D.M. Suflet et al.
/ Reactive & Functional Polymers 66 (2006) 1240–1249
Cell–P(O)(OH)2. The derivatives of pentavalent
phosphorus, i.e. H3PO4, P2O5, organic phosphates,
and POCl3 are most frequently used to obtain phos-
phorous containing cellulose derivatives.
Phosphorylation of cellulose was performed
either by reaction of the hydroxy groups of the par-
ent polymer, or by a second-hand derivatization of a
cellulose ether or ester already formed [3]. The syn-
thesis of phosphorylated cellulose was effectuated
by different systems of reactions: phosphoric acid
[3–5], P2O5 [6], PCl3 or dimethylphosphate [3], etc.
Original results were reported by Gospodinova
et al. about the phosphorylation of cellulose assisted
by microwaves which allowed achieving a degree of
substitution as high as 2.8 being at the same time
joined to the green context [7]. The data concerning
the synthesis and characterization of water-soluble
phosphorylated derivatives of cellulose are rather
scarce. The products obtained are frequently insolu-
ble due to crosslinking and rather ill-defined being
often characterized by their phosphorous content
only. Many of the reactions involved are not quite
clear yet regarding their course and mechanism.
Insoluble phosphorylated cellulose was mainly
employed as flame retardant additive for textiles
or plastics and as cation exchanger especially in bio-
chemical separation process. Water-soluble phos-
phorylated derivatives were used as thickeners in
aqueous systems when the lack of toxicity is an
important advantage. Another application is in the
biomedical topic. Cellulose phosphates substituted
in the secondary positions were reported to inhibit
the activation of detrimental blood proteins in hae-
modialysis after incorporation in haemodialysis
membranes [8]. A commercial product, Calci-Bind,
was proposed for the treatment of hypercalciuria
[9]. More recently Granja et al. synthesized highly
phosphorylated cellulose gels and demonstrated
their biocompatibility and ability to promote the
formation of calcium phosphates [10–12].
In this article we report new results on the syn-
thesis of water-soluble phosphorylated cellulose
and its characterization regarding the degree of sub-
stitution and the chemical structure as well as some
properties in solid state or in aqueous solution. The
effect of phosphorylated cellulose by the separation
of low soluble salts is also checked in order to
obtain useful information about the possibility of
application in the control of biomineralization pro-
cesses or as additive by the preparation of hydroxy-
apatite by hydrothermal method. These results are a
part of an extensive research regarding the phos-
1241
phorylation of different polysaccharides and the
properties of the phosphorylated polysaccharides
which will be the subject of subsequent articles.
2. Experimental
2.1. Materials
The parent polymer was microcrystalline cellu-
lose Avicel PH-101 obtained from Acros Organics,
Germany. Microcrystalline cellulose is highly crys-
talline particulate cellulose consisting primarily of
crystallite aggregates, obtained by removing amor-
phous (fibrous cellulose) regions of a purified cellu-
lose source material. The low molecular reactants
were phosphorous acid 99% from Sigma–Aldrich
Chemie GmbH Germany, urea 99.5% from Carl
Roth, Germany, and methanol from Chimopar,
Bucuresti, Romania. All chemicals were of research
grade purity and used without further purification.
2.2. Methods
2.2.1. Phosphorylation of cellulose
Reaction of cellulose with phosphorous acid was
performed according to the method described by
Inagaki et al. [13]. In a 250 ml, four-necked flask
equipped with a nitrogen inlet, a condenser, a ther-
mometer, and a stirrer, 12.48 g (0.208 mol.) urea
was added, heated at 140 °C and flushed with nitro-
gen. Two grams (0.0123 mol.) microcrystalline cel-
lulose and 10.28 g (0.1253 mol.) phosphorous acid
were added alternatively portionwise to the molten
urea in order to reduce the foaming. The reaction
was allowed to proceed at 150 °C for different peri-
ods of time. The reacted mixture was dissolved in
1 N aqueous sodium hydroxide and then precipi-
tated with methanol. This process was repeated
three times, in order to remove the residual
reagents. The modified cellulose was dried over
phosphorus pentoxide under reduced pressure, at
25 °C. The phosphorylated cellulose is a white,
water-soluble powder. Occasionally some sample
was further purified by diafiltration in a Millipore
cell equipped with a PM10 membrane up to the dia-
filtrate had a conductivity lower than 5 lS/cm. The
polymer was recovered by freeze-drying.
2.2.2. Analysis and characterization of
phosphorylated cellulose
Elemental analysis of phosphorous was
performed by Kjeldahl mineralization method
 Carbohydrate Polymers 82 (2010) 1271–1277

Contents lists available at ScienceDirect

Carbohydrate Polymers
journal homepage: www.elsevier.com/locate/carbpol

Phosphorylated polysaccharides. 2. Synthesis and properties of phosphorylated

dextran
Dana M. Suflet a , Gabrielle C. Chitanu a,∗ , Jacques Desbrières b
a
Petru Poni Institute of Macromolecular Chemistry, Aleea Grigore Ghica Voda 41A, 700487 Iasi, Romania
b
University of Pau, Equipe de Physique et Chimie des Polymères, IPREM - UMR 5254, Hélioparc, 2 avenue du Président Angot, 64053 Pau Cedex 09, France

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

Article history: New results regarding the synthesis and characterization of water-soluble phosphorylated dextran
Received 8 December 2009 (PDex) are presented. The phosphorylation was made by reaction of dextran with phosphorous acid in
Received in revised form 5 May 2010 molten urea. The structure of phosphorylated product was investigated by elemental analysis, by titration
Accepted 2 July 2010
of acidic groups and by FTIR and NMR spectroscopy. It was found that phosphorylated dextran prepared
Available online 21 July 2010
in these conditions is a monobasic dextran phosphate with (H)P(O)(OH) groups. The degree of polymer-
ization of PDex was found comparable to the parent dextran, suggesting that polysaccharide chain was
Keywords:
not degraded during phosphorylation. The polyelectrolyte behavior was studied by potentiometric and
Polysaccharides
Dextran
conductometric titration and by viscometric measurements.
Phosphorylation © 2010 Elsevier Ltd. All rights reserved.
Polyelectrolytes

1. Introduction In this article we report original results on the synthesis, charac-

Dextran is a family of neutral polysaccharides produced by bac-
terial enzymes, consisting of main chains from ␣-(1→6) linked
d-glucose units with different ratios of linkages and branches,
depending on the bacterial strains. The chemical structure of dex-
tran includes ␣-(1→6) linkages that can vary from 97 to 50% of total
glucosidic bonds. The branching is made in position ␣-(1→2), ␣-
(1→3) and/or ␣-(1→4) (Heinze, Liebert, Heublein, & Hornig, 2006;
Robyt, 1987). The hydroxyl groups in dextran are preferred for most
derivatization. The C-2 hydroxyl groups appear to be the first to
react (Allene, 1966) because the hydroxyl groups of C-6 are con-
cerned in ␣-1, 6 linkages. Until now various dextran derivatives
were obtained, such as: esters of aliphatic or aromatic organic acids,
of inorganic acids such as nitric, sulfuric, and phosphoric (Allene,
1966; Gekko, 1981, chap. 29; Heinze et al., 2006; Larsen, 1990;
Robyt, 1987; Whistler & Towle, 1969). The references regarding the
synthesis of phosphorylated derivatives are presented in Table 1.
Phosphorylated derivatives of dextran were characterized by
elemental analysis, potentiometric titration, 1 H, 31 P, and 13 C NMR
spectroscopy and high-performance exclusion chromatography
(Sacco, Klett-Zygmunt, & Dellacherie, 1988). Until now no mention
was made about their water solubility or polyelectrolyte behavior,
though these aspects are important in most of the applications.
∗ Corresponding author. Tel.: +40 232 217454; fax: +40 232 211299.
E-mail address: chita@icmpp.ro (G.C. Chitanu).
terization and properties of water-soluble phosphorylated dextran,
including polyelectrolyte behavior. These results are a part of an
extensive research regarding the phosphorylation of dissimilar
polysaccharides and the properties of the phosphorylated polysac-
charides the results of which will be the subject of subsequent
articles.
2. Experimental
2.1. Materials
The parent dextran (Mr ∼ 200.000, Leuconostoc mesenteroides)
was purchased from Fluka, Switzerland. The other reagents were
phosphorous acid 99% from Sigma–Aldrich, Germany, urea 99.5%
from Carl Roth, Germany, and methanol from Chimopar, Romania.
All chemicals were used without further purification.
2.2. Methods
Phosphorylation of dextran was performed by reaction of dex-
tran with phosphorous acid (Inagaki, Nakamura, Asai, & Katsuura,
1976; Suflet, Chitanu, & Popa, 2006). In a 250 mL, four-necked flask
equipped with nitrogen inlet, condenser, thermometer, and stir-
rer, 64 g (1.067 moles) urea was introduced and heated at 140 ◦ C
under flushing with N2 . 10 g (0.061 moles) dextran (Dex) and 51 g
(0.622 moles) phosphorous acid were added portionwise alterna-
tively to the molten urea. The reaction was allowed to proceed at
150 ◦ C for different times. The reaction mixture was cooled, dis-

0144-8617/$ – see front matter © 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carbpol.2010.07.007
1272 D.M. Suflet et al. / Carbohydrate Polymers 82 (2010) 1271–1277

Scheme 1. Reaction of dextran with phosphorous acid.

solved in 1 N aqueous NaOH and precipitated in methanol. The Table 2

Degree of substitution of phosphorylated dextran samples.
precipitation was repeated several times. The final product was
dried under reduced pressure, at 25 ◦ C, over phosphorous pentox- Sample code Reaction Dex/urea/ DSP DSt
ide. The phosphorylated dextran (PDex) is a white, water-soluble time (h) H3 PO3
powder. It was purified by diafiltration in a Millipore cell with PDex2 2 1/6/5 1.18 1.30
PM10 membrane up to the diafiltrate conductivity was lower than PDex5 5 1/6/5 1.78 1.90
5 ␮S/cm. Then the polymer was recovered from aqueous solution
by freeze-drying.
a sulfonic Dowex 50W×8, 20–50 mesh resin. Viscometric mea-
surements were performed on diluted solutions of Na salt of PDex
2.2.1. Analysis and characterization of phosphorylated dextran at 25 ± 0.1 ◦ C, using an Ubbelohde viscometer with 0a capillary
Elemental analysis of phosphorus was performed by Kjeldahl (K = 0.005319).
mineralization method combined with spectrophotometric assess-
ment at 740 nm after adding of ammonium molybdate. Elemental
3. Results and discussion
analysis of nitrogen was performed using Kjeldahl method. FTIR
spectra were recorded on KBr pellet using a Vertex 70 Bruker
3.1. Reaction of dextran with phosphorous acid
spectrometer. NMR spectra were recorded in D2 O using a Bruker
Avance 400.13 MHz instrument. The molar mass of PDex was
The degree of substitution (DS) of phosphorylated samples was
estimated by size exclusion chromatography using a Waters instru-
evaluated by phosphorus elemental analysis (DSP ) and by potentio-
ment equipped with two Ultrahydrogel columns (250 and 2000,
metric or conductometric titration (DST ) according to the methods
7.8 mm × 300 mm) and a refractive index detector. The eluent was
already verified (Suflet et al., 2006). The results are presented in
aqueous 0.1 M NaNO3 solution with 3% acetonitrile. The flow rate
Table 2. It can be noticed that the DSP and DST are close. Taking
was 0.8 mL/min and the feed was 150 or 50 ␮L of 1.0 or 3.0 mg/mL
into account the use of molten urea as reaction medium, nitrogen
polymer solution in aqueous 0.1 M NaNO3 . Poly(acrylic acid) sam-
analyses were also performed in order of checking the purity of the
ples from Waters were used as standard. The decomposition under
reacted polymer. All samples showed a very low N content indicat-
dynamic conditions of heating has been investigated with a Paulik-
ing a slight remanence of urea, even after diafiltration procedure.
Paulik-Erdey MOM-Budapest instrument on 50 mg samples, at
The reaction of dextran with phosphorous acid is expected to
12 ◦ C/min, in air.
proceed predominantly at the hydroxyl group of the C-2 atom
Polyelectrolyte behavior was studied by potentiometric and
(Heinze et al., 2006) according to Scheme 1.
conductometric titration and by viscometric measurements. Poten-
It is known that the derivatives containing mobile hydro-
tiometric titration was performed using a Metrohm all purposes
gen atoms directly attached to phosphor atom are in tautomeric
716 DMS Titrino apparatus equipped with a 6.0203.100 combined
equilibrium (Kabachnik & Mastryakova, 1953). Accordingly PDex
electrode. Monovalent bases LiOH and KOH were used for titra-
could present the tautomeric forms given in Scheme 2. In order
tion without or with added low molecular salt. Conductometric
to check which one of these structures is preferred, FTIR spec-
titrations were performed with same bases, with no added salt,
tra were registered and potentiometric/conductometric titrations
using a 712 Metrohm conductometer with 6.0908.110 cell. The
were performed. The characteristic bands of PDex2 and PDex5
potentiometric and conductometric titrations were made on the
phosphorylated samples are summarized in Table 3 compared to
acid form of PDex. As the phosphorylated dextran was obtained in
the parent dextran (Purama, Goswami, Khan, & Goyal, 2009; Hesse,
form of Na salt, the free acid form was prepared by passing diluted
Meier, & Zeeh, 1984).
solutions of Na salt through an ion exchange column filled with
In Figs. 1 and 2 are shown the potentiometric and conductomet-
ric titration curves of PDex samples with monovalent bases (0.0838
Table 1 or 0.0755 N KOH and 0.0947 N LiOH).
Methods of synthesis of phosphorylated dextran. One inflexion point is observed on the potentiometric curve
obtained by titration of PDex2 with DS = 1.3 (Fig. 1), which can be
Phosphoryiation agent Ref.

Polyphosphoric acid/tributylamine Heinze et al. (2006), Sacco,

(TBA)/dimethyl formamide (DMF)
H3 PO4 /faty acids esters
H3 PO4 /palmitic acid/tertiary amines
H3 PO4 /urea (the reaction results in
crosslinked dextran)
H3 PO4 /formamide
Klett-Zygmunt, Vigneron, and
Dellacherie (1990) and Spaltro and
Aronson (1992)
Suzuki, Suzuki, and Matsumoto (1975)
Heinze et al. (2006) and Suzuki, Suzuki,
and Mikami (1977)
Makarova et al. (1978)
Sato et al. (2004) Scheme 2. Tautomeric forms of phosphorylated dextran.
 Carbohydrate Polymers 84 (2011) 1176–1181

Contents lists available at ScienceDirect

Carbohydrate Polymers
journal homepage: www.elsevier.com/locate/carbpol

Phosphorylated polysaccharides. 3. Synthesis of phosphorylated curdlan and its

polyelectrolyte behaviour compared with other phosphorylated polysaccharides
Dana M. Suflet ∗ , Alina Nicolescu, Irina Popescu, Gabrielle C. Chitanu
Petru Poni Institute of Macromolecular Chemistry, Aleea Grigore Ghica Voda 41A, 700487 Iasi, Romania

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

Article history: A water-soluble phosphorylated curdlan (PCurd) was synthesized by reaction of curdlan with phospho-
Received 19 August 2010 rous acid in molten urea. The structure of PCurd was investigated by FTIR and NMR spectroscopy and the
Received in revised form substitution degree was calculated from potentiometric titration. It was found that the obtained curdlan
16 December 2010
derivative is a monobasic curdlan phosphate with –HP(O)(OH) groups. The polyelectrolyte behaviour
Accepted 10 January 2011
of PCurd, studied by electrochemical methods and by viscometry, was compared with other phospho-
Available online 15 January 2011
rylated polysaccharides. A slight influence of the polysaccharide chain flexibility on the polyelectrolyte
dissociation was observed.
Keywords:
Polysaccharide derivatives © 2011 Elsevier Ltd. All rights reserved.
Curdlan
Phosphorylation
Polyelectrolyte behaviour

1. Introduction In this study, we present the synthesis, characterization and

Curdlan, a bacterial polysaccharide formed by pure culture
fermentation of Agrobacterium biobar 1 (identified as Alcaligenes
faecalis var. myxogenes) is a polysaccharide with a linear struc-
ture composed entirely of d-glucose linked by ␤-glucosidic bonds
in (1 → 3) positions (Laroche & Michaud, 2007). Water insolubil-
ity of curdlan, generally attributed to the existence of extensive
intra/intermolecular hydrogen bonds, limited the biological appli-
cations. This disadvantage can be eliminated by derivatization
reactions of curdlan. The derivatives of curdlan with sulphate
(Wang, Zhang, Yu, & Cheung, 2009; Yoshida, Hatanaka, & Uryu,
1990), carboxymethyl (Jin, Zhang, Yin, & Nishinari, 2006), and
ammonium groups (Borjihan, Zhong, Baigude, Nakashima, & Uryu,
2003; Numata, Sugikawa, Kaneko, & Shinkai, 2008) were reported.
The data concerning the synthesis and characterization of water-
soluble phosphorylated derivatives of curdlan are rather scarce.
Only a phosphate derivative obtained by reaction of curdlan with
phosphoric acid in dimethyl sulphoxide – urea was presented in the
literature (Chen, Xu, Zhang, & Zeng, 2009; Williams et al., 1991). In
recent years, many papers have reported, for native curdlan and its
derivatives, an important biological activities such as anti-tumour,
anti-HIV (Borjihan et al., 2003; Yoshida et al., 1990), anti coagulant
(Alban & Franz, 2001), anti-oxidant activity and immunomodula-
tory effects (Chen & Seviour, 2007; Laroche & Michaud, 2007).
∗ Corresponding author. Tel.: +40 232 217454; fax: +40 232 211299.
E-mail address: dsuflet@icmpp.ro (D.M. Suflet).
properties of water-soluble phosphorylated curdlan obtained by
reaction with phosphorous acid in molten urea. The polyelectrolyte
behaviour of the curdlan derivative was compared with other phos-
phorylated polysaccharides (dextran and cellulose).
2. Experimental
2.1. Materials
Curdlan (Alcaligenes faecalis) from Wako Pure Chemical Ind.,
Japan was kindly provided by Prof. T. Uryu. The reagents: phospho-
rous acid 99% from Sigma–Aldrich, Germany, urea 99.5% from Carl
Roth, Germany, and methanol from Chimopar, Romania, were used
without further purification. The monobasic dextran phosphate
(PDex, with degree of substitution about 1.5), and the monoba-
sic cellulose phosphate (PCel, with degree of substitution about 1)
were synthesised in our laboratory (Suflet, Chitanu, & Desbrières,
2010; Suflet, Chitanu, & Popa, 2006).
2.2. Methods
2.2.1. Synthesis of phosphorylated curdlan
Phosphorylation of curdlan was performed by reaction of curd-
lan with phosphorous acid (Inagaki, Nakamura, Asai, & Katsuura,
1976). To obtain the reaction media, 20 g urea was heated at 140 ◦ C
into a flask equipped with nitrogen inlet, condenser, and stirrer. 2 g
curdlan (Curd) and 10 g phosphorous acid were added portion wise
alternatively to the molten urea. The temperature was raised to

0144-8617/$ – see front matter © 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carbpol.2011.01.010
 D.M. Suflet et al. / Carbohydrate Polymers 84 (2011) 1176–1181
Fig. 1. NMR spectrum of phosphorylated curdlan: (a) 1 H NMR and (b) 31 P NMR decoupled.
145 ◦ C and the reaction was allowed to proceed for 3, 4 or 5 h. After
cooling, the reaction mixture was dissolved in 1 N NaOH aqueous
solution and precipitated in methanol. The derivative was purified
by several reprecipitations and dried under reduced pressure for
48 h. The final product, PCurd in the sodium salt form, is a white
powder soluble in water. The advanced purification of PCurd was
performed by diafiltration in a Millipore cell with PM10 membrane.
The diafiltration was stopped when the filtrate conductivity was
lower then 5 ␮S/cm and the polymer was recovered from aqueous
solution by freeze-drying.
2.2.2. Analysis and characterization of phosphorylated curdlan
Elemental analysis of phosphorous was estimated by EDX anal-
ysis performed on the Quanta 200 (FEI) electron microscope
equipped with EDX system. FTIR spectra were recorded on KBr pel-
let using a Vertex 70 Bruker spectrometer. The NMR spectra have
been recorded on a Bruker Avance III 400 spectrometer, equipped
with a 5 mm multinuclear inverse detection probe, operating at
400.1 and 161.9 MHz for 1 H and 31 P nuclei, respectively. 1 H chemi-
cal shifts are reported in ı units (ppm) relative to the residual peak
of the solvent (ref.: H2 O, 1 H, 4.8 ppm). The molar mass of PCurd was
estimated by size exclusion chromatography using a Waters instru-
ment equipped with two Ultrahydrogel columns (250 and 2000,
7.8 × 300 mm) and a refractive index detector using aqueous 0.1 M
NaNO3 solution with 3% acetonitrile as eluent and poly(acrylic acid)
samples as standard. The decomposition under dynamic conditions
of heating has been investigated with a Paulik–Paulik–Erdey MOM-
Budapest instrument on 50 mg samples, at 12 ◦ C/min, in air.
Polyelectrolyte behaviour was studied by electrochemical meth-
ods and by viscometry. Potentiometric titration was performed
using a Metrohm all purposes 716 DMS Titrino apparatus equipped
with a 6.0203.100 combined electrode. A 712 Metrohm conduc-
tometer with 6.0908.110 cell was used for the conductometric
titrations. The potentiometric and conductometric titrations were
made on the acid form of phosphorylated polysaccharides. For this,
diluted solutions of phosphorylated polysaccharides in Na salt were
passed through a cationic exchange column filled with a sulfonic
Dowex 50 W × 8, 20–50 mesh resin. Viscometric measurements
were performed with an Ubbehlode viscometer with type 0a cap-
1177
illary, at 25 ± 0.02 ◦ C using an AVS 350 Schott automatic viscosity
measuring system.
3. Results and discussion
3.1. Chemical structure
The degree of substitution (DS) of PCurd was evaluated by
potentiometric or conductometric titration according to the meth-
ods already verified (Suflet et al., 2006) and compared with values
obtained by EDX analysis. The value of DS was 1 ± 0.11 regardless
of the reaction time.
In the FTIR spectrum of curdlan the following characteristic
bands are observed: a large band at 3400 cm−1 corresponding to
OH groups; a band at 2915 cm−1 attributed to CH2 groups; char-
acteristic band at 1652 cm−1 owing the associated water; bands
1436–1262 cm−1 for CH2 and C–OH from the glucosidic units; a
large band at 1164–993 cm−1 for C–O–C from the glucosidic units
and a band at 889 cm−1 for ␤-d-glucosidic bonds (Yang & Zhang,
2009). In the FTIR spectrum of PCurd several new bands appeared:
a band at 2410 cm−1 corresponding to the P–H bond, another one at
1216 cm−1 corresponding to the P O bond, a shoulder at 1054 cm−1
attributable to the P–OH bond, and a band at 836 cm−1 correspond-
ing to the P–O–C bonds (Hesse, Meier, & Zeeh, 1984).
The NMR data provide evidence for the formation of phospho-
rylated curdlan. In the 1 H NMR spectrum of PCurd, presented in
Fig. 1a, two peaks at 6.17 and 7.84 ppm are observed which cor-
respond to a doublet with a coupling constant JH–P of 668 Hz. This
value is characteristic for the P–H bond. The presence of the other
two peaks at 7.65 and 5.99 ppm indicate that –H2 PO3 groups occu-
pied not only the C-6 position in glucosidic unit, but also C-2 or C-4
positions. The two sharp peaks observed at 7.60 and 6.03 ppm can
be assigned to small traces of unreacted H3 PO3 .
The 31 P totally decoupled NMR spectrum of PCurd, presented
in Fig. 1b, exhibits a broad and complex signal between 2 and
12 ppm. It can be assigned to the phosphorus atoms from the
–H2 PO3 groups, present in different positions in the glucosidic
units. The sharp signal at 2.6 ppm corresponds to the free H3 PO3
which remained entrapped. These data in association with FTIR
48 H. Pan et al. / Polymer Degradation and Stability 106 (2014) 47e53
phosphorus and nitrogen compounds are more efficient flame re-
tardants owing to the synergistic effect [7e9].
In some reported works, flame retardant modification of various
biodegradable polymers and bio-composites was deeply studied by
the using ammonium polyphosphate (APP) combining with some
bio-based materials (lignin or starch) [10,11]. Casetta et al. studied
the flammability properties of PLA/APP/PER, PLA/APP/starch, and
PLA/APP/LIG. Compared to the PLA/APP/PER composite, materials
containing lignin and starch showed lower LOI values but obtained
V-0 ratings in vertical burning test. Because of their excellent me-
chanical properties, good chemical resistance, easy processability,
biodegradability and water solubility properties, poly(vinyl
alcohol) (PVA) have been widely used in home textiles, coating
materials and industrial materials, etc. [12e14]. However, wide-
spread applications are limited owing to the disadvantage of its
flammability. In our group previous works, phosphorylated chito-
san that contain nitrogen and nickel has been used as flame
retardant additives for PVA composites [15e17]. In this paper, a
novel biodegradable type flame retardant based on phosphorylated
cellulose derivatives was synthesized by the reaction of phos-
phorylated cellulose with ammonium hydroxide at room temper-
ature. The incorporation of PCA could improve effectively the flame
retardancy of PVA matrix. When the PCA content reached 15 wt%,
the PVA/PCA composites could meet the UL-94 V-0 classification
and got the LOI value of 30.0. Thermal and combustion properties of
PVA composites were investigated by TGA and MCC. The mechan-
ical property of PVA/PCA composites was also investigated by
DMTA. All results indicated that the novel flame retardant can
improve the thermal stability and flame retardancy of PVA matrix.
2. Experimental part
2.1. Materials
Poly(vinyl alcohol) (PVA) with the degree of polymerization of
1788  50 and the degree of saponification of 98%, was purchased
from Aldrich Chemical Corp. Microcrystalline cellulose (Avicel
PH101) delivered as a dry powder with 20 mm average particle size,
was used and purchased from ShangHai Gaoli Chemical Company.
Ethanol, phosphorous acid and ammonia solution were received
from Changzheng Chemical Reagent Corp. The water used in this
experiment was distilled.
2.2. Synthesis of PCA
Phosphorylated cellulose (PC) could be synthesized by the re-
action of microcrystalline cellulose with phosphorous acid in
molten urea [18e20]. In a 250 ml three-necked flask equipped with
a nitrogen inlet, a condenser and a stirrer, 12.48 g (0.208 mol) urea
was added, heated at 140
C and flushed with nitrogen atmosphere.
2 g (0.012 mol) microcrystalline cellulose and 10.28 g (0.125 mol)
phosphorous acid were added alternatively portionwise to the
molten urea. The reaction was allowed to proceed at 150
C for 5 h.
After that, the reacted mixture was dissolved in water, then
precipitated with ethanol. The process was repeated several times
until the supernatant become clarified. And then the phosphory-
lated cellulose was dried in vacuum oven at 65
C for 12 h in
presence of phosphorus pentoxide. Phosphorylated cellulose (PC)
was a white and water-soluble powder.
In a 250 ml, three-necked flask equipped with a nitrogen inlet, a
condenser and a stirrer, phosphorylated cellulose was added. And
then 60 ml water was introduced. The mixture was stirred at room
temperature until the solution was obtained. Then 16 g ammonia
water was added to the above solution. The reaction was continued
for 0.5 h. The product was washed with ethanol for three times and
then dried at vacuum oven at 65
C for 24 h in the presence of
phosphorus pentoxide. The final product was a white, water-
soluble powder. The synthetic route is shown in Scheme 1.
2.3. Preparation of PVA/PCA composites
PVA was dissolved in deionized water at 90
C in a three-neck
flask with mechanical stirring. After an aqueous solution (0.08 g/
ml) was formed, PCA was added to the above solution, stirring
slowly for 6 h. At last, the blend was cast into glass plates and dried
at 40
C for 2 days to form flat membranes. Then membranes were
peeled off and further heated at 80
C for another 2 days to remove
residual water. As a result, the thickness of obtained membranes
was 0.5  0.1 mm.
2.4. Characterization
FTIR spectra were recorded on a Nicolet MAGNA-IR 750 FTIR
spectrometer. KBr ground in a mortar with a pestle and enough
solid sample was ground with KBr to make a 1 wt.% mixture for
making KBr pellets. The mixture was pressed into a tablet, which
was then placed in a ventilated oven. The transition mode was used,
and the wavelength range was set from 4000 to 500 cm1.
The X-ray photoelectron spectroscopy measurement was car-
ried out using an ESCALAB MK II (VG Co., Ltd., England) spec-
trometer with Al K a excitation radiation (hn ¼ 1253.6 eV) in
ultrahigh vacuum conditions.
Heat release rate (HRR) and total heat release (THR) were
measured in a microscale combustion calorimeter (GOVMARK
MCC-2). About 5 mg samples were heated at a heating rate of 1 K/s
in a nitrogen stream flowing at 80 cm3/min. The degradation
products of the sample in nitrogen atmosphere were mixed with a
20 cm3/min stream of pure oxygen prior to entering a 900
C
combustion furnace.
The vertical burning test was conducted by a CZF-II horizontal
and vertical burning tester (Jiang Ning Analysis Instrument Com-
pany, China). The specimens used were 130  13  0.5 mm3 ac-
cording to ASTM D3801.
LOI was measured using an HC-2 oxygen index meter (Jiang
Ning Analysis Instrument Company, China) on sheets
100  6.5  0.5 mm3 according to the standard oxygen index test
GB/T 2406-93.
The TGA of samples was examined on a TGA-Q5000 apparatus
(TACompany, USA) from 50 to 700
C at a heating rate of 20
C/min.
The weight of all samples was kept within 3e5 mg in an open Al
pan.
Scheme 1. Synthesis route of PCA.
 M. Hassan et al. / Polymer Degradation and Stability 133 (2016) 8e15
structure of MCHP was confirmed by Fourier transform infrared
spectroscopy (FTIR). MCHP was incorporated into LLDPE at
different ratios. The flammability, combustion behavior, and ther-
mal stability of the prepared composites were addressed based on
the UL-94 test, cone calorimeter test, and thermogravimetric
analysis (TGA). The synergistic effect between OMMT and different
ratios of MCHP on LLDPE was studied. The char residue obtained
from cone test was studied by the scanning electron microscopy-
energy dispersive X-ray spectrometry (SEM-EDXS).
2. Experimental
2.1. Materials
LLDPE was produced by Sabik Company, Saudi Arabia. Melamine
with purity 99%, pentaerythritol with purity 98%, and montmoril-
lonite k10 were purchased from Alfa Aesar Company, Germany.
Phosphoric acid, Methanol HPLC, and chitosan with high molecular
weight and viscosity 800.000 cps were obtained from Sigma
Aldrich Co., Germany. CTAB was supplied by Merck Schuchardt Co.,
Germany. All materials and solvents were used without any further
purification processes.
2.2. Preparation of MCHP
MCHP was prepared by the same procedure which previously
reported [9,12e14], where 59 g of phosphoric acid and 3 g of chi-
tosan were placed in a 250 ml beaker fitted by stirrer. The stirring
was continued for 30 min and the temperature was gradually
increased from room temperature to 150
C. The flame retardant
intermediate, chitosan phosphate (CHP), was obtained with
yellowish color. In three necked flask 63 g of melamine was
dispersed into 500 ml methanol at 120
C. The synthesized CHP was
added into the above flask slowly for about 4 h. Then, the solution
was filtered, washed for three times with methanol and the solid
product was dried at room temperature. The solid product was
grinded to be fine powder. The final product yield was 92% with
pale yellow color. A schematic diagram for the reaction steps is
presented in Fig. 1 and the FTIR spectra of MCHP is shown in Fig. 2.
Fig. 1. A schematic representation for synthesis of MCHP.
9
2.3. Preparation of OMMT
In 500 ml beaker, 50 g of Na-MMT was swelled in methanol for
24 h. Then 20 g of CTAB was added to the mixture to swell for
another 12 h together at the room temperature [15]. The solid
residue was washed three times with methanol and was dried
under an infrared lamp. The FTIR spectra of OMMT is shown in
Fig. 3.
2.4. Preparation of flame retarded LLDPE composites
LLDPE composites were prepared by using twin screw extruder
(manufactured by Newplast Company, India) with three heating
zones at 170
C. The twin screw speed was maintained at 25 rpm.
IFRs with desired amounts were first mixed with LLDPE pellets and
then added to the extruder. The formulations of the prepared
samples are presented in Table 1. The polymer composite samples
were pressed under 15 MPa for 10 min at 160
C into sheets with
suitable thickness and size for UL-94 and cone calorimeter tests.
2.5. Characterization
FTIR analysis was performed on a Nicolet 380 spectrophotom-
eter. Each sample was mixed with KBr powder, and then pressed
into a tablet. The measurement was carried out in the optical range
of 4000e400 cm1. 1H NMR spectra were recorded on a varian
mercury VX-300 NMR spectrometer which run at 300 MHz in
dimethylsulphoxide (DMSO-d6). Chemical shifts were quoted in (d)
and were related to that of the solvents. The XRD experiments were
performed on Empyrean diffractometer from Panalytical Co.
(Netherland) using Cu Ka radiation corresponding to a wavelength
of 0.1542 nm. The scanning speed of the detector was 0.055
/s.
Thermogravimetric analysis (TGA) was conducted on a Shimadzu
DTG-60 with a linear heating rate 10
C/min. The temperature
range was starting from room temperature to 750
C under nitro-
gen atmosphere with flow rate 300 ml/min. Each sample was
placed in a platinum crucible and the samples weights were about
7e7.5 mg. The vertical burning test was carried out by UL-94 flame
chamber manufactured by Stanton Redcroft Ltd, UK, according to
 Chemical modification of chitosan by phosphorylation 1579

Acetylation of chitosan

Chitosan membranes were acetylated at room temperature [22, 23]. The membranes
were steeped in an aqueous methanolic acetic acid solution (water/methanol/acetic
acid = 50 : 200 : 1, v/v) and acetic anhydride (5.0 mol per glucosamine residue) was
added. After 12 h under stirring the membranes were rinsed in methanol, followed
by rinsing in ethanol, and then steeped in 0.5 M ethanolic KOH overnight to remove
any ester groups. The membranes were finally washed with ethanol, followed by
DDW, and then dried at 30◦ C for 24 h in a vacuum oven. The corresponding degree
of N -acetylation was estimated according to Shigemasa et al. [24], using the amide-
II band (ν = 1560 cm−1 ) as the analytical band and the C O stretching band
(ν = 1070 cm−1 ) as the internal reference band. In ATR–FT-IR spectra, the internal
reference band at 3450 cm−1 , previously used for determining the DA of chitosan
powder, presents low absorbance values when compared to transmission spectra.

Phosphorylation of chitosan

Surface phosphorylation was performed using the H3 PO4 /Et3 PO4 /P2 O5 /hexanol
method [15], substituting hexanol by 2-butanol, which has a higher water solubility.
Briefly, 1 g chitosan membrane strips was suspended in 40 ml 2-butanol in a round-
bottomed flask. The reaction mixture was prepared by adding 43 ml H3 PO4 to
37 ml Et3 PO4, followed by step-by-step addition of P2 O5 under constant stirring.
The resulting translucent solution was then gradually added to the flask. The
reaction was allowed to proceed under N2 atmosphere at 30◦ C, with constant stirring
in a thermostatised orbital shaker. Phosphorylation was carried out for different
time periods, up to 48 h. After chemical treatment, membranes were rinsed with
ethanol and suspended twice in this solvent for 30 min. Finally, they were dialyzed
against DDW for 24 h to remove free inorganic phosphate, and dried at 30◦ C for
24 h in a vacuum oven. Acetylated chitosan membranes, in which amines are
absent, were also phosphorylated for a period of 8 h. Alternatively, acetylated
and original chitosan membranes which were phosphorylated for 8 h were both
subjected to neutralization after the ethanol rinsing step. The neutralization step
was used in order to convert chitosan ammonium groups into uncharged amine
groups, with consequent release of ionically bound phosphates. The strips were
incubated in 0.5 M aqueous NaOH at 25◦ C for 30 min under stirring, thoroughly
washed with DDW until neutral pH, and finally dialyzed and dried as previously
described.

Surface characterization

X-ray photoelectron spectroscopy (XPS). Spectra were obtained on a VG Sci-

entific ESCALAB 200 A, using Mg Kα X-ray radiation as the excitation source
(15 kV), operated at 300 W. The analyzer was run in the constant analyzer trans-
 Polymer-Plastics Technology and Engineering, 52: 393–399, 2013
Copyright # Taylor & Francis Group, LLC
ISSN: 0360-2559 print=1525-6111 online
DOI: 10.1080/03602559.2012.754465
Preparation and Characterization of Chitosan-Based
Flame Retardant and Its Thermal and Combustible
Behavior on Polyvinyl Alcohol
Shuang Hu, Lei Song, and Yuan Hu
State Key Laboratory of Fire Science, University of Science and Technology of China, Hefei,
Anhui, PR China
Downloaded by [Universidad Autonoma de Barcelona] at 03:17 27 October 2014
A novel natural material-based flame retardant melamine chitosan
phosphate (MPCS) has been prepared. Its impact on thermal proper-
ties and flammability performance of polyvinyl alcohol (PVA) was
analyzed. Thermogravimetric analysis data exhibited that MPCS
enhanced the thermal stability of materials at high temperature.
The results of thermogravimetric analysis/infrared spectrometry
(TGA-IR) showed that MPCS decreased the yield of flammable vola-
tilized products greatly. Microscale combustion calorimetry (MCC)
data showed that peak heat release rate (PHRR), total heat release
(THR) and heat release capacity (HRC) all reduced, which indicated
that MPCS improved the flame retardancy of materials.
Keywords Combustion property; Flame retardant; Chitosan;
Thermal stability
INTRODUCTION
Polymer materials are used in ever more areas and under
ever more demanding environmental conditions. However,
fire hazard associated with the application of these poly-
meric materials lead to the loss of life and property, which
are of particular concern[1,2]. The intumescent flame retard-
ant system is composed of acid source, char forming source
and blowing source is a focus in recent research owing to its
efficiency and halogen-free characteristic[3,4]. The effective-
ness of the intumescent flame retardants is due to the
foamed char formed on the surface of the burning material,
which acts as a heat insulator, limiting the heat transfer
from the heat source to the polymer and the mass transfer
from the polymer to the flame resulting in the improvement
of fire retardant performances. A number of intumescent
flame retardants have been prepared, but most char forming
source used today are petroleum-based materials[5]. Because
of the crisis of petroleum and the deterioration of ecological
environment, it is necessary to search for the replacement of
petroleum-based materials.
Address correspondence to Yuan Hu, State Key Laboratory of
Fire Science, University of Science and Technology of China, 96
Jinzhai Road, Hefei, Anhui 230026, PR China. E-mail: yuanhu@
ustc.edu.cn
393
Function materials prepared from renewable nature
materials is an effective method to work out a series environ-
ment issues and depletion of conventional petroleum-based
resources[6–8]. Some work has been done on searching for
environmental-friendly and sustainable char former agent,
such as starch[9–13]. But in these researches, the nature mate-
rials were just applied as char former agent additives alone.
It is well known that flame retardant combining with an acid
source, a char forming source and a blowing source in one
molecule shows higher efficiency on fire resistance[14,15].
Hence, it is useful to prepare a flame retardant that contains
three components.
Chitosan, the fully or partially deacetylated form of
chitin, the principal component of living organisms such
as fungi and crustaceans, one of the most abundant bioma-
terials in the world, is considered as one of the most studied
materials in recent years due to the renewable property,
non-toxicity, high biocompatibility and so on[16,17]. It
should be noticed that chitosan possesses the structure of
multi-hydroxyl groups just like pentaerythrite and cyclo-
dextrins, which promote the formation of char during
burning[18,19]. The charring ability and thermal stability
enabled chitosan to be used as an effective char forming
source in intumescent flame-retardant system.
Herein, we have prepared a novel chitosan-based flame
retardant (MPCS) containing acid source, char forming
source and blowing source. MCC test was used to investi-
gate the flammability. Its effect on thermal properties was
evaluated through TGA, RT-IR and TGA-IR.
EXPERIMENT
Materials
Chitosan (viscosity 50–800 mPa.s, degree of deacetylation
80–95%), methanesulphonic acid, phosphorus pentoxide
and melamine were purchased from Sinopharm Chemical
Reagent Co., Ltd (China).
Synthesis of Melamine Chitosan Phosphate (MPCS)
The reaction between chitosan (CS, 2.0 g), and phos-
phorus pentoxide (10 g) occurred in methanesulphonic acid
 394
(30 ml). The mixture was stirred at 0–5
C for 3 h under
nitrogen. The pre-product (PCS) was precipitated with
ether and was washed with ether, acetone, methanol and
ether sequentially[20]. Then, the reaction between PCS
and melamine happened in glycol at 100
C for 4.5 h. The
product was washed with hot glycol several times to
remove the excessive melamine. At last, the yellowy powder
MPCS was obtained after it was dried at 80
C. Scheme 1
showed the reaction.
Preparation of MPCS=PVA Blended Membranes
MPCS mixed with PVA solution at room temperature
for 1 h. Then, it dried at 80
C for 2 h, 60
C for 10 h and
80
C for 2 h to get blended membrane.
Downloaded by [Universidad Autonoma de Barcelona] at 03:17 27 October 2014
Characterization
FTIR spectroscopy (Nicolet 6700, Nicolet Instrument
Company, USA) was employed to characterize samples
using KBr as the sample holder. 31P NMR analysis was per-
formed with Bruker AVANCE 400 WB spectrometer. The
X-ray photoelectron spectroscopy (XPS) spectrum was
recorded with a VG Escalab mark II spectrometer (VG
Scientific Ltd., UK), using Al KR excitation radiation
(1253.6 eV) to test the content of phosphorus and nitrogen.
Thermogravimetric analysis was examined on a TGAQ5000
apparatus (TA Co., USA) at the heating rate of 20
C=min
in air. Real time FTIR (RT-IR) method was used Nicolet
MAGNA-IR 750 spectrophotometer (Nicolet Instrument
Company, USA) to study the thermo-oxidative degradation
of the cured film. Powders of the cured sample were mixed
with KBr powders, and the mixture was pressed into a tab-
let, which was then placed into the oven.
The temperature of the oven was raised at a heating
rate of about 10
C=min. MCC test was carried out using
SCH. 1. Preparation of MPCS. (Color figure available online.)
S. HU ET AL.
a Govmak MCC-2 microscale combustion calorimeter;
samples were heated to 650
C at a heating rate of 1
C=s
in a stream of nitrogen flowing at 80 cm3=min. The volatile
anaerobic thermal degradation products in the nitrogen gas
stream were mixed with a 20 cm3=min stream of pure
oxygen prior to entering a 900
C combustion furnace.
Scanning electron microscopy (SEM) studies were per-
formed on the char residue using a Hitachi X650 scanning
electron microscope (Japan).
RESULTS AND DISCUSSION
Structural Characterization
Figure 1a showed the FTIR spectra of CS, PCS, and
MPS. Compared with CS, it can be observed from the curve
of PCS that there was a peak at 1200 cm1, which can be
attributed to the P¼O asymmetric stretching. Meanwhile,
the appearance of peaks at 1050 and 484 cm1 can be attrib-
uted to the P-O structure, such as P-O-C and P-OH group.
The absorption at 1600 cm1 was replaced by a new absorp-
tion at 1634 cm1, meaning that amino groups in chitosan
could change into ammonium groups in methanesulfonic
acid medium, yet some amino groups underwent the reac-
tion to form phosphorylamide groups[21].
After the reaction between PCS and melamine, the new
absorption peaks at 3360 and 3160 cm1 can be ascribed to
the symmetrical and asymmetrical stretching vibrations of
N–H bond; meanwhile the bending vibration of N–H
appears at 1693 cm1. Three peaks at 1510 cm1, 1390 cm1
and 1330 cm1 belonged to the stretching vibration of the
triazine ring. Compared with PCS, it should be noticed that
the P–OH group of MPCS shifted from 484 cm1 and
1050 cm1 to 509 cm1 and 1080 cm1 respectively, indicat-
ing that melamine has been grafted on PCS.
Figure 1b exhibited the 31P NMR spectrum of PCS. A
strong peak can be observed around 0 ppm. According
to the space hindrance and migration situation of
13
C NMR[22], this peak indicated that phosphorylation
occurred on C-6 hydroxyl groups of chitosan. The appear-
ance of a small peak at 1 ppm meant that part hydroxyl
groups on C-3 of chitosan took part in the reaction.
FIG. 1. Characterization of product (a: FTIR analysis, b: P NMR of
PCS).
ARTICLE WILEYONLINELIBRARY.COM/APP

Scheme 1. Synthetic route of HUMCS. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

accelerating agent to expand the solubility window of CS tioned solution was fed into a three-necked flask equipped with
because urea can also be used as a blowing agent in flame- a stirrer, a reflux condenser, and a glass stopper. Third, a solu-
retardant compounds. tion of HEDP (12.36 g, 0.06 mol) in distilled water (100 mL)
was added slowly to the three-necked flask, and the reaction
In this study, novel chitosan/urea compound based phosphonic
temperature was simultaneously increased to 120 C. The reac-
acid melamine salt (HUMCS) was prepared in aqueous solution
tion was carried out at 120 C for 2 h. Then, the reaction system
via 1-hydroxy ethylidene-1,1-diphosphonic acid (HEDP), urea,
temperature was set at 75 C, and the reaction was continued.
MA, and CS. Then, it was added to PP in addition to an IFR
When the system temperature was kept at 75 C, a solution of
system. The synergistic effect between HUMCS and IFR was
MA (7.56 g 0.06 mol), which was dissolved in hot water (200
studied. The tensile properties of the PP/IFR/HUMCS compo-
mL), was injected into the three-necked flask drop by drop
sites and the morphology of the residual char were also investi-
within 0.5 h. After further heating at reflux for 3 h, the reaction
gated. Through this investigation, we expected to make use of
was completed. After direct hot filtration, washing with warm
the CS/urea solution to prepare an efficient CS-based flame-
water, and drying at 90 C, the novel flame-retardant synergist,
retardant synergist in an aqueous solution.
HUMCS, was obtained as a light yellow powder with a 93.6 wt %
EXPERIMENTAL yield. Scheme 1 shows the synthetic route.
Materials
Preparation of the Flame-Retardant PP Samples
Commercial PP (TS30) was obtained from Fujian Petroleum
The IFR consisted of MPP and PER with a weight ratio of 2:1,
Chemical Co., Ltd. PER and MA were purchased from Fuchen
and the total dosage of flame retardant was kept at 25 wt % in
Chemical Reagent, (Tianjin, China). HEDP was provided by
the IFR–PP composites. Then, HUMCS was added systemati-
Shandong Taihe Water Treatment Co., Ltd. (Shandong China).
cally to the IFR–PP composites as a synergist from 1 to 5 wt %.
Urea was supplied by Zhiyuan Chemical Reagent (Tianjin,
The detailed formulations of the samples are listed in Table I.
China). CS was purchased from Aoxing Bio-Science Technology
All of the materials were dried in a vacuum oven at 80 C for 12
Co., Ltd. (Zhejiang, China). Melamine pyrophosphate (MPP)
h before use. Then PP, MPP, PER, and HUMCS were melt-
was prepared in our laboratory.19 All of these commercial mate-
mixed in a twin-roll mill (XK-160, Shanghai Rubber Machinery
rials were used directly without further purification.
Factory, China) for 15 min. The temperature of the mill was
Synthesis of HUMCS maintained at 175 C, and the roller speed was 40 rpm. The
First, CS (9.66 g) and urea (3.6 g, 0.06 mol) were dissolved in extruded strands were cut into pellets. Then, the composites
300 mL of deionized water, and the solution was placed in an were injected into standard testing bars with an injection-
icebox at 0 C for 24 h. Second, after unfreezing, the aforemen- molding machine (PL860 Haitian Plastic Machinery Co., Ltd.,

WWW.MATERIALSVIEWS.COM 40845 (2 of 8) J. APPL. POLYM. SCI. 2014, DOI: 10.1002/APP.40845

110 S. Hu et al. / Journal of Analytical and Applied Pyrolysis 97 (2012) 109–115
In this paper, we have prepared a novel chitosan based flame
retardant material (GPCS) containing phosphorus and acrylate
structure. Herein, phosphorus knowing as one of the most effective
flame retardant element was used as acid agent. Acrylate struc-
ture was introduced to graft on the backbone of polymer matrix to
enhance its compatibility. SEM was applied to reveal the dispersion
performance of GPCS in matrix. The characteristic of flammability
was presented by MCC. Thermal and mechanical properties were
showed by TGA, RT-IR, TGA-IR and DMA.
2. Experiment
2.1. Materials
Chitosan (viscosity 50–800 mPa s, degree of deacetylation
80–95%), phosphorus pentoxide (AR), methanesulfonic acid (99%)
were provided by Sinopharm Chemical Reagent Co., Ltd. Glycidyl
methacrylate (GMA), supplied from Dong-fang Chemical Co., Bei-
jing, China, was distilled before used.
2.2. Synthesis of GPCS
GPCS can be prepared as the following process. Firstly, phos-
phorylated chitosan (PCS) can be synthesized by the reaction of
chitosan (CS, 2 g) with phosphorous pentoxide (10 g) in methane-
sulfonic acid in ice water bath under nitrogen for 3 h. The residue
was washed with ether, acetone, methanol and ether, and then
dried to obtain PCS [36–38]. Then, PCS (2 g) was dispersed in 1,4-
Dioxane for 15 min before glycidyl methacrylate (20 g) was added.
The reaction occurred at 60 ◦ C for 12 h and 100 ◦ C for 3 h. After fil-
tration, the product was washed by 1,4-Dioxane several times to
remove the extra GMA. Finally, the white powder can be obtained,
after it was dried. Scheme 1 exhibited the reaction scheme.
2.3. Preparation of samples
The ultraviolet light source used for irradiation is a lamp
(80 W/cm; Lantian Co., Beijing, China) that emits light in the near UV
(characterized wavelength, 340–360 nm). The UV irradiation was
carried out in air atmosphere. The polymerization mixture was pre-
pared by adding 3 wt% photoinitiator (Darocur 1173, Ciba-Geigy,
Switzerland) into the GPCS/EA system.
2.4. Characterization
FTIR spectroscopy (Nicolet 6700, Nicolet Instrument Company,
USA) was employed to characterize samples using thin KBr as the
sample holder. Solid-state 31 P NMR was performed with Bruker
AVANCE 400 WB spectrometer. Atomic absorption spectrometry
(AAnalyst 800, Perkin Elmer) was used to test the content of phos-
phorus.
Scanning electron microscopy (SEM) (JSM—6700F, JEOL, Japan)
was used to study morphological features of the fracture face
(treated in liquid nitrogen) of GPCS/EA system, which was used
to evaluate the compatibility.
MCC test was carried out using a Govmak MCC-2 microscale
combustion calorimeter; samples were heated to 650 ◦ C at a heat-
ing rate of 1 ◦ C/s in a stream of nitrogen flowing at 80 cm3 /min. The
volatile anaerobic thermal degradation products in the nitrogen gas
stream were mixed with a 20 cm3 /min stream of pure oxygen prior
to entering a 900 ◦ C combustion furnace.
Thermogravimetric analysis (TGA) was carried out using a
Q5000IR thermoanalyzer instrument from 50 to 700 ◦ C at a lin-
ear heating rate of 20 ◦ C/min under an air flow of 60 cm3 /min.
Real time Fourier transform infrared (RT-IR) method was used
Fig. 1. FTIR spectra of CS, PCS and GPCS.
to study the thermo-oxidative degradation of the samples. Pow-
ders of the cured sample were mixed with KBr powders, and
the mixture was pressed into a tablet, which was then placed
into the oven. The temperature of the oven was raised at a heat-
ing rate of about 10 ◦ C/min. Thermogravimetric analysis/infrared
spectrometry (TGA-IR) was performed to analyze the volatilized
products after the pyrolysis of samples under a nitrogen flow of
35.0 cm3 /min. Thermomechanical properties of the sheets were
tested with a dynamic mechanical analyzer (DMA Q800, TA Instru-
ments, USA) using a double cantilever measuring mode. The sample
size was 50mm × 8mm × 3 mm. Tests were performed from room
temperature to 160 ◦ C at a heating rate of 5 ◦ C/min and an oscilla-
tion frequency of 10 Hz.
3. Results and discussion
3.1. Characterizations of GPCS
FTIR spectra of CS, PCS and GPCS were showed in Fig. 1. For CS,
characteristic peak at 1600 cm−1 was ascribed to N-H group. The
broad peak observed at 3500 cm−1 has the contribution of the OH
group. The other peaks at 2890 and 1380 cm−1 were assigned to
CH stretching and CH3 symmetric deformation, respectively. Phos-
phorylation led to an emerging shoulder at 1200 cm−1 , which can
be attributed to the P O asymmetric stretching. New peaks at 1050
and 484 cm−1 can be attributed to the P O structure, such as P O C
and P OH. The above phenomenon suggested phosphorylation of
hydroxyl groups on CS. The absorption at 1600 cm−1 was replaced
by a new absorption at 1634 cm−1 . Wang reported that lots of
amino groups in chitosan could change into ammonium groups in
methanesulfonic acid medium while some amino groups under-
went the reaction to form phosphorylamide groups [39]. Then, the
reaction happened with the addition of glycidyl methacrylate. It
can be observed from the FTIR curve of GPCS that a new peak at
1720 cm−1 was found, which was attributed to C O group of gly-
cidyl methacrylate. Moreover, the peak assigning to P O structure
shifted to higher wavenumber, increasing from 1050 and 484 cm−1
to 1070 and 513 cm−1 , respectively. Meanwhile, P O band moved
from 1200 to 1250 cm−1 . The above results indicated that glycidyl
methacrylate group has been grafted on the PCS.
Fig. 2(a) showed the solid state 31 P NMR spectrum of GPCS.
According to the space hindrance and migration situations [40], a
strong peak around −1 ppm meant that phosphorylation occurred
on C-6 hydroxyl groups of chitosan mainly. Besides, another small
peak appeared around −11 ppm implied that part hydroxyl groups
860 S. Hu et al.

Experimental sample holder. 31P NMR was conducted on Avance 300

Materials
Chitosan (viscosity 50–800 mPa s, degree of deacetylation
80–95 %), phosphorus pentoxide, methanesulfonic acid,
and urea were all provided by Sinopharm Chemical
Reagent Co., Ltd.
Synthesis of urea salt of chitosan phosphate (UPCS)
Chitosan (CS, 2.0 g), methanesulfonic acid (30 mL), and
phosphorus pentoxide (10 g) were put in a round bottom
flask. The mixture was stirred at 0–5 °C for 3 h with the
protection of nitrogen. Then, the pre-product (PCS) was
precipitated with ether and was washed with ether, acetone,
methanol, and ether sequentially, before it was dried [9, 10].
After that, the reaction between the pre-product and urea
happened in ethanol at 70 °C for 12 h. Then, the product was
washed with ethanol several times to remove the excessive
urea and the white powder UPCS was obtained after it was
dried. The reaction is shown in Scheme 1.
Preparation of UPCS/PVA blended membranes
spectrometer (Bruker Biospin, Switzerland, frequency) at
room temperature using CDCl3 as the solvent. Solid-state
13
C NMR analysis was performed with Bruker AVANCE
400 WB spectrometer. The XPS spectrum of UPCS was
recorded with a VG Escalab mark II spectrometer (VG Sci-
entific Ltd., UK), using Al KR excitation radiation
(1253.6 eV) to test the content of phosphorus and nitrogen.
MCC test was carried out using a Govmak MCC-2 micro-
scale combustion calorimeter; samples were heated to
650 °C at a heating rate of 1 °C s-1 in a stream of nitrogen
flowing at 80 cm3 min-1. The volatile anaerobic thermal
degradation products in the nitrogen gas stream were mixed
with a 20 cm3 min-1 stream of pure oxygen prior to entering
a 900 °C combustion furnace. Thermogravimetric analysis
was examined on a TGAQ5000 apparatus (TA Co., USA) at
the heating rate of 20 °C min-1 in air. Real time Fourier
transform infrared method was used to study the thermo-
oxidative degradation of the cured film. Powders of the cured
sample were mixed with KBr powders, and the mixture was
pressed into a tablet, which was then placed into the oven.
The temperature of the oven was raised at a heating rate of
about 10 °C min-1.

The blended membranes were prepared by mixing the
UPCS and PVA solution at room temperature for 1 h.
Then, the mixture was dried at 80 °C for 2 h, 60 °C for
10 h, and 80 °C for 2 h.
Characterization
FTIR spectroscopy (EQUINOX55, Bruker Co., Germany)
was employed to characterize samples using thin KBr as the
Results and discussion
Structural characterization
Figure 1a shows the FTIR spectra. Compared with CS, a
broad peak at 3,500 cm-1 in PCS weakened. The peaks
found at 1,050 and 484 cm-1 were ascribed to the P–OH
group. It implied that the reaction happened between the
–OH group and phosphorous pentoxide. The absorption at

Scheme 1 Preparation of UPCS O

OH
OH P
O OH
C6
O C5 O
O P2O5
O O
O C4
OH
NH2 O C2 C1
O C3
P NHR
OH
OH
CS PCS

CS CO(NH2)2
O

O
O H3N NH2

UPCS
R = -SO3H, -PO(OH)2

123