You are on page 1of 7

Full Paper

Aust. J. Chem. 2008, 61, 968–974

CSIRO PUBLISHING

www.publish.csiro.au/journals/ajc

Dynamics and Orientation of Parathion Dissolved in a Discotic Nematic Lyomesophase
Alejandra Vera,A Hernán Ahumada,B Victor Bahamonde,A Rodrigo Montecinos,C Ramiro Araya-Maturana,D Daniel Muñoz,A and Boris E. Weiss-LópezA,E
A Universidad

de Chile, Facultad de Ciencias, Departamento de Química, Casilla 653, Santiago, Chile. B Universidad de Santiago de Chile, Facultad de Química y Biología, Departamento Química de los Materiales Av. Lib. B. O’Higgins 3363, Santiago, Chile. C Universidad Andrés Bello, Facultad de Ecología y Recursos Naturales, Departamento de Ciencias Químicas, República 253, Santiago, Chile. D Universidad de Chile, Facultad de Ciencias Químicas y Farmacéuticas, Departamento de Química Orgánica y Fisicoquímica, Casilla 233, Santiago 1, Chile. E Corresponding author. Email: bweiss@uchile.cl

Parathion, an organophosphorous pesticide, presents serious hazards to the environment and health. It inhibits acetylcholinesterase, an enzyme incorporated in the cell membrane. A study on the behaviour of parathion in a lipid environment is interesting from environmental cleaning and biological perspectives. 2 H NMR quadrupole splittings ( νQ ) and longitudinal relaxation times (T1 ) of parathion-d4 , dissolved in a nematic discotic lyomesophase made of tetradecyltrimethylammonium chloride/decanol (10% 1,1-dideuterodecanol)/water (0.1% D2 O)/NaCl, have been measured. νQ and T1 from DHO and 1,1-dideuterodecanol were also obtained. For a detailed understanding of the experimental results, a 19 ns molecular dynamics (MD) simulation of a bilayer fragment including three parathion molecules was calculated. Parathion is strongly attached to the aggregate and the solubilization increases the alignment of the interface components. Calculated densities show that parathion is located in the hydrophobic core, near the interface, and experiences an electrostatic interaction with the ammonium headgroups. On average, the molecule orients with the ring plane containing the bilayer normal. Manuscript received: 15 May 2008. Final version: 25 September 2008.

Introduction With the massive development of agriculture, organophosphorous compounds have been widely used as pesticides for plague management and control. The use of these compounds represents serious hazards to mammals’ health and the environment.[1–3] For this reason they are forbidden in numerous countries, and when authorized it is under severe security conditions. The toxicity arises from the ability to inhibit acetylcholinesterase at cholinergic junctions in different sites of the nervous system, including certain synapses in the central nervous system.[4–6] Intoxication is due to the accumulation of acetylcholine and the symptoms are excessive salivation, local sweating, tearing, loss of coordination, and weakness. Severe poisoning may include paralysis of the body’s extremities and respiratory muscles, causing death.[7–11] Two of the most toxic chemicals used in the agriculture industry are parathion and methyl parathion (diethoxy(4-nitrophenoxy)sulfanilidene phosphorane and dimethoxy(4-nitrophenoxy)sulfanilidene phosphorane, respectively). Parathion itself is not an acetylcholinesterase inhibitor; instead it is metabolized at the liver to produce paraoxon, the actual

inhibitor.[12,13] In paraoxon the sulfur atom is replaced by an oxygen atom (see Fig. 1). Despite the precautions that should be taken when applying the pesticide, every year many people become seriously contaminated. The most common forms of intoxication are ingestion of contaminated food, inhalation, and dermal adsorption. In all these cases the pesticide must cross cell membranes to reach the liver and be transformed into paraoxon.[14] The conformations, interactions, and reactivity of parathion with different molecules have all been the subject of interest.[15–17] Of particular importance, from an environmental and toxicological point of view, is to study the solubilization of the pesticide in surfactant aggregates and its effects on the structure of synthetic and natural membranes.[18–22] In this context, a study on the average orientation, distribution, and dynamics of parathion dissolved in a nematic discotic lyotropic liquid crystal (NDLLC) provides valuable information about the behaviour of the insecticide inside the hydrophobic bilayer, as well as the effects produced on the integrity of the aggregate. The liquid crystal unit is made of an aggregate of a few hundred amphiphiles with average oblate symmetry.[23]
10.1071/CH08209 0004-9425/08/120968

© CSIRO 2008

Parathion Dissolved in Discotic Nematic Lyomesophase

969

In this paper we present a 2 H NMR and molecular dynamics (MD) study on the average orientation, distribution, and dynamics of parathion fully deuterated in the aromatic ring (see Fig. 1), dissolved in an NDLLC solution. The NDLLC unit is similar to a flat micelle made of tetradecyltrimethylammonium ion and decanol (DeOH). These aggregates form when mixing certain amounts of tetradecyltrimethylammonium chloride (TTAC)/DeOH/water/NaCl. Decanol contained 10% 1,1-dideuterodecanol and the solvent was enriched 0.1% D2 O. The aggregate orients spontaneously in a magnetic field, with the symmetry axis of the oblate perpendicular to the field direction, which provides an anisotropic medium to observe residual quadrupole splittings. 2 H NMR quadrupole splittings ( νQ ), as well as 2 H NMR longitudinal relaxation times (T1 ) were measured for all deuterium-enriched components. For a better

understanding of the experimental findings, a 19 ns MD trajectory calculation of the experimental system, represented as a bilayer fragment, was calculated using the program Gromacs v.3.0.[24] Results Two samples were prepared for the present work, one without parathion (sample A) and the other with parathion (sample B). Longitudinal 2 H NMR relaxation times of all deuterated species were measured using the T1IR experiment. A stacked plot of 2 H NMR spectra from sample B is provided in the Accessory Publication. νQ values of all deuterated species were measured directly from the fully relaxed spectrum. The largest splitting corresponds to 1,1-dideuterodecanol, the intermediate splitting arises from parathion-d4 , and the smallest splitting is from DHO. Integrals of the right-hand peaks were employed to obtain the values of T1 ; for DHO the complete signal was used. Only one splitting was resolved for parathion. The estimated errors in the splittings are ±1 Hz for DHO, ±5 Hz for parathion-d4 , and ±10 Hz for DeOH. Fig. 2 shows the 1 H NMR spectra of both liquid crystal solutions, the pure mesophase (sample A), and the mesophase with parathion (sample B). The broad signal arises from the dipolar-coupled protons of the magnetic field oriented aggregates. The width of this signal, νD , depends on the degree of alignment of the aliphatic chain protons with the magnetic field, i.e., the order degree of the aggregate components. The band width for sample B is ∼1100 Hz broader than for sample A, when measured at about half height. The peak at ∼3.9 ppm arises from the isotropic solvent, and the multiplet centred at 0.3 ppm is from the dipolar-coupled protons of the ethoxy chains of parathion. The signal at ∼2.2 ppm is an impurity. All experimentally determined properties of both samples are listed in Table 1. For a microscopic understanding of the experimental results, a 19 ns MD trajectory of the system, represented as a bilayer fragment, was calculated. Fig. 3 is a snapshot of the simulation after 15 ns of trajectory. TTA ions and decanol are grey, chlorides are orange, sodium ions are black, water oxygen is red, and hydrogen white. For parathion, a van der Waals representation is displayed; carbon is light blue, nitrogen is blue, sulfur is yellow, and deuterium is green. For clear visualization of Fig. 3, the

n0

O P O

O

S

θ2

O N θ1

O

Fig. 1. Schematic representation of parathion displaying the definition of angles θ1 and θ2 , necessary to completely describe the orientation of the ring.

Sample B

Sample A 18 16 14 12
1H

10

8

6

4

2

0

2

4

6

8

10

12

14

16 ppm

Fig. 2.

NMR spectra of sample A (without parathion) and sample B (with parathion).

970

A. Vera et al.

reader is referred to the online version of this article, where a colour version is presented. Fig. 4 shows a plot of the mass density profile of all components of the simulation along the z-axis of the box, the normal to the bilayer plane. The densities of the mesophase components are displayed in the left axis scale and the densities of the parathion molecules appear in the right axis scale. The surfactants were divided into three fragments each; for TTAC the first fragment, called Amn1 (grey solid trace), corresponds to the trimethylammonium headgroup and the next 2 methylene groups; the second fragment called Amn2 (grey dashed trace) is made of methylenes 3 to 7; and the last group, Amn3 (grey dotted trace), corresponds to the last seven carbon atoms. The first fragment of DeOH, Deo1 (black solid trace), contains the alcohol group and the first two methylenes, the second fragment, Deo2 (black dashed trace), includes methylenes 3 to 6, and the last portion, Deo3 (black dotted trace), contains the last four carbon atoms. Parathion densities correspond to the right axis scale and are represented using black line with +, ×, and ◦ signs for molecules 1, 2, and 3, respectively. The vertical dashed lines denote the limits of the interface, which have been previously defined.[25] For a clearer visualization of Fig. 4, the reader is referred to the online version of this article, where a colour version is presented. Fig. 5 shows the z-coordinate trajectory of the nitrogen and oxygen atoms directly attached to the aromatic rings of molecules 2 and 3.

Two angles are necessary to describe the orientation of the aromatic ring in the aggregate. Angle θ1 is defined as the angle between the symmetry axis of the ring and the z-axis of the box, and angle θ2 is defined between the normal to the ring plane and the z-axis of the box; both angles are depicted in Fig. 1. The histogram for the distribution of angle 1 of molecule 2 is shown in Fig. 6. Fig. 7 shows the radial distribution function of ammonium headgroups surrounding the aromatic ring, with the origin at the centre of mass of the ring (black trace), and the radial distribution

800

3]

600

Sol Amn1 Amn2 Amn3

Na Deo1 Deo2 Deo3

Cl

12 10 8

Density [kg m

400

Para1 Para2 Para3

6 4

200 2 0 0 1 2 3 z-axis [nm] 4 5 0

Table 1. 2 H NMR quadrupole splittings and relaxation times of all deuterated derivatives present in samples A and B. The width of the broad signal observed in the 1 H NMR spectra, νD , is also listed Parameter
2 H-T 2 H-T

Sample A 88 363 — 14 846 0.0 — 12 500

Sample B 78 345 13 17 200 8.4 15 444 13 500

Decanol [ms] DHO [ms] 2 H-T par [ms] 1 νQDeOH [Hz] νQDHO [Hz] νQpar [Hz] νD [Hz]
1 1

Fig. 4. Mass density profiles of all species along the z-axis of the box. The surfactants were divided in three fragments each; for TTAC the first fragment, Amn1 (grey solid trace), corresponds to the trimethylammonium headgroup and the next two methylene groups; the second fragment (Amn2, grey dashed trace) is made of methylenes 3 to 7; and the last group (Amn3, grey dotted trace) corresponds to the last seven carbon atoms. The first fragment of DeOH (Deo1, black solid trace) contains the alcohol group and the first two methylenes, the second fragment (Deo2, black dashed trace) includes methylenes 3 to 6, and the last portion (Deo3, black dotted trace), contains the last four carbon atoms. Parathion densities correspond to the right axis scale and are represented using a black line with +, ×, and ◦ signs for molecules 1, 2, and 3, respectively. The vertical dashed lines denote the limits of the interface.

z

y

Fig. 3. Snapshot of the MD simulation after 15 ns of trajectory. TTA ions and decanol are grey, chloride is orange, and sodium is black. Oxygen is red and hydrogen white. Carbon is light blue, nitrogen is blue, sulfur is yellow, and deuterium is green.

Parathion Dissolved in Discotic Nematic Lyomesophase

971

4.0 z-coordinate of the box [nm] 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 6000

Charge density [q m 3]

Para2 N atom Para2 O atom Para3 N atom Para3 O atom

1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0 0.2 0 1 2

0.003 Amn1 Para1 Para2 Para3 0.002 0.001 0.000 0.001 0.002 0.003 0.004 3 4 5 6 0.005

8000 10 000 12 000 14 000 16 000 18 000 Time [ps]

z-axis of the box [nm]

Fig. 5. Time evolution of the z-coordinate of the nitrogen and oxygen atoms directly attached to the aromatic ring of parathion 2 and 3.

Fig. 8. Charge density profiles of the nitro groups from the three parathion molecules (grey traces) and ammonium headgroups (black trace), along the z-axis of the box.

1.0 300 Para2 250 200 150 100 50 0 C [t] 0.6 0.4 0.2 0.0 0.2 0.8

0

1500

3000 4500 Time [ps]

6000

7500

0

20

40

60 80 100 120 140 Angle 1 distribution [°]

160

180

Fig. 9. Rotational correlation function of the ortho C–D bond of parathion 2, obtained from the trajectory (black trace) and the fit to a bi-exponential decay (white trace).

Fig. 6. Histogram of the values obtained for angle 1 of parathion molecule 2. The angle is defined in the text and in Fig. 1.

2.0 Ring 1.6 Nitro

1.2 g [r]

0.8

0.4

0.0 0.0

0.5

1.0 r [nm]

1.5

2.0

2.5

Fig. 7. Radial distribution functions of ammonium headgroups surrounding the nitro group (grey trace) and ammonium headgroups surrounding the aromatic ring (black trace).

function of ammonium headgroups around the nitro group, with the origin at the centre of mass of the nitro group (grey trace). The radial distribution of ammonium around the nitro group presents a fast growing behaviour and a shoulder for short distances. This suggests the existence of an attractive interaction between the negative portion of the electric dipole moment of the nitro group and the positively charged ammonium headgroup. The short range Coulomb energy for the parathion–ammonium interaction is calculated to be −10 kJ mol−1 . To explore the origin of this interaction, charge densities along the z-axis of the box of the three nitro groups (right scale) and the ammonium headgroups (left scale) appear in Fig. 8. Rotational correlation functions for all C–D bonds of parathion and decanol were obtained from the trajectory. Fig. 9 shows the rotational correlation function of the ortho C–D bond of parathion 2. It was impossible to obtain a good fit using a single exponential decay; instead a bi-exponential function was always necessary. This suggests the existence of at least two dynamic processes contributing to quadrupolar relaxation. For parathion, the fast and slow process can be characterized by τc = 92 ± 30 ps and τc = 835 ± 200 ps respectively, whereas for the C–D bonds of decanol the values are τc = 6 ± 2 ps and τc = 180 ± 60 ps.

972

A. Vera et al.

Diffusion coefficients of parathion in the bilayer plane (Dxy ) and perpendicular to the bilayer plane (Dz ) were calculated from the trajectory of molecule 2, using the Einstein relation.[26] The results are Dxy = 3.3 × 10−12 m2 s−1 and Dz = 1.4 × 10−10 m2 s−1 . Discussion 1,1-Dideuterodecanol is an integral component of the discotic aggregate, oriented with the hydroxy group towards the aqueous phase and the aliphatic chain to the interior. Therefore, quadrupole splitting from deuterium in carbon 1 provides information about the order degree of the interface components. Table 1 shows that the addition of parathion produces an increase in the quadrupole splitting of decanol. Because the mesophase is the same in samples A and B, the average orientation of decanol in the magnetic field has not been modified. This strongly suggests that the integrity of the aggregate interface of sample B has been altered, becoming more rigid than sample A. A similar conclusion is reached from the increase in T1 of water. Decanol also increases T1 with the addition of parathion. The broad band observed in the 1 H NMR spectrum of Fig. 2 arises from the residual dipolar couplings from the oriented aliphatic chains that form the aggregate. The width of this band is a measurement of the degree of alignment of the surfactant chains with the magnetic field. The 1100 Hz broadening observed in the 1 H NMR spectrum of sample B, with parathion, also evidences an increase in the alignment of the aggregate components becoming more rigid. Addition of parathion to sample B makes it possible to observe the quadrupole splitting of water. This may originate either from an increase in the order degree of DHO at the interface or because the interface can incorporate more solvent molecules. The amount of interfacial water depends on the width of the interface. For the simulation without parathion the width is ∼1.34 nm and the addition of parathion does not modify this value significantly. Therefore, it is very likely that the increase in the quadrupole splitting of DHO arises from an increase in the alignment of the solvent at the interface. The 2 H NMR quadrupole splitting of parathion-d4 is large (see Table 1), which suggests that the molecule is attached to the aggregate. All the experimental observations strongly suggest that parathion should be incorporated inside the aggregate, in agreement with an octanol/water partition coefficient of 8511,[27] and probably interacts with some of the mesophase components such as to increase the order degree of the interface. Similar findings regarding the location and mobility decrease were reported by Madeira and coworkers in a dipalmitoylphosphatidylcholine (DPPC) bilayer[21] and by Suwalsky and coworkers in multilayers of dimyristoylphosphatidylethanolamine (DMPE), dimyristoylphosphatidylcholine (DMPC), and vesicles of DMPC, as well as in cell membranes of isolated toad skin and human erythrocytes.[20] Regarding the MD simulation, the mass density distribution of the mesophase components in the present trajectory (Fig. 4) are the same as those observed in the pure phase. The distribution of parathion molecules strongly suggests that the pesticide is strongly attached to the aggregate, located near the interface. Fig. 5 shows the trajectories of O and N atoms of parathion directly attached to the aromatic rings of molecules 2 and 3. This figure shows that molecule 2 experiences significant rotational diffusion across the bilayer. From the broad distribution

of this molecule it is possible to infer that in the presence of a concentration gradient the pesticide should cross the bilayer. Because molecule 2 shows the largest displacement, their diffusion coefficients in the xy-plane and along the z-axis of the box were calculated. Based on the simulation, the calculated diffusion coefficient across the bilayer is around 50 times bigger than inside the bilayer plane (see Table 1). This is possibly a consequence of the orientation of parathion inside the bilayer. The observed anisotropic behaviour in the diffusion coefficient should be related to the two dynamic processes identified in the rotational correlation function, very likely a geometrical restriction to the accessible space imposed by the matrix. Therefore, the longer rotational correlation time should be associated to rotational diffusion in the xy-plane and the shorter rotational correlation time should be related to rotational diffusion along the normal to the bilayer. The histogram displayed in Fig. 6 shows that the distribution of angle 1 is significantly broad for molecule 2, with three most probable values, around 30◦ , 88◦ , and 170◦ . This is a consequence of the rotational diffusion across the bilayer experienced by molecule 2. Angle 2 shows a much narrower distribution, with maximum and average values around 90◦ . Angle distributions for molecules 1 and 3 show averages and most probable values around 90◦ for both angles. Therefore, parathion is preferentially oriented with the local C2 -axis of the aromatic ring perpendicular to the direction of the aliphatic chains, and the ring plane containing the bilayer normal, i.e. both angles near 90◦ . The results from the calculated radial distribution functions displayed in Fig. 7 indicate that the aromatic ring is surrounded by 4.5 ammonium headgroups in the first nearest neighbour shell, at a distance of 6.7 Å. At a much shorter distance, 4.2 Å, the nitro group is surrounded by 1.2 ammonium headgroups. The radial distribution of ammonium ions around the nitro group shows a fast growing behaviour and a shoulder for shorter distance. This suggests the existence of an electrostatic interaction between parathion and the positively charged ammonium headgroups. In effect, the short range Coulomb energy for the parathion–ammonium interaction is −10 kJ mol−1 . To explore the origin of this interaction, the charge densities along the z-axis of the box for the three nitro groups (right scale) and the ammonium headgroups (left scale) is shown in Fig. 8. It is observed that the negative charge of the nitro moiety is preferentially located near the positively charged ammonium headgroups, which confirms the existence of an electrostatic interaction strong enough to locate parathion near the interface. The same interaction might be responsible for the observed ordering effect on the headgroups of unsealed human erythrocyte membranes and DPPC bilayers, using fluorescence anisotropy measurements.[21,22] Conclusions Molecular dynamics and NMR provide valuable and complementary information about the structure and dynamics of oriented anisotropic aggregates, as well as guest molecules dissolved in these solutions. Parathion is strongly attached to the aggregate, preferentially located in the hydrophobic region near the interface. The incorporation of parathion into the TTAC mesophase increases the order degree of all components of the liquid crystal solution. It is oriented, on average, with the C2 -symmetry axis of the ring perpendicular to the bilayer normal and the ring plane containing this normal. It diffuses very slowly in the bilayer plane but more efficiently across the bilayer.

Parathion Dissolved in Discotic Nematic Lyomesophase

973

A stabilizing electrostatic interaction, of ∼10 kJ mol−1 , between the nitro fragment of parathion and the ammonium headgroup of TTAC, was detected from the simulation. This interaction may be partially responsible for the location of the pesticide into the aggregate and the fluidity decrease of the interface components. The observed in-vivo effects of parathion could be partially related to the existence of this interaction, which precludes the proper functioning of cell membranes. Experimental Sample Preparation and NMR Spectra TTAC was purchased from Anatrace (www.anatrace.com) and recrystallized three times from ethyl acetate/ethanol solutions before use. Anhydrous NaCl (Aldrich) was used as received. Decanol (Aldrich) was purified by fractional crystallization and enriched 20% with previously synthesized 1,1dideuterodecanol.[28] Parathion-d4 was prepared according to a previously described methodology,[29] starting with the nitration of phenol-d5 (Aldrich) with NaNO3 (Merck) in modified silica to obtain improved yields of p-nitrophenol-d4 .[30] The final product was purified by preparative TLC using a mixture of ethyl acetate/chloroform (2/1) as eluent. The liquid crystal was prepared by dissolving 551 mg of TTAC, 171 mg of NaCl, and 138 µL of DeOH (10% α-d2 ) in 1500 µL of H2 O (0.1% D2 O). Two identical volumes of 0.6 mL were separated from the solution. One was directly transferred to the NMR tube (sample A), and 10 µL of parathion-d4 was dissolved into the other (sample B). Both were allowed to equilibrate 1 day before being transferred to the NMR tube. All NMR measurements were performed on both samples under identical experimental conditions. 1 H and 2 H NMR spectra were obtained using a Bruker Avance 400 spectrometer, with a 5 mm inverse BB probe. The 90◦ pulses were 9 µs for 1 H and 19 µs for 2 H, and the spectroscopic windows were 16 and 25 kHz for 1 H and 2 H, respectively. All transients were accumulated and recorded in 32 kB files. Molecular Dynamics Simulation A bilayer fragment representation of the TTAC aggregate has been previously constructed and equilibrated for more than 10 ns.[31] To this box, we added three molecules of parathion, the first at the centre of the hydrophobic region (molecule 1), the second at the interface (molecule 2), and the third in the aqueous phase (molecule 3). The starting structure of parathiond4 was built with Hyperchem (Hypercube Inc.) and the charges were obtained from a 6-31G∗ ab-initio full geometry calculation followed by MK fitting.[32] The trajectory calculation and analysis were made using the software package Gromacs v.3.0.[24] For trajectory visualization and graphics the VMD program[33] was used. All interactions of parathion-d4 were calculated using the Gromos force field.[34,35] For the deuterium atoms in parathiond4 , the Gromos potential for hydrogen was used, except that the mass was doubled. The torsions were parameterized with the Rickaert–Belleman potential,[36] whereas LINKS was used to restrict the bond lengths.[37] A 1 nm cut-off was used for the Lennard–Jones potential and real space electrostatic potential. Long-range electrostatic interactions were calculated using particle mesh Ewald.[38,39] The neighbour list was updated every 10 time steps. The temperature and pressure were kept constant at 300 K (all species independently coupled) and 1 bar, using the weak coupling algorithm of Berendsen,[40] with time constants of 0.1 and 1.0 ps for temperature and pressure respectively.

A trajectory of 19 ns was calculated with a time step size of 2 fs. All the analysis was performed in the last 14 ns of trajectory. Accessory Publication A stacked plot of 2 H NMR spectra of sample B is available from the journal’s website. Acknowledgements
The authors are pleased to acknowledge financial assistance from FONDECYT (Grant no. 1050201) and Facultad de Ciencias Universidad de Chile.

References
[1] R. P. Moody, M.Akram, E. Dickson, I. Chu, J.Toxicol. Environ. Health, A 2007, 70, 985. doi:10.1080/15287390600870874 [2] U. Hoffmann, T. Papendorf, Intensive Care Med. 2006, 32, 464. doi:10.1007/S00134-005-0051-Z [3] M. Levario-Carrillo, A. Feria-Velasco, R. De Celis, E. RarnosMartinez, L. Cordova-Fierro, F. J. Solis, Gynecol. Obstet. Invest. 2001, 52, 269. doi:10.1159/000052989 [4] T. T. Sun, I. A. Paul, I. K. Ho, J. Biomed. Sci. 2006, 13, 515. doi:10.1007/S11373-006-9075-9 [5] P. Nisse, X. Forceville, C. Cezard, A. Ameri, M. Mathieu-Nolf, Vet. Hum. Toxicol. 1998, 40, 349. [6] R. E. Kramer, S. E. Wellman, H. Zhu, R. W. Rockhold, R. C. Baker, J. Biomed. Sci. 2002, 9, 140. doi:10.1007/BF02256025 [7] A. Muttray, U. Spelmeyer, M. Degirmenci, D. Jung, G. Backer, G. Hill, Environ. Toxicol. Pharmacol. 2005, 19, 477. doi:10.1016/ J.ETAP.2004.12.010 [8] R.A. Fenske, C. S. Lu, D. Barr, L. Needham, Environ. Health Perspect. 2002, 110, 549. [9] P. Z. Ruckart, K. Kakolewski, F. J. Bove, W. E. Kaye, Environ. Health Perspect. 2004, 112, 46. [10] A. Wasley, L. A. Lepine, R. Jenkins, C. Rubin, Environ. Health Perspect. 2002, 110, 1053. [11] C. Rubin, E. Esteban, S. Kieszak, R. H. Hill, B. Dunlop, R. Yacovac, J. Trottier, K. Boylan, T. Tomasewski, K. Pearce, Environ. Health Perspect. 2002, 110, 1047. [12] E. L. Abel, T. K. Bammler, D. L. Eaton, Toxicol. Sci. 2004, 79, 224. doi:10.1093/TOXSCI/KFH118 [13] K. Saffih-Hdadi, L. Bruckler, E. Barriuso, J. Environ. Qual. 2003, 32, 2207. [14] A. M. Butler, M. Murray, J. Pharmacol. Exp. Ther. 1997, 280, 966. [15] J. Ford-Green, D. Majumdar, J. Leszczynski, Int. J. Quantum Chem. 2006, 106, 2356. doi:10.1002/QUA.20977 [16] K. K. Ghosh, D. Sinha, M. L. Satnami, D. K. Dubey, A. Shrivastava, R. M. Palepu, P. R. Dafonte, J. Colloid Interface Sci. 2006, 301, 564. doi:10.1016/J.JCIS.2006.05.061 [17] R. A. Moss, S. Kanamathareddy, S. Vijayaraghavan, Langmuir 2001, 17, 6108. doi:10.1021/LA010453X [18] Q. R. Zeng, H. X. Tang, B. H. Liao, T. F. Zhong, C. Tang, Water Res. 2006, 40, 1351. doi:10.1016/J.WATRES.2006.01.036 [19] W. Chu, K. H. Chan, W. K. Choy, Chemosphere 2006, 64, 711. doi:10.1016/J.CHEMOSPHERE.2005.11.028 [20] M. Suwalsky, P. Ramos, F. Villena, H. Cardenas, B. Norris, F. Cuevas, C. P. Sotomayor, Pestic. Biochem. Physiol. 2001, 70, 74. doi:10.1006/PEST.2001.2539 [21] R. A. Videira, M. C. Antunes-Madeira, V I. C. F. Lopes, . V M. C. Madeira, BBA-Rev. Biomembranes 2001, 1511, 360. . doi:10.1016/S0005-2736(01)00295-4 [22] J. Blasiak, Pestic. Biochem. Physiol. 1993, 45, 72. doi:10.1006/ PEST.1993.1009 [23] R. Montecinos, H. Ahumada, R. Martinez, F. A. Olea, R. ArayaMaturana, M. P. Aliste, D. P. Tieleman, B. E. Weiss-Lopez, Langmuir 2004, 20, 5703. doi:10.1021/LA049801W [24] D. van der Spoel, A. R. van Buuren, E. Apol, P. J. Meulenhoff, D. P. Tieleman, A. L. T. M. Sijbers, B. Hess, K. A. Feenstra, et al.,

974

A. Vera et al.

[25] [26] [27] [28]

[29] [30] [31] [32]

Gromacs User Manual 2001 (Department of Biophisical Chemistry, University of Groningen: Groningen, The Netherlands). D. P. Tieleman, D. van der Spoel, H. J. C. Berendsen, J. Phys. Chem. B 2000, 104, 6380. doi:10.1021/JP001268F M. P. Allen, D. J. Tildesley, Computer Simulation of Liquids 1992, p. 60 (Oxford Science Publications: Oxford). G. G. Briggs, J. Agric. Food Chem. 1981, 29, 1050. doi:10.1021/ JF00107A040 R. Montecinos, H. Ahumada, R. Araya-Maturana, A. F. Olea, B. E. Weiss-López, J. Colloid Interface Sci. 2007, 316, 126. doi:10.1016/J.JCIS.2007.07.060 J. H. Fletcher, J. C. Hamilton, I. Hechenbleikner, E. I. Hoerberg, B. J. Sertl, J. T. Cassaday, J. Am. Chem. Soc. 1948, 70, 3943. M. A. Zolfigol, B. F. Mirjalili, A. Bamoniri, M. A. K. Zarchi, A. Zarei, L. Khazdooz, J. Noei, Bull. Korean Chem. Soc. 2004, 25, 1414. H. Ahumada, R. Montecinos, D. P. Tieleman, B. E. Weiss-Lopez, J. Phys. Chem. A 2005, 109, 6644. doi:10.1021/JP0441590 B. H. Besler, K. M. Merz, P. A. Kollman, J. Comput. Chem. 1990, 11, 431. doi:10.1002/JCC.540110404

[33] W. Humphrey, A. Dalke, K. Schulten, J. Mol. Graph. 1996, 14, 33. doi:10.1016/0263-7855(96)00018-5 [34] W. F. van Gunsteren, H. J. C. Berendsen, Angew. Chem. Int. Ed. Engl. 1990, 29, 992. doi:10.1002/ANIE.199009921 [35] W. F. van Gunsteren, H. J. C. Berendsen, Biomos Nijenborgh 4, 9747AG, 1995, Groningen, The Netherlands. [36] J. P. Ryckaert, A. Bellemans, Faraday Discuss. Chem. Soc. 1978, 66, 95. doi:10.1039/DC9786600095 [37] B. Hess, H. Bekker, H. J. C. Berendsen, J. G. E. M. Fraaije, J. Comput. Chem. 1997, 18, 1463. doi:10.1002/(SICI)1096987X(199709)18:12<1463::AID-JCC4>3.0.CO;2-H [38] T. Darden, D. York, L. Pedersen, J. Chem. Phys. 1993, 98, 10089. doi:10.1063/1.464397 [39] U. Essmann, L. Perera, M. L. Berkowitz, Langmuir 1995, 11, 4519. doi:10.1021/LA00011A056 [40] H. J. C. Berendsen, J. P. M. Postma, W. F. van Gunsteren, A. Dinola, J. R. Haak, J. Chem. Phys. 1984, 81, 3684. doi:10.1063/1.448118