A 1H NMR study is presented for the binding of rhodamine B (RB) to the polyanion containing aromatic
groups poly(sodium 4-styrenesulfonate) (PSS), which is also evidenced by diafiltration. 1H NMR spectra
showed an accentuated upfield shift of proton H6 of the benzoic ring of RB at pH 7, indicating the stacking
of RB onto PSS. The corresponding structure is proposed which is in accordance to Hunter and Sanders
rules. At pH 2, an upfield shift of the xanthene protons of RB would indicate a highly condensed state for
this molecule.
1. Introduction
Molecular association involving aromatic-aromatic interactions is attracting much attention nowadays.1-13 As a result of
the planar geometries of aromatic molecules, the molecular
surface/volume ratio is high compared to that of spherical
particles. Then, the aggregation of aromatic groups in water
may produce the release of higher amounts of surface-solvating
water molecules and, consequently, an increase in the favorable
entropic and enthalpic contributions to the free energy by means
of classical and nonclassical hydrophobic effects.1 In addition
to these solvent contributions, site-specific interactions such as
short-range electrostatic interactions, hydrogen bond formation,
- interactions, or cation- interactions may also contribute
to the free energy and define the geometry of the complexes.
These aromatic-aromatic interactions are found in water in
many systems such as nucleic acids, proteins, porphyrins,
semiconductors, molecular clips, and so forth1-13 and are one
of the principal noncovalent forces governing molecular recognition and biomolecular structure. In particular, they are
important in the stabilization of DNA and its association with
intercalators. A change in the hydrogen bonding capacity of
DNA bases as a consequence of - interactions has been
described in recent literature.4,5 The vast majority of X-ray
crystal structures of protein complexes with small molecules
reveal bounding interactions involving aromatic amino acid side
chains of the receptor and/or aromatic and heteroaromatic rings
of the ligand. A raise of the pKa of substrates by means of -
interactions with enzymes has been proposed as a mechanism
of enzymatic catalysis.6
We have recently found a smart system composed by poly(4-sodium styrenesulfonate) (PSS) and rhodamine B (RB).14 The
behavior of this system is pH dependent. Lowering the pH from
5 to 3, we have observed an abrupt change in the binding
* FAX: 56-63-221597. E-mail: imorenovilloslada@uach.cl.
Universidad Austral de Chile.
Universidad de Concepcio
n.
Materials Characterization Central Laboratory, Waseda University.
| Department of Applied Chemistry, Waseda University.
Figure 2. 1H NMR aromatic region spectra of solutions in D2O at pH 7.5 (1) and 2.2 (2) of different RB concentrations: (a) 110-2 M; (b) 510-3
M; (c) 110-3 M; (d) 510-4 M.
ratio between the volume in the filtrate and the constant volume
in the diafiltration cell, the concentration in the filtrate of the
low-molecular-weight species under study (LMWS) (cLMWSfiltrate), the concentration of free LMWS in the cell solution
(cLMWSfree), the concentration of LMWS reversibly bound to the
water-soluble polymer (WSP) (cLMWSrev-bound), the apparent
dissociation constant (KLMWSdiss-WSP), defined as the ratio
cLMWSfree/cLMWSrev-bound, the diafiltration parameters km, j, u, and
V, and the polymer concentration in moles per liter of monomeric
units (cP). km and j parameters (the absolute value of the slope
of the curve ln cLMWSfiltrate versus F in the absence and in the
presence of the WSP, respectively) are related to the strength
of the interaction, while V and u are related to the amounts of
LMWS reversibly or irreversibly bound to the polymer,
respectively. Results of diafiltration of a 10-4 M solution of
RB at pH 7 shown in Figure 3 and Table 1 indicate no
significant interaction of RB with the diafiltration cell components (km close to 1). On the other hand, the amounts of RB
irreversibly bound to PSS are negligible (u values close to 0),
and the fraction of RB reversibly bound to PSS increases with
Moreno-Villoslada et al.
TABLE 1: Results for Diafiltration of 10-4 M RB Solutions at pH 7 and Different PSS Concentrationsa
expt
cP (M)
RB-03
PSS -RB-01
PSS -RB-02
PSS -RB-03
PSS -RB-04
PSS -RB-05
PSS -RB-06
210-4
410-4
610-4
810-4
110-3
210-3
0.91
0.85
0.85
0.92
0.94
0.95
0.94
0.09
0.15
0.15
0.08
0.06
0.05
0.06
0.63
0.49
0.41
0.37
0.29
0.18
km
0.84
KRBdiss-PSS b
linear adjustments
for the experimental data
R2
2.4 ( 0.5
1.2 ( 0.2
0.8 ( 0.1
0.7 ( 0.8
0.5 ( 0.05
0.2 ( 0.03
y ) -0.84x - 9.5
y ) -0.63x - 9.6
y ) -0.49x - 9.9
y ) -0.41x - 10.0
y ) -0.37x - 10.1
y ) -0.29x - 10.4
y ) -0.18x - 10.9
1.00
0.99
0.99
0.99
0.99
1.00
1.00
a
For linear adjustments: y ) ln cRBfiltrate; x ) F; R2 ) linear regression factor. b KRBdiss-PSS is calculated following (j/1 - j) e KLMWSdiss-WSP
e (kmj/km - j).
Figure 4. 1H NMR aromatic region spectra of solutions in D2O at pH 7 of (a) RB 110-3 M; (b-d) RB 110-3 M in the presence of different PSS
concentrations: (b) 0.510-3 M, (c) 210-3 M, (d) 610-3 M; (e) PSS 110-2 M.
Figure 6. COSY spectrum of the aromatic region of a RB 110-3 M/PSS 210-3 M solution in D2O at pH 7.
Moreno-Villoslada et al.
Figure 8. 1H NMR aromatic region spectra of solutions in D2O of RB 3.410-3 M and PSS 210-2 M at different pHs: (a) 6.9; (b) 5.2; (c) 4.6;
(d) 4.1; (e) 2.2.