Accepted Manuscript

Title: Surface tension and dilation rheology of DNA solutions

in mixtures with azobenzene-containing cationic surfactant

Author: N. Moradi Yuriy Zakrevskyy A. Javadi E.V.

Aksenenko V.B. Fainerman Nino Lomadze Svetlana Santer R.
Miller

PII: S0927-7757(16)30253-9
DOI: http://dx.doi.org/doi:10.1016/j.colsurfa.2016.04.021
Reference: COLSUA 20578

To appear in: Colloids and Surfaces A: Physicochem. Eng. Aspects

Received date: 25-8-2015

Revised date: 6-4-2016
Accepted date: 7-4-2016

Please cite this article as: {http://dx.doi.org/

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 Surface tension and dilation rheology of DNA solutions in mixtures with azobenzene-
containing cationic surfactant

N. Moradi1*, Yuriy Zakrevskyy2,3, A. Javadi1,4, E.V. Aksenenko5, V.B. Fainerman6,

Nino Lomadze2, Svetlana Santer2 and R. Miller1

1
MPI of Colloids and Interfaces, Potsdam, Germany
2
Experimental Physics, Institute of Physics and Astronomy, University of Potsdam, Germany
3
Institute of Physics, Cologne University of Applied Sciences, Germany
4
Chemical Engineering Department, University of Tehran, Tehran, Iran
5
Institute of Colloid Chemistry and Chemistry of Water, Kyiv (Kiev), Ukraine
6
Donetsk Medical University, Donetsk, Ukraine
Graphical Abstract

2
 Highlights
Surface tension and dilation rheology of DNA solutions in mixtures with azobenzene-
containing cationic surfactant
N. Moradi, Yuriy Zakrevskyy, A. Javadi, E.V. Aksenenko, V.B. Fainerman, Nino
Lomadze, Svetlana Santer and R. Miller

DNA in aqueous solution has no significant surface activity.

Due to its negative net charge DNA forms complexes with cationic surfactant such as
AzoTAB.
Mixed DNA+AzoTAB solutions show high surface activity
The surface activity of the complexes depends on the cationic surfactant concentration.

3
Abstract
The surface tension and dilational surface visco-elasticity of the individual solutions of the
biopolymer DNA and the azobenzene-containing cationic surfactant AzoTAB, as well as their
mixtures were measured using the drop profile analysis tensiometry. The negatively charged DNA
molecules form complexes with the cationic surfactant AzoTAB. Mixed DNA+AzoTAB solutions
exhibit high surface activity and surface layer elasticity. Extremes in the dependence of these
characteristics on the AzoTAB concentration exist within the concentration range of 3106 to
5105 M. The surface tension of the mixture shows a minimum with a subsequent maximum. In
the same concentration range the elasticity shows first a maximum and then a subsequent minimum.
A recently developed thermodynamic model was modified to account for the dependence of the
adsorption equilibrium constant of the adsorbed complex on the cationic surfactant concentration.
This modified theory shows good agreement with the experimental data both for the surface tension
and the elasticity values over the entire range of studied AzoTAB concentrations.

Keywords: Mixed adsorption layers, polymer/surfactant interaction, water/air interface,

thermodynamics of adsorption, dilational rheology, drop profile analysis tensiometry

* Corresponding author: Narges.Moradi@mpikg.mpg.de

4
1. Introduction
The interaction between polymers and surfactants is an interesting field of research. Their mixtures
are used in various modern technologies and medicine. Recently, there has been interest on complex
formation between DNA and different kind of surfactants and particularly cationic surfactants e.g.
CnTAB. The majority of these studies focus on investigating the bulk properties e.g. the binding
isotherms, mechanism of binding, structural transitions, thermodynamics and
compaction/decompaction of DNA using different spectroscopic and electrophoresis methods [1-6].
It has been discovered that the adsorption of DNA on different surfaces such as lipids, polypeptides
and phospholipids is very important in biological processes that lately became more applicable also
in medicine. Therefore, investigations of the DNA adsorption at different interfaces (liquid/air,
liquid/liquid or liquid/solid) became an interesting field of research for interface scientists.
It has been shown experimentally that DNA alone does not have a strong tendency to adsorb at
liquid interfaces [7]. It has been suggested that no measurable changes in surface pressure or surface
potential could be detected for DNA at the water/air interface [8]. This can be explained by the
special structure of DNA. DNA is a highly negatively charged molecule and the hydrophobic
groups are buried in the interior of the molecule and are in little contact with the solvent.
In an aqueous solution of DNA and cationic surfactants, complexes may form in the bulk and at the
interface. It has been demonstrated that solutions of DNA mixed with DTAB show a strong
reduction in surface tension [9, 10]. However, there are only very few studies on the behaviour of
mixed DNA+cationic surfactants at the solutions/air interface in literatures [9-12].
Recently, azobenzene-containing cationic surfactants e.g. AzoTAB are commonly used to make
complexes with DNA [13-16]. AzoTAB is a photosensitive molecule that upon exposure with UV
light, undergoes to a trans-cis isomerization. Under visible light, AzoTAB has trans conformation.
Trans isomer is highly hydrophobic and surface active, while the cis isomer which occurs under UV
light, is hydrophilic. This particular properties of AzoTAB was used in DNA+AzoTAB mixtures to
control DNA compaction (DNA condensation) by applying light. DNA/ AzoTAB mixtures were
studied in order to understand this process and eventually find a novel method for gene delivery and
gene regulation [13-16].
Controlled compaction of DNA by AzoTAB is mainly followed by the reduction of DNA charges.
Interestingly, it was demonstrated by Zakrevskyy and co-workers in [17-19] that a complete DNA
compaction with the AzoTAB occurs already at a 20% compensation of the DNA charges. This
work gave the motivation for us to study the interfacial behaviour of DNA/AzoTAB solutions
which to the best of our knowledge has not been investigated before.

5
The interaction between AzoTAB and DNA is still not fully understood, in particular additional
arguments are required to support the fact that a much lower amount of surfactant is required for the
compaction of DNA, i.e. already at 20% charge compensation. Therefore, attention is primarily paid
to the mechanism of complex formation of DNA with the cationic AzoTAB in the solution bulk and
at the solution/air surface. This should help to optimize the use of surfactant in compaction in
general, and controlling the completely reversible light-induced DNA compaction process by the
special light-sensitive surfactant AzoTAB.
There are several works trying to study the formation, interaction and structure of DNA+AzoTAB
complexes in the bulk [13-19]. In the presence of the cationic AzoTAB complexes are formed both
in the bulk and at the interface, but to the best of our knowledge there has not been made any
attempt of experimental or quantitative analysis on DNA+AzoTAB complexes at water/air
interface.
In this work, we try to understand the behaviour of DNA and AzoTAB at the solution/air surface by
measuring the surface tension of individual and mixed DNA/AzoTAB solutions. In addition the
surface dilational visco-elasticity of adsorbed DN/AzoTAB complexes is measured by using the
oscillating drop profile tensiometry at different perturbation frequencies. It is known that dilational
surface viscoelasticity data provide excellent insight into molecular interactions of polymers and
biopolymers such as proteins with surfactants in the bulk and essentially at the interface [20]. The
experimental results are analysed using a thermodynamic model, which was developed for
describing mixtures of ionic surfactants with proteins. The model is modified such that it takes into
account the dependence of the adsorption equilibrium constant of the adsorbed complex on the
cationic surfactant concentration.

2. Material and methods

In this work, calf thymus DNA, type I, containing 6% sodium salt, from Sigma (no. D1501) was
used. The molecular weight of the DNA is 10.8106 Da (16.4 kbp) determined using intrinsic
viscosity measurements [17]. The DNA was dissolved in MilliQ water in presence of 5 mM NaCl,
and its concentration was determined spectroscopically. Azobenzene containing
trimethylammonium bromide surfactant (C4-Azo-OC6TMAB) summarized as AzoTAB with the
molecular weight of 476.22 g/mol was synthesized according to the procedure described in Ref.
[17]. Fig. 1 shows the chemical structure of the surfactant.
The surfactant was dissolved in 5 mM aqueous NaCl. The concentration of the surfactant stock
solution was determined spectroscopically [17]. In all experiments the DNA monomer
concentration was fixed to 5105 M.

6
Fig. 1. Chemical structure of the surfactant C4-Azo-OC6TMAB

The surface tension of DNA, azobenzene surfactant and their mixed solutions at the air-solution
surface was measured using the drop profile analysis tensiometer PAT-1 (SINTERFACE
Technologies, Germany) [20]. The instrument consists mainly of a capillary to form the drop, a
video camera with an objective and a frame grabber to transfer the drop image into a PC. The drop
profile analysis tensiometer can also be applied for studies of the surface dilational rheology of
interfacial layers. For this purpose, a drop is subjected to harmonic volume oscillations and the
resulting area changes lead to compressions and expansions of the adsorbed layer at the drop
surface. The surface dilational modulus is defined by an expression originally proposed by Gibbs as
the increase in surface tension due to a small increase of surface area: E = d/dlnA, where is the
surface tension, and A is the drop surface area. After equilibration, harmonic oscillations of the
surface area with an amplitude of 6% and at frequencies in the range between 0.01 and 0.2 Hz
were generated. For amplitudes less than 10% the measured rheological quantities are within the
linear rheological regime, i.e. the parameters are independent of the oscillation amplitude. The
visco-elasticity is a complex quantity E = Er + iEi, where Er and Ei are its real (elasticity) and
imaginary (viscosity) components, respectively. The phase angle between stress (d) and strain
(dA) is defined as = arctg (Ei/Er). The dilational elasticity and viscosity are obtained finally via a
Fourier transformation of the measured harmonic surface tension changes provided by the
instruments software [21]. For a quantitative interpretation, equations of state and adsorption
isotherms for the single components and for the mixture must be available. These relationships are
the same for the interpretation of the equilibrium as well as dynamic surface tension data.

3. Theory
The surface behaviour of solutions containing a protein (biopolymer) mixed with an ionic surfactant
was studied for example in [22, 23]. For the theoretical analysis of the data a thermodynamic model
was used based on the analysis of the chemical potentials of the components in the bulk solution
and in the surface layer assuming a non-ideality of enthalpy and entropy. When a biopolymer
molecule which possesses free (unbound) charges at a concentration of cP interacts with counter-
charged ionic surfactant molecules of concentration cS, the Coulomb interaction leads to the
formation of complexes with the association number m. Here we use the subscript P for the polymer
molecule or the polymer/surfactant complex, and S for the surfactant molecule, respectively. The
7
adsorption of these complexes at the interface is determined by the average activity of ions

c c
m
P S
1 /(1 m )
participating in the reaction. In this study we assume that this model applies also to
biopolymer solutions containing counter-charged surfactants, the polyelectrolyte DNA and the
cationic surfactant AzoTAB.
The model assumes that the polymer molecule (and also the polymer-surfactant complex) can exist
at the surface in n states, each with its own molar area P,i = P,min + (i 1)P,0, where P,0 is the
area occupied by one segment of the polymer molecule (the area increment). The maximum molar
area covered by the polymer molecule is P,max. A corresponding equation of state of mixed
adsorbed layer was derived in [22, 23]:

0*
ln(1 P S ) P (1 P ,0 / P ) aP P2 aS S2 2aPS P S (1)
RT
Here is the surface pressure, R is the gas law constant, T is the temperature, S is the surface
coverage by surfactant molecules; aP and aS are the parameters which describe the interactions
between complexes and between non-associated surfactant molecules, respectively, while aPS is the
parameter which accounts for the interactions between the polymer-surfactant complexes and

surfactant molecules. The total polymer adsorption in all n states is defined by P i1 P,i where
n

P,i is the adsorption of the polymer in the i-th state, P P P i1 P,i P,i is the total surface
n

coverage by polymer molecules, and P is the average molar area of the adsorbed polymer. In
Eq. (1) *0 is the averaged molar area which accounts for a small difference between the molar area
of the solvent and that occupied by one segment of a polymer molecule:

P ,0 P S ,0 S
0* (2)
P S
where it is assumed (see e.g. [23]) that the molar area of the solvent is approximately equal to that
of the surfactant. The corresponding adsorption isotherms for the polymer-surfactant complexes in
state j = 1 (similar isotherms can be obtained for any of the other possible n states) and for the
unbound surfactant read:

PP,1
cPm /(1 m)c1S /(1 m) BP ,1 exp 2 aP P ,1 P aPS S (3)
1 P S P ,1 / P
P
S
bS (cS cC )1 / 2 exp 2aS S 2aPS P (4)
1 P S
where BP,1 and bS are the adsorption activities of the polymer-surfactant complex and the surfactant,
respectively; and cC is the surfactant counter-ion concentration. Note that at high concentrations of
inorganic electrolyte the ordinary Frumkin equation of state is applicable, which implies cS =
8
[24]. The Eqs. (1) to (4) describe the co-adsorption of the components of a mixture and the
increasing surface activity of the polymer-surfactant complexes as compared to the original
individual polymer. This, however, is not sufficient for an appropriate description of some systems,
for example the present DNA/AzoTAB mixtures, where DNA itself is not surface active. Therefore,
Eq. (3) was modified in [25] to introduce the dependence of the complex surface activity BP,1 on cS.
In the present study, this dependence is considered in more details. The parameter BPS,1 in the left
hand side of Eq. (3) is determined by the expressions:

BP,1 b P,1
at cS < c0 (5)
BP,1 bP,11 a X cS c0 at c0 < cS < cm (5b)

BP,1 bP,11 aX cm c0 at cS >cm (5c)

where a is an adjustable parameter which accounts for the influence of the surfactant concentration
on the activity of the polymer. Therefore, at low surfactant concentrations corresponding to
condition (5a) the parameter BP,1 is equal to the adsorption equilibrium constant of the individual
polymer or the polymer/surfactant complex bP,1 with = 0. The condition (5b) corresponds to the
variation of this constant with the increase in surfactant concentration, and the condition (5)
implies the fixed value of the complex adsorption activity at surfactant concentrations above a
critical value cm. The constant ax can be positive for opposite charges of polymer and surfactant
ions: this electrostatic attraction leads to a hydrophobization of the complex and an increasing
adsorption activity. However, negative ax could also be possible, e.g. due to surfactant aggregation
in the solution bulk; this would decrease the hydrophobization of the complex and a decrease in its
adsorption activity.
Following [25] we assume that the intrinsic compressibility of surfactant molecules exists in the
surface layer, i.e. the surfactant molar area S and the corresponding adsorption S are dependent
on the surface pressure and the total surface coverage = P + S:

S S 0 1 , and S SS SS 0 1 (6)
For surfactant molecules, this parameter () can be interpreted for example by changes in the
average tilt angle of adsorbed molecules upon surface layer compression, accompanied by an
increase in the surface layer thickness [25]. The idea of compressibility of adsorbed molecules takes
into account that with increasing surface coverage or surface pressure the molar area can change
and this has been used in different theoretical adsorption models [23]. The set of equations (1)-(6) is
sufficient to simulate the adsorption behaviour of mixed solutions of a polymer and a surfactant, i.e.
of polymer/surfactant complexes.

9
The surface elasticity of mixed surface layers was extensively discussed in [26], where a procedure
was developed to calculate the rheological characteristics of mixed adsorption layers. To perform
these calculations, it is sufficient to know the dependencies of surface pressure and adsorptions of
the system P and S as functions of the bulk concentrations cP and cS. The expression for the visco-
elasticity is E presented here as a complex quantity:

1 i i
E aSS aSP P aSS aPP aPS aSP
B ln S P DS DP S DS DP
, (7)
1 i i
aPS S aPP aSS aPP aPS aSP
B ln P DS P DP DS DP
S

where E = Er + iEi, a SS (S / cS )cP , a SP (S / c P ) cS , a PS (P / cS ) cP , a PP (P / c P ) cS ,

i i
and B 1 a SS a PP a SSa PP a PSa SP ; Er and Ei are the real (elasticity) and
DS DP DS D P

imaginary (viscosity) constituents of the visco-elasticity, respectively; = 2f, where f is the

frequency in Hz.

4. Results and Discussion

4.1. Surface tension

The experimental values of dynamic surface tension for an individual DNA solution are shown in
Fig. 2. The DNA concentration (monomers) was 5105 M. It is seen that the surface tension
decreases during the experimental run by only 0.35 mN/m, which is of the order of the experimental
error. This effect could be ascribed just to the presence of traces of surface active impurities. The
results confirm the data reported in [7, 8], evidencing the negligible adsorption activity of individual
DNA solutions without any presence of cationic surfactants which can influence their structure. The
dynamic surface tension of DNA solutions at the same concentration but with different additions of
AzoTAB is illustrated in Fig. 3. The addition of the cationic surfactant results in a decrease of the
surface tension as compared to the individual solution; however, the particular features of this
dependence are quite irregular. For example, the curve for the AzoTAB concentration of
1.5105 M is located below all other curves, also those which correspond to higher surfactant
concentrations in the solution. Note, the small jumps in the curves at higher AzoTAB
concentrations are experimental artefacts the origin of which is not known yet.

10
Fig. 2. Dynamic surface tension of an individual DNA solution for concentration of 5105 .

Fig. 3. Dynamic surface tension of mixed 5105 DNA solutions with added AzoTAB
concentrations between 106 and 5105 .
11
For the analysis of this behaviour, the equilibrium surface tension isotherms of the individual
AzoTAB solutions and its mixtures with DNA at the adsorption time of 14000 s are used below.
Fig. 4 illustrates the dependence of surface tension on the AzoTAB concentration for individual
AzoTAB solutions () and AzoTAB/DNA mixtures () at a fixed monomer DNA concentration
of 5105 . The theoretical isotherm of individual AzoTAB (black solid curve 1) was calculated
using the Frumkin model equation of state and adsorption isotherm assuming an intrinsic
compressibility of the adsorption layer, Eqs. (1), (4) and (6) with P = 0. As an excess of NaCl is
present in the AzoTAB solutions, the salt concentration could be disregarded, which implies cS =
in Eq. (4) (see [25] for further explanations). The best fit of the theoretical curve to the experimental
data yields the adsorption parameters for AzoTAB: S0 = 2.4105 m2/mol, S = 1.3, bS = 23.9
m3/mol, = 0.003 m/mN. These values are close to those found for CnTAB in [27], however, with
the bS constant approximately by a factor of 8 higher than that for C16TAB.

Fig. 4. Equilibrium surface tension isotherms of individual AzoTAB solutions () and

DNA+AzoTAB mixtures () at a fixed monomer DNA concentration of 5105 vs the AzoTAB

12
bulk concentration; black curve 1 - values calculated for the individual AzoTAB solutions using the
Frumkin model; blue curve 2 - values calculated for the mixtures with aX = 0; red dashed and dotted
curves 3 and 4 - values calculated with aX 0 using the values of the model parameters listed in the
text.

It was noted before (see Fig. 2) that the DNA adsorption activity in its individual solution is
negligible. The equilibrium isotherm for the AzoTAB+DNA mixtures with the same DNA
concentration as in Fig. 3, is shown in Fig. 4. The decrease of the surface tension of mixed solutions
becomes apparent at an AzoTAB concentration as low as 106 M. At this concentration the surface
tension of the individual AzoTAB solution is 71 mN/m, while for the mixture this value is
69 mN/m, i.e. by 2 mN/m lower.
Based on the surface tension values at AzoTAB concentrations of 106 and 3106 M, the model
adsorption parameters of the DNA+AzoTAB complex were calculated. As the DNA molecules
possess a globular structure, and noting that the DNA molecular weight is for example by a factor
of 600 higher than that of -lactoglobulin (BLG) [28, 29]; the molecular areas of DNA were taken
to be by a factor of 6002/3 70 larger than those of BLG. The DNA model parameters at polymer
concentrations below a critical value were found to be: aP = 1.3; P,0 = 3.5105 m2/mol;
P,min = 5108 m2/mol; P,max = 109 m2/mol; bP,1 = 7.5102 m3/mol; aPS = 0.5. Note that the
adsorption equilibrium constant of the DNA+AzoTAB complex is quite low, approximately by a
factor of 40 lower than that for BLG.
The surface tension isotherm calculated using Eqs. (1)-(6) and aX = 0 is shown in Fig. 4 by the blue
curve 2. It is seen that in the AzoTAB concentration range of 3106 to 5105 , the theoretical
model for the co-adsorption of biopolymer+surfactant complexes and unbound surfactant yields
values which are essentially different from the experimental data. This can be ascribed to the fact
that for the estimation of the value for bP,1 data at very low AzoTAB concentrations were used. It
can be supposed that the aggregation of cationic surfactant with the biopolymer not only leads to the
increase of the adsorption activity of the complex due to the increase of the effective concentration
c mP /(1m) c1S/(1m) value, but also affects the adsorption activity due to changes in the hydrophobicity of

the complex.
The influence of the cationic surfactant concentration on the coefficient BP,1 can be taken into
account using the coefficient aX in Eqs. (5a)-(5c); this is illustrated by the red curves in Fig. 4. To
describe the section of the curve where a decrease of the surface tension is observed, the values of
the coefficients were taken to be 0 = 6106 , cm = 2105 , and aX = 6.5105 1/M. The values
thus calculated are shown by the red dashed curve 3. It follows from Eq. (5c) that the solution of the
set of Eqs. (1)-(6) with these parameter values is confined to the cS values below the maximum
13
concentration cm = 2105 . The red dotted curve 4 was obtained for the values 0 = 3106 ,
cm = 2.2105 , and aX = 4.9105 1/M. The fitting for these two sets of parameter values resulted
in an increase of the BP,1 value which corresponds to values of the limiting concentration cm by a
factor of 9.1 and 9.3 as compared with the value bP,1 = 7.5102 m3/mol obtained above when any
dependence of the adsorption activity of the biopolymer+surfactant complex was disregarded. It
should be noted that in [25] the increase of the adsorption activity of the polyelectrolyte
polyallylamine hydrochloride+SDS complexes as a function of the SDS concentrations essentially
above the polyelectrolyte concentration was shown to be even higher. The section of the curve
where the surface tension temporarily increases (AzoTAB concentrations above 2.5105 ) was
calculated with the values 0 = 2.5105 , cm = 4105 , and aX = 5.9104 1/M; note that the
negative aX value corresponds to a decrease of the activity of the adsorbed complex. The minimum
BP,1 value for the DNA corresponds to the concentration cm = 4.0105 M; for this concentration the
expression in the square brackets in Eq. (5c) becomes 0.88. This means that the BP,1 value at this
concentration becomes by a factor of 8.7 lower (as compared with its value at cS = 2.2105 M at
which the BP,1 exceeds bP,1 approximately by a factor of 9), and becomes virtually equal to bP,1.
A similar shape of the surface tension isotherm for the DNA/cationic DTAB was reported in [9]: at
low DTAB concentrations the surface tension decreases, attains its minimum at a DTAB
concentration of 103 M, exhibits a short plateau, followed by a maximum and then decreases with
further concentration increase. This behaviour was analysed in [9] using various methods
(ellipsometry and Brewster angle microscopy). The authors proposed an explanation of this
behaviour that is stated below:
For low concentrations, the majority of surfactant molecules are free in the solution and form
complexes with DNA at the surface. This leads to a decrease in surface tension. The breakpoint
corresponds to a surfactant concentration where a cooperative binding occurs in the bulk. Above
this concentration, the surfactant gradually becomes cooperatively bounded to the DNA in the bulk.
Adsorption of the preformed hydrophobic DNA/surfactant aggregates is now possible at the
surface. This quickly leads to the formation of a thick surface layer. At still higher concentrations of
surfactant, the surface tension increases due to the competition with bulk aggregates. Finally, bulk
aggregates become less soluble and large amounts are adsorbed forming a surface layer which is
solid-like, and can be fractured.
This very similar shape of the observed surface tension behaviour, leads us to similar
explanation. In a mixed solution, at low AzoTAB concentrations (1106 to 2105 ) free
surfactant molecules start to form complexes with DNA at the surface. This occurs when surfactants
adsorb at the interface and form a positively charged monolayer. DNA can bind to this layer by
electrostatic interaction. The formation of complexes at the interface leads to a decrease in surface

14
tension. There is a minimum in the surface tension dependence on concentration at 2105 where
the surfactants gradually start to cooperatively bind to the DNA in the bulk. Above this
concentration, the surface tension starts to increase until it reaches a maximum at the AzoTAB
concentration of 4105 . This could be due to surfactant interaction competition between bulk
and interface. We may suggest that in the maximum region, the bulk association is more preferred
than a surface association. However, this reduction in surface activity of the complex causes a
negative value of the model parameter ax (ax = 5.9104 1/M). As said before, this parameter is
adjusted to reflect the influence of the surfactant concentration on the surface activity of the
polymer/surfactant complex. At very high surfactant concentrations (above 4105 ) the surface
tension decreases again due to the adsorption of the bulk aggregates.
Thus, the complicated nature of the experimental dependence of the surface tension for the
DNA+DTAB mixtures (and probably DNA+AzoTAB mixtures) exhibiting two extremes, is
described satisfactorily by the theoretical model developed in [22, 23, 25] which assumes the
influence of the cationic surfactant on the adsorption equilibrium constant of the DNA+AzoTAB
complex.

4.2. Dilational interfacial elasticity

The experimental dependencies of the elasticity of the DNA+AzoTAB mixed surface layers at
oscillation frequencies of 0.01 Hz () and 0.2 Hz () on the AzoTAB bulk concentration after
attaining the adsorption equilibrium are shown in Fig. 5. It is seen that above a concentration of
3106 a significant increase of the surface elasticity is observed. It is interesting that the course
of the experimental elasticity values agree well with the surface tension isotherm shown in Fig. 4:
the minimum of the surface tension at the AzoTAB concentration of 2105 corresponds to a
maximum elasticity in Fig. 5, while the maximum surface tension at a concentration of 4105
coincides with the minimum elasticity. This interrelation is physically reasonable, because for the
saturated adsorption layer the increase of the adsorption and the decrease of surface tension should
be accompanied by an increase in the elasticity [26, 30, 31].

15
Fig. 5. Dilational elasticity of the surface layer of mixed DNA+AzoTAB solutions at a fixed
monomer DNA concentration of 5105 vs the AzoTAB concentration at surface area oscillation
frequencies 0.01 () and 0.2 Hz (); dashed lines are guides for eye; the theoretical values (black
curve 1 and red curve 2 are for the oscillation frequencies 0.01 and 0.2 Hz, respectively) were
calculated from Eqs. (1)-(7) with the same parameters values as the red curves in Fig. 4; the values
of diffusion coefficients used here are given in the text.

The theoretical curves calculated using Eqs. (1)-(7) are seen to agree well with the experimental
data. The parameters values (including aX) used in the calculations were the same as those used for
the isotherms shown in Fig. 4. The diffusion coefficients for DNA and AzoTAB were taken to be
1011 and 1010 m2/s, respectively, which are physically reasonable values. Note that the decrease of
the DNA diffusion coefficient by one order of magnitude, and the three-fold decrease or increase of
the AzoTAB diffusion coefficient virtually do not affect the elasticity values shown in Fig. 5. This
is because of the small value of the phase angle, which did not exceed few degrees at concentrations
below 5105 . Note also, that the individual AzoTAB solutions have a surface elasticity of the
order of 10 mN/m at the given low frequencies so that a separate discussion is not suitable.

16
Conclusions
The dynamic and equilibrium surface tension and surface dilational characteristics of the individual
solutions of the biopolymer DNA, the cationic surfactant AzoTAB, and their mixtures were
measured by using drop profile analysis tensiometry. The experiments with the DNA solutions have
confirmed that this biopolymer does not exhibit any significant surface activity at the solution/air
interface. However, the negatively charged DNA molecules form complexes with the cationic
surfactant AzoTAB. The cationic surfactant AzoTAB has trans and cis isomers and is
photosensitive. The trans form of the molecule is more hydrophobic than the cis form and it can
adsorb at water/air interfaces. Mixed DNA+AzoTAB solutions exhibit high surface activity and
surface layer elasticity. The dependence of these characteristics show extremes on the AzoTAB
concentration within the range between 3106 and 5105 M. Subsequent minimum and maximum
in the surface tension dependence goes parallel to a maximum and minimum in the dilational
elasticity. A thermodynamic model developed earlier was used to explain the adsorption behaviour
of the DNA+AzoTAB complex. The values calculated with a classical co-adsorption model of
DNA+AzoTAB mixtures and unbound AzoTAB molecules disagree with the experimental data in
this AzoTAB concentration range. Therefore, the theoretical model was modified to account for the
dependence of the adsorption equilibrium constant of the adsorbed complex on the cationic
surfactant concentration. This modification resulted in a good agreement between the theoretical
predictions and the experimental data both for the surface tension and the elasticity values
throughout the entire range of studied AzoTAB concentrations.

Acknowledgements
The work was supported by projects of the DFG (Mi418/20-2) and the two COST actions CM1101
and MP1106.

References
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