Food Hydrocolloids 121 (2021) 107033

Contents lists available at ScienceDirect

Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Carrageenan molecule conformations and electrokinetic properties in

electrolyte solutions: Modeling and experimental measurements
Aneta Michna *, Wojciech Płaziński, Dawid Lupa, Monika Wasilewska, Zbigniew Adamczyk
Jerzy Haber Institute of Catalysis and Surface Chemistry, Polish Academy of Sciences, Niezapominajek 8, PL-30239 Krakow, Poland

A R T I C L E I N F O A B S T R A C T

Keywords:
Carrageenan molecule conformations
Electrokinetic charge of carrageenan
Hydrodynamic diameter of carrageenan
Molecular dynamics of carrageenan
Viscosity of carrageenan solutions
Physicochemical properties of κ–carrageenan (KC) molecules and their aqueous solutions were determined using
molecular dynamics modeling and experimental techniques. Chain conformations, the end-to-end distance, the
gyration radius and the density of the molecule were theoretically calculated. Experimentally, the diffusion
coefficient and electrophoretic mobility of the molecules were measured for ionic strength range 10− 4 to 0.15 M
and different pHs. Using these data, the hydrodynamic diameter, the electrokinetic charge and the effective
ionization degree of KC molecules were determined. The dynamic viscosity measurements for dilute KC solutions
enabled to determine of its molecule intrinsic viscosity for various ionic strengths. The viscosity attained 50,000
for dilute electrolyte solutions compared to the Einstein value for spheres equal to 2.5. This confirmed that KC
molecules assumed extended conformations in accordance with theoretical modeling. It is concluded that the
application of complementary theoretical and experimental methods furnishes reliable information about KC
molecule conformations in electrolyte solutions.

1. Introduction
Carrageenans are non-toxic, biocompatible macroions of large vis­
cosity and excellent solubility even in cold water. Thus, they are wildly
applied in drug delivery systems (Bonferoni et al., 1993); (Bonferoni
et al., 1994); (Hariharan, Wheatley, & Price, 1997), for controlled drug
release (Li, Ni, Shao, & Mao, 2014), as potent inhibitors of viruses
(Gonzalez, Alarcón, & Carrasco, 1987); (Rodríguez et al., 2014);
(Nakashima et al., 1987); (Rodríguez et al., 2014), and bacterial in­
fections (Briones, Sato, & Bigol, 2014) as well as as potential therapeutic
agents against SARS-CoV-2 (Pereira & Critchley, 2020). It is also re­
ported that λ-carrageenan exhibits anti-tumour and immunomodulation
properties (Harding, 2017); (Zhou et al., 2004); (Luo et al., 2015). In
addition to these applications, they are used in cosmetics and food
products as efficient thickeners, gelling agents or stabilizers (Abd Karim
et al., 1999); (Saha & Bhattacharya, 2010), (Harding, 2017).
These macroions belong to a group of linear polysaccharides pos­
sessing backbones formed by α(1 → 3) and β(1 → 4)-linked galactose
residues with repeating sulphate half-ester groups and 3,6-anhydro-
bridges (Imeson, 2009). There are six main forms of carrageenans: λ
(lambda), κ (kappa), ι (iota), μ (mu), θ (theta) and ν (nu), which differ in
the position and the number of sulphate groups within the disaccharide
repeat units and in the content of 3,6-anhydrogalactose residues. The
forms are mainly obtained by extraction from red seaweeds (Montolalu,
Tashiro, Matsukawa, & Ogawa, 2008); (Bono, Anisuzzaman, & Ding,
2014). However, one should notice that the red seaweed extract is a
mixture of various forms of carrageenans, and the composition of the
mixture depends on the algal source, life stage and even extraction
procedure (Reis et al., 2008). For example, κ and ι form is mainly
extracted from Kappaphycus alvarezii and Eucheuma denticulatum, and
λ-carrageenan - from Gigartina skottsbergi and Sarcothalia crispate (Alba
& Kontogiorgos, 2019). The κ and λ forms of carrageenans are especially
interesting in terms of their structure and possible applications.
Oppositely to λ-carrageenan (LC), the κ-carrageenan (KC) possesses
only one sulphate group per building block of disaccharide. The sulphate
content is in the range of 25%–30% (Harding, 2017). It is constructed
from alternating α(1 → 3)-D-galactose-4-sulphate and β(1 → 4)-3,
6-anhydro-D-galactose (Harding, 2017); (Reis et al., 2008). KC is soluble
in hot water, forms gels in the presence of K+ due to the helical form
(Reis et al., 2008), which is additionally stabilized by hydrogen bonds
(Reis et al., 2008). It also belongs to a group of “reversibly
soluble-insoluble polymers” because of precipitating with 0.2% KCl at

* Corresponding author.
E-mail addresses: aneta.michna@ikifp.edu.pl (A. Michna), wojciech.plazinski@ikifp.edu.pl, wojtek_plazinski@o2.pl (W. Płaziński), dawid.lupa@ikifp.edu.pl
(D. Lupa), monika.wasilewska@ikifp.edu.pl (M. Wasilewska), zbigniew.adamczyk@ikifp.edu.pl (Z. Adamczyk).

https://doi.org/10.1016/j.foodhyd.2021.107033
Received 3 March 2021; Received in revised form 17 June 2021; Accepted 11 July 2021
Available online 13 July 2021
0268-005X/© 2021 Published by Elsevier Ltd.
A. Michna et al.
38 ◦ C and dissolving the precipitate in distilled water (Reis et al., 2008);
(Roy & Gupta, 2003). The molar mass of κ-carrageenan, determined by
analytical centrifugation and light scattering, is of the order of 300 kg
mol− 1 (Slootmaekers, van Dijk, Varkevisser, van Treslong, & Reynaers,
1991); (Slootmaekers, Mandel, & Reynaers, 1991); (Harding, 2017). KC
is applied in the whole-cell and enzyme immobilization, for wastewater
treatment and beer and in ethanol production (Reis et al., 2008).
Because of its significance, the properties of carrageenan solutions
have been studied with the aim to evaluate their molar mass distribution
(Slootmaekers, van Dijk et al., 1991); (Thành et al., 2002); (Almutairi
et al., 2013); (Schuck et al., 2014), radii of gyration, contour lengths
(Slootmaekers, Mandel, & Reynaers, 1991), persistence lengths (Berth,
Vukovic, & Lechner, 2008), hydrodynamic diameters and second virial
coefficients (Thành et al., 2002).
Several works have been also focused on the determination of the
intrinsic viscosity of carrageenan solutions in various electrolytes
comprising multivalent ions (Zabik & Aldrich, 1965), (Zabik & Aldrich,
1967) for large ionic strength (Chronakis, Doublier, & Piculell, 2000).
Interesting results were obtained by Berth et al. who analysed LC and
KC solutions by static light scattering (multi-angle light scattering,
MALS) in 0.1 M NaNO3, which allowed to determine the molar mass of
the carrageenans (1400 kg mol− 1 for LC and 596 kg mol− 1 for KC), their
radius of gyration (102 nm for LC and 104 nm for KC) and the second
virial coefficient (10− 4 mol ml g− 2 for LC and 9.9 × 10− 4 mol ml g for
KC) (Berth et al., 2008). The obtained data were interpreted in terms of
the wormlike chain model using the Skolnik-Odijk-Fixman approach.
The intrinsic persistence length of 3–4 nm and expansion factor of 1.5
were also calculated.
Physicochemical characteristics of LC involving the molar mass,
intrinsic viscosity and sedimentation coefficient (at pH 7.0 and ionic
strength of 0.1 M) were acquired by Almutairi et al. (Almutairi et al.,
2013) using size exclusion chromatography coupled to MALS, capillary
viscometry, and analytical ultracentrifugation. An extended and flexible
conformation for LC molecules is confirmed by these investigations.
However, despite of extensive experimental effort, no comprehen­
sive characteristics of KC molecules were acquired simultaneously
applying theoretical modeling and experimental techniques, which can
yield information about the electrokinetic properties and the molecule
conformations under various ionic strengths and pHs.
Therefore, the goal of this work is to perform thorough characteris­
tics of KC solutions using the all-atom molecular dynamics (MD)
modeling and complementary experimental methods such as the dy­
namic light scattering (DLS), micro-electrophoresis (LDV), the optical
waveguide lightmode spectroscopy (OWLS), and the dynamic viscosity
measurements. The previously unexplored range of dilute supporting
electrolyte (NaCl) concentration is studied in order to acquire reliable
information about the extended chain length. These investigations yield
quantitative information about the KC molecule properties, such as its
effective charge, ionization degree, chain length and its cross-section
area for a broad ionic strength range. This is significant for predicting
and controlling the adsorption of KC molecules at solid/electrolyte in­
terfaces, which is often applied to produce multilayer shells for targeted
drug delivery systems. Our results also facilitate a proper interpretation
of experimental data derived from the quartz crystal microbalance or the
streaming potential measurements and other experimental techniques
that require calibration. Additionally, the flexibility of adjusting the
viscosity of KC samples by ionic strength change has practical implica­
tions for the regulation of the thickening ability of carrageenan
solutions.
2. Materials and methods
2.1. Materials
Carrageenan sodium salt was supplied by Sigma Aldrich (Poland) in
the form of crystalline powder.
2
Food Hydrocolloids 121 (2021) 107033
Analytical grade (pure p. a.) NaCl was purchased from Avantor
Performance Materials Poland S.A.
Eight generation of poly (amidoamine) dendrimers, PAMAM8,
(8,33% w/w aqueous solution, dispersity index below 1.04 (Steche­
messer & Eimer, 1997)), of the molar mass of 233 kg mol− 1, was pur­
chased from Dendritech, Inc. (the USA). The PAMAM8 solutions were
applied for the formation of the anchoring layer, which was used to
determine the carrageenan adsorption kinetics in OWLS experiments.
The obtained results were described in Supplementary Material.
The solutions of carrageenan were prepared by dissolving a proper
amount of the powder in NaCl solutions of a precisely controlled con­
centration and pH. Deionized water was obtained using Milli-Q Elix &
Simplicity 185 purification system from Millipore SAS Molsheim,
France.
The temperature of all experiments was fixed of 298 K.
2.2. Methods
The chemical composition of the sample, especially the sulfur con­
tent was determined by elemental analysis (Thermo Scientific FlashS­
mart Elemental Analyzer) and Fourier Transform Infrared Spectroscopy
(FTIR) (FTIR Nicolet 6700 spectrometer, Thermo Scientific), whereas its
molar mass was acquired using size exclusion chromatography coupled
to multi-angle light scattering (SEC-MALS) analysis. SEC-MALS analysis
was performed using high-performance liquid chromatography (HPLC)
instrument (1260 Infinity LC, Agilent Technologies) equipped with a UV
detector, MALS detector (DAWN HELEOS II, Wyatt Technology) and a
differential refractometer (Optilab T-rEX, Wyatt Technology).
The electrophoretic mobility and diffusion coefficients of carra­
geenan molecules were measured for the broad ionic strength range
(10− 4 to 0.15 M at pH 5.8), using a combination of the micro-
electrophoresis with Laser Doppler Velocimetry (LDV) and the dy­
namic light scattering (DLS) (Malvern Zetasizer Nano ZS apparatus). The
Henry (Ohshima, 2012) and Stokes-Einstein (Einstein, 1908) equations
were applied for determining the KC zeta potentials and hydrodynamic
diameters for the above conditions. The carrageenan bulk concentra­
tions were equal to 300 and 500 mg L− 1 in DLS and 500 mg L− 1 in LDV
measurements.
Mass loss measurements were performed by thermogravimetric
analysis (TGA) and differential scanning calorimetry (DSC) measure­
ments using Netzsch STA 409 PC microcalorimeter. Additionally, the
dry mass of the carrageenan powder was determined using static ther­
mogravimetry, where KC mass changes on time were monitored at
constant temperature (378 K). The detailed protocol for static ther­
mogravimetry analysis and obtained TGA/DSC curves can be found in
Supplementary Material.
The density of carrageenan solutions was determined using the high
precision Anton Paar DMA 5000 M densitometer.
The dynamic viscosity of the carrageenan solutions of defined mass
concentrations was determined using the Anton Paar rolling-ball
viscometer Lovis 2000 M/ME equipped with a short capillary tube.
This allowed determining the viscosity of samples with a large precision
(0.05%) using relatively small volumes of carrageenan solutions (0.1
mL).
Three sensors, situated in the viscometer capillary, measure a run­
time of a steel ball in the capillary filled by the sample (pure electrolyte
or carrageenan solutions of various concentrations). From the measured
runtimes, the viscometer software yields the dynamic viscosity of the
solution using the following equation:
η = K(ρb − ρs )Δt (1)
where: η is dynamic viscosity [mPa s], K [m2 s− 2] is the calibration
constant, ρb is the ball density [kg m− 3], ρs is sample density [kg m− 3],
and Δt is the runtime [s].
The measurements were carried out for the KC volume fraction not
A. Michna et al.
exceeding 5 × 10− 4 (dilute macroion concentration limit) and at a fixed
NaCl concentration varied between 10− 4 and 0.15 M.
The kinetics of carrageenan adsorption from dilute solutions on
supporting PAMAM8 layers was determined using the OWLS method
according to the procedure described elsewhere (Wasilewska, Adamc­
zyk, Pomorska, Nattich-Rak, & Sadowska, 2019); (Michna, Pomorska,
Nattich-Rak, Wasilewska, & Adamczyk, 2020). The OWLS 210 instru­
ment (Microvacuum Ltd., Budapest, Hungary) was used. The apparatus
is equipped with a laminar slit shear flow cell comprising a silica-coated
waveguide (OW2400, Microvacuum). The adsorbing substrates were
planar optical waveguides made of a glass substrate (refractive index
1.526) covered by a film of Si0.78Ti0.22O2 of the thickness 170 nm, and
the refractive index 1.8. A grating embossed in the substrate enables the
light to be coupled into the waveguide layer. The sensor surface was
coated with an additional layer (10 nm) of pure SiO2 (Wasilewska et al.,
2019).
2.3. Theoretical modeling
A series of carrageenan chains of various lengths, composed of 4, 10,
20 and 40 monosaccharide residues (equivalent to 2, 5, 10 and 20
‘monomers’, in accordance to the notation accepted in this work) was
considered in the theoretical modeling. The initial configurations of the
systems, including the solvation of the KC chains as well as the addition
of ions were created using the CHARMM-GUI online server (Park et al.,
2019); (Lee et al., 2016). The modeling system consisted of cubic boxes
of the initial edge dimensions varying from 4.2 to 14.7 nm, depending
on the system. The corresponding number of water molecules varied
from 2300 to 110000, respectively. The appropriate number of Na+ and
Cl− ions was added to each system, accounting for its neutral charge and
the desired ionic strength value (10− 2 M).
The all-atom molecular dynamics (MD) simulations were carried out
using the GROMACS 2016.4 package (Abraham et al., 2015). The
CHARMM36 force field (Guvench et al., 2011); (Guvench et al., 2008)
was used to describe the interactions involving KC molecules, accom­
panied by the CHARMM-compatible explicit TIP3P water model. The
parameters describing the anhydro residues were prepared manually
and relied on the parameters for unfunctionalized galactose units. The
introduced changes include: (i) defining new covalent bond between O6
and C3; (ii) conversion of the O3 and H3 atoms into ‘dummy’ atoms,
non-interacting with any other part of the system by non-bonded in­
teractions; (iii) adjusting the partial charges on C6 and O6 to maintain
the nautral charge of the residue; (iv) changing the atom types accord­
ingly to their altered chemical character in the case of O6, C6 and C3.
The modeling was carried out applying periodic boundary conditions
and in the isothermal-isobaric ensemble. The temperature was main­
tained close to its reference value (298 K) by applying the V-rescale
thermostat (Bussi, Donadio, & Parrinello, 2007), whereas for the con­
stant pressure (1 bar, isotropic coordinate scaling) the
Parrinello-Rahman barostat (Parrinello & Rahman, 1981) was used with
a relaxation time of 0.4 ps. The equations of motion were integrated with
a time step of 2 fs using the leap-frog scheme (Hockney, 1970). The
hydrogen-containing solute bond lengths were constrained by the
application of the LINCS procedure with a relative geometric tolerance
of 10− 4 (Hess, Bekker, Berendsen, & Fraaije, 1997); (Hess, 2008). The
full rigidity of the water molecules was enforced by application of the
SETTLE procedure (Miyamoto & Kollman, 1992). The electrostatic in­
teractions were modeled by using the particle-mesh Ewald method
(Darden, York, & Pedersen, 1993) with cut-off set to 1.2 nm, while van
der Waals interactions (LJ potentials) were switched off between 1.0 and
1.2 nm. The translational center-of-mass motion was removed every
timestep separately for the solute and the solvent.
The systems were subjected to geometry minimization and MD-based
equilibrations in the NPT ensemble, lasting 2–20 ns, depending on the
system size. After this step we checked that the geometry of the rings in
the anhydro residues was in accordance to the expectations, i.e. 1C4.
3
Food Hydrocolloids 121 (2021) 107033
After equilibration, production simulations were carried out for a
duration of 200 ns and the data were saved to trajectory every 2 ps. The
end-to-end distance and gyration radius values were calculated by using
the GROMACS routines gmx polystat and gmx mindist.
3. Results and discussion
3.1. Theoretical modeling results
The calculation presented hereafter were performed for κ-carra­
geenan (KC) molecules composed of 2, 5, 10 and 20 monomers, each
comprising one –OSO-3 group (see Fig. 1). The molar mass of fully
ionized monomer calculated from its chemical composition is equal to
0.385 kg mol− 1, whereas the molar mass of sodium ion compensated
monomer is equal to 0.408 kg mol− 1. Let us note that in the accepted
nomenclature ‘monomer’ is equivalent to the two consecutive residues
in the KC chain (i.e. disaccharide building block).
Primarily, in the MD simulations, the molecule conformation, the
end-to-end distance and the gyration radius were determined as a
function of the monomer number, denoted by Nm. Exemplary snapshots
of KC chain conformations obtained for NaCl concentration of 10− 2 M
and different monomer numbers are shown in Fig. 2. Qualitatively, one
can observe that the chains are quasi-rigid, exhibiting a rod-like shape
with no sharp bending. The most frequently occurring pattern of solute-
solute hydrogen bonding included interactions between the –SO-3 and
–OH groups of the two consecutive monosaccharide residues. Apart
from the conformationally restricted mutual orientation of the neigh­
bouring residues, no tendency to the formation of regular, helical shapes
in a larger dimensional scale was observed.
The semi-rigid conformation of the KC chain was confirmed by the
additional, enhanced sampling simulations and the resulting free energy
maps (FEMs) calculated with respect to the glycosidic dihedral angle
values (see details in Supplementary Material). Inspecting FEMs (Sup­
plementary Material), one can observe that the FEM area corresponding
to the free energy levels < 5 kJ/mol covers a relatively significant
fraction of the map calculated for the β(1 → 4) type of linkage, on the
contrary to that calculated for the α(1 → 3) linkage. This indicates that
the semi-rigid conformation of the KC chain is distorted mainly by the
rearrangments within the β(1 → 4) linkage. Furthermore, the alternative
minima of the free energy, not associated with the shapes sampled
during standard MD simulations, are located at approx. 12 kJ/mol (β(1
→ 4) linkage) above the most favorable conformation or essentially
absent (α(1 → 3) linkage). Thus, the population of the corresponding
chain geometries is negligible (<1%).
Independent MD runs confirmed that the monomer length was
practically independent of the chain length, adopting an average value
of 0.90 ± 0.03 nm (denoted later as lm). Interestingly, this agrees within
error bounds with the experimental value reported by (Slootmaekers,
Mandel, & Reynaers, 1991) for λ-carrageenan, which was equal to 0.89
nm.
The MD modeling also allowed to determine the average end-to-end
distance of the molecule Le and the average gyration radius Rg as a
function of the number of monomers Nm. These dependencies are
illustrated in Fig. 3. As seen, both the end-to-end distance and the gy­
ration radius were well approximated by linear functions with the slope
slightly smaller than predicted for the fully extended chain where one
has:
Le = lm Nm (2)
Also, as seen in Fig. 3, the values of the gyration radius derived from
MD slightly deviate from the values of gyration radius predicted for a
straight cylinder of the length equal to Le, i.e., Rg = lm Nm /121/2 . This
confirms that the carrageenan molecules assume extended rod-like
conformations for Nm up to 20.
The MD-originating descriptors were applied to determine the KC
A. Michna et al.
Fig. 1. A schematic representation of the chemical structure of the KC molecule with the marked monomer unit, corresponding to the two consecutive residues of
3,6-anhydrogalactose (3,6-AnGal) and galactopyranose-4-sulphate (Gal4S).
Fig. 2. Snapshots of KC chain conformations for 20 and 40 residues derived from MD modeling for 10−
respectively). Hydrogen atoms and solvent molecules are omitted for clarity.
molecule density using the method developed in previous work
(Adamczyk, Morga, Kosior, & Batys, 2018). Accordingly, the size of the
simulation boxes, where a single KC molecule was confined, was sys­
tematically increased that resulted in a decrease in the KC mass fraction
from 0.01 to 0. The density of these systems ρs, as well as that of the pure
solvent ρe, were determined in the independent MD runs. Then, the
dependence of ρe/ρs on wp was plotted and fitted by a straight line
characterized by the slope sp and the density was calculated from the
formula:
ρe
ρp = (3)
1 + sp
In this way, one obtains, at the temperature of 298 K, three different
ρp values, dependent on the assumption inherent in the calculation
scheme: (i) ρp = 2.24 × 103 kg m− 3 for bare, negatively charged chain
without either hydration or co-cations contributing to the KC mass; (ii)
ρp = 2.10 × 103 kg m− 3
when considering the sodium co-cations
contributing to the mass of the KC chain but no hydration; (iii) ρp =
2.01 × 103 kg m− 3 in the case where sodium co-cations contribute to the
mass of the KC chain and, additionally, the chain hydration is assumed
(1 water molecule per monomer). Using these values one can calculate
the volume of a monomer considering its molar mass M1 from the
dependence v1 = M1 /(ρp Av), which was equal to 0.29, 0.30 and 0.32
nm3 for the cases: (i), (ii) and (iii), respectively. Consequently, the
equivalent monomer diameter assuming its cylindrical shape calculated
as d1 = (4v1 /π lm )1/2 was equal to 0.64, 0.66 and 0.67 nm, respectively.
For the sake of convenience, the theoretical data obtained from
modeling are collected in Table 1.
It should be mentioned the extrapolation of these results to a larger
4
Food Hydrocolloids 121 (2021) 107033
2
M NaCl concentration (equivalent to 10 and 20 monomers,
molar mass of KC furnishes useful data that are inaccessible for direct
theoretical modeling because of excessive time of computations. In
order to assess the validity of this extrapolation, throughout experi­
mental studies were performed to determine the diffusion coefficient,
the hydrodynamic diameter, the electrokinetic charge, the ionization
degree, and intrinsic viscosity of KC molecules.
3.2. Experimental characteristics of carrageenan solutions
To evaluate the composition of carrageenan sample, a comprehen­
sive physicochemical analysis was performed using the FTIR and the
elemental analysis methods. The details of performed experiments and
their results were discussed in Supplementary Material (sections: 1.2
and 1.3). Using these methods it is shown that the molar fraction of KC in
sample is equal to 0.944, whereas the second present component is LC.
The number average molar mass of KC was equal to 475 ± 0.08 kg
mol− 1, as found using SEC-MALS. This value is close to the mass average
molar mass of KC, which was determined to be equal to 480 ± 0.08 kg
mol− 1. Therefore, one can conclude that KC sample exhibits low dis­
persity (Mw/Mn = 1.01).
The dry mass of the carrageenan powder used for preparing its so­
lutions used in the experiments was determined via the TGA and static
thermogravimetry experiments described in Supplementary Informa­
tion. The static thermogravimetry revealed that the water content in the
carrageenan sample was equal to 10.5%, whereas the TG/DTA/DSC
measurements showed 13.5%. For further calculations, a mean value of
water content equal to 12% was used. The determined water content is
comparable with the value of 14% reported by Dul et al. (Dul et al.,
2015).
A. Michna et al. Food Hydrocolloids 121 (2021) 107033

Fig. 3. Part (a): The average KC molecule end-to-

end distance Le vs. the number of monomers
derived from MD modeling. The solid line denotes
the linear fitting of theoretical data. The dashed line
shows the results predicted for the fully extended
chain, i.e., Le = Nm lm, Part (b): The gyration radius
of KC vs. the number of monomers. The solid line
denotes linear fitting of theoretical data derived
from MD simulations and the dashed line shows the
results predicted for the fully extended chain of a
cylindrical shape, i.e., Rg = Nmlm/121/2. Vertical
bars denote the fluctuations of the given quantity
found during MD simulations and expressed as
standard deviation values. The NaCl concentration
was equal to 10− 2 M.

Primarily, in the experiments, the diffusion coefficient of KC mole­ mg L− 1 were extrapolated to the infinite dilution using the correction
cules for various ionic strengths was determined by DLS as described function proposed by Slootmaekers et al. (Slootmaekers, Mandel, &
above. It was equal to (4.9 ± 0.4) × 10− 12 and (6.7 ± 0.3) × 10− 12 m2 Reynaers, 1991).
s− 1 at the ionic strength of 10− 4 and 0.15 M, respectively (see Table 2). It
should be mentioned, however, that the precision of these measure­ D0 = D(1 + kD cb )− 1
(4)
ments decreases for ionic strength below 10− 3 M due to volume exclu­
where: D is the diffusion coefficient determined by DLS, D0 is the cor­
sion effects enhanced by the long-range electrostatic interactions among
rected diffusion coefficient and kD is the constant equal to 4.3 × 10− 4 L
charged KC molecules. In order to account for this effect, the primary
mg− 1.
experimental data obtained for the KC bulk concentration of 300–500
The corrected values are listed in Table 2. Interestingly, for 0.15 M

5
A. Michna et al. Food Hydrocolloids 121 (2021) 107033

Table 1 coefficient D data one can determine the electrokinetic charge at the KC
Primary physicochemical characteristics of the KC molecule. molecule applying the Lorentz–Stokes relationship (Adamczyk, Bratek,
Quantity [unit], symbol Value Remarks Jachimska, Jasiński, & Warszyński, 2006); (Michna, Adamczyk, Sofiń­
ska, & Matusik, 2017):
Monomer molar mass [kg 0.385 fully ionized chain
mol − 1], M1 0.408 sodium salt kT
Monomer contour length 0.90 ± this work, MD modeling qe = μ = 3πηdH μe (5)
D e
[nm], lm 0.03
Molecule density [kg m− 3], 2.24 ± 0.2 this work, MD modeling, Consequently, the number of elementary charges Nc per one KC
ρp × 103 no hydration molecule can be calculated as:
2.10 ± 0.2 this work, MD modeling,
/
× 103 contributing mass of Na+ kT
2.01 ± 0.2 this work, MD modeling, contributing Nc = qe e = 6.25 × 1010 μe (6)
D
× 103 mass of Na+, with hydration
Monomer volume [nm3], ν1 0.286 no hydration, calculated as v1 = M1 /
(ρp Av) where e is the elementary charge (equal unity), D is expressed in m2 s− 1,
0.305 contributing mass of Na + kT is expressed in J (kg m2 s− 2) and μe is expressed in μm cm (V s)− 1.
0.318 contributing mass of Na+, with Eq. (6) is valid for an arbitrary charge distribution and the shape of
hydration molecules. However, its accuracy decreases for larger ionic strengths
Monomer equivalent 0.64 ± no hydration, calculated as where the double-layer thickness κ− 1 = (εkT/2e2I)1/2 (where ε is the
cylinder diameter [nm], 0.04 (4v1 /πlm )1/2 electric permittivity of the solvent, and I is the ionic strength) becomes
dc
0.66 ± contributing mass of Na+ comparable with the molecule diameter.
0.03
It should be emphasized that the electrokinetic charge which is
0.67 ± contributing mass of Na+, with
0.03 hydration located below the flow shear plane (Hunter, 1981), is an important
parameter controlling macromolecule interactions among themselves, i.
* hydration for KC: 0.5 H2O per residue.
e., their solution stability and their interactions with interfaces, i.e.,
their adsorption capacity.
NaCl the corrected value of the diffusion coefficient is equal to (5.6 ± Using the experimental diffusion coefficient and the electrophoretic
0.3) × 10− 12 m2 s− 1, which matches the value obtained by Slootmaekers mobility data one can calculate from Eq. (6) that KC molecule charge
et al. (Slootmaekers, Mandel, & Reynaers, 1991), albeit for the LC
sample of an average molar mass of 607 kg mol− 1 and ionic strength of
0.1 M.
As previously discussed (Adamczyk et al., 2018) the increase of the
diffusion coefficient with the supporting electrolyte concentration is
caused by the change in the macromolecule conformation, which as­
sumes a less extended form.
In order to determine the effective (compensated) charge of KC
molecules in NaCl solutions, the electrokinetic method exploiting the
LDV technique (Ohshima, 2012) was applied. This method yields the
absolute values of the ratio of the molecule migration velocity to the
applied electric field, defined as the electrophoretic mobility and
denoted byμe . The dependence of the electrophoretic mobility on the
NaCl concentration is shown in Fig. 4 and these data are also collected in
Table 2. As can be noticed, the mobility increases from − 6.3 to − 2.9 μm
cm (V s)− 1 for the NaCl concentration (ionic strength) equal to 10− 4 and
0.15 M, respectively (pH 5.8). These electrophoretic mobility values
correspond to the zeta potential of the molecule (calculated from the
Debye-Hückel-Henry model) equal to − 120 and − 56 mV, respectively.
Negative values of the electrophoretic mobility and the zeta potential
confirm that the electrokinetic charge of the KC molecule is negative for Fig. 4. The dependence of the electrophoretic mobility of the KC molecule
this range of NaCl concentrations. (bulk concentration of 500 mg L− 1) on the NaCl concentration, pH 5.8, the solid
Using the experimental electrophoretic mobilityμe and the diffusion line is the guide for the eyes.

Table 2
Experimental characteristics of KC solutions at pH 5.8 in NaCl electrolyte of various concentrations [M], T = 298 K.
1
cNaCl [M] κ− [nm] D [m2 s− 1] D0* [m2 s− 1] dH [nm] dH0 [nm] dH0*** [nm] μe [μm cm (Vs)− 1] ζ [mV] Nc [1] α** [1]
− 12 − 12
10–4
30.5 4.9 × 10 4.0 × 10 100 ± 20 120 ± 20 _ − 6.3 ± 0.3 − 120 ± 10 − 400 ± 80 0.34
12 12
10–3 9.63 5.6 × 10− 4.6 × 10− 88 ± 20 106 ± 20 110 ± 20 − 5.2 ± 0.3 − 100 ± 12 − 290 ± 60 0.25
12 12
10–2 3.05 5.8 × 10− 4.8 × 10− 85 ± 20 102 ± 20 110 ± 20 − 4.6 ± 0.3 − 89 ± 12 − 250 ± 70 0.21
12 12
0.05 1.36 5.9 × 10− 4.9 × 10− 83 ± 20 100 ± 20 _ − 4.4 ± 0.1 − 89 ± 6 − 230 ± 70 0.20
12 12
0.15 0.786 6.7 × 10− 5.6 × 10− 73 ± 20 88 ± 20 80 ± 10 − 2.9 ± 0.2 − 56 ± 6 − 130 ± 30 0.11
− 1
D0 = D(1 + kD cb ) ;
− 4 − 1
* kD = 4.3 × 10 L mg ;
dH = kT/3πηD;
dH0 = kT/3πηD0 ; T = 298K
kT kT
α = Nc /Nm ; Nc = μ = 6.25 × 1010 μe , Nm = Mp /M1
** D0 e e D0

***determined from OWLS measuremetns.

6
A. Michna et al.
varies between − 400 and − 130 for NaCl concentration varying between
10− 4 and 0.15 M, respectively, see Table 2. Given that the number of
monomers in the KC molecule Nm = Mp/M1 is equal to 1160, its nominal
charge stemming from the dissociation of ionogenic groups should be
equal to − 1160 e. This comparison indicates that the electrokinetic
charge is only a small fraction of the nominal molecule charge, i.e., the
effective dissociation degree is equal to 34 and 11% for NaCl concen­
tration equal to 10− 4 and 0.15 M, respectively. Physically, such a sig­
nificant charge compensation is the result of the counterion
accumulation in the diffuse part of the electric double-layer caused by
the large electrostatic field generated by the presence of the surface
charges. In the macroion oriented literature, it is referred to as the
Manning ion condensation (Manning, 1979). It is also interesting to
mention that such behaviour was previously reported for PDADMAC
(Adamczyk, Jamroży, Batys, & Michna, 2014), and for PLL (Adamczyk
et al., 2018) macroions.
In the further analysis of the KC molecule conformations is useful to
define the hydrodynamic diameter, which in contrast to diffusion co­
efficient, is independent of the temperature and the solvent viscosity. It
can be directly calculated using the Stokes-Einstein relationship (Ein­
stein, 1908):
dH =
kT
(7)
3πηD
where η is the dynamic viscosity of the solvent.
For the NaCl concentration of 10− 4 and 0.15 M one obtains from Eq.
(7) the value of dH (calculated using the corrected diffusion coefficients
listed in Table 2) equals to 120 ± 20 and 88 ± 20 nm, respectively (see
Table 2). One can notice that the dH values obtained from the OWLS
measurements for the NaCl concentration range of 10− 3 to 0.15 M agree
with the data derived from DLS. The method for calculating dH using the
OWLS measurements is described in the Supplementary Material.
It is interesting to compare hydrodynamic diameter derived from
experiments with the theoretical model pertinent to slender body hy­
drodynamics characterized by the length to width (aspect ratio)
parameter, denoted by λ, much larger than unity (Brenner, 1974). For
such a case the hydrodynamic diameter can be expressed in the
following form (Mansfield & Douglas, 2008); (Adamczyk et al., 2012):
Le λ (where ηe is the supporting electrolyte viscosity) on the KC volume
d H0 = = dc = f1 (λ) (8)
(c1 ln 2 λ + c2 ) c1 ln 2 λ + c2
where Le can be treated as the contour length, and c1, c2 are the
dimensionless constants depending on the shape of the body.
It should be mentioned that Eq. (8) is valid for monodisperse mac­
roion solutions.
For prolate spheroids one has c1 = 1, c2 = 0; for blunt cylinders: c1 =
1, c2 = − 0.11 (Brenner, 1974); for linear chain of touching beads c1 = 1,
c2 = − 0.25, for the chain of beads forming a semi-circles: c1 = 0.95, c2 =
0.02 (Adamczyk et al., 2006).
In our case Le is equal to 1040 nm, which is calculated considering
that the number of monomers is equal to 1160 and that the length of the
monomer derived from MD modeling is equal to 0.90 nm. This value
corresponds to the limiting length of the KC molecule in a fully expanded
state. Additionally, considering the chain diameters given in Table 1 one
obtains λ = 1460, for dc = 0.71. In consequence, one can calculate from
Eq. (8) that the hydrodynamic diameter is equal to 130 nm, for the cy­
lindrical molecule shape. Interestingly, for the linear touching bead
shape, one obtains almost identical values of 133 nm. As can be seen,
these values agree within experimental error bounds with experimental
hydrodynamic diameter in the limit of low ionic strength, which was
equal to 120 ± 20 nm (see Table 2).
It is interesting to estimate the influence of the dispersity of the
molecule molar mass distribution on the precision of the hydrodynamic
diameter determination. As shown in (Adamczyk et al., 2018) the
correction to the hydrodynamic diameter is given by:
7
Food Hydrocolloids 121 (2021) 107033
( )− 1
dH0 = dH 1 + CH σ 2M (9)
where:dH0 is the hydrodynamic diameter for a monodisperse system, dH
is the experimental value of the hydrodynamic diameter for a disperse
system exhibiting the same average molar mass, σ M is the relative
standard deviation of the size distribution and CH is the dimensionless
constant given by
0.5 c21
CH = − c 1 + (10)
c1 ln 2 λ + c2 (c1 ln 2 λ + c2 )2
For our case taking λ = 1460, c1 = 1, and c2 = − 0.11 (this corre­
sponds to cylindrical shape of the molecule) one obtains CH = − 0.05.
Thus, even for a disperse system where σM = 0.5, the correction is less
than 1%, which is practically negligible compared to other experimental
errors.
However, given the low sensitivity of the hydrodynamic diameter to
the aspect ratio parameter predicted by Eq. (8) a more reliable infor­
mation about the KC conformation can be derived from the intrinsic
viscosity data obtained via the dynamic viscosity measurements.
3.3. Dynamic viscosity measurements
A proper interpretation of the viscosity measurements in terms of the
volume fraction requires the density of the KC molecule to be known.
This parameter was experimentally determined by the solution dilution
method as described in the Supplementary Material. At the temperature
of 298 K, the density was equal to ρp = (2.03 ± 0.13) × 103 kg m− 3 for
the NaCl concentration of 10− 4 - 10− 2 M that agrees with error bounds
with previously reported by Slootmaekers et al. (Slootmaekers, Mandel,
& Reynaers, 1991) and the above theoretical value of (2.01 ± 0.2) × 103
kg m− 3.
Initially, in these measurements, the dependencies of the dynamic
viscosity of dilute KC solutions (characterized by bulk concentration cb
up to 1000 mg L− 1) was measured using the above-described method.
The zero shear viscosity of the solution denoted by ηs was calculated by
an efficient extrapolation procedure. These primary experimental results
were expressed as the dependence of the normalized viscosity ηs/ηe
fraction Φv = cb/ρp for various NaCl concentrations in the range 10− 4 to
0.15 M. The isotonic electrolyte solutions required for the lower NaCl
concentration were prepared considering the above-estimated KC
molecule ionization degree using the following formula:
/
1
I * = × 10− 3 Nc cb Mp (11)
2
where I* is the excess ionic strength due to the KC molecule ionization
and cb is expressed in mg L− 1.
One can calculate from Eq. (11) that for cb = 100 mg L− 1 and Nc
equal to − 400, I* = 4 × 10− 5 M.
The dependencies of the relative viscosity, ηs/ηe on the volume
fraction Φv of KC molecules acquired for various NaCl concentrations are
presented in Fig. 5. The slopes of these dependencies in the limit of low
volume fractions give directly the intrinsic viscosity [η], which varied
between (2.2 ± 0.2) × 103 and (5.0 ± 0.3) × 104 for the NaCl concen­
tration of 0.15 and 10− 4 M, respectively (at pH = 5.6), see Table 3. It
should be mentioned that the intrinsic viscosity for the PBS buffer so­
lutions (ionic strength 0.15 M, pH 7.4) is equal to that determined for
pure 0.15 M NaCl solutions.
It is also interesting to mention that the value of the intrinsic vis­
cosity for 0.15 M NaCl obtained in this work is comparable with that
reported by (Almutairi et al., 2013) who determined 1080 mL g− 1 for
λ-carrageenan of the average molar mass of 870 kg mol− 1 and NaCl
concentration of 0.15 M. This corresponds to the dimensionless value of
the intrinsic viscosity used in our work [η] = 2160.
A. Michna et al. Food Hydrocolloids 121 (2021) 107033

Fig. 5. a) The dependence of the relative viscosity ηs/ηe on the volume fraction of KC Φv, determined for various concentrations of NaCl. Hollow squares show the
results obtained for 0.15 M solution of the PBS buffer (pH 7.4). The solid line denotes a linear fit of experimental data. b) Dependence of ηi/Φv (where ηi = ηs/ηe) on
the volume fraction of KC in its solutions, Φv. Dashed lines denote the linear fit of experimental data.

Given that the obtained intrinsic viscosity values exceed by orders of where c1v = 3/15, c2v = 1/15 and cv is equal to 14/15 for blunt cylinders.
magnitude the Einstein results for spherically shaped macromolecules Eq. (12) is valid for λ ≫ 1 and for monodisperse macroion solutions.
where [η] = 2.5, one can assume that KC molecules assume extended As shown in (Adamczyk et al., 2018), using Eq. (12) the following for­
conformation for the entire NaCl concentration range, which corre­ mula can be derived for converting the experimental intrinsic viscosity
sponds to the slender body regime. Therefore, the experimental data can data obtained for disperse system to the corresponding values pertinent
be quantitatively interpreted exploiting the theoretical results pertinent to monodisperse system:
to the zero shear rate intrinsic viscosity for slender bodies (Brenner,
( )− 1
1974): [η]0 = [η] 1 + Cv σ 2M (13)
λ2 λ2
[η]* = c1 v + c2v + cv (12)
ln 2 λ − 0.5 ln 2 λ − 1.5 where [η]0 is the corrected intrinsic viscosity for the monodisperse sys­
tem and the dimensionless Cv constant is given by

8
A. Michna et al. Food Hydrocolloids 121 (2021) 107033

Table 3 pertinent to the extended chain derived from MD modeling. For ionic
The intrinsic viscosity [η], the λ parameter, the equivalent cylinder chain strength 0.15 M, the predicted chain length of KC molecule is equal to
diameter dc, the cylinder lengths Lc and the cross-section area in the side-on 520 nm, which confirms its largely elongated shape. One can argue that
orientation Sc of the KC molecules for various ionic strengths derived from these data have significance for interpreting KC molecule adsorption,
DLS and viscometry. especially for producing macroion films in the layer-by-layer processes.
cNaCl [η][1] λ [1] dc [nm] Lc [nm] Sc [nm2]
[M]
4. Conclusions
0 7.9 × 104* 1460* 0.71 1040* 740
10–4 5.0 ± 0.4 × 1140 ± 0.81 ± 920 ± 750 ± 150
Physicochemical properties of κ–carrageenan (KC) were thoroughly
104 50 0.15 200
10–3 2.4 ± 0.2 × 760 ± 40 1.0 ± 0.2 760 ± 760 ± 150 determined by molecular dynamics (MD) modeling, the SEC-MALS,
104 150 FTIR, DLS, OWLS, LDV techniques and the dynamic viscosity measure­
10–2 1.0 ± 0.1 × 520 ± 30 1.3 ± 0.25 690 ± 970 ± 200 ments. The modeling confirmed that the KC molecule exhibits a flexible-
104 150 rod shape with no sharp bending. There appear intramolecular hydrogen
0.15 2.2 ± 0.2 × 210 ± 10 2.5 ± 0.4 520 ± 70 1300 ±
bonding between –SO3 and –OH groups within the monomer. No ten­
103 250
dency to the formation of a large-scale helical conformation was pre­
*Calculated from the MD modeling using the monomer length of 0.90 nm. dicted. Additionally, several parameters of primary significance were
determined such as the monomer length equal to 0.90 nm and the chain
3 β21 + 3β22 β31 + 3β32 diameter equal to 0.71 nm. This enabled to calculate the maximum
Cv = 1 − + (14)
2 β1 + 3β2 β1 + 3β2 extended (contour) length of a KC molecule of arbitrary molar mass.
From experimental measurements, the diffusion coefficient, the hy­
where β1 = ln 2λ−1 0.5β2 = ln 2λ−1 1.5. drodynamic diameter and the electrophoretic mobility of molecules
From Eq. (14) one can calculate that for λ = 1460 pertinent to hy­ were determined for the NaCl concentration of 10− 4 to 0.15 M (at pH 5.6
drated and fully extended chain, Cv = 0.85, which indicates that for and 7.4). This allowed to calculate the electrokinetic charge of KC
macroion molar mass distribution characterized by σM = 0.1 the molecules, which was considerably smaller (in absolute terms) than the
correction is below 1%. However, for samples of large dispersity typical nominal charge calculated assuming full dissociation of ionogenic
to LC (Slootmaekers, Mandel, & Reynaers, 1991) where σ M = 0.3 the groups. This resulted in the low effective ionization degree of the
correction is equal from 7 to 10%. molecule amounting to 31 and 11% for NaCl concentration range of
Using these data, one can calculate the aspect ratio parameter of an 10− 4 to 0.15 M, respectively.
equivalent cylinder by a numerical inversion of Eq. (12) Additionally, the viscometric measurements for dilute KC solutions
( ) confirmed that the intrinsic viscosity attains large values between (2.2
λ = fv− 1 [η]0 (15) ± 0.1) × 103 and (5.0 ± 0.3) × 104 for the NaCl concentration of 0.15
One can also calculate λ using the iterative scheme, which produces and 10− 4 M, respectively, which confirmed that the molecules assume
the following formula (Adamczyk et al., 2018) very extended conformations. Using the intrinsic viscosity and the hy­
( ) drodynamic diameter experimental data the equivalent cylinder diam­
λ=
15[η]0 1
2 (16) eter, the length of the KC chain and the geometrical cross-section area
f1 (λ1 ) were explicitly calculated. For NaCl concentration of 10− 4 M the
experimental equivalent cylinder lengths was equal to 920 nm. For NaCl
where concentration of 0.15 M, the experimental chain length of KC molecule
3 1 was still vary large and equal to 520 nm. These data unequivocally
f1 (λ1 ) = + (17) confirm that the KC molecule behaviour in electrolyte solutions can be
(ln 2λ1 − 0.5) (ln 2λ1 − 1.5)
adequately described in terms of the slender body hydrodynamic model.
( )12 One can argue that these newly acquired data have significance for
andλ1 = 15[η]0 . The precision of Eq. (17) for is ca. 1% for the
predicting and properly interpreting carrageenian molecule thickening
intrinsic viscosity exceeding 100. properties and for producing macroion films in the layer-by-layer pro­
Using Eq. (16) one can calculate that the λ parameter for KC mole­ cesses at various substrates, comprising carrier nano- and
cules and NaCl concentration of 10− 4 M is equal to 1140, which ap­ microparticles.
proaches the value pertinent to the fully extended chain of 1460 derived
from MD modeling. For NaCl concentration of 0.15 M, the λ parameter
significantly decreases to 210. These results unequivocally confirm that Declaration of competing interest
for this broad range of NaCl concentration the KC molecules exhibit an
extended conformation, hence the assumption of the slender body The authors declare no conflict of interest.
regime is fulfilled.
Additionally, knowing λ one can calculate the equivalent cylinder Acknowledgments
diameter of the KC chain using Eq. (8) transformed to the form
c1 ln 2 λ + c2 This work was financially supported by the National Science Centre,
dc = dH0 (18) Poland, Opus Project, UMO-2018/31/B/ST8/03277.
λ

where dH0 is previously determined hydrodynamic diameter of KC References

molecules listed in Table 2.
Consequently, the equivalent cylinder length can be calculated as Lc
= λdc and the geometrical cross-section area in the side-on orientation is
given by Sc = dc Lc.
Values of dc and Lc calculated in this way are given in Table 3. It is
interesting to mention that the equivalent cylinder lengths for NaCl
concentration of 10− 4 M is equal to 920 nm, which approaches the value
Abd Karim, A., Sulebele, G. A., Azhar, M. E., & Ping, C. Y. (1999). Effect of carrageenan
on yield and properties of tofu. Food Chemistry, 66(2), 159–165. https://doi.org/
10.1016/S0308-8146(98)00258-1
Abraham, M. J., Murtola, T., Schulz, R., Páll, S., Smith, J. C., Hess, B., et al. (2015).
GROMACS : High performance molecular simulations through multi-level
parallelism from laptops to supercomputers. SoftwareX, 1–2, 19–25. https://doi.org/
10.1016/j.softx.2015.06.001
Adamczyk, Z., Bratek, A., Jachimska, B., Jasiński, T., & Warszyński, P. (2006). Structure
of poly(acrylic acid) in electrolyte solutions determined from simulations and

9
A. Michna et al.
viscosity measurements Z. Journal of Physical Chemistry B, 110(45), 22426–22435.
https://doi.org/10.1021/jp063981w
Adamczyk, Z., Cichocki, B., Ekiel-Jezewska, M. L., Słowicka, A., Wajnryb, E., &
Wasilewska, M. (2012). Fibrinogen conformations and charge in electrolyte
solutions derived from DLS and dynamic viscosity measurements. Journal of Colloid
and Interface Science, 385(1), 244–257. https://doi.org/10.1016/j.jcis.2012.07.010
Adamczyk, Z., Jamroży, K., Batys, P., & Michna, A. (2014). Influence of ionic strength on
poly(diallyldimethylammonium chloride) macromolecule conformations in
electrolyte solutions. Journal of Colloid and Interface Science, 435, 182–190. https://
doi.org/10.1016/j.jcis.2014.07.037
Adamczyk, Z., Morga, M., Kosior, D., & Batys, P. (2018). Conformations of poly- l -lysine
molecules in electrolyte solutions: Modeling and experimental measurements.
Journal of Physical Chemistry C, 122(40), 23180–23190. https://doi.org/10.1021/
acs.jpcc.8b07606
Alba, K., & Kontogiorgos, V. (2019). Seaweed polysaccharides ( agar , alginate
carrageenan ). In L. Melton, F. Shahidi, & P. Varelis (Eds.), Encyclopedia of food
chemistry (pp. 240–250). Academic Press. https://doi.org/10.1016/B978-0-08-
100596-5.21587-4.
Almutairi, F. M., Adams, G. G., Kök, M. S., Lawson, C. J., Gahler, R., Wood, S., et al.
(2013). An analytical ultracentrifugation based study on the conformation of lambda
carrageenan in aqueous solution. Carbohydrate Polymers, 97(1), 203–209. https://
doi.org/10.1016/j.carbpol.2013.04.027
Berth, G., Vukovic, J., & Lechner, M. D. (2008). Physicochemical characterization of
Carrageenans— A critical reinvestigation. Journal of Applied Polymer Science, 110,
3508–3524. https://doi.org/10.1002/app.28937
Bonferoni, M. C., Rossi, S., Tamayo, M., Pedraz, J. L., Dominguez-Gil, A., & Caramella, C.
(1993). On the employment of λ-carrageenan in a matrix system . I . Sensitivity to
dissolution medium and comparison with Na carboxymethylcellulose and xanthan
gum. Journal of Controlled Release, 26, 119–127.
Bonferoni, M. C., Rossi, S., Tamayo, M., Pedraz, J. L., Dominguez-Gil, A., & Caramella, C.
(1994). On the employment of λ-carrageenan in a matrix system . II . λ-Carrageenan
and hydroxypropylmethylcellulose mixtures. Journal of Controlled Release, 30,
175–182.
Bono, A., Anisuzzaman, S. M., & Ding, O. W. (2014). Effect of process conditions on the
gel viscosity and gel strength of semi-refined carrageenan (SRC) produced from
seaweed (Kappaphycus alvarezii). Journal of King Saud University - Engineering
Sciences, 26(1), 3–9. https://doi.org/10.1016/j.jksues.2012.06.001
Brenner, H. (1974). Rheology of a dilute suspension of axisymmetric Brownian particles.
International Journal of Multiphase Flow, 1(2), 195–341. https://doi.org/10.1016/
0301-9322(74)90018-4
Briones, A. V., Sato, T., & Bigol, U. G. (2014). Antibacterial activity of polyethylenimine/
carrageenan multilayer against pathogenic bacteria. Advances in Chemical Engineering
and Science, 4(2), 233–241. https://doi.org/10.4236/aces.2014.42026
Bussi, G., Donadio, D., & Parrinello, M. (2007). Canonical sampling through velocity
rescaling. The Journal of Chemical Physics, 126. https://doi.org/10.1063/1.2408420,
014101.
Chronakis, I. S., Doublier, J., & Piculell, L. (2000). Viscoelastic properties for kappa- and
iota-carrageenan in aqueous NaI from the liquid-like to the solid-like behaviour.
International Journal of Biological Macromolecules, 28(1), 1–14. https://doi.org/
10.1016/S0141-8130(00)00141-0
Darden, T., York, D., & Pedersen, L. (1993). Particle mesh Ewald : An N ⋅ log ( N ) method
for Ewald sums in large systems in large systems. The Journal of Chemical Physics, 98,
10089. https://doi.org/10.1063/1.464397
Dul, M., Paluch, K. J., Kelly, H., Healy, A. M., Sasse, A., & Tajber, L. (2015). Self-
assembled carrageenan/protamine polyelectrolyte nanoplexes-investigation of
critical parameters governing their formation and characteristics. Carbohydrate
Polymers, 123, 339–349. https://doi.org/10.1016/j.carbpol.2015.01.066
Einstein, A. (1908). Elementare theorie der brownschen 1) bewegung. Zeitschrift Für
Elektrochemie Und Angewandte Physikalische Chemie, 14(17), 235–239. https://doi.
org/10.1002/bbpc.19080141703
Gonzalez, M. E., Alarcón, B., & Carrasco, L. (1987). Polysaccharides as antiviral Agents :
Antiviral activity of carrageenan. Antimicrobial Agents and Chemotherapy, 31(9),
1388–1393.
Guvench, O., Greene, S. N., Kamath, G., Brady, J. W., Venable, R. M., Pastor, R. W., et al.
(2008). Additive empirical force field for hexopyranose monosaccharides. Journal of
Computational Chemistry, 29(15), 2543–2564. https://doi.org/10.1002/jcc.21004
Guvench, O., Mallajosyula, S. S., Raman, E. P., Hatcher, E., Vanommeslaeghe, K.,
Foster, T. J., et al. (2011). CHARMM additive all-atom force field for carbohydrate
derivatives and its utility in polysaccharide and carbohydrate À protein modeling.
Journal of Chemical Theory and Computation, 7(10), 3162–3180. https://doi.org/
10.1021/ct200328p
Harding, S. E. (2017). In An introduction to polysaccharide biotechnology (2nd ed.). CRC
Press. https://doi.org/10.1016/s0300-9084(98)80079-5.
Hariharan, M., Wheatley, T. A., & Price, J. C. (1997). Controlled-release tablet matrices
from Carrageenans : Compression and dissolution studies. Pharmaceutical
Development and Technology, 2(4), 383–393.
Hess, B. (2008). P-LINCS : A parallel linear constraint solver for molecular simulation.
Journal of Chemical Theory and Computation, 4(1), 116–122. https://doi.org/
10.1021/ct700200b
Hess, B., Bekker, H., Berendsen, H. J. C., & Fraaije, J. G. E. M. (1997). LINCS : A linear
constraint solver for molecular simulations. Journal Of Computational Chemistry, 18
(12), 1463–1472. https://doi.org/10.1002/(SICI)1096-987X(199709)18.
:12<1463::AID-JCC4>3.0.CO;2-H.
Hockney, R. W. (1970). The potential calculation and some applications. Methods in
Computational Physics, 9, 135–211.
10
Food Hydrocolloids 121 (2021) 107033
Hunter, R. J. (1981). In R. Ottewill, & R. Rowell (Eds.), Zeta potential in colloid science.
Principles and applications. Academic Press.
Imeson, A. P. (2009). Carrageenan and furcellaran. In Handbook of hydrocolloids.
Woodhead Publishing Limited. https://doi.org/10.1533/9781845695873.164.
Lee, J., Cheng, X., Swails, J. M., Yeom, M. S., Eastman, P. K., Lemkul, J. A., et al. (2016).
CHARMM-GUI input generator for NAMD, GROMACS, AMBER, OpenMM, and
CHARMM/OpenMM simulations using the CHARMM36 additive force field. Journal
of Chemical Theory and Computation, 12(1), 405–413. https://doi.org/10.1021/acs.
jctc.5b00935
Li, L., Ni, R., Shao, Y., & Mao, S. (2014). Carrageenan and its applications in drug
delivery. Carbohydrate Polymers, 103, 1–11. https://doi.org/10.1016/j.
carbpol.2013.12.008
Luo, M., Shao, B., Nie, W., Wei, X. W., Li, Y. L., Wang, B. L., et al. (2015). Antitumor and
adjuvant activity of λ-carrageenan by stimulating immune response in cancer
immunotherapy. Scientific Reports, 5, 11062. https://doi.org/10.1038/srep11062
Manning, G. S. (1979). Counterion binding in polyelectrolyte theory. Accounts of
Chemical Research, 12(12), 443–449. https://doi.org/10.1021/ar50144a004
Mansfield, M. L., & Douglas, J. F. (2008). Transport properties of rodlike particles.
Macromolecules, 41(14), 5422–5432. https://doi.org/10.1021/ma702839w
Michna, A., Adamczyk, Z., Sofińska, K., & Matusik, K. (2017). Monolayers of poly(amido
amine) dendrimers on mica – in situ streaming potential measurements. Journal of
Colloid and Interface Science, 485, 232–241. https://doi.org/10.1016/j.
jcis.2016.09.007
Michna, A., Pomorska, A., Nattich-Rak, M., Wasilewska, M., & Adamczyk, Z. (2020).
Hydrodynamic solvation of poly(amido amine) dendrimer monolayers on silica.
Journal of Physical Chemistry C, 124(32), 17684–17695. https://doi.org/10.1021/
acs.jpcc.0c04638
Miyamoto, S., & Kollman, P. A. (1992). Settle: An analytical version of the SHAKE and
RATTLE algorithm for rigid water models. Journal of Computational Chemistry, 13(8),
952–962. https://doi.org/10.1002/jcc.540130805
Montolalu, R. I., Tashiro, Y., Matsukawa, S., & Ogawa, H. (2008). Effects of extraction
parameters on gel properties of carrageenan from Kappaphycus alvarezii
(Rhodophyta). Journal of Applied Phycology, 20(5), 521–526. https://doi.org/
10.1007/s10811-007-9284-2
Nakashima, H., Kido, Y., Kobayashi, N., Motoki, Y., Neushul, M., & Yamamoto, N.
(1987). Purification and characterization of an avian myeloblastosis and human
immunodeficiency virus reverse transcriptase inhibitor , sulfated polysaccharides
extracted from sea algae. Antimicrobial Agents and Chemotherapy, 31(10), 1524–1528.
Ohshima, H. (Ed.). (2012). Electrical phenomena at interfaces and biointerfaces:
Fundamentals and applications in nano-, bio-, and environmental sciences. John Wiley &
Sons, Inc. https://doi.org/10.1002/9781118135440.
Park, S., Lee, J., Qi, Y., Kern, N. R., Lee, H. S., Jo, S., et al. (2019). CHARMM-GUI Glycan
Modeler for modeling and simulation of carbohydrates and glycoconjugates.
Glycobiology, 29(4), 320–331. https://doi.org/10.1093/glycob/cwz003
Parrinello, M., & Rahman, A. (1981). Polymorphic transitions in single crystals: A new
molecular dynamics method. Journal of Applied Physics, 52(12), 7182–7190. https://
doi.org/10.1063/1.328693
Pereira, L., & Critchley, A. T. (2020). The COVID 19 novel coronavirus pandemic 2020:
Seaweeds to the rescue? Why does substantial, supporting research about the
antiviral properties of seaweed polysaccharides seem to go unrecognized by the
pharmaceutical community in these desperate times? Journal of Applied Phycology,
32(3), 1875–1877. https://doi.org/10.1007/s10811-020-02143-y
Reis, R. L., Neves, N., Mano, J. F., Gomes, M. E., Marques, A. P., & Azevedo, H. S. (Eds.).
(2008). Natural-based polymers for biomedical applications (1st ed.). CRC Press.
Rodríguez, A., Kleinbeck, K., Mizenina, O., Kizima, L., Levendosky, K., Jean-Pierre, N.,
et al. (2014). In vitro and in vivo evaluation of two carrageenan-based formulations
to prevent HPV acquisition. Antiviral Research, 108, 88–93. https://doi.org/10.1016/
j.antiviral.2014.05.018
Roy, I., & Gupta, M. N. (2003). Smart polymeric Materials : Emerging biochemical
applications. Chemistry & Biology, 10, 1161–1171. https://doi.org/10.1016/j.
chembiol .2003.12.004
Saha, D., & Bhattacharya, S. (2010). Hydrocolloids as thickening and gelling agents in
food: A critical review. Journal of Food Science & Technology, 47(6), 587–597.
https://doi.org/10.1007/s13197-010-0162-6
Schuck, P., Gillis, R. B., Besong, T. M. D., Almutairi, F., Adams, G. G., Rowe, A. J., et al.
(2014). SEDFIT-MSTAR: Molecular weight and molecular weight distribution
analysis of polymers by sedimentation equilibrium in the ultracentrifuge. Analyst,
139, 79–92. https://doi.org/10.1039/c3an01507f
Slootmaekers, D., Mandel, M., & Reynaers, H. (1991). Dynamic light scattering by κ- and
λ-carrageenan solutions. International Journal of Biological Macromolecules, 13(1),
17–25. https://doi.org/10.1016/0141-8130(91)90005-F
Slootmaekers, D., van Dijk, J. A. P. P., Varkevisser, F. A., van Treslong, C. J. B., &
Reynaers, H. (1991). Molecular characterisation of κ- and λ-carrageenan by gel
permeation chromatography, light scattering, sedimentation analysis and
osmometry. Biophysical Chemistry, 41(1), 51–59. https://doi.org/10.1016/0301-
4622(91)87209-N
Stechemesser, S., & Eimer, W. (1997). Solvent-dependent swelling of poly(amido amine)
starburst dendrimers. Macromolecules, 30(7), 2204–2206. https://doi.org/10.1021/
ma9614914
Thành, T. T. T., Yuguchi, Y., Mimura, M., Yasunaga, H., Takano, R., Urakawa, H., et al.
(2002). Molecular characteristics and gelling properties of the carrageenan family, 1:
Preparation of novel carrageenans and their dilute solution properties.
Macromolecular Chemistry and Physics, 203(1), 15–23. https://doi.org/10.1002/
1521-3935(20020101)203:1<15::AID-MACP15>3.0.CO. ;2-1.
Wasilewska, M., Adamczyk, Z., Pomorska, A., Nattich-Rak, M., & Sadowska, M. (2019).
Human serum albumin adsorption kinetics on silica: Influence of protein solution
A. Michna et al.
stability. Langmuir, 35(7), 2639–2648. https://doi.org/10.1021/acs.
langmuir.8b03266
Zabik, M. E., & Aldrich, P. J. (1965). The effect of selected anions of potassium salts on
the viscosities of lambda-carrageenan dispersions. Journal of Food Science, 30(1),
111–117. https://doi.org/10.1111/j.1365-2621.1965.tb00272.x
11
Food Hydrocolloids 121 (2021) 107033
Zabik, M. E., & Aldrich, P. J. (1967). The effect of cations on the viscosity of Lambda-
Carrageenan. Journal of Food Science, 32(1), 91–97. https://doi.org/10.1111/j.1365-
2621.1967.tb01966.x
Zhou, G., Sun, Y., Xin, H., Zhang, Y., Li, Z., & Xu, Z. (2004). In vivo antitumor and
immunomodulation activities of different molecular weight lambda-carrageenans
from Chondrus ocellatus. Pharmacological Research, 50(1), 47–53. https://doi.org/
10.1016/j.phrs.2003.12.002