins
adsorption were measured by Fourier transform infrared (FT-IR)
spectroscopy (Bruker, ALPHA, Germany). The sample was mixed
with dry KBr in a ratio of 2mg:200mg and then the mixture was
ground and compressed. The FT-IR spectra of samples were taken
at 4004000cm
1
wavenumber range.
2.7. Microscopic analysis
For the analysis of chitin nanobers by transmission electron
microscopy (TEM) (JEM-2100, Japan), a droplet of a dilute suspen-
sion of nano-chitin was deposited on a carbon-coated grid and
allowed to dry. The accelerating voltage was 80kV.
2.8. Adsorption study
Adsorption experiments were performed in a 15mL vial using
0.05g of adsorbents with 10mL of melanoidins at various concen-
trations in the range of 203000mg/L. The mixtures were shaken
at 150rpmat 20

C, 40

C or 60

C at an equilibriumtime for 1h and

then centrifuged at 4500rpmfor 10min. The pHof the supernatant
was measured using a pH meter (Mettler Delta 340, England) and
the color was measured for absorbance at a maximumwavelength
at 420nm by spectrophotometer (UNICO-2100, USA). Melanoidins
adsorption was analyzed using the Langmuir isotherm Eq. (1).
q
e
=
q
max
bC
e
1 +bC
e
(1)
where q
e
is the amount of melanoidins adsorbed at equilibrium
(mg/g); C
e
is the concentration of melanoidins at equilibrium
Table 1
Surface area, pore volume and pore size of chitin. chitin nanobers, modied chitin
and chitosan.
Sample types Multipoint BET
(m
2
/g)
Total pore
volume (cc/g)
Average pore
diameter (nm)
Chitin 6.04 0.0209 13.82
Chitin nanobers 8.76 0.0220 10.03
Modied chitin 3 01 0.0052 6.98
Chitosan 6.86 0.0078 4.57
(mg/L); q
max
is the maximum adsorption capacity of melanoidins
per g of adsorbent (mg/g); and b is the Langmuir constant related
to the energy of adsorption (L/mg)
Sugar syrup was passed through packed columns containing
chitin nanobers at 1%, 5% and 10% dosage at a controlled ow
rate of 10mL/min for 1h. Samples were then analyzed for pH,
ICUMSA color remaining [23] and decolorization efciency using
Eqs. (2)(4). All experiments were conducted twice.
Concentration (g/100 mL) = 0.00002 (Brix
3
) +0.0036 (Brix
2
)
+1.0013 (Brix) (2)
ICUMSA color
=
Abs (420 nm) 100, 000
Cell length (1 cm) Concentration (g/100 mL)
(3)
Decolorization efciency (%)
=

1
OD
420
color of permeate
OD
420
color of sugar syrup

100 (4)
2.9. Elution study
Suspensions of 1% (w/v) of melanoidin-adsorbed adsorbents
were addedto a syringe, whichwas connectedto a peristaltic pump
with a ow rate of 2.0mL/min, and eluted with distilled water
until no color appeared in the solution. 0.5M NaOH was then used
as a second eluent followed by 80% methanol, and the color was
analyzed by spectrophotometer at 420nm.
3. Results and discussion
3.1. Physical properties of chitin nanobers
Table 1 shows the surface area, total pore volume and average
pore diameter of chitin and chitin nanobers measured by surface
area analysis. The surface area of chitin nanobers is slightly higher
thanchitin, but the average pore diameters of bothchitinandchitin
nanobers are in the mesopore range (250nm).
After hydrolysis with 3M HCl, the morphology of chitin
nanobers was observed by transmission electron microscopy
(TEM) (Fig. 1). The results showthat these chitin fragments consist
of slender rods with sharp points and a broad size distribution with
lengths and widths of 150800nm and 1050nm, respectively.
The FT-IR spectrum of chitin (Fig. 2a) is typical of chitin,
with characteristic peaks at 3449cm
1
(NH), 1651cm
1
for
amide I (C O), 1379cm
1
(OH), 1255cm
1
(CNH) and 1083cm
1
(CO). Accordingly, sodiumhypochlorite affectedthe alcohol group
(CH
2
OH) of chitin and transformed it to CH
2
OCl [24], modied
chitin (Fig. 2b) shows a signicant increase peaks at 1649cm
1
and 1561cm
1
and a newpeak at 698cm
1
(Cl) appeared. Chitosan
(Fig. 2c) shows peaks at 1655cm
1
(amide I, C O) and 1560cm
1
(combination amide II, combination of CN STR and NH bend).
892 R. Dolphen, P. Thiravetyan / Chemical Engineering Journal 166 (2011) 890895
Fig. 1. Transmission electron micrograph (TEM) of chitin nanobers.
Table 2
Parameter of the Langmuir constants for the adsorption of melanoidins by chitin
nanobers at various temperatures.
Adsorbent Temperature (

C) Langmuir constants
Qmax (mg/g) R
2
b (L/mg)
Chitin nanobers 20 131 6.35 0.9774 0.0417
40 331 3.32 0.9793 0.0146
60 353 2.90 0.9803 0.0137
Obviously, chitin nanobers (Fig. 2d) show sharp characteristic
peaks as compared to the starting chitin, which is due to the highly
crystalline nanobers; respectively the peaks at 1158, 1115, 1072
and 1028cm
1
(CO).
However, after acid hydrolysis, chitin was transformed from
polymeric to monomeric structures (Fig. 3), resulting in shifts in
the absorption wavenumber and an increased intensity of peaks in
chitin nanobers.
3.2. Adsorption isotherm study
The adsorption isotherms (Fig. 4) show that adsorption of
melanoidins by chitin nanobers increased with increasing tem-
peratures. The Langmuir constants are presented in Table 2, and
the high value of R
2
indicates a good t to the Langmuir model, and
hence homogeneous surface or monolayer adsorption.
Fig. 2. FT-IR spectra of chitin (a), modied chitin (b), chitosan (c) and chitin
nanobers (d).
The maximum adsorption capacities of melanoidins by chitin
nanobers were 131, 331 and 353mg/g at 20

C, 40

C, and
60

C, respectively. However, maximum adsorption capacity at

60

C was higher than at lower temperatures. The b values were

decreased when increasing the temperature were 0.0417, 0.0146
and 0.0137L/mg at 20

C, 40

C, and 60

C, respectively. It implied
that at lowtemperature (20

C) chitinnanobers hada higher afn-

ity to adsorb melanoidins than at high temperature (Table 2).
From the preparation of chitin nanobers, the pH obtained
was 3.5. The decrease in pH is probably due to the release of
some deacetylated amino-groups, which complex with a proton
from within the crystallites to give NH
3
+
at the crystallite surfaces
[17]. Moreover, the melanoidins behave as anionic hydrophilic
polymers, which are able to bind with metal cations [25,26]. The
mechanism of adsorption may involve the amine groups in the
chitin. A proportion of the amine groups will be protonated and
Fig. 3. Structure of chitin and chitin nanobers after acid hydrolysis.
R. Dolphen, P. Thiravetyan / Chemical Engineering Journal 166 (2011) 890895 893
Fig. 4. Adsorption isotherm of melanoidins by 0.5% chitin nanobers at various
temperatures, 20

C (), 40

C () and 60

C ().
hence the main mechanism of melanoidins adsorption will be the
electrostatic attraction between the melanoidins and the R-NH
3
+
of amine groups [2729].
3.3. IR analysis
The IR spectra of melanoidins show peaks at 3260cm
1
(OH,
stretching), 2950cm
1
(CH, symmetric stretching), 1603cm
1
(C O, stretching), 1383cm
1
and 1307cm
1
(OH, deformation),
1220cm
1
(CN, stretching) and 1035cm
1
(CO, stretching)
(Fig. 5).
The results of FT-IR spectra of chitin nanobers before and
after melanoidins adsorption are shown in Fig. 5a and b. The func-
tional groups are changed, the peak at 1627cm
1
(amide II, NH
2
)
appeared and the peaks 1425cm
1
(CN) and 1204cm
1
(CN)
were shifted to 1415cm
1
and 1254cm
1
while the other peaks
were slightly shifted. It indicated that the interaction between
melanoidins and chitin nanobers involved both electrostatic and
chemical adsorption.
3.4. Desorption studies
After adsorption of melanoidins by chitin nanobers, an elu-
tion test was performed in order to investigate the adsorption
mechanisms. Cammerer and Kroh propose the general structure
Fig. 5. FT-IR spectra of chitin nanobers before (a) and after (b) melanoidin adsorp-
tion.
of the melanoidins polymer as anionic hydrophilic polymers that
containing ketone and hydroxyl groups act as donor groups in
melanoidins [30]. Therefore, desorption of the color-adsorbed on
chitin nanobers using distilled water followed by 0.5M NaOH
and 80% methanol. Table 3 shows that distilled water, 0.5M NaOH
and 80% methanol could elute 2%, 89% and 6% of the melanoidins,
respectively. These results conrmed that the melanoidins adsorp-
tionontochitinnanobers involvedbothelectrostatic andchemical
adsorption.
Fig. 6. Comparison of the adsorption of melanoidins by using 0.5% of various adsorbents at a temperature of 30

C; chitin (a), modied chitin (b), chitosan (c), and chitin

nanobers (d).
894 R. Dolphen, P. Thiravetyan / Chemical Engineering Journal 166 (2011) 890895
Table 3
Thepercentageof melanoidinelutionfromchitinnanobers byusingdistilledwater,
0.5M NaOH and 80% methanol as eluents.
Step Eluent % Elution pH
Initial System
1 Distilled water 2 0.82 8.50 8.45
2 0.5M NaOH 89 5.23 13.49 13.58
3 80% CH
3
OH 6 0.59 12.32 10.81
Total elution (%) 97
Table 4
Langmuir constants for the adsorption of melanoidins by using 0.5% of various
adsorbents at 30

C.
Adsorbents Qmax (mg/g) System pH R
2
b (L/mg)
Chitin nanobers 242 0.82 3.83 0.9935 0.0173
Chitosan 99 2.34 5.93 0.9931 0.0017
Chitin 61 2.55 3.92 0.9458 0.0017
Modied chitin 8 0.89 3.87 0.9225 0.0027
3.5. Use of other chitin-derived adsorbents
Melanoidins adsorption by chitin nanobers was higher than
by other adsorbents, such as chitin, modied chitin and chitosan
(Table 4, Fig. 6); the Langmuir isotherms obtained by using 0.5%
of various adsorbents at 30

C show values for q

max
of 242, 99,
61 and 8mg/g for chitin nanobers, chitosan, chitin and modied
chitin, respectively. The b values showed that chitin nanobers
was 0.0173L/mg while chitosan, chitin and modied chitin were
0.0017, 0.0017 and 0.0027L/mg, respectively. It suggested that
chitin nanobers had a high afnity and strength for melanoidins
adsorption at low concentration higher than other chitin-derived
adsorbents. However, the system pH of chitin nanobers, chi-
tosan, chitin and modied chitin were 3.83, 5.93, 3.92 and 3.87,
respectively. Therefore, it suggests that the functional groups of
adsorbents had affected to the adsorption of melanoidins.
The functional groups of chitinandchitosanwere OH, NHCOCH
3
and NH
2
. The higher efciency of chitosan compared with chitin
can be explained by the accessibility of the amine functional in
chitosan. The pK
a
of primary amino groups was 6.3, at low pH,
the positive charge on the NH
3
+
groups converts chitosan to a
water-soluble cationic polyelectrolyte [31]. This may be due to the
increase in the number of positive charges on the sorbent sur-
face that leads to greater interaction to the negatively charged
melanoidins molecules. All amine groups of chitosan and chitin
were ionized and the melanoidins anion was pulled in strongly by
the electrostatic attraction.
Moreover, chitin nanobers was hydrolyzed from polymer to
monomer form with HCl. The physical properties include surface
area and the particle sizes of adsorbent were changed; it was very
ne powdered formthat may be easily suspended in the solution to
be treated. It affects an increasing dissociation of (R-NH
2
) into (R-
NH
3
+
) [17,2729]. Theamount of melanoidins adsorptionincreased
with increasing numbers of binding sites of chitin nanobers.
Furthermore, adsorption with modied chitin was lower than
other adsorbents, probably because the actionof sodiumhypochlo-
rite (NaOCI) transforming (CH
2
OH) to CH
2
OCl [20] may have
affectedthe properties of modiedchitinandresultedina decrease
in adsorption.
3.6. Application of chitin nanobers with sugar syrup
The decolorization efciency of sugar syrup by chitin nanobers
at various dosages is shown in Table 5. The decolorization ef-
ciency increased with increasing adsorbent dosage. 1%, 5% and 10%
Table 5
Decolorization of sugar syrup by various dosages of chitin nanobers.
Sample System pH ICUMSA
color
Decolorization
efciency (%)
Initial sugar syrup 6.98 0.18 598 16.87
Initial sugar syrup at
various system pH
6.49 0.15 582 3.42
5.39 0.09 545 2.76
4.49 0.11 485 5.01
Sugar syrup+1% chitin
nanobers
6.49 0.10 373 2.60 39 0.15
Sugar syrup+5% chitin
nanobers
5.39 0.12 184 1.47 65 0.32
Sugar syrup+10% chitin
nanobers
4.49 0.15 143 9.71 73 1.40
of chitin nanobers could reduce the color of sugar syrup to 39%,
65%and73%, respectively. Inaddition, color fromtheinitial ICUMSA
of sugar syrup after adjusted systempHwas decreased from598 to
582, 545 and 485 at system pH of 6.49, 5.39 and 4.49, respectively.
While ICUMSA of sugar syrup by chitin nanobers was decreased
from 598 to 373, 184 and 143 at a 1%, 5% and 10% dosage, respec-
tively.
4. Conclusion
The results of this study showed that chitin nanobers pre-
pared from shrimp shell waste is very promising for adsorption
of melanoidins especially in Thailand, shrimp shell waste is read-
ily available and cheap. After acid hydrolysis, chitin nanobers
was hydrolyzed from polymer to monomer form; it was very ne
powdered form that easily suspended in the melanoidins solution,
the NH
3
+
groups has affected to increase of the adsorption of
melanoidins. The maximum adsorption capacities of melanoidins
by chitin nanobers were 131, 331 and 353mg/g at 20

C, 40

C,
and 60

C, respectively. Chitin nanobers had a higher afnity for

melanoidins whencomparedwithother chitin-derivedadsorbents.
Desorption of the color-adsorbed on chitin nanobers showed that
distilled water, 0.5M NaOH and 80% methanol could elute 2%, 89%
and 6% of melanoidins, respectively. The results fromFTIR and elu-
tion studies conrmed that the interaction between melanoidins
and chitin nanobers involved both electrostatic and chemical
adsorption. For application in sugar industry, chitin nanobers can
be used to adsorb melanoidins and other pigments from sugar
syrup.
Acknowledgements
This research was funded by The National Research University
Project of Thailands ofce and the Commission of Higher Educa-
tion.
References
[1] M. Plavsic, B. Cosovic, C. Lee, Copper complexing properties of melanoidins and
marine humic material, Sci. Total Environ. 366 (2006) 310319.
[2] A. Arnoldi, E.A. Corain, L. Scaglioni, J. Ames, Newcoloured compounds fromthe
Maillard reactions between xylose and lysine, J. Agric. Food Chem. 45 (1997)
650655.
[3] B. Cmmerer, L.W. Kroh, Investigation of the inuence of reaction conditions
on the elementary composition of melanoidins, Food Chem. 53 (1995) 5559.
[4] S.I.F.S. Martins, M.A.J.S. van Boekel, Melanoidins extinction coefcient in the
glucose/glycine Maillard reaction, Food Chem. 83 (2003) 135142.
[5] J.M. Silvan, J.V.D. Lagemaat, A. Olano, M.D.D. Castillo, Analysis and biological
properties of amino acid derivates formed by Maillard reaction in foods, J.
Pharm. Biomed. Anal. 4 (2006) 15431551.
[6] R.C. Borrelli, C. Mennella, F. Barba, M. Russo, G.L. Russo, K. Krome, H.F. Erbers-
dobler, V. Faist, V. Fogliano, Characterization of coloured compounds obtained
by enzymatic extraction of bakery products, Food Chem. Toxicol. 4 (2003)
13671374.
R. Dolphen, P. Thiravetyan / Chemical Engineering Journal 166 (2011) 890895 895
[7] V.P. Migo, M. Matsumara, E.J.D. Rosario, H. Kataoka, Decolorization of molasses
wastewater using an inorganic occulant, J. Ferment. Bioeng. 75 (1993)
438442.
[8] S.B. Kim, F. Hayase, H. Kato, Decolourisation and degradation products of
melanoidins on ozonolysis, Agric. Biol. Chem. 49 (1985) 785792.
[9] H. Tamon, T. Saito, M. Kishimura, M. Okazaki, R. Toei, Solvent regeneration of
spent activated carbon in wastewater treatment, J. Chem. Eng. Jpn. 23 (1990)
426432.
[10] E.C. Bernardo, R. Egashira, J. Kawasaki, Decolorization of molasses wastew-
ater using activated carbon prepared from cane bagasse, Carbon 35 (1997)
12171221.
[11] A. Simaratanamongkol, P. Thiravetyan, Decolorization of melanoidin by acti-
vated carbon obtained from bagasse bottom ash, J. Food Eng. 96 (2010) 1417.
[12] M. Ruiz, C. Rolz, Activated carbons from sugar cane bagasse, Ind. Eng. Chem.
Prod. Res. 4 (1971) 429432.
[13] P. Kumar, R. Chandra, Decolourisation and detoxication of synthetic molasses
melanoidins by individual and mixed cultures of Bacillus spp., Biores. Technol.
7 (2006) 20962102.
[14] V. Kumar, L. Wati, P. Nigam, I.M. Banat, G. MacMullan, D. Singh, R. Marchant,
Microbial decolourisation and bioremediation of anaerobically digested
molasses spent wash efuent by aerobic bacterial culture, Microbios 89 (1997)
8190.
[15] S. Ohmomo, Y. Kaneko, S. Sirianantapiboon, P. Somachi, P. Atthasampunna, I.
Nakamura, Decolourisation of molasses waste water by a thermophilic strain
Aspergillus fumigatus G-2-6, Agric. Biol. Chem. 52 (1987) 33393346.
[16] I. Aoshima, Y. Tozawa, S. Ohmomo, K. Udea, Productionof decolourizing activity
for molasses pigment by Coriolus versicolor Ps4a, Agric. Biol. Chem. 49 (1985)
20412045.
[17] R.H. Marchessault, F.F. Morehead, N.M. Walter, Liquid crystal systems from
brillar polysaccharides, Nature 184 (1959) 632633.
[18] K.G. Nair, A. Dufresne, Crab shell chitin whisker reinforced natural rubber
nanocomposites. 1. Processing and swelling behavior, Biomacromolecules 4
(2003) 657665.
[19] M. Paillet, A. Dufresne, Chitin whisker reinforced thermoplastic nanocompos-
ites, Macromolecules 34 (2001) 65276530.
[20] A. Morin, A. Dufresne, Nanocomposites of chitin whiskers fromRiftia Tubes and
poly (caprolactone), Macromolecules 35 (2002) 21902199.
[21] J. Sriupayo, P. Supaphol, J. Blackwell, R. Rujiravanit, Preparation and character-
ization of -chitin whisker-reinforced chitosan nanocomposite lms with or
without heat treatment, Carbohyd. Polym. 62 (2005) 130136.
[22] S.L. Wang, I.L. Shih, T.W. Liang, C.H. Wang, Purication and characterization of
twoantifungal chitinases extracellularlyproducedbyBacillus amyloliquefaciens
V656ina shrimpandcrabshell powder medium, J. Agric. FoodChem. 50(2001)
22412248.
[23] ICUMSA Colour Determination Method, Method 2. International Commission
for Uniform Methods of Sugar Analysis, 1994.
[24] E. Sadov, M. Korchagin, A. Matetsky, Chemical Technology of Fibrous Mate-
rials: Bleaching of Cellulosic Fibrous Materials, Mir Press, Moscow, 1973, pp.
127157.
[25] T. Gomyo, M. Horikoshi, On the interaction of melanoidin with metallic ions,
Agric. Biol. Chem. 40 (1976) 3340.
[26] V.P. Migo, E.J. Del Rosario, M. Matsumura, Flocculation of melanoidins induced
by inorganic ions, J. Ferment. Bioeng. 83 (1997) 287291.
[27] F.C. Wu, R.L. Tseng, R.S. Juang, Kinetic modeling of liquid-phase adsorption of
reactive dyes and metal ions on chitosan, Water Res. 35 (2001) 613618.
[28] N. Sakkayawong, P. Thiravetyan, W. Nakban, Adsorption mechanism of syn-
thetic reactive dye wastewater by chitosan, J. Colloid Interface Sci. 286 (2005)
3642.
[29] R.S. Juang, R.L. Tseng, F.C. Wu, S.W. Lee, Adsorption behavior of reactive dyes
from aqueous solutions on chitosan, J. Chem. Technol. Biotechnol. 70 (1997)
391399.
[30] B. Cammerer, L.W. Kroh, Investigation of the inuence of reaction condi-
tion on the elementary composition of melanoidin, Food Chem. 53 (1995)
5559.
[31] C.K.S. Pillai, W. Paul, C.P. Sharma, Chitin and chitosan polymers: chemistry,
solubility and ber formation, Progr. Polym. Sci. 34 (2009) 641678.