Synthetic Metals 195 (2014) 286293

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Synthetic Metals
journal homepage: www.elsevier.com/locate/synmet

Purication of a conducting polymer, polyaniline, for biomedical

applications
Jaroslav Stejskal a, , Milena Hajn a , Vera Kasprkov b,c , Petr Humpolcek b,d ,
Alexander Zhigunov a , Miroslava Trchov a
a

Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, 162 06 Prague 6, Czech Republic
Centre of Polymer Systems, Polymer Centre, Tomas Bata University in Zlin, 760 05 Zlin, Czech Republic
c
Department of Fat, Surfactant and Cosmetics Technology, Faculty of Technology, Tomas Bata University in Zlin, 762 72, Zlin, Czech Republic
d
Polymer Centre, Faculty of Technology, Tomas Bata University in Zlin, 762 72, Zlin, Czech Republic
b

a r t i c l e

i n f o

Article history:
Received 18 March 2014
Received in revised form 2 June 2014
Accepted 23 June 2014
Available online 15 July 2014
Keywords:
Biocompatibility
Conducting polymer
Polyaniline
Reprecipitation
Toxicity

a b s t r a c t
A conducting polymer, polyaniline, was prepared in globular and nanotubular morphologies. The protonated forms were converted to the corresponding bases and both types of samples were tested for
cytotoxicity. The polyanilines were then suspended in N-methylpyrrolidone or in concentrated sulphuric
acid, and the soluble parts were precipitated into methanol acidied with sulphuric acid. Such a dissolution/precipitation cycle was tested as a purication procedure for polyaniline, which would remove the
potential low-molecular-weight components. The original morphology of polyaniline was destroyed in
soluble part (1824 wt.%) but maintained in the fraction insoluble in N-methylpyrrolidone. The fraction
soluble in sulphuric acid was higher (5664 wt.%). The original morphology converted to fragments after
reprecipitation, and the samples became amorphous. The conductivity was reduced on average by two
orders of magnitude. FTIR spectroscopy was used to assess the molecular structure, hydrogen bonding,
and their changes. The cytotoxicity of polyaniline salt determined on mouse embryonic broblast cell
line NIH/3T3 was reduced after reprecipitation from N-methylpyrrolidone when compared to the initial
polymer and showed the absence of cytotoxicity at the extract concentration of 5 and 10% in the case
of globular and nanotubular polymer, respectively. A corresponding positive effect was not observed for
polyaniline reprecipitated from sulphuric acid.
2014 Elsevier B.V. All rights reserved.

1. Introduction
Polyaniline (PANI) [1] is one of the most frequently studied conducting polymers. In addition to its electrical, optical, or responsive
properties, scientists have also been attracted by the variety of
nanostructures this polymer produces [2,3]. In addition to standard
globular PANI, which is obtained by the oxidation of aniline in
strongly acidic aqueous media, nanotubes are produced in solutions of moderate acidity [47], and nanobres typically under a
diluted regime [3,8,9].
Medicine and biology are promising areas, in which conducting
polymers, such as polyaniline or polypyrrole, could be used for
monitoring vital function in living organisms, as well as for their
stimulation [10]. The mixed electronic and proton conductivity

Corresponding author. Tel.: +420 296 809 351; fax: +420 296 809 410.
E-mail address: stejskal@imc.cas.cz (J. Stejskal).
http://dx.doi.org/10.1016/j.synthmet.2014.06.020
0379-6779/ 2014 Elsevier B.V. All rights reserved.

of conducting polymers makes them unique materials that are

expected to mediate the interfacial transfer between ionic and
electronic charge transport [1115] at the interface of biological
objects and electrodes. The interaction with living tissue is of interest especially with respect to cardiac [1618] and neural tissues
[1921] or brain and other cells [22]. Many potential applications
of conducting polymers, however, are not based on conductivity. Among them, the antimicrobial compositions [2325],
cell-proliferation supports [26,27], and the use in photothermal
tumour destruction [28], may serve as examples.
The biocompatibility of PANI has recently been tested [22,29,30]
and, for some applications, improvement is still needed. The
deprotonationreprotonation cycles performed on PANI led to
reduction in cytotoxicity. Polyaniline, being completely insoluble
and stable in aqueous media, can hardly be toxic. Thus it seems that
prevailing hazards with respect to cytotoxicity on biological objects
are associated with low-molecular-weight components. These are
of two types: (1) reaction by-products and aniline-based oligomers

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287

[31,32], and (2) the acids that constitute the salts with PANI. For that
reason, the purication of PANI with respect to the former group is
of importance.
The reprecipitation of polymers is a routine method used in
polymer science for the removal of residual monomers and any
other low-molecular-weight compounds. In this procedure, a polymer is dissolved in a suitable solvent and added drop-wise to a
large excess of non-solvent. The polymer precipitates but lowmolecular-weight compounds stay dissolved and can be separated.
So far, for PANI, this method has not been tested due to the small
selection of solvents, which is limited to N-methylpyrrolidone and
concentrated sulphuric acid and, even in these solvents, the solubility is not complete. The present study tests these reprecipitation
procedures as the means for the purication of PANI for applications in medicine or biosciences.
An additional question of interest is connected with the morphology and conductivity changes caused by the reprecipitation.
Globular and nanotubular PANI have therefore been selected for
this study, and their fate in this procedure is reported.

Spectrometer with a DTGS TEC detector. Samples were dispersed

in potassium bromide and compressed into pellets. Raman spectra,
excited with a HeNe 633 nm laser, were collected on a Renishaw
inVia Reex Raman spectroscope. The conductivity was measured
by a four-point van der Pauw method using a current source SMU
Keithley 237 and a Multimeter Keithley 2010 voltmeter with a
2000 SCAN 10-channel scanner card on pellets compressed at
540 MPa with a manual hydraulic press.
Wide-angle X-ray scattering (WAXS) experiments were performed using a pin-hole camera (Molecular Metrology System)
attached to a microfocused X-ray beam generator (Osmic MicroMax 002) operating at 45 kV and 0.66 mA (30 W). The camera was
equipped with a removable and interchangeable Imaging Plate
23 25 cm2 (Fujilm). Experimental setup covered the momentum transfer range q = (4/) sin  of 0.253.5 A 1 , where  = 1.54 A
is the wavelength and 2 is the scattering angle. Calibrations
of the centre and sample-to-detector distance were made by
using silicon powder. Samples were measured in the transmission
mode.

2. Experimental

2.4. Cytotoxicity test

2.1. Polyaniline preparation

Prior to in-vitro cytotoxicity testing, the samples were sterilized

by a dry heat at 121 C for 20 min, homogenized in a mortar, and
subsequently extracted, according to ISO 10993-12, at 0.2 g of the
PANI per 1 mL of culture medium. The extraction was performed
in chemically inert closed containers using aseptic techniques at
37 1 C under stirring for 24 1 h. The parent extracts (100%) were
then diluted in a culture medium to obtain a series of dilutions with
concentrations of 50, 25, 10, 5 and 1%. All extracts were used within
24 h. The ability of cells to respond to cytotoxic substances was
veried by application of sodium dodecyl sulphate solution (SDS;
Sigma, Czech Republic).
Cytotoxicity testing was conducted in accordance with EN
ISO 10993-5 using mouse embryonic broblast cell line NIH/3T3
(American Type Culture Collection, HB-8065), cultivated according to the protocol recommended by the supplier. Cells were
pre-cultivated for 24 h and the culture medium was subsequently
replaced with PANI extracts. As a reference experiment providing 100% cell proliferation, the pure extraction medium was used.
To assess cytotoxic effect, the MTT assay (Invitrogen Corporation,
USA) was performed after one-day cell cultivation in extracts at
37 0.1 C. All the tests were performed in quadruplicates. The
absorbance was measured at 570 nm by Innite M200 PRO (Tecan,
Switzerland). Dixons Q test was used to remove outlying values
and means were calculated. The cell viability, expressed as percentage of cells present in the corresponding PANI extract relatively
to cells cultivated in pure extraction medium (100% viability) was
determined. The morphology of the cells was assessed after their
cultivation in extracts for 24 h. The cells from each culture plate
were observed by an inverted Olympus phase contrast microscope
(Olympus IX81, Japan) at 40 magnication.

Standard globular PANI (PANI-S) was prepared by the oxidation of 0.2 M aniline hydrochloride (Lachner, Czech Republic)
with 0.25 M ammonium peroxydisulphate (APS; Lachner, Czech
Republic) in water [1] at room temperature. The nanotubular PANI
(PANI-NT) was similarly synthesized by the oxidation of 0.2 M aniline (Fluka, Switzerland) with 0.25 M APS in 0.4 M acetic acid [4].
Polyaniline powders were separated by ltration, rinsed with acetone, and dried under ambient conditions. They were subsequently
deprotonated to the PANI bases by suspension in excess of 1 M
ammonium hydroxide and dried as above.
2.2. Reprecipitation
Polyaniline bases (10 g) were suspended in 250 mL of Nmethylpyrrolidone (NMP) or concentrated sulphuric acid (96 wt.%).
The suspensions were occasionally shaken. After 30 days, the insoluble part was separated by ltration. The insoluble part was well
rinsed with methanol containing sulphuric acid, and dried in an
ambient atmosphere. The solutions containing the soluble part of
the PANI base were added drop-wise into 1.5 L of methanol containing 15 mL of concentrated sulphuric acid. The precipitate, PANI
sulphate, was collected on a lter and dried in air at room temperature.
2.3. Characterization
Infrared spectra in the range 4004000 cm1 were recorded
using a fully computerized Thermo Nicolet NEXUS 870 FTIR

Table 1
The conductivity of globular and nanotubular polyanilines and of their respective components soluble and insoluble in N-methylpyrrolidone or concentrated sulphuric acid.
Type

Solvent

Soluble
fraction [wt.%]

Conductivity, soluble
part [S cm1 ]

Globular

Original PANI base

NMP
Sulphuric acid
Original PANI base
NMP
Sulphuric acid

18.6
64.0

24.4
56.0

1.3
0.028
0.073
0.029
4.2 105
8.8 104

Nanotubular

Degree of crystallinity of original or insoluble parts. All soluble parts were amorphous.

Conductivity insoluble
part [S cm1 ]
0.035
0.14
1.3 104
5.0 104

Crystallinitya [%]
15
18
0
35
35
0

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J. Stejskal et al. / Synthetic Metals 195 (2014) 286293

Fig. 1. Illustration of (a) globular and (b) nanotubular forms of PANI. For more information, see Ref. [6].

3. Results and discussion

3.1. Soluble and insoluble parts of PANI
The rst important observation is based on the fact that the
dissolution of PANI in both solvents was not complete. This is
probably not surprising to most readers working in this eld [33]
but the insoluble fraction is, in this particular case, higher than
most of them would expect (Table 1). Molecular weights of PANI
determined by gel-permeation chromatography have already been

reported in the literature [3436]. It has therefore to be stressed

that they refer only to the soluble part of the polymer, and not to
the sample as the whole.
The fraction soluble in NMP is higher for nanotubular PANI than
for globular PANI (Table 1). This may be due to the higher content of
aniline oligomers that accompany PANI [37] and are preferentially
produced in the reaction media of low acidity, such as 0.4 M acetic
acid [2,4,6], used for the preparation of nanotubes.
There is also the second point to be considered. Polyaniline base
dissolved in NMP is not precipitated when poured into excess of

Fig. 2. Soluble (left) and insoluble fractions (right) of globular (top) and nanotubular PANI (bottom) in N-methypyrrolidone.

J. Stejskal et al. / Synthetic Metals 195 (2014) 286293

289

Fig. 3. Soluble (left) and insoluble fractions (right) of globular (top) and nanotubular polyaniline (bottom) in concentrated sulphuric acid.

methanol. This may have important consequences for the solution

processing of PANI base. Only when sulphuric acid was added to
methanol, the PANI sulphate quantitatively precipitated.
No substantial improvement of solubility has been observed
after addition of lithium chloride, even at high concentrations,
although many papers claim that the presence of this salt reduces
the hydrogen bonding in PANI [36,38]. Other papers reported no
improvement [33], in the accordance with the present study.

3.2. Conductivity
It has earlier been reported that the conductivity of the globular form of PANI is higher that of its nanotubular analogue [6],
because of the larger fraction of non-conducting aniline oligomers
in the latter sample [37]. This is consistent with the fact that the
conductivity of PANI increased after reuxing with tetrahydrofuran
[39], which may have led to the reduction of the non-conducting
oligomeric fraction.
Reprecipitation of PANI reduces the conductivity by about two
orders of magnitude. This is caused by the disordering of orderedchain regions after the dissolution, or swelling of the insoluble
parts. A similar reduction in conductivity was observed after
reprecipitation of a PANI fraction soluble in dimethylformamide
followed by reprotonation [40]. The model of highly-ordered conducting islands distributed in chain-disordered matrix is used in the

discussion of PANI conductivity [11,41,42]. The swelling of islands

leads to local chain-disordering, and the overall conductivity of the
material becomes reduced.
The insoluble fraction has generally a higher conductivity, probably due to the higher fraction of residual ordered parts. This is
supported by the observation that organized structures, such as
nanotubes, constitute an insoluble and partly crystalline fraction
(Fig. 2). In contrast to the soluble parts, which are amorphous, the
insoluble part is partly crystalline (Table 1). The difference in the
conductivity, however, is not large [3] (Table 1). The conductivity
of samples processed from sulphuric acid is somewhat higher compared with PANI precipitated from NMP solution. This means that
the molecular structure of PANI was not damaged by suspension in
concentrated sulphuric acid. This is also supported by the reports
of good stability of PANI in concentrated sulphuric acid, even in the
presence of strong oxidants such as ammonium peroxydisulphate
or hydrogen peroxide [43].

3.3. Morphology
The globular and nanotubular morphology of the original PANI
(Fig. 1) is completely destroyed in the fraction soluble in NMP but
partly preserved in the insoluble part (Fig. 2). It is tempting to
assume that the part preserving the original morphology is chemically crosslinked. In concentrated sulphuric acid, however, both the

J. Stejskal et al. / Synthetic Metals 195 (2014) 286293

soluble and insoluble fractions have a similar morphology (Fig. 3).

This means that the part controlling the morphology is insoluble in
NMP due to hydrogen bonding interactions, which are destroyed
in sulphuric acid.
We conclude that both PANI samples are composed of (1) macromolecules that are weakly hydrogen-bonded and are soluble in
both NMP and sulphuric acid (ca 20 wt.%), (2) strongly hydrogenbonded fraction, which is insoluble in NMP but soluble in sulphuric
acid (ca 40 wt.%), and (3) a fraction, which is insoluble in both solvents (ca 40 wt.%) and is represented, most likely, also by covalently
crosslinked polymer chains.
This conclusion is conrmed by follow-up results: (1) the fraction which was originally soluble in NMP is still soluble in NMP.
(2) The fraction which was soluble in sulphuric acid is completely
insoluble in NMP but it is still soluble in sulphuric acid. There seem
to be two types of hydrogen bonding, weak and strong ones.

FTIR in KBr

a
3434

1566

1300
1240
800
572
1034
989

3235

G- NMP- S

618

1653

G- NMP- IS

3.4. Infrared spectroscopy

G- H2SO4- S
G- H2SO4- IS
4000

3000

2000

1000
1

Wavenumbers, cm

FTIR in KBr

1151
1497
1575 1310
1240
1038
817
572

NT

3434

Absorbance

The molecular structures of the original sample and fractions

soluble and insoluble in NMP were studied with infrared spectroscopy. The main bands of globular PANI salt (spectrum G in
Fig. 4a) have been interpreted earlier [44]. A broad absorption band
at wavenumbers higher than 2000 cm1 corresponds to absorption
of free charge-carriers in the protonated polymer, and the bands
at 1566 and 1476 cm1 are due to quinonoid (Q) and benzenoid
(B) ring-stretching vibrations, respectively. An absorption band at
1300 cm1 corresponds to -electron delocalization induced in
the polymer by protonation, while the band characteristic of the
conducting protonated form observed at 1248 cm1 has been interpreted as corresponding to a C N+ stretching vibration in the
polaron structure. The strong and broad band centred at 1132 cm1 ,
has been assigned to a vibration mode of the NH+ structure of
protonated imine nitrogens, and also to strong interchain NH+ N
hydrogen bonding. The asymmetric SO3 stretching vibration in the
hydrogen sulphate counter-ion may also contribute to this band.
The shoulder observed at 1032 cm1 is attributable to the symmetric SO3 stretching in the hydrogen sulphate counter-ion. The
band at 800 cm1 is associated with C H out-of-plane bending
vibrations of two adjacent hydrogen atoms on a 1,4-disubstituted
benzene ring. The band observed at 572 cm1 is due to the hydrogen
sulphate or sulphate counter-ion.
The main bands of nanotubular PANI salt (spectrum NT in Fig. 4b)
have been interpreted earlier [44]. The sample is composed of two
components, aniline oligomers and true PANI [45]. The differences
between the spectra of globular (spectrum G in Fig. 4a) and nanotubular PANI (spectrum NT in Fig. 4b), thus reect the presence
of aniline oligomers. The main peaks are slightly shifted in the
spectrum of nanotubular PANI salt. The additional weak peaks in
the substitution region 900650 cm1 are due to the C H outof-plane bending vibrations and indicate the presence of aniline
oligomers bearing mono-substituted phenyl rings as terminal units
in oligomers and/or more pronounced branching of the chains in
the nanotubular sample.
Both the soluble and insoluble fractions of the globular and
nanotubular samples were very difcult to disperse in potassium
bromide pellets because of their compact stone-like structure (see
Figs. 2 and 3). Consequently, the absorption of samples was very
small and the recorded spectra contained relatively strong absorption bands in the region of the stretching vibrations of water
molecules at about 3434 cm1 . The spectra of all fractions were
multiplied by factor 5 in Fig. 4 for better resolution. The higher
degree of aggregation after precipitation of the emeraldine salt
in aqueous acidic solution was observed by Colomban et al. [46].
The creation of the strong hydrogen bonds between the amine
and the O S group in the sulphonic acid has been described [47].

1476 1132

Absorbance

290

3235

NT- NMP- S

1653

989 687

NT- NMP- IS

NT- H2SO4- S
NT- H2SO4- IS

4000

3000

2000

Wavenumbers, cm

1000
1

Fig. 4. Infrared spectra of globular G (a) and nanotubular NT (b) fractions soluble
(S) or insoluble (IS) in N-methylpyrrolidone (NMP) or sulphuric acid (H2 SO4 ).

A shoulder which can be distinguished at about 3235 cm1 belongs

most probably to different types of intra- and inter-molecular
hydrogen-bonded N H stretching vibrations of secondary aromatic amines [45]. We assume the presence of intramolecular
hydrogen bonding in N H O, where the oxygen atoms belong
to the carbonyl group or to the oxygen atom of sulphate counterions. This is supported by the presence of the peak at 1653 cm1
in the spectra of the fraction soluble in N-methylpyrrolidone,
and by the peak observed at 10341038 cm1 of the symmetric
SO3 stretching vibrations. In all fractions we observe bands with

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291

Table 2
Cytotoxicity of PANI extracts of various concentrations presented as average absorbance standard deviation (Abs) and as a relative value compared to reference (RV)
according to ISO 10 993-5 standard.a
Extract

100%

Globular
0.40 C
Original PANI saltb
PANI base
0.20 0.02
Abs
0.37 D
RV
Fraction soluble in NMP
0.20 0.02
Abs
RV
0.37 D
Fraction soluble in sulphuric acid
0.55 0.05
Abs
RV
0.98 A
Nanotubular
0.72 Bc
Original PANI salt
PANI base
0.18 0.01
Abs
0.33 D
RV
Fraction soluble in NMP
0.29 0.01
Abs
RV
0.52 C
Fraction soluble in sulphuric acid
0.41 0.03
Abs
0.74 B
RV

50%

25%

10%

5%

1%

0.40 C

0.40 C

0.54 C

0.56 C

0.90 A

0.43 0.03
0.78 B

0.53 0.02
0.94 A

0.66 0.03
1.17 A

0.59 0.01
1.06 A

0.58 0.03
1.04 A

0.27 0.04
0.49 C

0.29 0.03
0.52 C

0.37 0.04
0.67 B

0.50 0.02
0.89 A

0.54 0.01
0.98 A

0.42 0.05
0.76 B

0.27 0.03
0.48 C

0.26 0.04
0.46 C

0.26 0.02
0.48 C

0.50 0.04
0.90 A

0.76 Bc

0.79 Bc

0.67 Bc

0.70 B

0.85 A

0.21 0.01
0.38 D

0.37 0.02
0.66 B

0.52 0.02
0.93 A

0.56 0.04
1.00 A

0.64 0.03
1.14 A

0.27 0.01
0.50 C

0.32 0.03
0.57 C

0.51 0.02
0.91 A

0.51 0.00
0.92 A

0.59 0.04
1.05 A

0.41 0.02
0.74 B

0.27 0.00
0.49 C

0.24 0.04
0.43 C

0.21 0.01
0.39 C

0.58 0.02
1.03 A

a
Cytotoxicity in relative values equal to 1 corresponds to 100% cell survival compared to reference. Values >0.8 are assigned to no cytotoxicity (A), 0.60.8 mild cytotoxicity
(B), 0.40.6 moderate cytotoxicity (C), and <0.4 severe cytotoxicity (D).
b
Results presented in Ref. [30].
c
The formation of a gel-like substance was observed.

a maximum at about 989 cm1 belonging to the vibrations of

sulphate counter-ions resulting from sulphuric acid used in the
precipitation medium.
The samples soluble in sulphuric acid are hydrogen-bonded
more strongly than those after dissolution in N-methylpyrrolidone.
This means that the dissolution is promoted by competitive hydrogen bonding between PANI and N-methylpyrrolidone, and this
solvent remains in the samples after their separation.
On the basis of FTIR spectra we conclude that (1) the molecular
structure of PANI was not damaged by the reprecipitation procedure, and (2) there is no indication that some polymeric component
was removed from the sample.
3.5. Cytotoxicity
According to a previous study [30], both PANI hydrochloride
and PANI base prepared by the standard procedure according
to IUPAC [1] showed cytotoxicity, which was notably higher for
PANI hydrochloride compared to PANI base. In that work, the
efciency of the purication step, consisting of the deprotonation/reprotonation, cycle was tested. It was concluded that such
a procedure reduces the toxicity of the samples but not to the
desired level. In the current work, dissolution/precipitation was
applied as an alternative purication method for removal of residual monomers and potential low-molecular-weight contaminants
(Table 2). The cytotoxicity data are reported, according to ISO
10 993, in relative values by comparison with reference sample
corresponding to 100% cell survival after application of pure culture medium. Values of >0.8 are then labelled as no cytotoxicity,
0.60.8 mild cytotoxicity, 0.40.6 moderate cytotoxicity, and <0.4
as severe cytotoxicity.
Simple comparison of the original globular PANI base from
current work and PANI base reported earlier [30] conrms that
the cytotoxicity of both preparations, at comparable concentrations, is roughly similar. This fact proves good reproducibility of
both PANI syntheses and the procedure used for biocompatibility
testing. Compared to globular sample, the cytotoxic level of PANI

nanotubes in the base form is higher with a toxic extract concentration of 10% compared to 25%.
Reprecipitated samples can be compared with original PANI
hydrochloride, as they were precipitated in acidied methanol and
obtained thus as PANI sulphate. Here, improvement relative to asprepared PANI globular salt is observed after reprecipitation from
NMP with shift of the cytotoxicity level to higher concentrations of
PANI extracts. In this case, the effect of the dissolution/precipitation
(=reprecipitation) cycle was nevertheless comparable to the purication efcacy of the deprotonation/reprotonation procedure [30],
with no cytotoxicity observed for 5% extracts in both cases
(compared to 1% cytotoxic extract of original PANI hydrochloride). On the other hand, the reprecipitation has no signicant
effect on the cytotoxicity of PANI dissolved in sulphuric acid.
The efcacy of purication through reprecipitation is, in this particular case, lower in comparison with reprotonation (1% vs 5%
extract).
It might be speculated that the acid reaction of PANI salt
originating from hydrolysis can contribute to cytotoxicity. This
view is, however, relevant only for the highest extract concentrations, as the buffering effect of the culture medium after dilution
suppresses sample acidity and cells are grown under physiological conditions. The non-uniform impact of precipitation can be
observed between globular and nanotubular forms of PANI. Globular samples processed from NMP display higher cytotoxicity of
extracts (5%) relatively to extracts from the nanotubular sample
(10%). The performance of samples reprecipitated from sulphuric
acid is, however equal (1%). Moreover, in the case of samples
dissolved in sulphuric acid, the astonishing effect of decreased
cytotoxicity for 100% and 50% extracts was observed. This atypical effect was conrmed in the case of both PANI forms, and
it can be connected to the hormetic effect (dose-response relationship in which there is a stimulatory response at low doses,
but an inhibitory response at high doses) or possible aggregation
of toxic substances with medium components, mainly proteins.
More comprehensive research is needed to explain this unexpected
effect.

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J. Stejskal et al. / Synthetic Metals 195 (2014) 286293

4. Conclusions
Dissolution in a suitable solvent followed by precipitation in a
non-solvent is a routine method for the purication of polymers.
With globular and nanotubular polyaniline, only 18 and 24 wt.%,
respectively, could be dissolved in N-methylpyrrolidone. The features of morphology and degree of crystallinity are maintained in
the insoluble fraction. The morphology of the soluble part after precipitation of polyaniline is different, fragmentary. In concentrated
sulphuric acid, the solubility is higher, 64 and 56 wt.%, respectively,
for the globular and nanotubular sample. Both the soluble and insoluble parts have fragmentary morphology and they are amorphous.
The conductivity becomes reduced by two orders of magnitude
after the purication steps. FTIR spectra suggest that intramolecular
hydrogen bonding between polyaniline chains and bonds produced
between polyaniline and N-methylpyrrolidone are responsible for
the observed behaviour. The most severe cytotoxic effect has been
observed on parent polyaniline salts, whether it was globular or
nanotubular polymer, with the results being slightly worse for
the globular form. Correspondingly, differences between initial
polyaniline bases have been recorded as well, showing thresholds
for cytotoxic concentration of extracts being of 10 and 25% for globular and tubular polymer, respectively. The behaviour of both NMPand sulphuric acid-soluble fractions illustrated that reprecipitation
procedure enables only limited improvement of the cytotoxicity,
when compared with the initial polyaniline salts. This improvement was, however, notable only for the NMP-soluble fraction and
was absent in the case of fractions soluble in sulphuric acid. With
respect to the fact that both soluble fractions showed a notable
reduction in conductivity, the applied procedure does not seem
to be a straightforward solution for PANI purication in terms of
the removal of low-molecular-weight substances. The biologically
safe application of reprecipitated as well as parent polymer is thus
limited to mixtures or blends with other polymers.
Acknowledgments
Authors thank the Czech Science Foundation (13-08944S) for
nancial support. Thanks are also due to Dr. Jan Prokes from the
Charles University in Prague for conductivity measurements.
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