Sum Frequency Generation Intensity Research Articles

DFT-Based Calculation of Molecular Hyperpolarizability and SFG Intensity of Symmetric and Asymmetric Stretch Modes of Alkyl Groups.

Vibrational sum frequency generation (SFG) spectroscopy has been extensively used for obtaining structural information of molecular functional groups at two-dimensional (2D) interfaces buried in the gas or liquid medium. Although the SFG experiment can be done elegantly, interpreting the measured intensity in terms of molecular orientation with respect to the lab coordinate is quite complicated. One of the main reasons is the difficulty of determining the hyperpolarizability tensors of even simple molecules that govern their SFG responses. The single-bond polarizability derivative model has been proposed to estimate the relative magnitude of SFG-active hyperpolarizability by assuming that the perturbation associated to each vibration is negligible. In this study, density functional theory was used to calculate the polarizability and dipole derivative tensors of the CH3 stretch mode of CH3I, CH3CH2I, CH3OH, and CH3CH2OH. Then, the hyperpolarizability tensors of symmetric and asymmetric vibration modes were calculated considering the Boltzmann distribution of representative conformers, which allowed us to theoretically calculate their SFG intensities at all polarization combinations as a function of the tilt angle of the CH3 group with respect to the surface normal direction. Then, the ratios of the calculated SFG intensities for the CH3 symmetric and asymmetric stretch peaks used in experimental studies for the CH3 tilt angle determination were compared. This comparison clearly showed that the effect of vibrational coupling among neighboring functional groups is significant and cannot be assumed to be negligible. This study presents new parameters that can be used in determining the average tilt angle of the CH3 group at the 2D interface with SFG measurements as well as limitations of the method.

Sum-frequency generation at interfaces: A Fresnel story. II. Analytical expressions for multilayer systems.

The well-known formalism for Sum-Frequency Generation (SFG) reflected or transmitted by a three-layer system involves three equations defining the emitted SFG intensity, the effective nonlinear susceptibility, and a set of Fresnel factors specific to the three-layer system. We generalize the equations to an N-layer system, where all media have non-vanishing thicknesses, by leaving the first two equations unchanged and modifying only the Fresnel factors. These universal Fresnel factors bear all the complexity of light propagation and interference in the system, in amplitude and phase. They are analytically known anywhere in the N-layer system, either at any interface or in any of the bulks, and share common expressions for the three beams, incoming or emitted, of the SFG process in reflection. Enclosing an ultrathin layer (e.g., a molecular monolayer) in the system does not modify the Fresnel factors except for boundary conditions at this layer, as in the three-layer case. Specific rules are elaborated to simplify systems containing macroscopic layers. Equations for the four- and five-layer systems are explicitly provided. Simulations in the four-layer system allow for the recovery of the results of the transfer matrix formalism at a lower complexity cost for SFG users. Finally, when several interfaces in the system produce SFG signals, we show that it is possible to probe only the most buried one by canceling all the SFG responses except at this last interface, generalizing the results of the three-layer system.

Electric double layer contribution to sum frequency generation signal from Au electrode.

Understanding the electric double layer (EDL) of the metal electrode-electrolyte interface is essential to electrochemistry and relevant disciplines. In this study, potential-dependent electrode Sum Frequency Generation (SFG) intensities of polycrystalline gold electrodes in HClO4 and H2SO4 electrolytes were thoroughly analyzed. The potential of zero charges (PZC) of the electrodes was -0.06 and 0.38V in HClO4 and H2SO4, respectively, determined from differential capacity curves. Without specific adsorption, the total SFG intensity was dominated by the contribution from the Au surface and increased similar to that of the visible (VIS) wavelength scanning, which pushed the SFG process closer to the double resonant condition in HClO4. However, the EDL contributed about 30% SFG signal with specific adsorption in H2SO4. Below PZC, the total SFG intensity was dominated by the Au surface contribution and increased with potential at a similar slope in these two electrolytes. Around PZC, as the EDL structure became less ordered and the electric field changed direction, there would be no EDL SFG contribution. Above PZC, the total SFG intensity increased much more rapidly with potential in H2SO4 than in HClO4, which suggested that the EDL SFG contribution kept increasing with more specific adsorbed surface ions from H2SO4.

Lamellar orientation at the surface of isotactic polystyrene thin films analyzed by sum frequency generation spectroscopy.

Analyzing the orientation of polymeric crystalline lamella at the surface of thin films can be challenging. Even though atomic force microscopy (AFM) is often sufficient for this analysis, there are cases when imaging is not sufficient to confidently determine lamellar orientation. Here, we used sum frequency generation (SFG) spectroscopy to analyze the lamellar orientation at the surface of semi-crystalline isotactic polystyrene (iPS) thin films. The SFG orientation analysis indicated that the iPS chains are oriented perpendicular to the substrate (flat-on lamellar orientation), which was confirmed by AFM. By analyzing the evolution of the SFG spectral features with the progress of crystallization, we demonstrated that the ratios of the SFG intensities of the phenyl ring resonances are a good indication of the surface crystallinity. Furthermore, we explored the challenges associated with SFG measurements of heterogeneous surfaces, which is commonly present in many semi-crystalline polymeric films. To the best of our knowledge, this is the first time that the surface lamellar orientation of semi-crystalline polymeric thin films was determined by SFG. Also, this work pioneers in reporting the surface conformation of semi-crystalline and amorphous iPS thin films by SFG and in linking the SFG intensity ratios to the progress of the crystallization and the surface crystallinity. This study demonstrates the potential applicability of SFG spectroscopy in the conformational analysis of polymeric crystalline structures at interfaces and opens the way to the investigation of more complex polymeric structures and crystalline arrangements, especially for the case of buried interfaces, where AFM imaging is not an option.

Electrostatic charges and their distribution on the charged surfaces probed by sum-frequency generation spectroscopy

We examined the electrostatic charging states of insulating polymer surfaces using sum-frequency generation (SFG) spectroscopy. For the negatively charged polypropylene, the SFG peak amplitudes increased significantly with increasing surface potential, indicating that the electric-field formed by the electrostatic charges directly affects the SFG signal intensities. In the organic thin films stacked on top of PMMA, an increase in the SFG signal of buried PMMA is observed, indicating that the electrostatic field formed by the electrical charges is extended into the bulk direction. In addition, visualization of the location and distribution of the charges is demonstrated using the SFG intensity variations.

Molecular Signature of an Unexpected Hydrophobization of Silica Surfaces in Lead Nitrate Solutions as Detected by Sum Frequency Generation Spectroscopy

Responsive silica surfaces empower their smart applications for catalysis, mass transfer, and self-cleaning. Here, we reveal the molecular signature of an unexpected hydrophobization of silica surfaces as their pH response in a lead nitrate solution. Strikingly, the pH-dependent sum frequency generation (SFG) spectra of silica surfaces show two profound minima of the intensity at pH = 7.2 and 10.2, respectively. Our in-depth analysis of SFG spectra using the maximum entropy method show emerging vibrational bands at >3600 cm–1 for the surface-bound water at this pH range, which are similar to the stretching modes of the dangling OH at the air–water surface. The observed minima in SFG intensity and the occurrence of the “hydrophobic-like” stretching modes of interfacial water provide a molecular fingerprint of the hydrophobicity of the silica surfaces, which is confirmed by contact angle measurements. These step-change findings help better deal with silica-related processes in nature and technologies.

Surface Properties of Saponin—Chitosan Mixtures

The surface properties of saponin and saponin-chitosan mixtures were analysed as a function of their bulk mixing ratio using vibrational sum-frequency generation (SFG), surface tensiometry and dilational rheology measurements. Our experiments show that saponin-chitosan mixtures present some remarkable properties, such as a strong amphiphilicity of the saponin and high dilational viscoelasticity. We believe this points to the presence of chitosan in the adsorption layer, despite its complete lack of surface activity. We explain this phenomenon by electrostatic interactions between the saponin as an anionic surfactant and chitosan as a polycation, leading to surface-active saponin-chitosan complexes and aggregates. Analysing the SFG intensity of the O-H stretching bands from interfacial water molecules, we found that in the case of pH 3.4 for a mixture consisting of 0.1 g/L saponin and 0.001 g/L chitosan, the adsorption layer was electrically neutral. This conclusion from SFG spectra is corroborated by results from surface tensiometry showing a significant reduction in surface tension and effects on the dilational surface elasticity strictly at saponin/chitosan ratios, where SFG spectra indicate zero net charge at the air-water interface.

Numerical Simulation of Vibrational Sum Frequency Generation Intensity for Non-Centrosymmetric Domains Interspersed in an Amorphous Matrix: A Case Study for Cellulose in Plant Cell Wall.

Vibrational sum frequency generation (SFG) spectroscopy can specifically probe molecular species non-centrosymmetrically arranged in a centrosymmetric or isotropic medium. This capability has been extensively utilized to detect and study molecular species present at the two-dimensional (2D) interface at which the centrosymmetry or isotropy of bulk phases is naturally broken. The same principle has been demonstrated to be very effective for the selective detection of non-centrosymmetric crystalline nanodomains interspersed in three-dimensional (3D) amorphous phases. However, the full spectral interpretation of SFG features has been difficult due to the complexity associated with the theoretical calculation of SFG responses of such 3D systems. This paper describes a numerical method to predict the relative SFG intensities of non-centrosymmetric nanodomains in 3D systems as functions of their size and concentration as well as their assembly patterns, i.e., the distributions of tilt, azimuth, and rotation angles with respect to the lab coordinate. We applied the developed method to predict changes in the CH and OH stretch modes characteristic to crystalline cellulose microfibrils distributed with various orders, which are relevant to plant cell wall structures. The same algorithm can also be applied to any SFG-active nanodomains interspersed in 3D amorphous matrices.

Adsorption of CTAB on Sapphire-c at High pH: Surface and Zeta Potential Measurements Combined with Sum-Frequency and Second-Harmonic Generation.

The adsorption of cetyltrimethylammonium bromide (CTA+Br-) on sapphire-c surfaces was studied at pH 10 below the surfactants' critical micelle concentration. The evolution of interfacial potentials as a function of CTAB concentration was characterized by surface and zeta potential measurements and complemented by molecular dynamic (MD) simulations as well as by second-harmonic (SHG) and vibrational sum-frequency generation (SFG) spectroscopy. The changes in interfacial potentials suggest that the negative interfacial charge due to deprotonated surface aluminols groups is neutralized and can be even overcompensated by the presence of CTA+ cations at the interface. However, SFG intensities from strongly hydrogen-bonded interfacial water molecules as well as SHG intensities decrease with both increasing CTAB concentration and the magnitude of the surface potential. They do not suggest a charge reversal at the interface, while the change in zeta potential is actually consistent with an apparent charge inversion. This can be qualitatively explained by results from MD simulation, which reveal adsorbed CTA+ cations outside a first strongly bound hydration layer of water molecules, where they can locally distort the structural order and replace some of the interfacial water molecules adjacent to the first layer. This is proposed to be the origin for the significant loss in SFG and SHG intensities with increasing CTAB concentration. Moreover, we propose that CTA+ can act as a counterion and enhance the occurrence of deprotonated surface aluminols that is consistent with the decrease in surface potential.

Role of imidazolium cations on the interfacial structure of room‐temperature ionic liquids in contact with Pt(111) electrodes

AbstractRoom‐temperature ionic liquids (ILs) have gained considerable attention as an important addition to conventional electrolytes because they exhibit large electrochemical windows and can reduce existing overpotentials in electrocatalysis. For the interfacial electrochemistry of ILs, a comprehensive understanding of molecular ions and the resulting electric double‐layer structures as a function of electrode potential is mandatory, but the structures are largely different from conventional electrolytes. For that reason, we have studied the interfaces of Pt(111) in contact with ILs using 1‐butyl‐3‐methylimidazolium [BMIM] and 1‐butyl‐2,3‐dimethylimidazolium [BMMIM] cations as well as bis(trifluoromethylsulfonyl)imide [NTf2] anions. We applied vibrational sum‐frequency generation (SFG), where we interrogate vibrational bands from interfacial cations, anions, as well as interfacial water in situ and under potential control. Structuring of [NTf2] anions and H2O with electrode potential show hysteresis while a strong Stark tuning was absent. This indicates that the IL ions are oriented in the vicinity of the interface, without being directly adsorbed to the Pt(111) surface. Using the C‐H stretching band from CH groups at the imidazolium ring, the ring reorientation with electrode potential was qualitatively determined. The imidazolium ring reorients as a function of potential from a more parallel orientation to an upright orientation with respect to the interfacial plane. This leads to the formation of voids in the layered structure of ions at the interface, which can be then filled with H2O as evidenced by an increased SFG intensity from O‐H stretching modes that are attributable to hydrogen‐bonded interfacial water. Comparing the responses of the ILs, particularly of [BMMIM][NTf2], shows a compact structure and a significantly pronounced rearrangement of the imidazolium ring that can also facilitates better incorporation of H2O and significantly affects the reorientation of [NTf2] anions and, thus, causes a pronounced hysteresis with electrode potential.

Hydrolysis and Condensation of Tetraethyl Orthosilicate at the Air–Aqueous Interface: Implications for Silica Nanoparticle Formation

The silica (SiO2) nanoparticles of a well-known silica precursor tetraethyl orthosilicate (TEOS) are generally synthesized via a promising solution-gelation inorganic polymerization process. The monodisperse silica nanoparticles have potential applications ranging from the formation of dental nanocomposites to antireflective coatings. In the present study, we have systematically investigated the in situ interfacial molecular structure of TEOS and its impact on the interfacial water structure during the processes of hydrolysis and condensation at the air–aqueous interface using sum-frequency generation (SFG) vibrational spectroscopy. With the presence of water, a gradual decrease in the SFG intensity in the CH-stretch region for each concentration of TEOS with time reflects the elimination of ethoxy groups, which is a signature of the hydrolysis process. Further, the impact of the hydrolysis process is revealed from the significant enhancement in the SFG signal in the OH-stretch region. The hydrolysis of TEOS is then followed by condensation in which the ≡Si─O– charged species are replaced by forming the ≡Si─O─Si≡ bridging network. The signature of the condensation process is reflected with the gradual decrease in the observed enhanced SFG signal in the OH-stretch region. The formation of monodispersed silica nanoparticles as an end product of size variation from 1.75 to 4.67 nm with the increase in TEOS concentration is confirmed with the DLS measurements. We have also probed the pH-dependent SFG studies at three different pH values (2.0, 5.8, and 9.0). The dominant pH-dependent hydrolysis process is revealed from the observed molecular structure of TEOS at the air–aqueous interface.

Elucidating molecular mechanisms of two-dimensional chemical reactions

Elucidating molecular mechanisms of two-dimensional chemical reactions

PH effects on the molecular structure and charging state of β-Escin biosurfactants at the air-water interface

Saponins like β-escin exhibit an unusually high surface activity paired with a remarkable surface rheology which makes them as biosurfactants highly interesting for applications in soft matter colloids and at interfaces. We have applied vibrational sum-frequency generation (SFG) to study β-escin adsorption layers at the air-water interface as a function of electrolyte pH and compare the results from SFG spectroscopy to complementary experiments that have addressed the surface tension and the surface dilational rheology. SFG spectra of β-escin modified air-water interfaces demonstrate that the SFG intensity of OH stretching vibrations from interfacial water molecules is a function of pH and dramatically increases when the pH is increased from acidic to basic conditions and reaches a plateau at a solution pH of > 6. These changes are attributable to the interfacial charging state and to the deprotonation of the carboxylic acid group of β-escin. Thus, the change in OH intensity provides qualitative information on the degree of protonation of this group at the air-water interface. At pH < 4 the air-water interface is dominated by the charge neutral form of β-escin, while at pH > 6 its carboxylic acid group is fully deprotonated and, consequently, the interface is highly charged. These observations are corroborated by the change in equilibrium surface tension which is qualitatively similar to the change in OH intensity as seen in the SFG spectra. Further, once the surface layer is charge neutral, the surface elasticity drastically increases. This can be attributed to a change in prevailing intermolecular interactions that change from dominating repulsive electrostatic interactions at high pH, to dominating attractive interactions, such as hydrophobic and dispersive interactions, as well as, hydrogen bonding at low pH values. In addition to the clear changes in OH intensity from interfacial H2O, the SFG spectra exhibit drastic changes in the CH bands from interfacial β-escin which we relate to differences in the net molecular orientation. This orientation change is driven by tighter packing of β-escin adsorption layers when the β-escin moiety is in its charge neutral form (pH < 4).

Orientation analysis of sum frequency generation spectra of di-chain phospholipids: Effect of the second acyl chain

Sum Frequency Generation (SFG) spectroscopy is widely used for studying the di-chain phospholipid monolayers incorporated in model cell membranes. In this context, it is frequently assumed, without justification, that the chains are identical, so their individual contributions to the SFG spectra are indistinguishable. However, the combination of both attractive and repulsive Van der Waals interactions between the chains results in a finite angle between their two terminal methyl groups, resulting in non-equivalent contributions to the non-linear susceptibility. This work describes the application of the underlying non-linear theory required to produce the accurate SFG spectral simulations needed to test this assertion and therefore provides the necessary quantitative validation. For phospholipids comprising two identical saturated chains, which typically have small angles of divergence, these simulations predict only small deviations in the SFG intensities from those calculated assuming a single methyl orientation. Non-identical tails, however, with differences in the degree or type of chain unsaturation, or in the parity of the chain lengths, show much larger discrepancies than the assumption of a single chain. In these cases, the two tails must be treated as separate entities, and their structural relationship must be incorporated into the interpretation of their SFG spectra. A second important result from the simulations arises from the systematic nature of the deviations, which shows that even small intensity changes should not be quickly dismissed on the basis of being subsumed by the uncertainties associated with spectral noise.

Role of Ions on the Surface-Bound Water Structure at the Silica/Water Interface: Identifying the Spectral Signature of Stability.

Isolating the hydrogen-bonding structure of water immediately at the surface is challenging, even with surface-specific techniques like sum-frequency generation (SFG), because of the presence of aligned water further away in the diffuse layer. Here, we combine zeta potential and SFG intensity measurements with the maximum entropy method referenced to reported phase-sensitive SFG and second-harmonic generation results to deconvolute the SFG spectral contributions of the surface waters from those in the diffuse layer. Deconvolution reveals that at very low ionic strength, the surface water structure is similar to that of a neutral silica surface near the point-of-zero-charge with waters in different hydrogen-bonding environments oriented in opposite directions. This similarity suggests that the known metastability of silica colloids against aggregation under both conditions could arise from this distinct surface water structure. Upon the addition of salt, significant restructuring of water is observed, leading to a net decrease in order at the surface.

Polarization- and Wavelength-Dependent Shell-Isolated-Nanoparticle-Enhanced Sum-Frequency Generation with High Sensitivity.

Sum-frequency generation (SFG) spectroscopy is a highly versatile tool for surface analysis. Improving the SFG intensity per molecule is important for observing low concentrations of surface species and intermediates in dynamic systems. Herein, Shell-Isolated-Nanoparticle-Enhanced SFG (SHINE-SFG) was used to probe a model substrate. The model substrate, p-mercaptobenzonitrile adsorbed on a Au film with SHINs deposited on top, provided an enhancement factor of up to 10^{5}. Through wavelength- and polarization-dependent SHINE-SFG spectroscopy, the majority of the signal enhancement was found to come from both plasmon enhanced emission and chemical enhancement mechanisms. A new enhancement regime, i.e., the nonlinear coupling of SHINE-SFG with difference frequency generation, was also identified. This novel mechanism provides insight into the enhancement of nonlinear coherent spectroscopies and a possible strategy for the rational design of enhancing substrates utilizing coupling processes.

Dual Resonant Sum Frequency Generations from Two-Dimensional Materials.

We propose dual resonant optical sum frequency generation (SFG), where the two most singular resonances could be selected, and report for the monolayer (1L-) WSe2 when one (ω1) of two excitation pulses is resonant to A exciton and their sum frequency (ω1 + ω2) to D exciton. The dual resonant SFG confirms that, under an irradiation of ω1 and ω2 pulses with the same fluence of ∼1.4 × 1010 W/m2, its signal intensity could be enhanced about 20 times higher than the resonant SHG (i.e., 2ω1 to the D excitonic absorption). Further, the dual resonant SFG intensity of 1L-WSe2 is found to be 1 order of magnitude higher than the single resonant SFG intensity of 1L-WS2 under the same condition of two-pulse irradiation. Finally, observations of the dual resonant SFG are thoroughly examined using real-time time-dependent density functional theory (rt-TDDFT), and the relevant nonlinear optical characteristics are scrutinized using the Greenwood-Kubo formalism.

Anomalous pH-Dependent Enhancement of p-Methyl Benzoic Acid Sum-Frequency Intensities: Cooperative Surface Adsorption Effects.

Vibrational sum-frequency generation (SFG) spectroscopy is used to determine the surface pKa of p-methyl benzoic acid (pMBA) at the air-water interface by monitoring the carbonyl and carboxylate stretching modes over the pH range of 2 to 12. The SFG intensities of pMBA and its conjugate base, p-methyl benzoate (pMBA-), exhibit an anomalously large enhancement over a narrow pH range (∼0.5) centered at pH 6.3 near the SFG-determined surface pKa, 5.9 ± 0.1. The increase in the surface pKa relative to the bulk value of 4.34 is consistent with the trend previously observed for long chain carboxylic acids in which the surface pKa is higher than the bulk solution pKa. SFG polarization studies help distinguish the orientation and number density contributions to this observed anomalous surface phenomenon. The large SFG intensity increase is attributed to an increase in the pMBA and pMBA- surface concentrations in this narrow pH range due to a cooperative adsorption effect between pMBA and pMBA-. This cooperativity is manifested only on the 2D air-water interface, where the interactions between the acid and base are not as dielectrically screened as in the aqueous bulk phase. Surface effects are critical to understanding and controlling the reactivity, solubility, and behavior of organic acids at interfaces and can have an impact on biomedical applications.

Sum frequency generation, calculation of absolute intensities, comparison with experiments, and two-field relaxation-based derivation

The experimental sum frequency generation (SFG) spectrum is the response to an infrared pulse and a visible pulse and is a highly surface-sensitive technique. We treat the surface dangling OH bonds at the air/water interface and focus on the absolute SFG intensities for the resonant terms, a focus that permits insight into the consequences of some approximations. For the polarization combinations, the calculated linewidths for the water interface dangling OH SFG band at 3,700 [Formula: see text] are, as usual, too large, because of the customary neglect of motional narrowing. The integrated spectrum is used to circumvent this problem and justified here using a Kubo-like formalism and theoretical integrated band intensities rather than peak intensities. Only relative SFG intensities are usually reported. The absolute integrated SFG intensities for three polarization combinations for sum frequency, visible, and infrared beams are computed. We use molecular dynamics and the dipole and the polarizability matrix elements obtained from infrared and Raman studies of [Formula: see text]O vapor. The theoretical expressions for two of the absolute susceptibilities contain only a single term and agree with experiment to about a factor of 1.3, with no adjustable parameters. The Fresnel factors are included in that comparison. One of the susceptibilities contains instead four positive and negative terms and agrees less well. The expression for the SFG correlation function is normally derived from a statistical mechanical formulation using a time-evolving density matrix. We show how a derivation based on a two-field relaxation leads to the same final result.

Interfacial water at synthetic and natural lipid bilayers probed by vibrational sum-frequency generation spectroscopy

Background: At lipid interfaces, water plays a crucial role in carrying biological processes, so that there is a huge interest in unravelling the behaviour of water close to membranes. At charged bio-interfaces, water dipoles form an organized layer. Probing such an interfacial thin layer buried between macroscopic bulk environments is a real challenge. Vibrational sum frequency generation (SFG) spectroscopy is intrinsically specific to interfaces, and has already proven to be an ideal tool to investigate model membranes and their surrounding water. Objectives: The goal of this work is to measure the vibrational SFG response of interfacial water around different model membranes — from easiest synthetic lipids to more complex natural lipids, — in order to use it as diagnostic signal able to distinguish the lipid bilayer interface by its charge properties. Materials and methods: Lipid bilayers made either of synthetic or natural lipids (Avanti Polar Lipids) were physisorbed on CaF2 prisms (Crystran), by using the method of the spontaneous fusion of lipid vesicles, to form so called solid-supported lipid bilayers (SSLBs). The model membranes were investigated by SFG spectroscopy at the solid/water interface. Results: The SFG response was measured between 3600 cm-1 and 2800 cm-1, where OH stretching vibrations of water molecules show-up. The SFG intensity of the OH peak maximum at 3125 cm-1 was recorded during the adsorption of lipid vesicles on the surface, and provided knowledge of the changes of the charge properties of the interface due to the adsorption of the model membranes. The SFG signal indicated that the organization of water was larger at negatively charged than at positively lipid interfaces, and reached the highest value with natural E. сoli сardiolipin layers. Moreover, when the full composition of natural lipids was unknown, the behaviour of the SFG response enabled establishing the charge characteristics of the corresponding lipid interfaces. Conclusion: The SFG response of water enabled estimating average charge behaviour of synthetic and natural lipid bilayers in pure water, thus paving the way to use the SFG signal of water as new diagnostic tool to identify lipid interfaces.