Cellulose (2007) 14:543–552
DOI 10.1007/s10570-007-9122-3
PEGylation of chitosan for improved solubility and fiber
formation via electrospinning
Jian Du Æ You-Lo Hsieh
Received: 18 January 2007 / Accepted: 6 April 2007 / Published online: 10 June 2007

Springer Science+Business Media B.V. 2007
Abstract PEG-N-chitosan and PEG-N,O-chitosan
were synthesized via reductive amination and acyl-
ation of chitosan, respectively. The structures were
confirmed by FTIR and H1NMR. The extents of
PEGylation increased with reducing chain lengths of
either chitosan (Mv = 137–400 kDa) or poly(ethye-
lene glycol) (PEG, Mn = 500–2 kDa). Water solubil-
ity were easily achieved at degree of substitution
(DS) as low as 0.2 for either derivtive whereas the
PEG-N,O-chitosan at DS = 1.5 was soluble in
organic solvents, including CHCl3, DMF, DMSO and
THF. None of the aqueous solutions of PEG-N-
chitosan or PEG-N,O-chitosan alone could be elec-
trospun into fibers. Electrospinning of PEG550-N,O-
chitosan145 at 25% in DMF produced fibrous struc-
ture intermixed with beads. The efficiency of fiber
formation and the uniformity of fibers were improved
by increasing the solution viscosity using a cosolvent
or reducing the solution surface tensions with a non-
ionic surfactant. Ultra-fine fibers with diameters
ranging from 40 nm to 360 nm and an average
diameter of 162 nm were efficiently generated from
electrospinning of 15% PEG550-N,O-chitosan145 in
75/25 (v/v) THF/DMF cosolvents with 0.5% Triton
X-100TM.
J. Du
Y.-L. Hsieh (&)
Fiber and Polymer Science, University of California,
Davis, CA 95616, USA
e-mail: ylhsieh@ucdavis.edu
Keywords Chitosan
Acylation
Amination

Poly(ethylene glycol) graft
Water soluble: Organic
soluble
Electrospinning from organic solutions
Introduction
Chitosan, a partial N-deacetylated derivative of chitin,
is a copolymer of N-acetyl-D-glucosamine and D-
glucosamine. The molecular weights and degree of
deacetylation (DDA) of chitosan are typically over
1,000 kDa and 70%, respectively, and vary greatly
with the chitin source and the method of hydrolysis.
The inherent biological and physicochemical charac-
teristics of chitosan are unique among polymers and,
couple with its abundance, have attracted increasingly
attention for a wide range of biological and techno-
logical applications (Jayakumar et al. 2005).
One major challenges in utilizing chitosan as a
material feedstock is its limited processibility. Its
rigid D-glucosamine structure, ability to hydrogen
bond intermolecularly and high crystallinity contrib-
ute to its insolubility in common organic solvents and
non-thermoplastic behavior. Reducing the molecular
weight and lowering crystallinity of chitosan by
random deacetylation generally improve its solubility
in dilute acids and enable its solution processing into
various forms of beads (Guo et al. 2005), membranes
(Devi et al. 2006), and fibers (Binias et al. 2005).
While many challenges remain regarding the
dissolution and processing of chitosan in general,
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544
few have ventured into making much smaller chito-
san fibers by electrospinning where a high voltage is
applied to a capillary droplet of polymer solutions or
melts to overcome the liquid surface tension.
To date, electrospinning of chitosan, either alone
or with another polymer, has only been possible from
acid solutions of trifluoroacetic acid (Ohkawa et al.
2004), formic acid (Park et al. 2004), and acetic acid
(Spasova et al. 2004; Duan et al. 2004; Bhattarai et al.
2005a; Geng et al. 2005; Li and Hsieh 2006).
Electrospinning of chitosan alone has only been
reported on relatively low molecular weight
(Mv = 210 kDa, 78% DDA) from trifluoroacetic acid
(Ohkawa et al. 2004) and (Mv = 106 kDa, 54% DDA)
from 90% acetic acid (Geng et al. 2005). Others have
generated fibers by electronspinning chitosan with
another polymer from acid solutions (Park et al.
2004; Spasova et al. 2004; Duan et al. 2004; Bhattarai
et al. 2005a; Geng et al. 2005; Li and Hsieh 2006).
These studies dealt mostly with relatively low
molecular weight chitosan that was often a minor
component in the mixture. A 3/7 chitosan
(Mv = 220 kDa, 86% DD)/silk fibroin mixture in
formic acid was electrospun to 130 nm diameter
fibers (Park et al. 2004). Electrospinning of 45 wt%
chitosan (Mv = 210 kDa, 78% DD) mixed with
polyvinyl alcoho (PVA) (Mn = 88 kDa) in 0.2 M
acetic acid produced 120 nm diameter fibers (Ohkawa
et al. 2004). Fibers with 40–290 nm diameters have
been electropsun from mixtures of up to 90%
chitosan (Mw = 190 kDa, 85% DD) and poly(ethylene
oxide) (PEO) (Mw from 800 kDa to 900 kDa) in
acetic acid (Spasova et al. 2004; Duan et al. 2004;
Bhattarai et al. 2005). We have generated much finer
20 nm diameter fibers from chitosan (82.5% DDA) at
Mv = 1,600 kDa, the highest molecular weight
reported, by mixing with 50% or more PVA
(Mw = 186 kDa) (Li and Hsieh 2006). Upon
dissolution of PVA from these bicomponent fibers,
nanoporous chitosan fibers were created.
To improve solubility of chitosan in a broader
range of solvents, chemical modification through
reactions of the amine and/or hydroxyl groups should
be promising as demonstrated by cellulose deriva-
tives. Poly(ethylene glycol) (PEG) is an ideal graft-
forming polymer because it is soluble in both water
and organic solvents and its low toxicity, good
biocompatibility and biodegradability (Harris 1992);
and approved use in food, cosmetics, personal care
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Cellulose (2007) 14:543–552
products and pharmaceutical (Tirelli et al. 2002).
Indeed, PEGylation of chitosan has been shown to
improve the affinity to water or organic solvents,
most commonly by reductive amination of chitosan
with PEG-aldehyde in aqueous organic acid (Sugim-
oto et al. 1998; Gorochovceva et al. 2005; Bhattarai
et al. 2005b). Grafting PEG onto very short chitosan
(Mn = 28 kDa) produced water-soluble PEGylated
chitosan (Sugimoto et al. 1998), at a very low DS of
0.05. PEGylation of chitosan (Mn = 150 kDa) with
PEG (Mn = 2 kDa) by carbodimide hydroxybenzo-
triazole coupling with the 6-O-triphenylmethyl-chito-
san derivative also yielded water-soluble products at
a low DS of 0.1 (Ouchi et al. 1998). Etherification of
N-phthaloyl chitosan (Mn = 400 kDa) with PEG
monomethyl ether iodide (Mn = 2 kDa) produced
water-soluble derivative with DS  0.15 and
organic-soluble (acetone, ethanol, chloroform, DMF
and toluene at 40–508C) at 1.97 DS (Gorochovceva
and Makuška 2004). These PEGylation works either
produced very low substitutions or, in order to reach
high DS, involved complex, multiple-step reactions
to protect or generate intermediates, and/or catalysts
that are difficult to remove.
This study aims to improve solubility of chitosan
by molecular weight control and PEG modification
and to generate ultra-fine fibers by electrospinning of
PEGylated chitosan in non-acidic solutions. The
focus is to understand how the chain length of
chitosan and PEG as well as the degree of side chain
modification affect the solubility of the PEGylated
chitosan. Reductive amination and acylation of
chitosan are targeted for N-PEGylation and N,O-
PEGylation of chitosan, respectively. Furthermore,
fiber formation of various PEGylated chitosan via
electrospinning from aqueous and organic solutions is
investigated.
Experimental
Materials
The starting chitosan (Aldrich) had a viscosity
molecular weight (Mv) of 405 kDa and 84.7% DDA
(CS400). The molecular weight of CS400 was
reduced by alkaline and acid hydrolysis. Alkaline
hydrolysis (40% NaOH, ambient temperature, 4d) of
CS400 reduced Mv to 190 kDa and raised DDA to
Cellulose (2007) 14:543–552 545

92.1% (CS190). Further alkaline hydrolysis of CS190
at 608C for 8 h yielded CS137 (Mv = 137 kDa, DDA
= 98.4%). CS145 (Mv = 145 kDa, DDA = 89.3%)
was obtained by hydrolysis of CS400 in 2% acetic
acid with 2% hydrogen peroxide at 608C for 30 min.
The molecular weight was derived from the intrinsic
viscosity [g] measured in a 0.1 M acetic acid-0.2 M
NaCl mixture using a Cannon–Fenske viscometer at
38 ± 0.58C. The Mv was calculated by the Mark–
Houwink equation for chitosan (Roberts and Domszy
1982): ½g ¼ KMva , where K and a are 1.81 · 103 and
0.93, respectively. Poly(ethylene glycol) (PEG) at Mn
of 550, 2000, and 5000 Da was purchased from
Aldrich. Anhydrous dimethylsulfoxide (DMSO), tol-
uene, and pyridine were provided by Fisher. All other
reagents and chemicals were of analytical grade or
higher and used without further purification.
was mixed in a 2% acetic acid and methanol (2/1 v/v)
solution (pH 6), while aqueous cyanoborohydride
(NaCNBH3) (10% v/v) was added dropwise to a 1.5/1
NaCNBH3/PEG-aldehyde molar ratio. The resultant
mixture was dialyzed with a dialysis membrane (MW
12,000–14,000 cut) against distilled water and
0.05 M aqueous NaOH solution, then concentrated.
N-PEGylated chitosan was finally precipitated as
white powder by adding excess acetone.
Synthesis of PEG-N,O-chitosan by acylation of
chitosan (Scheme 2)
The terminal hydroxyl of PEG was converted to
carboxyl group by adding a pyridine solution of
O
O

Synthesis of PEG-N-Chitosan by reductive CH3O-(CH2CH2O)n-OH

O

amination of chitosan (Scheme 1) Pyridine/Toluene

SOCl2 , toluene
CH3O-(CH2CH2O)n-COOH CH3O-(CH2CH2O)n-COCl
PEG-aldehyde was prepared by oxidizing PEG with Refluxing
10-fold molar excess of acetic anhydride in anhy- OH
OOCR
drous DMSO/chloroform (90/10 v/v) under nitrogen
O
and constant stirring at 228C for 9 h, following * RCOCl
O

O
precipitation with excess diethyl ether. The precipi- Pyridine
O
NH2
tate was separated, reprecipitated twice with diethyl OH
n OOCR
NHCOR
n
ether and vacuum dried to white PEG-aldehyde
powder. Equal molar of PEG-aldehyde and chitosan Scheme 2 Synthesis route of PEG-N,O-Chitosan

OH
O
CH3O(CH2CH2O)mCH2CH2OH
*
O
Ac2O DMSO,CHCl3
HO NH2
n

CH3O(CH2CH2O)mCH2CHO
AcOH/MeOH
pH5-6
OH OH
O O
* NaBH3CN *
O O
HO AcOH/MeOH, pH6 HO
N NH
CH3O(CH2CH2O)mCH2CH n CH3O(CH2CH2O)mCH2CH2
n

Scheme 1 Synthesis route of PEG-N-Chitosan

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546
succinic anhydride (4-fold molar excess) dropwised
into 10% PEG toluene solution under anhydrous
conditions and refluxed for 6 h. The product was
precipitated with ether addition, filtered, reprecipitat-
ed twice from chloroform solution with diethyl ether,
then vacuum dried. The carboxylate-terminated PEG
was combined with 2-fold molar excess of thionyl
chloride under nitrogen atmosphere to obtain PEG-
acyl chloride. The reaction was kept refluxing for 6 h
followed by degassing under vacuum to remove
residual SO2 gas and excess thionyl chloride. Chito-
san was homogenized by soaking in dry pyridine for
3 days, and then added to the PEG acyl chloride
under nitrogen at a 0.1 chitosan/PEG-acyl chloride
molar ratio. The mixture was stirred for 2 h and
refluxed at boil for 24 h. The cooled product was
precipitated twice with ethyl ether, and vacuum dried.
Fiber formation from PEGylated Chitosan by
electrospinning
The chitosan derivatives solution for electrospinning
was fed into a 5 ml syringe fitted with a pipette tip of
0.5 mm in diameter. The solution feed was fed by the
tilt angle of the syringe. A DC voltage (Gamma High
Voltage Supply, ES 30-0.1 P) of 10–15 kV was
applied between the syringe tip and grounded alumi-
num foil collector, separated by a distance of 17–
20 cm. Upon applying an electric charge, the pendant
droplet at the syringe tip was split by the repulsion
force to form a cone-shape jet that traveled towards
the collector and deposited on the collector in form of
a fibrous mat upon evaporation of the solvent. The as-
spun fibers were further dried under vacuum at room
temperature.
Characterization
Fourier Transform Infra-Red (FTIR) spectra were
collected using the KBr method on a Nicolet Magna-
IR 560 spectrometer with the wavenumber range
from 500–2000 cm-1. 1H NMR measurements were
performed on a Bruker Avance DMX500 spectrom-
eter under a static magnetic field of 500 MHz. The
morphology of Au/Pd sputter-coated nanofibers was
examined with a SEM (XL30-SFEG, FEI/Philips) at
an accelerating voltage of 5 kV. The diameters of
electrospun nanofibers were determined on 100
locations of SEM images using the ImageJ software.
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Cellulose (2007) 14:543–552
The fiber diameters were grouped for size distribution
and averaged.
Results and discussion
Synthesis and characterizations of PEGylation of
chitosan
PEGylation of chitosan proceeded with two reactions:
reductive amination of the amino group (Scheme 1)
and acylation reaction of the amino and hydroxyl
groups (Scheme 2). Both reactions required convert-
ing the hydroxyl-terminal groups of PEG to more
reactive end groups. In reductive amination, the
hydroxyl terminal was converted to aldehyde by
oxidation with acetic anhydride in DMSO. The
aldehyde-terminated PEG was coupled to the chito-
san amino group via Schiff base formation, then to
PEG-N-Chitosan by reductive amination. The extent
of this reaction was found to be low with DS = 0.12–
0.44 (Table 1). One reason is that PEG-aldehyde
oxidzes readily and aldol condensation may emerge
during the reaction, resulting in polymerization of
PEG-aldehyde (Bentley et al. 1998). In addition, the
excess NaCNBH3 can turn the solution to more alkali
in which chitosan becomes less soluble. Precipitation
is expected to suppress the reduction of Schiff-base
(Sugimoto et al. 1998). To avoid these drawbacks and
to improve the selectivity of this reaction, the
hydroxyl end group on the PEG was converted to
acyl chloride. The acyl chloride is more reactive than
the corresponding aldehyde while also capable of
reacting with not only amino but also hydroxyl groups
of chitosan in the esterification process (Scheme 2).
The structures of the PEGylated chitosan deriva-
tives were confirmed by FTIR and 1HNMR analyses.
The PEG characteristic peaks of das(CH) and c(CH2)
appeared at 1461 cm1 and 946, 838 cm1, respec-
tively. The characteristic amide bands of chitosan at
1651 cm1 (amide I), 1593 cm1 (amide II) and
1382 cm1 (amide III) were clearly evident. PEGy-
lated chitosan exhibited the characteristic absorption
bands of both chitosan and PEG. PEG grafting,
however, shifted the chitosan amino and amide peaks
slightly to the lower wavenumbers and significantly
reduced their intensities. By contrast, the amide I
band at 1651 cm-1 became less intense and was
shifted to 1648 cm-1, possibly due to hydrogen
bonding between amide I carbonyl with PEG
Cellulose (2007) 14:543–552 547

Table 1 Solubility of PEG-modified chitosan

Sample PEG Chitosan PEG Solubilityc
Mn (Da) Mv (kDa) DSb Mass%

CHCl3 DMF DMSO THF Water

PEG-N-CSa 550 400(DP = 2400) 0.39 56.3 ±  ±  +

2000 0.27 76.4     ±
5000 0.20 85.7     +
550 190(DP = 1160) 0.48 56.7 ±  ±  +
2000 0.32 79.6     +
5000 0.31 90.4 ±    +
550 137(DP = 850) 0.12 29.0     
5000 0.44 93.1 ±  ±  +
PEG-N,O-CS 550 145 (DP = 880) 1.50 83.0 + + + + +
2000 0.68 89.2 ±  ±  +
Chitosan – – – – –
a
PEG550-N-CS137: –CHO of PEG/-NH2 of Chitosan feed molar ratio was 1:3, others situation were 1:1
b
DS of PEG-N-CS and PEG-N,O-CS were determined by potentiometric titration (Avigad 1983) and H1NMR, respectively. And
mass% means the weight ratio of PEG in PEG -chitosan calculated by DS and Mn of PEG
c
5 mg/ml for 24 h later. +, soluble; ±, partially soluble or swelling; –, insoluble

hydroxyl. For PEG550-N,O-Chitosan145 with DS
= 1.5, the –NH2 peak at 1593 cm1 disappeared,
indicating that amino group of chitosan was fully
reacted with PEG. A new shoulder at 1730 cm1 is
attributed to the ester bond formed between PEG acyl
chloride and chitosan hydroxyl, indicating low sub-
stitution of the hydroxyl with PEG. Fig. 1
The 1HNMR spectra of the N-PEGylated chitosan
showed sharp singlets of PEG –OCH3 at 3.3 ppm (a)
and –NH–CH2CH2O– at 2.7 ppm (c), and a strong
broad PEG –OCH2CH2O– peaks at 3.5–3.8 ppm (b)
(Fig. 2a). N,O-PEGylated chitosan exhibited the
H-proton of –OCOCH 2– and –NHCOCH 2 – at
1730
1556
T%
2.3 ppm (a) and 2.2 ppm (c), respectively (Fig. 2b).
In PEGylated chitosan spectrum, peaks in the range
of 3.6–4.0 ppm were not well separated due to the
overlapping of a more intense PEG methylene peak with
the peaks from the chitosan saccharine backbone.
Solubility of PEGylation of chitosan
Reductive amination targets only the amino groups of
chitosan and generates N-PEGylated chitosan or
PEG-N-CS. Chitosan at 137, 190 and 400 kDa was
reductive aminated with PEG grafts of varied lengths.
Generally, the extent of amination or DS of
N-PEGylation increased with shorter chitosan chains
d
as well as PEG grafts except for PEG550-N-CS137
c that feeded by 1/3 molar ratio –CHO of PEG/-NH2 of
chitosan, which was different from others 1/1 molar
b ratio because of the gel situation at higher molar ratio
a (Table 1). At any given chitosan chain length, the

1593

1651
extents of amination or DS, increased with decreasing
PEG chain lengths. On the original chitosan (CS400),
1382
1461 the DS increased from 0.20 to 0.39 when reacted with
946 838 reducing PEG molecular weights from 5 kDa to
550 Da. Similarly, the extents of reaction with PEG at
2000 1500 1000 500 a fixed molecular weight improved with decreasing
Wavenumbers (cm-1)
chitosan molecular weights. Reductive amination
Fig. 1 IR spectrum of: (a) PEG, (b) chitosan, (c) PEG5k-N- with 5 kDa PEG, a 0.44 DS was reached on the
CS137, (d) PEG550-N,O-CS145 137 kDa chitosan, more than double of the 0.20 DS

123
548
Fig. 2 (a) 1HNMR spectrum of PEG5k-N-CS137 in D2O; (b)
1
HNMR spectrum of PEG550- N, O-CS145 in CDCl3
with on 400 kDa chitosan. These results indicate that
reductive amination of chitosan can be optimized by
reducing steric hindrance between the main (chito-
san) and side (PEG) chains with lower molecular
weight of either chitosan or PEG.
Acylation of chitosan, which could occur on either
the amine or the hydroxyl groups, was studied with
PEG at shorter PEG chain lengths of 550 and
2000 Da. Similar to the observations made on
reductive amination, higher acylation was achieved
with PEGs of shorter chain lengths, i.e., acylation
reached a DS of 1.50 with the 550 Da PEG in
comparison to the 0.68 DS with the 2000 Da PEG. At
DS > 1, the more reactive amine is likely to be fully
reacted while the hydroxyls are partially acylated.
Acylation of the C6 hydroxyl is thought to be more
likely than the C3 hydroxyl because its higher
123
Cellulose (2007) 14:543–552
reactivity as well as lower steric hindrance due to
proximity of C3 to the C2 amino group.
All PEGylated chitosan became water soluble
because of the high PEG contents (>50%), even with
the PEG-N-CS at a low DS of 0.2. Some of the PEG-
N-CS with DS  0.31 were swellable or partially
soluble in CHCl3, and DMSO, but not soluble in the
organic solvents studied. Both PEG-N,O-CS had
higher DS than the reductive aminated chitosan and
were water soluble. At DS = 1.50, PEG-N,O-CS also
became soluble in CHCl3, DMF, DMSO and THF.
The improved solubility indicates the significance of
DS on the solubility of the PEGylated chitosan
derivatives. Poor solubility of chitosan has been
attributed to it partially crystalline structure and very
tight hydrogen bonding between amino and hydroxyl
groups (Nishimura et al. 1991). PEGylation of
chitosan reduces the ordered structure by separating
the chitosan backbones as well as lowering hydrogen
bonding capacity. Both improve accessibility and
affinity for water and organic solvents.
Electrospinning of PEGylated chitosan
Electrospinning of reductive aminated and acylated
chitosan was investigated under a common electros-
pinning condition. Electrospinning of all aqueous
solutions of PEGylated chitosan produced only beads
(Fig. 3). Sprayed droplets were observed with aqueous
solutions of all PEG-N-chitosan via reductive amina-
tion, even at the highest DS of 0.44 (Fig. 3a). Both
acylated PEG-N,O-chitosan (DS = 0.68 and 1.50) were
soluble in water to higher concentrations than any of
the PEG-N-chitosan, yet their electrospun products
consisted only large beads (Fig. 3b, c). These three
PEGylated chitosans had similar main chain lengths,
but varied levels of PEGylation (DS) and PEG lengths.
The larger beads appear to be associated with either
higher DS, lower PEG lengths or simply higher
PEGylated chitosan concentrations, all related to better
water solubility. The main challenge of fiber formation
from PEGylated chitosan is most likely due to the
limitation in inter-molecular chain entanglement. Very
fine fibers linking the beads were observed with those
having lower DS, but longer PEG chains (Fig. 3a, b).
This enhanced chain entanglement among PEGylated
chitosan with longer but not higher extent of PEG
grafts suggest the role of long PEG side-chains on
chain entanglement.
Cellulose (2007) 14:543–552 549

Fig. 3 Electrospinning of PEG modified chitosan from aqueous solutions: (a) 4 wt% PEG5k-N-CS137 (DS = 0.44); (b) 13 wt%
PEG2k-N,O–CS145 (DS = 0.68); (c) 15 wt% PEG550-N,O–CS145 (DS = 1.50)

In electrospinning of any aqueous solution, the
high surface tension of water demands stronger
electric fields or higher voltages. Therefore, Triton
X-100 was added to reduce the solution surface
tension. The voltage required was indeed lowered,
however, still only beads were observed. Electros-
pinning of organic solutions was therefore studied
employing PEG550-N,O-chitosan145, the PEGylated
chitosan that is soluble in all organic solvents chosen.
DMF was selected as the solvent because its high
dielectric constant, a desirable characteristic for
electrospinning. Spraying of large beads with 1-
3mm diameters was observed from the 10% DMF
solution, similar to that from the 15% aqueous
solution (Fig. 3c). Increasing the concentration to
15% produced smaller beads with a few incipient
fibers (Fig. 4a). Much more fibrillated structure
intermixed with beads were observed as the concen-
tration increased to 25%, the highest possible
(Fig. 4b). The enhance chain entanglement with
higher concentrations played the major role in this
series. At low polymer concentrations, the solutions

Fig. 4 Electrospinning of
PEG550-N,O-CS145 from
DMF at concentrations of:
(a) 15 wt%; (b) 25 wt%

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550 Cellulose (2007) 14:543–552

Table 2 Properties of solvents used for electrospinning molecules in the solution increases rapidly, reaching
Solvent Surface tension Dielectric Vapor pressure
a critical level of chain entanglement to sustain fiber
(mN m1) constant (Torr) formation. It is important to point out that the same
PEG550-N,O-chitosan145 was not fiber forming (or
Water 72.8 80.1 17.54 least fiber-forming in comparison to those with longer
DMF 33.92 36.71 2.7 PEG, but lower DS) from aqueous solutions. The
CHCl3 27.12 4.81 158.4 difference could be due to the ability of PEG grafts to
THF 26.4 7.58 142 hydrogen bond with water molecules, preventing or
Source: Handbook of organic solvent properties, Smallwood, reducing their entanglement.
I.M., Elsevier (1996). All data were measured at 208C In the attempt to further improve fiber formation of
do not contain sufficient entanglement to produce PEG550-N,O-chitosan145 from DMF, THF and
fibers. With increasing polymer concentration, the CHCl3, cosolvents were added (Table 2). Accelerated
number of interchain associations among chitosan solvent evaporation has shown to increase surface

Fig. 5 Electropsinning of
15 wt% PEG550-N,O-
CS145 in co-solvents: (a)
50:50 (v/v) DMF:CHCl3;
(b) 50:50 (v/v) DMF:THF;
(c) 25:75(v/v) DMF:THF;
and (d) 25:75(v/v)
DMF:THF with 0.5%
Triton X-100; (e) fiber size
distribution of image d

123
Cellulose (2007) 14:543–552
charge density, thus forces, to promote fiber formation
(Shawon and Sung 2004). As poorer solvents for
PEGylated chitosan, these cosolvents should also
increase the solution viscosities, which should favor
fiber formation. At 15% PEG550-N,O-chitosan145,
equal volume of either THF or CHCl3 cosolvent was
added to DMF. Both increased the extents of fibers
(Fig. 5a, b vs 4a), while addition of THF produced
more continuous fibers with fewer beads as well as
increased fiber efficiency than CHCl3. The lower
efficiency with CHCl3 as a cosolvent is due to its high
vapor pressure, causing too rapid solvent evaporation
and plugging of the syringe injection nozzle during
electrospinning. Further improvement in fiber forma-
tion was achieved by raising the THF content to 75%.
Much finer fibers with diameters around tens nano-
meter, were generated (Fig. 5c). However, the fibers
were irregularly shaped and with few elongated beads.
Yet another approach to improve fiber formation
was by reducing the solution surface tension with
added surfactant. In 75/25 (v/v) THF/DMF cosolvent
with 0.5% Triton X-100TM, the beaded structure was
essentially eliminated and the fibers became more
uniform (Fig. 5d). However, the fibers became larger
with diameters ranging from 40 nm to 360 nm and an
average diameter of 162 nm (Fig. 5e). The improved
fiber uniformity and enlarged sizes are attributed to
the enhanced chain entanglement from greater inter-
actions between the surfactant and PEG side chains.
Strong interactions between PEG and TX-100
micelles have been depicted as a strong binding of
the surfactant to the polymer segment and micelliza-
tion of surfactant molecules along the polymer chain
(Wang et al. 2004).
Conclusions
PEG-N-chitosan and PEG-N,O-chitosan were synthe-
sized via reductive amination and acylation of
chitosan (Mv = 137 kDa to 400 kDa), respectively.
The extent of PEGylation with the activated PEG
(Mn = 550, 2000, and 5000 Da), in each case,
generally increased with reduced PEG lengths and
chitosan molecular weights. All PEGylated chitosan
became water-soluble, even with a DS as low as 0.2
in the case of PEG-N-CS. Solubility was further
improved by increased DS of PEG grafts. Some of
the PEG-N-chitosan with DS  0.31 were swellable
551
or partially soluble in CHCl3 and DMSO. At a 1.50
DS, PEG-N,O-CS was completely soluble in CHCl3,
DMF, DMSO and THF. The improved solvent
compatibility and solubility indicate that DS has the
significant effect on the solubility of the grafted
chitosan derivatives.
Electrospinnig of various PEG-N,O-chitosan from
aqueous solutions produced only beads. The associ-
ation of longer PEG lengths and lower PEGylation
with reduced bead sizes suggests further work on
PEGylation remains promising toward successful
fiber electrospinning from aqueous media. In DMF,
PEG550-N,O-chitosan145 (DS = 1.50) could be
electrospun into fibers and fiber uniformity was
improved by increasing the PEG-N,O-chitosan con-
centrations, using a cosolvent and adding a surfactant.
For instance, electrospinning of 15% PEG550-N,O-
chitosan145 from 75/25(v/v) THF/DMF coslovents
with 0.5% Triton X-100TM produced uniform nanof-
ibers with diameters ranging from 40 nm to 360 nm
and an average diameter of 162 nm. Findings of this
study show that chitosan nanofibers can be efficiently
generated from organic solutions, enabling engineer-
ing of new chitosan nanofibrous membranes. Contin-
uing investigations are underway in our laboratory to
further explore new chitosan derivatives to allow
electrospinning of aqueous systems to generate
chitosan nanofibers.
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