Applications of Raman and Surface-Enhanced Raman Scattering Techniques to Humic Substances

This paper highlights the use of Raman, FT-Raman and surface-enhanced Raman scattering (SERS) techniques for the study of humic substances. In contrast to other technologies which reveal information only about the average compositions and the kinds of functional groups present in humic substances, Raman and especially FT-Raman spectroscopies characterize the building blocks of humic substances and their changes in derivation and separation processes. Furthermore, surface-enhanced Raman scattering (SERS) techniques are able to readily detect humic substances and co-existing organic species at low concentrations typically found in natural environments and reveal definitive information about the specific groups in humic substances that bind on metal electrodes. Further applications of both Raman and SERS techniques can be extended to complicated systems as well as real environmental samples. Experiments have demonstrated: (1). the backbones of humic substances are structurally disordered carbon networks in most cases; (2). The backbones of humic substances from different sources and types are similar to each other; (3). Normal Raman spectroscopic study of humic substances should concentrate on the use of near-IR laser(s) resulting from strong fluorescence background and self-adsorption under the excitation with visible laser irradiation; (4). FT-Raman spectroscopy is the required analytical method to assess the effectivity of fractionation methods; (5). SERS spectra of humic substances on metal colloids and films are in most aces very similar to the corresponding Raman spectra of neutralized samples; (6). SERS techniques are very sensitive and highly selective, also both visible lasers and near-IR lasers can be used for SERS study; (7). SERS spectra on metal electrodes may provide additional information about the binding sites and adsorption mechanisms of humic substances on metal surfaces.     

Some aspects of SERS temporal fluctuations: analysis of the most intense spectra of hydrogenated amorphous carbon deposited on silver

Surface‐enhanced Raman scattering (SERS) spectra of hydrogenated amorphous carbon (a C:H) deposited on silver substrates have been recorded with a confocal Raman microscope. When scattered radiation is collected during a short time from an area of a few square micrometres, the subsequently measured SERS spectra often exhibit strong temporal changes (fluctuations). In this paper we present examples of spectra for which the intensity maxima of the fluctuating narrow Raman bands are significantly higher than that of the background (the background is usually dominated by two broad Raman bands centred at about 1350 and 1590 cm⁻¹). In a series of successively measured spectra, one can find spectra with noticeably different total integral intensity. This suggests that the results of averaging the spectra revealing strong and weak fluctuations may be different (at least in intensity). The influence of some electrolytes on the SERS spectral fluctuations is also analysed. Our experiments revealed that the efficiencies of quenching of the SERS spectral fluctuations by various electrolytes are significantly different. We suggest that only anions directly interacting with the metal surface quench strong SERS fluctuations, and that the large differences between chloride and perchloride solutions are caused by differences in the strength of interaction of Cl⁻ and ClO₄⁻ anions with the silver surface. Copyright © 2007 John Wiley & Sons, Ltd.     

Surface‐enhanced Raman scattering on colloid gels originated from low molecular weight gelator

Surface‐enhanced Raman scattering (SERS) on silver and gold colloid gels formed by a low molecular weight organic gelator, bis‐(S‐phenylalanine) oxalyl amide, was obtained. Strong Raman signals dominate in the SERS spectra of hydrogels containing silver nanoparticles prepared by citrate and borohydride reduction methods, whereas broad bands of low intensity are detected in the spectra of gold colloid gels. Resemblance between Raman spectrum of the crystalline substance and the SERS spectra of the silver nanoparticle–hydrogel composites implies the electromagnetic nature of the signal enhancement. A change in Raman intensity of the benzene and amide II bands caused by an increase in temperature and concentration indicates that the gelling molecules are strongly attached through the benzene moieties to the metal nanoparticles while participating in gel formation by intermolecular hydrogen bonding between the adjacent oxalyl amide groups. Transmission electron microscopy reveals a dense gel structure in the close vicinity of the enhancing metal particles for both silver colloid gels. Copyright © 2008 John Wiley & Sons, Ltd.     

Raman,infrared, SERS and theoretical study of 3‐(1‐phenylpropan‐2‐ylamino) propanenitrile,fenproporex

Infrared, Raman and surface‐enhanced Raman scattering (SERS) spectra of 3‐(1‐phenylpropan‐2‐ylamino)propanenitrile (fenproporex) have been recorded. Density functional theory (DFT) with the B3LYP functional was used for optimizations of ground state geometries and simulation of Raman and SERS vibrational spectra of this molecule. Bands of the vibrational spectra were assigned in detail. The comparison of SERS spectra obtained by using colloidal silver and gold nanoparticles with the corresponding Raman spectrum reveals enhancement and shifts in bands, suggesting a possible partial charge‐transfer mechanism in the SERS effect. Information about the orientation of fenproporex on the nanometer‐sized metal structures is also obtained. Copyright © 2011 John Wiley & Sons, Ltd.     

Adjustment and control of SERS activity of metal substrates by pressure

Metal pellets of silver and copper for surface‐enhanced Raman scattering (SERS) spectroscopy were prepared by compression with different pressures. It was found that the SERS activity of the pellet could be controlled by pressure. Enhanced Raman scattering properties of the metal pellets in the presence of adsorbed 4‐mercaptobenzoic acid (4‐MBA) with excitation at 632.8 or 514 nm could be obtained by choosing proper pressure of pellatization. The SERS peak intensity of the band at ∼1584 cm⁻¹ of 4‐MBA adsorbed on the metal pellets varies as a function of applied pressure, and which is about 1.2–32 times greater than when it is adsorbed on silver and copper particles. The calculated results of three‐dimensional finite‐difference time‐domain method (3D‐FDTD) are in good agreement with the experimental data. Moreover, no spurious peaks appear in the SERS spectra of the samples because no other chemicals are involved in the simple preparation process of the metal pellets, which will facilitate its use as an SERS‐active substrate for analytical purposes. In summary, SERS‐active metal pellets can be produced simply and cost effectively by the method reported here, and this method is expected to be utilized in the development of SERS‐based analytical devices. Copyright © 2009 John Wiley & Sons, Ltd.     

Raman and SERS study of metoclopramide at different pH values

Conjugate acid–base forms of the drug metoclopramide were investigated by Raman spectroscopy in aqueous solutions and by surface‐enhanced Raman scattering (SERS), when the molecules were adsorbed on colloidal silver surfaces. Raman spectra were recorded at pH values below 8, metoclopramide being poorly water soluble at higher pH values. The SERS spectra of metoclopramide were recorded in the 3–11 pH range, even in spite of its low solubility at basic pH values. The Raman and SERS spectra were assigned by means of density functional theory (DFT) calculations. By monitoring several SERS marker bands, the protonated, neutral or the coexistence of both molecular species adsorbed on the colloidal silver particles could be evidenced. The adsorbate orientation was deduced to be perpendicular to the metal surface for the protonated molecular species and tilted for the neutral metoclopramide molecular species. Copyright © 2009 John Wiley & Sons, Ltd.     

Understanding tip‐enhanced Raman spectra of biological molecules: a combined Raman,SERS and TERS study

Although conventional Raman, surface‐enhanced Raman (SERS) and tip‐enhanced Raman spectroscopy (TERS) have been known for a long time, a direct, thorough comparison of these three methods has never been carried out. In this paper, spectra that were obtained by conventional Raman, SERS (on gold and silver substrates) and TERS (in ‘gap mode’ with silver tips and gold substrates) are compared to learn from their differences and similarities. Because the investigation of biological samples by TERS has recently become a hot topic, this work focuses on biologically relevant substances. Starting from the TER spectra of bovine serum albumin as an example for a protein, the dipeptides Phe–Phe and Tyr–Tyr and the tripeptide Tyr–Tyr–Tyr were investigated. The major findings were as follows. (1) We show that the widely used assumption that spectral bands do not shift when comparing SER, TER and conventional Raman spectra (except due to binding to the metal surface in SERS or TERS) is valid. However, band intensity ratios can differ significantly between these three methods. (2) Marker bands can be assigned, which should allow one to identify and localize proteins in complex biological environments in future investigations. From our results, general guidelines for the interpretation of TER spectra are proposed. Copyright © 2012 John Wiley & Sons, Ltd.     

Engineering of SERS Substrates Based on Noble Metal Nanomaterials for Chemical and Biomedical Applications

Abstract

Surface-enhanced Raman scattering (SERS) has attracted great interest due to its remarkable enhancement, excellent sensitivity, and the “fingerprinting” ability to produce distinct spectra for detecting various molecules. Noble metal nanomaterials have usually been employed as SERS-active substrates because of their strong SERS enhancement originated from their unique surface plasmon resonance (SPR) properties. Because the SPR property depends on metal material's size, shape, morphology, arrangement, and dielectric environment around metal nanostructures, the key to wider applications of SERS technique is to develop plasmon-resonant structures with novel geometries to enhance Raman signals and to control the periodic ordering of these structures over a large area to obtain reproducible Raman enhancement. This review presents a general view on the theory background of SERS effect and several basic concepts and focuses on recent progress in engineering metallic nanostructures with various morphologies using versatile methods for improving SERS properties. Their potential applications in the field of chemical detection and biological sensing are overviewed.     

The role of surface electronic states in surface enhanced raman scattering

The dependence of the surface enchanced Raman scattering (SERS) intensity on the electrode potential is considered in terms of the nonresonant adiabatic charge-transfer mechanism. Surface electronic states of a substrate metal are proposed to be responsible for the observed potential dependence of the SERS signal. The contribution of surface states to the nonlinear metal polarizability was found from electroreflectance spectra of silver single-crystals. Good agreement between the measured and calculated potential behaviour of the SERS signal was found for pyridine absorbed on the silver electrode.     

Metal–organic frameworks: a novel SERS substrate

We report the direct observation of surface‐enhanced Raman scattering (SERS) effect using metal–organic frameworks (MOFs) as substrates. Without the aid of any metal colloids or enhancing agents, the SERS signals of methyl orange (MO) adsorbed in MOFs were observed and even remained active if the organic linkers in MOFs were completely removed by high temperature and O₂ plasma treatments. It implies that the SERS active site is at the metal oxide clusters. The ultraviolet‐visible spectra of MO, MOFs, and MO–MOF complexes show that absorption peaks are far from laser excitation line. Thus, conventional resonance enhancement effect should be ruled out, and charge‐transfer mechanism is the most likely scenario responsible for the observed SERS effect. Density functional theory (DFT) was used to interpret the chemical enhancement mechanism and the adsorption orientation‐dependent SERS spectra in our observation. The preferred adsorption orientations calculated by DFT method are consistent with the observed SERS results. Copyright © 2013 John Wiley & Sons, Ltd.     

Vibrational spectroscopic studies and computational study of methyl(2‐methyl‐4,6–dinitrophenylsulfanyl)ethanoate

FT‐IR and FT‐Raman spectra of methyl(2‐methyl‐4,6–dinitrophenylsulfanyl)ethanoate (MDIE) were recorded and analyzed. Surface‐enhanced Raman scattering (SERS) spectra were recorded in silver colloid and silver electrode. The vibrational wavenumbers were computed using HF/6‐31G* and B3LYP/6‐31G* basis. The data obtained from vibrational wavenumber calculations are used to assign vibrational bands obtained in infrared and Raman spectroscopies as well as in SERS of the studied molecule. The first hyperpolarizability and infrared intensities are reported. The geometrical parameters of the title compound are in agreement with the reported similar derivatives. The presence of new bands at 1045 and 948 cm⁻¹ in the SERS spectrum in silver electrode is related to the change in orientation of the molecule with respect to the metal surface. In silver colloid SERS spectrum, the methyl group attached to the methoxy carbonyl group is close to the metal surface, whereas on silver electrode the methyl group attached to the phenyl ring is close to the metal surface. Copyright © 2009 John Wiley & Sons, Ltd.     

Identification of species formed after pyridine adsorption on iron,cobalt, nickel and silver electrodes by SERS and theoretical calculations

The adsorption of pyridine (py) on Fe, Co, Ni and Ag electrodes was studied using surface‐enhanced Raman scattering (SERS) to gain insight into the nature of the adsorbed species. The wavenumber values and relative intensities of the SERS bands were compared to the normal Raman spectrum of the chemically prepared transition metal complexes. Raman spectra of model clusters M₄(py) (four metal atoms bonded to one py moiety) and M₄(α‐pyridil) where M = Ag, Fe, Co or Ni were calculated by density functional theory (DFT) and used to interpret the experimental SERS results. The similarity of the calculated M₄(py) spectra with the experimental SERS spectra confirm the molecular adsorption of py on the surface of the metallic electrodes. All these results exclude the formation of adsorbed α‐pyridil species, as suggested previously. Copyright © 2009 John Wiley & Sons, Ltd.     

树枝状银微米材料的制备及其表面增强拉曼光谱研究

为使表面增量拉曼散射(SERS)衬底的制备方法简单快速且提高的基底增强效果,采用置换反应的制备方法,用锌片和硝酸银反应制备出微米银结构SERS活性基底,其具有稳定性好,易保存,制备方法简单,过程快速等特点。用扫描电子显微镜观察得银微米材料表面形貌呈均匀对称的树枝状结构。实验中控制置换反应的时间分别为40,50,60 s时,得到的树枝状银微米材料的长度分别为3,5,10 μm左右,分支分别为700 nm,2 μm,3 μm,发现随着置换反应的时间的增长,微米银树枝及分枝的长度越长,且树枝分枝上逐渐长出纳米级“树叶”结构, 使得微米级银树枝表面具有纳米结构。并且将微米银材料置于硅片上作为SERS衬底,并以罗丹明6G为探针分子,用激发波长为1 064 nm的傅里叶变换拉曼光谱仪检测,研究其在表面增强拉曼光谱中的应用,结果表明树枝状银微米材料有很好的SERS特性,其中置换反应时间为40 s时制备的微米银树枝的增强效果最佳,其增强因子可达到103左右,并且采用用表面活性剂PVP处理硅片的方法后,保持其他条件不变,微米银衬底的SERS增强效果得到进一步加强,增强因子达到104左右。此外,将树枝状银微米材料用水可封存数月,且实验结果的重复性较好。     

Nanoporous Ag–GaN thin films prepared by metal‐assisted electroless etching and deposition as three‐dimensional substrates for surface‐enhanced Raman scattering

Three‐dimensional (3D) nanoporous gallium nitride (PGaN) scaffolds are fabricated by Pt‐assisted electroless hydrofluoric acid (HF) etching of crystalline GaN followed by in situ electroless deposition of Ag nanostructures onto the interior surfaces of the nanopores, yielding a large surface area substrate for surface‐enhanced Raman scattering (SERS). The resulting 3D SERS‐active substrates have been optimized by varying reaction parameters and starting material concentration, exhibiting enhanced Raman signals 10–100× more intense than either (1) sputtered Ag‐coated porous GaN or (2) Ag‐coated planar GaN. The increase in SERS signal is attributed to a combination of the large surface area and the inherent transparency of PGaN in the visible spectral region. Overall, Ag‐decorated PGaN is a promising platform for high sensitivity SERS detection and chemical analysis, particularly for reaction and metabolic products that can be trapped inside the highly anisotropic nanoscale pores of PGaN. The potential of this sampling mode is illustrated by the ability to acquire Raman spectra of adenine down to 5 fmol. Additionally, correlated SERS and laser desorption/ionization mass spectrometry spectra can be acquired from same sample spot without further preparation, opening new possibilities for the investigation of surface‐bound molecules with substantially enhanced information content. Copyright © 2012 John Wiley & Sons, Ltd.     

Raman and surface‐enhanced Raman spectroscopic studies of the 15‐mer DNA thrombin‐binding aptamer

Aptamers are single‐stranded oligonucleotides that selectively bind to their target molecules owing to their ability to form secondary structures and shapes. The 15‐mer (5′‐GGTTGGTGTGGTTGG‐3′) DNA thrombin‐binding aptamer (TBA) binds to thrombin following the formation of a quadruplex structure via the Hoogsten‐type G–G interactions. In the present study, Raman and SERS spectra of TBA and thiolated TBA (used to facilitate covalent bonding to metal nanoparticle) in different conditions are investigated. The spectra of the two analogs exhibit vibrations, such as the C8N7 H2 deformation band at ∼1480 cm⁻¹ of the guanine tetrad, that are characteristic of the quadruplex structure in the presence of K⁺ ions or at low temperature. Interestingly, SERS spectra of the two analogs differ markedly from their respective normal Raman spectra, possibly due to changes in the conformation of the aptamer upon binding, as well as to the specific interaction of individual vibrational modes with the metal surface. In addition, the SERS spectra of the thiolated aptamer show significant changes with different concentrations, which may be due to different orientation of the molecule with respect to the metal surface. This study provides useful information for the development of label‐free aptamer‐based SERS sensors and assays. Copyright © 2009 John Wiley & Sons, Ltd.     

Raman and SERS study on cimetidine–metal complexes with biomedical interest

Cimetidine (cim) is one of the most potent histamine H₂‐receptor antagonists for inhibiting excessive acid secretion caused by histamine; it has been hypothesized that the therapeutic effects can be related to its interactions with metal ions. Raman spectra of the solid cim with Co(II), Cu(II), Ni(II) and Zn(II) metal complexes show that they can adopt two different structures: one is octahedral and the other, with Zn(II), is probably tetrahedral. The octahedral structure appears to be distorted both by the different metal ions as well as by the different anion present. The study was extended to very dilute solutions (ppm range) by using the surface‐enhanced Raman scattering (SERS) technique, mimicking the physiological concentrations of cim and its metal complexes. SERS spectra suggest that, upon the binding of cim to silver colloids, the formation of stable 1:2 cim–metal complexes is excluded, the formation of 1:1 adduct appearing more probable; in this product the metal reaches its total coordination shell by complexion with water molecules. To better explain the binding mechanism of cim to a metal (Ag) surface, we performed theoretical B3LYP calculations on cim alone as well as on cim bonded to an Ag₂ metal cluster in presence of water, observing a sufficiently good agreement between experimental and theoretical wavenumbers. Copyright © 2010 John Wiley & Sons, Ltd.     

Biocompatible gold/silver nanostars for surface‐enhanced Raman scattering

Raman‐enhancing properties of chitosan (CS)‐coated gold/silver nanostars (Au/AgNSs) were demonstrated by using them as a surface‐enhanced Raman scattering (SERS) probe. Based on the energy‐dispersive X‐ray spectroscopy element distribution maps and highly enhanced SERS spectra, we suggest that the incorporation of silver into the NS tips leads to a stronger SERS behavior. The SERS spectra of the proteins adsorbed on the NS surface greatly differ from their respective Raman spectra in both the band positions and relative intensities, indicating that the protein molecules penetrate through the CS coating layer and interact closely with the NS surface. Raman and SERS spectra of Chlamydia trachomatis protease/proteasomelike activity factor are reported for the first time, demonstrating the potential of these NSs for the development of a diagnosis method for Chlamydia based on SERS. The results showed a good SERS performance of the Au/AgNSs and their potential for SERS detection of biomolecules. Copyright © 2016 John Wiley & Sons, Ltd.     

SERS study of methylated and nonmethylated ribonucleosides and the effect of aggregating agents

We present surface‐enhanced Raman scattering (SERS) spectra of the RNA nucleosides adenosine, cytidine, guanosine and uracil and the methylated derivatives 5‐methylcytidine and 7‐methylguanosine, a class of important nucleic acid components that has not previously been well characterised using SERS spectroscopy. Our work shows that the selection of aggregating agent plays a crucial role for SERS analysis of these nucleosides with K₂SO₄ generating immediate enhancement while NaCl only gave immediate enhancement for the pyrimidine nucleosides. The SERS spectra contain a number of marker bands that are highly sensitive to structural differences between these nucleosides, in particular methylation, and at lower concentration ranges than are possible for conventional Raman scattering. Finally, spectral analyses and assignments of the vibrational modes responsible for these marker bands are also presented, and the effect of the aggregating agent on these modes is discussed in terms of interactions between each nucleoside and the metal surface. Copyright © 2011 John Wiley & Sons, Ltd.     

聚甲基丙烯酸甲酯间隔的金纳米立方体与金膜复合结构的表面增强拉曼散射研究

金属纳米颗粒与金属薄膜的复合结构由于其局域表面等离子体和传播表面等离子体间的强共振耦合作用,可作为表面增强拉曼散射(SERS)基底,显著增强吸附分子的拉曼信号.本文提出了一种聚甲基丙烯酸甲酯(PMMA)间隔的90 nm金纳米立方体与50 nm金膜复合结构的SERS基底,通过有限元方法数值模拟,得到PMMA的最优化厚度为15 nm.实验制备了PMMA间隔层厚度为14 nm的复合结构,利用罗丹明6G (R6G)为拉曼探针分子, 633 nm的氦氖激光器作为激发光源,研究了复合结构和单一金纳米立方体的SERS效应,发现复合结构可以使探针分子产生比单一结构更强的拉曼信号.在此基础上,研究了不同浓度金纳米立方体水溶液条件下复合结构中R6G的拉曼光谱.结果表明,当金纳米立方体水溶液浓度为5.625μg/mL的条件下复合结构中R6G的拉曼信号最强,且可测量R6G的最低浓度达10~(–11) mol/L.     

Surface‐enhanced Raman spectroscopy for trace‐level detection of explosives

The detection of explosives and their associated compounds for security screening is an active area of research and a wide variety of detection methods are involved in this very challenging area. Surface‐enhanced Raman scattering (SERS) spectroscopy is one of the most sensitive tools for the detection of molecules adsorbed on nano‐scale roughened metal surface. Moreover, SERS combines high sensitivity with the observation of vibrational spectra of species, giving complete information on the molecular structure of material under study. In this paper, SERS was applied to the detection of very small quantities of explosives adsorbed on industrially made substrates. The spectra were acquired with a compact Raman spectrometer. Usually, a high signal‐to‐noise (S/N) spectrum, suitable for identification of explosive molecules down to few hundreds of picograms, was achieved within 30 s. Our measurements suggest that it is possible to exploit SERS using a practical detection instrument for routine analysis. Copyright © 2010 John Wiley & Sons, Ltd.