Polarization Modulation Infrared Reflection Absorption Research Articles

H/D Isotope Effects in the Electrochemistry of Electrochromic Iron Hexacyanoruthenate

AbstractThe electrochemical deposition of iron hexacyanoruthenate (Fe−HCR) on gold electrodes was studied in electrolytes prepared with light and heavy water. Cyclic voltammetry of the material during preparation and after transfer to a precursor‐free solution exhibits two reductions peaks in H2O‐based electrolytes but only one reduction peak in D2O‐based electrolytes. The voltammetric behavior changes reversibly upon transfer of the material between D2O‐based and H2O‐based 1 mol L−1 KCl solutions. No clear structural differences between samples prepared in D2O and H2O were detected by means of X‐ray photoelectron spectroscopy (XPS) and polarization modulation infrared reflection absorption spectroscopy (PM IRRAS). We noted a relatively slow exchange of coordinated water and a fast exchange of zeolitic water. Using voltammetric experiments we could rule out simple effects of solution conductivity for K+, participation of H+/D+ in the charge compensation and surface effects on the observed dependence of the peak splitting on the isotopic composition of the solvent. The most likely reason for the observed behavior is the different structure of the H‐bonded water network of coordinated H2O and zeolitic H2O/D2O which is supported by the PM IRRAS data.

In-situ polarization modulation IRRAS investigation of ammonia electrooxidation on Pt-Ir and Pt-Ru nanoparticles prepared on engineered catalyst supports

The catalytic activity and surface reactivity of monometallic Pt and bimetallic Pt-Ir and Pt-Ru nanoparticles, supported on two distinct Engineered Catalyst Supports (ECSs), were investigated for the Ammonia Electrooxidation Reaction (AmER) in alkaline media. XRD measurements confirmed alloy formation between Pt-Ir and Pt-Ru nanoparticles, as indicated by the shift of the (111) reflection to higher 2θ values. Cyclic voltammetry, linear sweep voltammetry, and chronoamperometry experiments were conducted to assess the catalytic activity of the Pt, Pt-Ir, and Pt-Ru electrocatalysts. All bimetallic catalysts exhibited lower onset potentials compared to Pt. The differing Tafel slopes between Pt (74 mV dec⁻¹), Pt-Ir (152 mV dec⁻¹), and Pt-Ru (118–197 mV dec⁻¹) suggest that alloying Pt with Ir or Ru alters the reaction mechanisms. Furthermore, the bimetallic Pt-Ir and Pt-Ru catalysts demonstrated greater tolerance for concentrated ammonia solutions relative to Pt.In-situ Polarization Modulation Infrared Reflection Absorption Spectroscopy (PM-IRRAS) provided insights into the formation of N-H species, azide anions (N₃⁻), and N-O compounds. For the Pt-Ru catalyst, an additional peak around ∼3600 cm⁻¹ was observed, corresponding to OH⁻ species. The PM-IRRAS results align with the Gerischer–Mauerer mechanism, indicating that partially dehydrogenated ammonia adsorbates act as active intermediates in the oxidation of ammonia over Pt-Ir and Pt-Ru catalysts.

Analysis of a film-forming amine response in impedance spectra

In this study, a detailed analysis of the dielectric response of a film-forming amine (1,3 N-oleyl propanediamine (OLDA)) in impedance spectra is presented, addressing both “bulk” OLDA and adsorbed OLDA layer on a carbon steel surface. The dielectric response of the “bulk” OLDA in the absence of electrolyte over an extensive temperature range (-150 °C to 50 °C) was explored by broadband dielectric spectroscopy (BDS). Subsequently, the response of the OLDA adsorbed layer on the steel surface was analysed by electrochemical impedance spectroscopy (EIS), for various immersion times (2 min to100 min) and temperatures (25 °C to 75 °C) in an aqueous NaCl solution.Impedance spectra (BDS and EIS), interpreted through the complex conductivity formalism, showed a resistive behaviour in the mid-frequency range and a capacitive behaviour at higher frequencies. The high frequency capacitive response in EIS spectra was shown to contain contributions from the OLDA layer and from the electrochemical double layer, as well. The temperature dependency of the OLDA conductivity was found to be similar to the temperature dependency of the electrolyte resistance, suggesting that the electrolyte (water and ions) crossed the OLDA layer to reach the steel surface. The presence of gauche defects in the OLDA alkyl chain, revealed by polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) analyses, may be a contributing factor to the disorder in the adsorbed layer, facilitating electrolyte infiltration.

Adhesion promotion and corrosion resistance of mixed phosphonic acid monolayers on AA 2024

Mixed monolayers were used to prepare surfaces of AA 2024 with precisely controlled corrosion protective and adhesive properties. In this study, the corrosion inhibition and adhesion promotion as a function of the composition of mixed phosphonic acid monolayers on AA 2024 was investigated. Mixed monolayers were formed by competitive adsorption of n-dodecylphosphonic acid (DPA) and 12-aminododecylphosphonic acid (ADPA) from ethanolic solution. Their compositions were determined by X-ray photoelectron spectroscopy. Atomic force microscopy studies and polarization modulation infrared reflection–absorption spectroscopy as well as contact angle studies proved the formation of monolayers with varying surface chemistry. The wet-adhesion strength was tested by 90° peel tests after application of an epoxy-amine adhesive. Linear sweep voltammetry was performed to evaluate the corrosion inhibition of the monolayers, and the corrosion resistance of the metal/adhesive composite was investigated by means of its corrosive delamination behavior. The electrochemical measurements demonstrated that reducing the surface concentration of ADPA improved the corrosion inhibition of the monolayer. However, the corrosive delamination experiments revealed that the improved corrosion inhibition by the self-organized DPA is of minor importance for the corrosion behavior of the metal/adhesive composite, as the adhesive properties dominate.

Adsorption kinetics and inhibition mechanisms of a film-forming amine on carbon steel surfaces

In the present study, the inhibition mechanisms and the adsorption kinetics of a film-forming amine (N-oleyl-1,3-propanediamine, OLDA) were investigated on a carbon steel surface in various corrosive environments, relevant to industrial water/steam circuits. In situ electrochemical characterizations including Electrochemical Impedance Spectroscopy (EIS) and polarization curves were combined with ex situ surface analysis, such as Polarization Modulation Infrared Reflection Absorption Spectroscopy (PM-IRRAS) and Raman Spectroscopy. OLDA acts as a mixed inhibitor for all the studied conditions. In a deaerated medium, OLDA adsorption is temperature-independent (25 °C–50 °C) and PM-IRRAS analyses reveal the formation of a monolayer (thickness of about 1.6 nm) on the steel surface. In aerated media, mixed OLDA/corrosion products layers are formed exceeding the monolayer thickness (about 20 nm). Finally, the presence of a well-defined time constant in the high frequency range in impedance spectra is correlated with the accumulation of OLDA molecules with corrosion products.

How do Antimicrobial Peptides Interact with the Outer Membrane of Gram-Negative Bacteria? Role of Lipopolysaccharides in Peptide Binding, Anchoring, and Penetration.

Gram-negative bacteria possess a complex structural cell envelope that constitutes a barrier for antimicrobial peptides that neutralize the microbes by disrupting their cell membranes. Computational and experimental approaches were used to study a model outer membrane interaction with an antimicrobial peptide, melittin. The investigated membrane included di[3-deoxy-d-manno-octulosonyl]-lipid A (KLA) in the outer leaflet and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) in the inner leaflet. Molecular dynamics simulations revealed that the positively charged helical C-terminus of melittin anchors rapidly into the hydrophilic headgroup region of KLA, while the flexible N-terminus makes contacts with the phosphate groups of KLA, supporting melittin penetration into the boundary between the hydrophilic and hydrophobic regions of the lipids. Electrochemical techniques confirmed the binding of melittin to the model membrane. To probe the peptide conformation and orientation during interaction with the membrane, polarization modulation infrared reflection absorption spectroscopy was used. The measurements revealed conformational changes in the peptide, accompanied by reorientation and translocation of the peptide at the membrane surface. The study suggests that melittin insertion into the outer membrane affects its permeability and capacitance but does not disturb the membrane's bilayer structure, indicating a distinct mechanism of the peptide action on the outer membrane of Gram-negative bacteria.

Statin Action Targets Lipid Rafts of Cell Membranes: GIXD/PM-IRRAS Investigation of Langmuir Monolayers.

Lipid rafts are condensed regions of cell membranes rich in cholesterol and sphingomyelin, which constitute the target for anticholesterolemic drugs - statins. In this work, we use for the first time a combined grazing-incidence X-ray diffraction (GIXD)/polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS)/Brewster angle microscopy (BAM) approach to show the statin effect on model lipid rafts and its components assembled in Langmuir monolayers at the air-water interface. Two representatives of these drugs, fluvastatin (FLU) and cerivastatin (CER), of different hydrophobicity were chosen, while cholesterol (Chol) and sphingomyelin (SM), and their 1:1 mixture were selected to form condensed monolayers of lipid rafts. The effect of statins on the single components of lipid rafts indicated that both the hydrophobicity of the drugs and the organization of the layer determined the drug-lipid interaction. For cholesterol monolayers, only the most hydrophobic CER was effectively changing the film structure, while for the less organized sphingomyelin, the biggest effect was observed for FLU. This drug affected both the polar headgroup region as shown by PM-IRRAS results and the 2D crystalline structure of the SM monolayer as evidenced by GIXD. Measurements performed for Chol/SM 1:1 models proved also that the statin effect depends on the presence of Chol-SM complexes. In this case, the less hydrophobic FLU was not able to penetrate the binary layer at all, while exposure to the hydrophobic CER resulted in the phase separation and formation of ordered assemblies. The changes in the membrane properties were visualized by BAM images and GIXD patterns and confirmed by thermodynamic parameters of hysteresis in the Langmuir monolayer compression-decompression experiments.

Film-forming amines adsorption and corrosion kinetics on carbon steel surface in neutral solution investigated by EIS and PM-IRRAS analysis

N-oleyl-1, 3-propanediamine (OLDA) is one of the most used film-forming amines (FFAs) for the corrosion protection of carbon steel circuits of a wide range of industrial plants. The present work focused on the study of OLDA adsorption kinetics onto the carbon steel surface by electrochemical impedance spectroscopy (EIS). The diagrams were obtained in a 200 mg.kg−1 NaCl with and without OLDA for immersion times ranging from 2 min to 140 min and for two temperatures (25°C and 50°C). It was shown that, from the beginning of immersion (2 min), OLDA adsorption and corrosion occurred simultaneously on the carbon steel surface. With increasing immersion time, a dynamic and competitive process between both phenomena was established until a steady-state was reached and the corrosion was slowed down. Ex situ Raman spectroscopy analyses revealed a heterogeneous distribution of the OLDA molecules on the surface with a preferential adsorption on the corrosion products, mainly γ-FeOOH. Ex situ Polarization Modulation Infrared Reflection Absorption Spectroscopy (PM-IRRAS) analyses confirmed the formation of mixed barrier layers (OLDA molecules/corrosion products) whose thickness increased with time and temperature.

Current Response Enhancement According to the Doping Anion’s Nature in Redox Polyelectrolyte─Enzyme Assemblies

High-power density output in enzymatic fuel cells is a key feature to reduce the size of self-powered implantable medical devices. Electron transfer mediated through redox polyelectrolytes allows the transport of electrons from enzymes away from the electrode, improving the current output. It is known that doping ions in polyelectrolytes introduce relevant characteristics in the generation of assemblies regarding mass adsorption and stiffness. In this work, binary 1:1 sodium salts (NaX; X = F–, Cl–, Br–, NO3–, ClO4–) were studied as doping ions of two redox polyelectrolytes (osmium-based branched polyethyleneimine and osmium-based linear polyallylamine) to enhance the adsorption and electron transfer process in glucose oxidase/redox polyelectrolyte assemblies. Cyclic voltammetry, polarization modulation infrared reflection absorption spectroscopy, quartz crystal microbalance with dissipation, and atomic force microscopy were used to understand the growth mechanism of these films and their performance. Ion hydrophobicity plays a key role, bromide being the one that generates the greater absorption and the best electron transfer efficiency for both redox polyelectrolytes. Branched polyethyleneimine doped with bromide was the best combination for the construction of bioanodes. Its application on an O2–glucose enzymatic fuel cell yields a power density output of 2.5 mW cm–2, achieving state-of-the-art performance.

Low Temperature Multilayer Adsorption of Methanol and Ethanol on Platinum.

Adsorption of methanol and ethanol on the clean Pt (111) surface was studied at temperatures between 80 and 130 K using polarization-modulation infrared reflection absorption spectroscopy (PM-IRRAS). It was shown that adsorption of methanol at 80 K leads to the formation of amorphous solid methanol, and fast crystallization of the amorphous phase occurs upon warming at 100 K. Vapor deposition of methanol at 100 K directly leads to the formation of well-crystallized layers of solid methanol. According to PM-IRRAS, these crystalline layers consist of chains of hydrogen-bonded methanol molecules lying in a plane oriented close to the normal to the platinum surface. Adsorbed methanol is removed completely from platinum after heating to 120 K. Vapor deposition of ethanol at 80 K also leads to the formation of amorphous solid ethanol. However, subsequent warming does not lead to ordering of the adsorption layers, and at 130 K, ethanol is also completely desorbed.

The Ca2+ response of a smart forisome protein is dependent on polymerization.

Forisomes are giant self‐assembling mechanoproteins that undergo reversible structural changes in response to Ca2+ and various other stimuli. Artificial forisomes assembled from the monomer MtSEO‐F1 can be used as smart biomaterials, but the molecular basis of their functionality is not understood. To determine the role of protein polymerization in forisome activity, we tested the Ca2+ association of MtSEO‐F1 dimers (the basic polymerization unit) by circular dichroism spectroscopy and microscale thermophoresis. We found that soluble MtSEO‐F1 dimers neither associate with Ca2+ nor undergo structural changes. However, polarization modulation infrared reflection absorption spectroscopy revealed that aggregated MtSEO‐F1 dimers and fully‐assembled forisomes associate with Ca2+, allowing the hydration of poorly‐hydrated protein areas. A change in the signal profile of complete forisomes indicated that Ca2+ interacts with negatively‐charged regions in the protein complexes that only become available during aggregation. We conclude that aggregation is required to establish the Ca2+ response of forisome polymers.

Electrochemical Characterization of Redox Probes Confined in 3D Conducting Polymer Networks.

In this manuscript we present a versatile platform for introducing functional redox species into tailor‐made 3D redox polymer networks. Electrochemical characterization based on cyclic voltammetry is applied to verify the immobilization of the redox species within the conducting networks. Ultimately this strategy shall be extended to (photo)electrocatalytic applications which will profit from the conducting polymer matrix. Soluble precursor copolymers are synthesized via radical copolymerization of vinyltriphenylamine (VTPA) with chloromethylstyrene (CMS) in different ratios, whereas CMS is subsequently converted into azidomethylstyrene (AMS) to yield poly(VTPA‐co‐AMS) copolymers. Spin‐coating of poly(VTPA‐co‐AMS) on gold electrodes yields thin films which are converted into stable polymer network structures by electrochemical crosslinking of the polymer chains via their pendant triphenylamine groups to yield N,N,N′,N′‐tetraphenylbenzidine (TPB) crosslinking points. Finally, the resulting redox‐active, TPB‐crosslinked films are functionalized with ethynylferrocene (EFc) as a representative redox probe using a click reaction. Main experimental tools are polarization modulation infrared reflection absorption spectroscopy and scan rate dependent cyclic voltammetry. Especially the latter proves the successful conversion and the immobilization of redox probes in the polymer matrix. The results are compared with the reference system of azide‐terminated self‐assembled monolayers on gold substrates, allowing to distinguish between free and immobilized EFc species.

Cover Feature: Effect of Ectoine on the Conformation and Hybridization of dsDNA in Monolayer Films: A Spectroelectrochemical Study (ChemElectroChem 20/2021)

The Cover Feature shows double strand DNA monolayers on the gold electrode surface stored for a long time in aqueous solutions in the absence and presence of ectoine, compatible solute. The use of polarization modulation infrared reflection absorption spectroscopy allowed to follow both the electric potential and time dependent changes in the conformation, orientation and hybridization state of DNA helices. Our results demonstrate that in monolayer films, the addition of compatible solutes, such as ectoine, to the electrolyte solution may have a protective effect on dsDNA stability over time. More information can be found in the Article by J. Wittmar et al.

Mixed monolayer of a nucleolipid and a phospholipid has improved properties for spectroelectrochemical sensing of complementary nucleobases

A mixed monolayer of 1,2-dipalmitoyl-sn-glycero-3-(cytidine diphosphate) nucleolipid (DG-CDP) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was transferred from the air–water interface onto an gold (1 1 1) electrode surface using the Langmuir-Schaeffer (horizontal touch) technique. Compression isotherms were recorded for the monolayer spread at the air–water interface. These measurements demonstrated that the mixture composition 7:3 of DG-CDP:DPPC displayed reduced repulsive interactions between polar heads and ideal packing of acyl chains. Chronocoulometric experiments were performed to demonstrate stability of this film at the gold–solution interface. Photon polarization modulation infrared reflection absorption spectroscopy showed that the tilt angle of the acyl chains is slightly higher, and the chains are less twisted in the mixed than in the pure nucleolipid monolayer. In the mixed film, the tilt of the cytosine moiety (the sensing element of the monolayer) assumes a large angle with respect to the surface normal, even when the monolayer becomes detached from the gold surface. The mixed monolayer has improved properties for future applications for detection of guanine, its complementary base.

Adsorption of lysozyme on gold surfaces in the presence of an external electric potential

Adsorbed protein films consist of essential building blocks of many biotechnological and biomedical devices. The electrostatic potential may significantly modulate the protein behaviour on surfaces, affecting their structure and biological activity. In this study, lysozyme was used to investigate the effects of applied electric potentials on adsorption and the protein structure. The pH and the surface charge determine the amount and secondary structure of adsorbed lysozyme on a gold surface. In-situ measurements using polarization modulation infrared reflection absorption spectroscopy indicated that the concentration of both the adsorbed anions and the lysozyme led to conformational changes in the protein film, which was demonstrated by a greater amount of aggregated β-sheets in films fabricated at net positive charges of the Au electrode (Eads > Epzc). The changes in secondary structure involved two parallel processes. One comprised changes in the hydration/hydrogen-bond network at helices, leading to diverse helical structures: α-, 310- and/or π-helices. In the second process β-turns, β-sheets, and random coils displayed an ability to form aggregated β-sheet structures. The study illuminates the understanding of electrical potential-dependent changes involved in the protein misfolding process.

CO and H2 adsorption on Au-Ni bimetallic surfaces: a combined experimental and DFT theoretical study

Au-Ni bimetallic thin films were grown on refractory metal substrates. CO and H2 adsorption on Au-Ni bimetallic surfaces have been studied by a combination of in situ polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS), temperature-programmed desorption (TPD), and density functional theory (DFT) calculations. It is found CO desorption peak shifts from 413 K on pure Ni surfaces to 293 K on the isolated Ni atoms formed by alloying with Au atoms. The sharp decrease of CO desorption temperature on Au-Ni surfaces with increasing Au coverage is caused by the change of the favored CO adsorption sites from bridge/hollow sites on pure Ni surfaces to Ni top sites on Au-Ni bimetallic surfaces. In situ PM-IRRAS shows two CO adsorption bands on Au sites at 2119 cm−1 and 2103 cm−1 on Au-Ni surfaces at 80 K, which are due to CO bound on under-coordinated Au atoms and electron negatively charged Au sites modified with nearby Ni atoms, respectively. Even with the Au-Ni surface temperature at as low as 100 K, CO adsorption induced Ni surface segregation has been observed by in situ PM-IRRAS. Furthermore, DFT calculation results discover the adsorption energy of CO on Ni top sites continues to decrease with increasing Au coverage due to the geometric ensemble effect and the lowered d-band center after Ni alloying with Au. H2 desorption temperature decreases from 363 K on pure Ni thin films to 302 K with increasing Au coverage to 0.6 ML. A new H2 peak appears at around 170 K on the Au-Ni surfaces with Au coverages between 0.6 ML and 0.9 ML. This new H2 TPD peak is assigned to H2 desorption from the totally isolated Ni sites. With Au coverage above 1.5 ML, there is no any H2 desorption detected. The combined surface science studies and DFT calculations provide new insights into the surface structure-activity correlation of Ni-base bimetallic surface alloys.

How the replacement of cholesterol by 25-hydroxycholesterol affects the interactions with sphingolipids: The Langmuir Monolayer Study complemented with theoretical calculations.

In this paper, a representative of chain-oxidized sterols, 25-hydroxycholesterol (25-OH), has been studied in Langmuir monolayers mixed with the sphingolipids sphingomyelin (SM) and ganglioside (GM1) to build lipid rafts. A classical Langmuir monolayer approach based on thermodynamic analysis of interactions was complemented with microscopic visualization of films (Brewster angle microscopy), surface-sensitive spectroscopy (polarization modulation-infrared reflection-absorption spectroscopy) and theoretical calculations (density functional theory modelling and molecular dynamics simulations). Strong interactions between 25-OH and both investigated sphingolipids enabled the formation of surface complexes. As known from previous studies, 25-OH in pure monolayers can be anchored to the water surface with a hydroxyl group at either C(3) or C(25). In this study, we investigated how the presence of additional strong interactions with sphingolipids modifies the surface arrangement of 25-OH. Results have shown that, in the 25-OH/GM1 system, there are no preferences regarding the orientation of the 25-OH molecule in surface complexes and two types of complexes are formed. On the other hand, SM enforces one specific orientation of 25-OH: being anchored with the C(3)-OH group to the water. The strength of interactions between the studied sphingolipids and 25-OH versus cholesterol is similar, which indicates that cholesterol may well be replaced by oxysterol in the lipid raft system. In this way, the composition of lipid rafts can be modified, changing their rheological properties and, as a consequence, influencing their proper functioning.

Ion transport mechanism in gramicidin A channels formed in floating bilayer lipid membranes supported on gold electrodes

The ion channel properties of gramicidin A (gA) in a model floating bilayer lipid membrane were studied by electrochemical impedance spectroscopy (EIS) and polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS). To determine if the transport of Na+ across the membrane is coupled with the transport of the counterion, fluoride and perchlorate anions were explored due to their large differences in hydration energy. The combined EIS and IR experiments demonstrated that in contrast to valinomycin, the mechanism of Na+ transfer through the gA channel is independent of the accompanying anion present in the supporting electrolyte. These studies also demonstrated that the conductivity of the gA sodium channel depends on the transmembrane potential and the IR measurements indicated that more gA molecules insert into the bilayer with the decrease of the transmembrane potential. The results of this present study demonstrate that the fBLM system is a suitable model bilayer for the in situ investigations of membrane-active peptides.

Towards the systematic design of multilayer O/W emulsions with tannic acid as an interfacial antioxidant.

This work discusses the possibility of designing multilayer oil-in-water emulsions to introduce the maximum possible amount of an antioxidant at the droplet interfaces for the optimal protection of a linseed oil core against oxidation, using a systematic three-step colloidal procedure. An antioxidant (here Tannic Acid – TA) is chosen and its interactions with a primary emulsifier (here Bovine Serum Albumin – BSA) and several polysaccharides are first examined in solution using turbidity measurements. As a second step, LbL deposition on solid surfaces is used to determine which of the polysaccharides to combine with BSA and tannic acid in a multilayer system to ensure maximum presence of tannic acid in the films. From UV-vis and polarization modulation infrared reflection–absorption (PM-IRRAS) spectroscopic measurements it is suggested that the best components to use in a multilayer emulsion droplet, together with BSA and TA, are chitosan and pectin. BSA, chitosan and pectin are subsequently used for the formation of three-layer linseed oil emulsions, and tannic acid is introduced into any of the three layers as an antioxidant. The effect of the exact placement of tannic acid on the oxidative stabilization of linseed oil is assessed by monitoring the fluorescence of Nile red, dissolved in the oil droplets, under the attack of radicals generated in the aqueous phase of the emulsion. From the results it appears that the three-stage procedure presented here can serve to identify successful combinations of interfacial components of multilayer emulsions. It is also concluded that the exact interfacial placement of the antioxidant plays an important role in the oxidative stabilization of the valuable oil core.

Influence of the Molecular Orientation and Ionization of Self-Assembled Monolayers in Biosensors: Application to Genosensors of Prostate Cancer Antigen 3

Electrochemical sensing depends on the immobilization of biorecognition elements and charge transfer to the electrode, which can be tuned and sometimes controlled using self-assembled monolayers (SAMs) as matrices. In this study, we show that an efficient immobilization of an ssDNA probe to detect the prostate cancer antigen 3 biomarker is achieved even for less organized SAMs provided that the terminal groups were ionized. These conclusions were reached by comparing the biosensors made with 11-mercaptoundecanoic acid (11-MUA) prepared in ethanol and thioglycolic acid (mercaptoacetic acid: MAA) prepared in ethanolic and aqueous solvents. The highest sensitivity with electrochemical impedance spectroscopy was observed for sensors assembled over MAA monolayers on gold prepared in ethanol and water (at pH 12.5), with limits of detection of 1.2 × 10–9 and 1.7 × 10–9 mol L–1, respectively. The biosensors made on MAA films prepared in water at pH 7.0 and 2.5 were not efficient because they are mostly terminated by protonated carboxylic acid groups, according to sum-frequency generation vibrational and polarization modulation-infrared reflection-absorption (PM-IRRAS) spectroscopies, which prevent an adequate attachment of the biomarker. Well-organized SAMs of 11-MUA also showed good detection performance, but the limit of detection was not as low, 2.1 × 10–9 mol L–1, probably due to its long chains that decrease the charge transfer. Therefore, the matrix fabrication method can control the molecular assembly of biosensors and hence their performance.