Journal of Molecular Liquids 287 (2019) 111008

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Journal of Molecular Liquids

journal homepage: www.elsevier.com/locate/molliq

Chain conformation of xanthan in solution as influenced by temperature

and salt addition
Cristina-Eliza Brunchi ⁎, Mihaela Avadanei, Maria Bercea, Simona Morariu
“Petru Poni” Institute of Macromolecular Chemistry, 41-A Grigore Ghica Voda Alley, 700487 Iasi, Romania

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

Article history: The capillary viscometry and Fourier transform infrared spectroscopy in attenuated total reflection (ATR-FTIR)
Received 23 April 2019 were used to investigate the conformation of xanthan gum (XG) chains in aqueous solutions, in the absence/pres-
Received in revised form 15 May 2019 ence of NaCl and in the temperature range from 25 to 90 °C. The content of acetate and pyruvate groups per side
Accepted 19 May 2019
of XG was determined by 1H NMR spectroscopy. The viscometric parameters (the intrinsic viscosity, [η], and the
Available online 22 May 2019
hydrodynamic interaction parameter, B) were determined by modeling the experimental data according to the
Keywords:
new Wolf model. The viscosity for XG solution in the absence/presence of NaCl as temperature rises has an ascen-
Xanthan gum dent evolution which takes place with slope variations. These changes were attributed to conformational transi-
Conformational change tion of the chains from the ordered to disordered form affecting the polymer-polymer and polymer-solvent
Viscosity interactions. Salt addition diminishes the electrostatic repulsions between COO− groups, the ordered conforma-
NMR tion is stabilized and the temperature at which conformation changes occur is shifted to higher values (above 55
FTIR °C for XG solution with 10−2 mol/L NaCl). The ordered-disordered transition of the chains was also observed in
ATR–FTIR spectra. Significant shifts in the regions 1750–1550 cm−1, corresponding to vibration of COO− groups,
indicate changes in the hydrogen bonding network.
© 2019 Elsevier B.V. All rights reserved.

1. Introduction degree of substitution with acetate and pyruvate groups depends on

Xanthan gum (XG), an extracellular microbial anionic polysaccha-
ride produced by the bacterium Xanthomonas campestris, is continu-
ously in the attention of researchers looking for the improvements of
the existing materials properties or the obtaining others for new poten-
tial applications. In this context, the researches on XG focus on its chem-
ical modification or on the finding of optimal conditions to obtain the
materials with desired properties. The chemical structure can be
changed even from the production process by controlling the tempera-
ture [1,2], pH [1], stirring speed [2,3], air flow rate [4], fermentation time
[5], bacterial strain [6,7] or by subsequent degradation [8–16],
functionalization [12,17–19] or oxidation [20,21].
The primary structure of this polysaccharide consists of repeating
glucose units as main chain and side chains glycosidically linked at
each second glucose residue in C-3 position (Fig. 1) [22].
The side chains consist of one glucuronic acid residue located be-
tween two mannose units. Predominantly, the inner mannose is
substituted stoichiometrically with an acetyl group (at C-6) and the ter-
minal one has variable degrees of pyruvate substitution (at C-4 and C-6)
but recently, other substitution patterns have been identified [23]. The
⁎ Corresponding author.
E-mail address: brunchic@icmpp.ro (C.-E. Brunchi).
the fermentation process and post-fermentation treatments. In aqueous
media, the XG chains can adopt a helical ordered conformation (rigid
structure) or a disordered coil (flexible structure) as a function of tem-
perature, ionic strength, pH, shear rate [24–27], solvent quality (which
is varied by using solvent/non-solvent mixtures) [28], or other environ-
mental conditions. In ordered conformation the side chains are folded
and associated with the main chain by hydrogen bonds (i.e., methyl
group of acetate residues binds to hemiacetal oxygen atoms of alternate
D-glucose residues from the main chain) [29]. In disordered conforma-
tion, the side chains are no longer associated as a result of the repulsion
electrostatic interactions between COO− groups from pyruvate resi-
dues. Since the difference in rigidity between the two conformations
leads to distinct properties in solution, a variety of techniques have
been involved in studying the helix-coil transition of xanthan chains
(such as nuclear magnetic resonance, circular dichroism, optical rota-
tion, electron microscopy, differential scanning calorimetry, viscometry,
light scattering, potentiometric titration [30,31], atomic force micros-
copy [32], dynamic mechanical thermal analysis and phase transition
analysis [33]). Another method able to provide valuable information
about chains conformation is FTIR spectroscopy. In literature there are
studies related to the chains conformation of some polysaccharide
based on shifts in the absorption frequency and/or changes in the ab-
sorptivity of particular absorption bands [34–36] but for xanthan are

https://doi.org/10.1016/j.molliq.2019.111008
0167-7322/© 2019 Elsevier B.V. All rights reserved.
2 C.-E. Brunchi et al. / Journal of Molecular Liquids 287 (2019) 111008
Fig. 1. Primary structure of xanthan gum.
missing. To our knowledge, this method was used to identify the
xanthan structure obtained from different carbon sources [37–40] or
modified by ultrasonic degradation [41] and interactions with other
(macro)molecules [17,42–45]. The aim of this study is to investigate
how the characteristics of XG chains and intermolecular interactions
change with increasing the temperature or change of ionic strength,
useful information for food, medicine, cosmetic, agriculture, water
treatment or other applications. The conformations adopted of the XG
chains in aqueous solution mediated by salt and temperature were an-
alyzed through Fourier transform infrared spectroscopy (ATR–FTIR)
corroborated with viscometric data.
2. Experimental
2.1. Materials
Commercial xanthan gum (XG) and sodium chloride (NaCl) from
Sigma Aldrich were used as received. According to relation proposed
by Milas et al. [46] the viscometric molecular weight of XG was
determinated as being 1.165 × 106 g/mol [47] and by X-ray fluorescence
measurements potassium was identified as the metallic ion [48].
2.2. Sample preparation
XG aqueous solutions, salt free and with different salt concentrations
(cNaCl = 10−5 and 10−2 mol/L), were prepared in highly purified deion-
ized water from Milli-Q PF (Millipore, Switzerland) at room tempera-
ture under gentle stirring. For viscosity measurements, solutions with
polymer concentrations, c, of 2.1 × 10−2 g/dL and 1 g/dL for ATR–FTIR
spectroscopy, respectively, were used. Proton nuclear magnetic reso-
nance (1H NMR) measurements were performed for XG aqueous solu-
tion (with XG concentration of 0.5 g/dL) with sodium acetate (as
internal standard with concentration of 3 × 10−3 mol/L) prepared in
D2O. The concentration of the salt solution was selected taking into ac-
count the conformational peculiarity and segments mobility of xanthan
chains which have an important impact on the interactions in aqueous
solution. Measurements were conducted on homogeneous solution
equilibrated overnight at room temperature.
2.3. Methods
2.3.1. 1H NMR spectroscopy
1
H NMR spectrum for XG aqueous solution was obtained with a
Bruker Avance DRX 400 spectrometer. The signal corresponding to
methyl protons in specific groups from XG (i.e., acetate and pyruvate
groups) were expressed in δ (ppm) relative to the protons from CH3
of sodium acetate (AcNa). Spectrum acquisition was performed at 25,
50 and 80 °C.
2.3.2. Viscosity measurements
Viscosity measurements were performed in temperature range be-
tween 27.5 and 75 °C by using an Ubbelohde capillary viscometer con-
nected to a LMV 830 (Lauda, Germany). The temperature was
controlled by a circulating-water bath (ECO ET 155). Polymer solutions
with increasingly low concentrations were prepared directly inside the
viscometer by sequentially dilution of the stock polymer solution. For
each concentration of XG aqueous solution, the flow time taken into ac-
count represents an average value of five measurements.
2.3.3. ATR–FTIR spectroscopy
The FTIR spectral measurements were performed by using the Ver-
tex 70 spectrometer (Bruker, Germany), in the attenuated total reflec-
tion configuration (ATR) on a Golden Gate accessory (single bounce
diamond crystal, Specac Ltd.). The measurements were carried out in
the temperature range of 26–90 °C with a heating rate of 1 °C/min.
Millipore water and solutions with cNaCl of 10−5 and 10−2 mol/L were
used as reference. The raw data and the reference were collected as
128 co-added scans at a 4 cm−1 resolution. The spectra for analysis
were obtained by subtracting the reference from the raw spectra. Each
sample was subjected to temperature scanning in two separate mea-
surements, in the same experimental conditions. Processing and manip-
ulation of data were made with the OPUS 6.5 software (Bruker Optics,
Germany).
3. Results and discussion
3.1. 1H NMR analysis
The content of acetate and pyruvate groups determines the con-
formation of XG chains in different media, influencing the solution
properties (e.g., viscosity, rheological properties, phase behavior,
etc.). The acetate groups favor the intramolecular attractive interac-
tions stabilizing the ordered conformation of XG while the pyruvate
groups prevent the formation of a compact helix due to intramolec-
ular electrostatic repulsion interactions [29,49]. These groups, in or-
dered conformation, cannot be detected in 1H NMR spectra under
high-resolution condition and room temperature due to the dipole-
dipole strong interactions between hydrogen or carbon nuclei
which determine the broadening of resonance line [50] (as exempli-
fied in Fig. 2 for XG solution). At 25 °C the protons corresponding to
CH 3 groups from acetate and pyruvate units located at 2.16 and
1.46 ppm, respectively, show very low signals due to the ordered
and rigid conformation of XG chains in solution.
Increasing temperature (at 50 and 80 °C, respectively) weakens
the dipole-dipole interactions between proton nuclei and the
strength of the signals becomes stronger (Fig. 2). The best signals
for protons corresponding to acetate and pyruvate groups were ob-
tained at 80 °C when the XG chains have a random coil conformation.
 C.-E. Brunchi et al. / Journal of Molecular Liquids 287 (2019) 111008 3

Fig. 2. 1H NMR spectra of XG in D2O (400 MHz) at different temperatures.

In addition, the signals for the protons from carboxyl (ionizable) or from 1H NMR spectrum were used to calculate the number of the ace-
hydroxyl groups overlap and change their position from 4.76 ppm tate (Nac) and pyruvate (Npyr) groups per gram of polysaccharide:
(at 25 °C) to 4.11 ppm (at 80 °C). This shift is due to the existence
of intramolecular H-bonds in which OH groups are implicated as Aac =AAcNa
donor to an alkoxy or hydroxy group [51]. A distinct peak at nac ¼ ð1Þ
c=cAcNa
5.3 ppm corresponding to the equatorial alpha anomeric proton of
mannopyranosic unit carbon and the peaks between 3.4 and
Apyr =AAcNa
3.9 ppm which can be attributed to the hydrogen from CH or CH2 npyr ¼ ð2Þ
c=cAcNa
groups are, also, shown in 1H NMR spectra.
A better quantification of the data provided by 1H NMR spectrum for
deuterated aqueous solution of xanthan in the presence of AcNa at 80 °C where c and cAcNa are the concentrations of XG (g/L) and AcNa (mol/L).
allowed us to add new information on the polysaccharide structure. So, The number of acetate and pyruvate groups (nac, npyr) corresponding
an acetate/pyruvate ratio of 6/5 was determined for the investigated XG to one side chain of XG was determined with the following relationships
considering the number of acetate and pyruvate groups per side chain of [17]:
xanthan.
For this purpose the integrals of the protons signals from substituent M∙Nac
nac ¼ ð3Þ
groups (Aac, Apyr) with reference to the protons from AcNa estimated 1−M pyr ∙N pyr −M ac ∙Nac
4 C.-E. Brunchi et al. / Journal of Molecular Liquids 287 (2019) 111008

M∙Npyr
npyr ¼ ð4Þ of state and, so, its changes can be described by a total differential
1−Mpyr ∙Npyr −Mac ∙Nac
as follows [53]:
     
∂lnη ∂lnη ∂lnη
d lnη ¼ dc þ dT þ dp
Here, M is the molar mass of repeating unit of XG excluding acetate ∂c T;p;γ_ ∂T c;p;γ_ ∂p c;T;γ_
 
and pyruvate groups, Mac and Mpyr are the molar masses of acetate and ∂lnη
þ dγ_ ð6Þ
pyruvate groups, respectively. ∂γ_ c;p;T

3.2. Viscometric investigations

Viscometry is a simple method able to investigate the behavior of
macromolecular chains in dilute solution by means of different visco-
metric parameters. Between them, the intrinsic viscosity (related to
the hydrodynamic dimensions, conformation and flexibility of polymer
chains) and a viscometric dimensionless parameter (quantifying the
hydrodynamic interactions that occur between the polymer chain and
solvent molecules) are the most used.
A commonly used method in determination of the intrinsic viscosity
([η]) for uncharged polymers and polyelectrolites in solvents containing
an adequate amount of salt is that of extrapolation. For example, the
Huggins extrapolation method described in Eq. (5) can be applied
when exists a linear dependence between reduced viscosity, ηsp/c, and
polymer concentration, c.
ηsp
¼ ½η þ kH ½η2 c
c
Fig. 3a exemplifies, for XG solution with 10−2 mol/L NaCl, the lin-
ear evolution of ηsp/c as a function of c at 70 °C; the same allure of ηsp/
c - c curve was observed for all temperatures. So, [η] can be obtained
as intercept of the linear fit of ηsp/c vs. c. As the salt concentration de-
creases, XG solution exhibits the characteristics of polyelectrolyte
due to the repulsive Coulombic interactions between charged groups
located along the main chains. The upturn of reduced viscosity in the
field of low polymer concentrations describes the expanded confor-
mation of the chains and prevents the determination of [η] by the
above mentioned method. Therefore, we have chosen to evaluate
the experimental viscosity data by a more adequate model [53]
which demonstrates its ability to describe the viscosity behavior
for different polyelectrolytes [47,52–54], uncharged polymers
[55–57], polymer mixtures [44,58], polyelectrolyte complexes [59].
This model was proposed by Wolf and starts from the assumption
that the viscosity of the dilute polyelectrolyte solutions is a variable
If the first partial differential, Eq. (6), represents the specific hydro-
dynamic volume of polymer, {η}, at a certain polymer concentration
(c), for a given value of temperature (T), pressure (p) and shear rate (γ_
), in the limit of infinite dilution and absence of shear rate, {η} becomes
identical with the intrinsic viscosity.
lim fηg ¼ ½η
c→0
_
γ→0
According to this model, [η] can be determined both from initial
slope of relative viscosity, lnηr, dependence on polyelectrolyte concen-
tration (which is independent of any model assumption) and by using
the following equation:
c½η þ Bc2 ½η½η
ln ηr ¼ ð7Þ
1 þ Bc½η
ð5Þ
where B, and [η]* are system specific constants representing viscometric
interaction parameters.
In the present study, the results of viscometric measurements were
modeled with Eq. (7).
The dependence of relative viscosity, lnηr, on the polymer concentra-
tion for XG solution in the absence/presence of NaCl is presented in
Fig. 3b only for 70 °C in order to avoid cluttering the chart; for all studied
temperatures, the evolution of lnηr - c dependences for XG solution in
the absence/presence of NaCl are the same as at 70 °C.
In Fig. 3b it can be observed the agreement between experimental
data and the curves (line) which demonstrates that Eq. (7) is appropri-
ate to calculate the [η], hydrodynamic interaction parameter B and char-
acteristic specific hydrodynamic volume [η]*; in all cases, the errors
were b10 %. The values of [η] and B obtained as a function of tempera-
ture and cNaCl are presented in Fig. 4.
Fig. 4a shows that increasing temperature from 25 to 75 °C leads to
high values of intrinsic viscosity for XG solution both in the absence
and presence of NaCl; the low values of [η] indicate an ordered

Fig. 3. Dependences of ηsp/c (a) and lnηr (b) on the polymer concentration, c, for XG aqueous solutions in the absence/presence of NaCl at 70 °C. The lines in the plot (a) are only to guide the
eye, while in the plot (b) they result from modeling the experimental data with Eq. (7).
 C.-E. Brunchi et al. / Journal of Molecular Liquids 287 (2019) 111008
Fig. 4. Variation of [η] (a) and B parameter (b) determined with Eq. (7) as a function of temperature for XG aqueous solutions in the absence/presence of NaCl. The plot (a) includes
previous data on [η] for aqueous solutions of XG in the absence/presence of NaCl at 25 °C [47] and in salt free solutions at 37 °C [44]. The lines in both plots are only to guide the eye.
conformation of the chains while the high ones reveal a disordered
conformation.
Starting from the assertion that at 25 °C in aqueous solution the
xanthan chains exist in disordered and partly ordered conformation
[24], we consider that up to 37 °C the intermolecular associations
based on hydrogen bonds are destroyed and so, the solution viscosity
decreases. As the temperature increases (from 37 to 50 °C), the intramo-
lecular hydrogen bonds between the side chains and backbone are bro-
ken, the side chains are free to move and initiates the transition of the
main chain conformation (but also of theirs) to a disordered form. The
disordered conformation is extended due to the repulsion between car-
boxylate groups [24,60] and explains the high viscosity of the XG solu-
tion. Fitzpatrick and co-workers [27] have found that the xanthan
chains (in native state) in salt free solution undergo a conformational
transition over a wide range of temperatures (35–55 °C) while, a pre-
transition attributed to the aggregates disruption, located around 45
°C, was evidenced by rheological investigations [61]. The high reactivity
of XG chains in disordered form as compared with the ordered ones is
responsible for the development of new intermolecular interactions
(such as the repulsion electrostatic interactions as a results of the kinetic
energy growth of the side chains or non-specific interactions favored by
the temperature increase or energetic and geometric conditions)
[51,60,62] which increase the viscosity at temperatures above 60 °C.
The presence of salt counteracts the repulsion electrostatic interactions
between charged groups and helps to stabilize the ordered conforma-
tion and strengthens the intermolecular interactions. Consequently,
the xanthan chains with such conformation need a higher temperature
to generate remarkable changes of viscosity (i.e., around 55 °C for solu-
tion with cNaCl = 10−2 mol/L).
Conformational modification of polymer chains in solution can
influence the thermodynamic preference of contacts between poly-
mer segments against those of the polymer chains and solvent mol-
ecules or vice versa. In Eq. (7), B parameter (as the Huggins
constant) quantifies the interactions between polymer segments
and gives the shape of lnηr - c plot. For uncharged polymers (i.e.,
XG solution with cNaCl = 10−2 mol/L, Fig. 3b) the lnηr dependence
of c is linear while for polyelectrolytes the Coulombic interactions
between charged groups lead to a curved dependence (i.e., XG solu-
tion in the absence of NaCl and with cNaCl = 10−5 mol/L, Fig. 3b). In
the case of polyelectrolytes, B counts the effect of Coulombic and
non-Coulombic interactions (B = BCoulomb + Bnon-Coulomb); BCoulomb
represents the positive contribution of Coulombic interaction and
Bnon-Coulomb is the negative contribution of non-Coulombic interac-
tions to the hydrodynamic interaction parameter [63].
5
As can be seen in Fig. 4b, the values of B both in absence/pres-
ence of NaCl are positive on the entire temperature range typical
for the behavior of polymers in thermodynamically good solvents;
in unfavorable thermodynamic conditions the of B are negative
[47,52]. In the absence of salt and for low concentration NaCl, the
positive values of B are higher than those obtained in solution
with cNaCl = 10−2 mol/L. In the range of temperatures between
25 and 55 °C, the B values in these solvents are close and reach a
maximum value at the same temperature (about 50 °C). Up to
≈40 °C, the Coulombic and non-Coulombic interactions counter-
balance each other, but above it the polymer-polymer interactions
became strong, the chain conformation expands and the macro-
molecule responds different to temperature rise. By adding more
salt (cNaCl = 10−2 mol/L), the value of B decreases and approaches
to zero as a result of shielding repulsion electrostatic interactions
and macromolecular chains adopt a shrink coiled conformation
similar those adopted in poor thermodynamic solvents. In determi-
nation of B values, for XG solution with cNaCl = 10−2 mol/L, for the
best fit with Eq. (7), [η]* was set zero. [η]* is the characteristic spe-
cific hydrodynamic volume only required of polyelectrolytes to
model the maxima of η sp/c - c plot in the domain of extremely
low c and in the absence of electrostatic repulsion interactions it
becomes zero.
3.3. Infrared spectral analysis
In order to improve the information resulted from the viscometric
behavior of XG chains in aqueous solution, the infrared spectroscopy
was used. The xanthan gum aqueous solutions in the absence/presence
of salt were subjected to temperature increase between 26 and 90 °C,
with an increment of 1 °C/min.
Fig. 5 shows the modification at temperature increasing from 26 to
90 °C of the FTIR spectra, for the XG solution at three salt concentrations.
The asymmetric stretching vibrations of carboxylate groups in the glu-
curonic acid and pyruvate units are observed in the 1750–1550 cm−1
region [64]. The ν(CO) vibrations of the alcoholic COH groups, the
rings skeletal ones and those characteristic to the glycosidic COC link-
ages are all covered in the broad 1170–970 cm−1 band.
At room temperature (26 °C), the profile of the bands is influenced
by the ionic strength of the solution. The νasym(COO−)glu vibration is
quite stable, being positioned at 1586 cm−1 in water and 1587 cm−1
in 10−2 mol/L NaCl, and dominates almost the entire spectrum. The
νasym(COO−)pyr, with a finer vibronic structure than νasym(COO−)glu,
have the maximum around 1673 cm−1 in water and around
6 C.-E. Brunchi et al. / Journal of Molecular Liquids 287 (2019) 111008
Fig. 5. Representative temperature–dependent ATR-FTIR in the fingerprint region spectra for XG solution in the absence/presence of NaCl. ΔT = 26–90 °C.
1670 cm−1 in the salt solutions, and the most important shoulder is
around 1690 cm−1. At room temperature, the νasym(COO−)pyr band be-
comes lower as the ionic strength of solution increased. When the tem-
perature was gradually increased, the most spectacular spectral changes
occurred at the glucuronic and pyruvate carboxylates, as expected, and
the effect of temperature was stronger in the saline solutions. In the so-
lutions with no salt or with trace of salt, the position of the νasym(COO−)
−
glu shifted to the blue, while νasym(COO )pyr shifted to the red, accom-
panied by the simultaneous switching in their whole intensity. At 90
°C, νasym(COO−)glu is greatly diminished in intensity and is observed
at 1598 cm−1 in water, and at 1611 cm−1 when cNaCl = 10−5 mol/L.
In the solution with 10−2 mol/L NaCl, at 90 °C the νasym(COO−)glu prac-
tically became buried in the more intense band of νsym(COO−)pyr that is
centered on 1570 cm−1.
The two starting components of the νasym(COO−)pyr band, around
1670 and 1690 cm−1, were selectively growing up upon temperature
increase as a function of the ionic strength: the sub-band at
1670 cm−1 increased in pure water, and that at 1690 cm−1 increased
when cNaCl was 10−2 mol/L. For solution with cNaCl = 10−5 mol/L, the
two sub-bands increased in a concerted way. As a results, at 90 °C the
maximum of νsym(COO−)pyr was red-shifted to 1664 cm−1 for XG solu-
tion in the absence of NaCl, but greatly displaced to the blue, at
1691 cm−1, when cNaCl = 10−2 mol/L. In parallel, the temperature in-
crease led to the development of the bands around 1024, 1050–1070,
1100–1110, 1350 or 1468 cm−1, belonging to ν(CO), δ(COH), δ(CH)
and δ(CH2) vibrations in new structures.
As representative features of the movement of the lateral trisaccha-
ride chains, the νasym(COO−)glu and νasym(COO−)pyr bands can give a
hint about the onset temperature of the main phase transition. The
changes in the bands profile with temperature is the sum of the growths
and/or decays of some of their components, which shifts the maximum
of the whole band. As the νasym(COO−)pyr band is isolated in compari-
son with νasym(COO−)glu, its evolution under the impact of temperature
increasing was easiest to follow and interpret. The temperature depen-
dence of the position of νasym(COO−)pyr maximum, exemplified in
Fig. 6a, shows that the temperature of the main transition increases
from ≈46 °C in pure water to ≈52 °C in the low salt solution. When
cNaCl = 10−2 mol/L, the sub-band at 1690 cm−1 blueshifts constantly
up to ≈48 °C (reaching 1693 cm−1), where it remains until the temper-
ature is around 70 °C. After 70 °C, νasym(COO−)pyr shifts clearly to lower
wavenumbers.
From the intensity variations presented in Fig. 6b (intensity that
was normalized to a reference band, 968 cm−1 , a skeletal C-O-C
mode [65]), the enhancement of the 1690 cm−1 component when
 C.-E. Brunchi et al. / Journal of Molecular Liquids 287 (2019) 111008
Fig. 6. Temperature – dependent variation of the spectral characteristics of νsym(COO−)pyr: (a) position, given by wavenumber (cm−1); (b) ATR absorbance.
cNaCl = 10−2 mol/L, started after 45 °C and is extended over approx.
25 °C. This temperature domain roughly corresponds to the plateau
domain in the wavenumber plot in Fig. 6a and the ascending part
of the solution viscosity from Fig. 4a. For the low salt concentration
or no-salt solutions, the dependences of the normalized intensity of
the whole νasym(COO−)pyr band on temperature are very similar to
each other (Fig. 6b). They present the same small plateau region be-
tween 30 and 42 °C and identified above with the reorganization of
hydrogen bonds in the chains entanglements and disordered re-
gions. The increase in intensity of νasym(COO−)pyr took place up to
≈55 °C in water solution and ≈50 °C for 10−2 mol/L NaCl, followed
by a relative decrease, which was counterbalanced by an increase
in the bandwidth.
The sudden redshift of νasym(COO−)pyr at a certain temperature
(Fig. 6a) is an indicative of a change in the micro-environment of the py-
ruvate (and glucuronic acids) units, in terms of the strength and nature
of the hydrogen bonds they establish. The two main types of pyruvate
carboxylates (corresponding to the 1690 and 1670 cm−1 components)
that were selectively evidenced in the higher salt concentration and in
the no-salt solution, respectively, are engaged in hydrogen bonding of
different strengths. They exist from the beginning, as the xanthan in so-
lution has the chains in both the folded and unfolded structures, where
the terminal carboxylates form intra- and intermolecular hydrogen
bonds. The detaching of the trisaccharide chains from the main cellulose
backbone is connected with an increase in viscosity [66] and with the
sudden shift and intensity of the carboxylate groups' bands in the termi-
nal pyruvate and glucuronic units.
4. Conclusions
Xanthan gum chain conformation in aqueous solution was discussed
by using capillary viscometry as well as 1H NMR and ATR–FTIR spectros-
copy at different temperatures and salt contents. According to 1H NMR
spectra of XG solution in D2O and AcNa, the xanthan chains exist in or-
dered conformation at 25 °C (with low signals of protons corresponding
to CH3 groups from acetate and pyruvate units), while above 50 °C the
random coils prevail (the signals of protons from the two groups be-
come stronger). By heating the XG solutions, the order-disorder confor-
mational transitions occur and led changes in the balance between
Coulombic and non-Coulombic electrostatic interactions and, conse-
quence, the viscosity increases. The viscometric parameters (intrinsic
viscosity and hydrodynamic interaction parameter) were determined
according to Wolf model which describes the viscometric behavior of
macromolecules in various conditions of temperature and ionic
7
strength. Remarkable changes noticed in ATR–FTIR spectra of XG aque-
ous solutions with temperature and salt concentration are closely re-
lated to structural organization of its macromolecule and indicate the
complexity of this process. The movement of the lateral trisaccharide
chains, by means of the carboxylate groups from pyruvate as terminal
units, indicated the values of the 46 and 52 °C as main temperature tran-
sition points in the no-salt and low-salt solutions.
There is a good agreement between the spectral behavior and the
observations from viscometry measurements, especially for the salt
free XG aqueous solution.
Acknowledgement
This work was supported by a grant of the Ministry of National Edu-
cation, CNCS–UEFISCDI, project number PN–III–P2–2.1–PED–2016-
1536 (contract 130PED/2017).
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