Raman Optical Activity Measurements Research Articles

Chirality transfer observed in Raman optical activity spectra

Chirality transfer (also called induced chirality) is a phenomenon present in chiroptical spectra that manifests itself as a new band or bands of an achiral molecule interacting with a chiral one. In the Raman optical activity (ROA) spectroscopy, the bands of achiral solvents have been recently observed, but the latest papers have shown that they corresponded to the new ECD-Raman (eCP-Raman) effect. Here, we show an unambiguous example of chirality transfer observed in the ROA spectra. The spectra registered for the (1:1) mixtures of achiral benzonitrile with the enantiomers of 2,2,2-trifluoro-1-phenylethanol, 1-phenylethanol, and 1-phenylethylamine exhibited the v(CN) vibration band at about 2230 cm−1. The ROA measurements were repeated several times to ensure the reliability of the phenomenon. Calculations revealed the CN···HO or CN···HNH hydrogen bond formation accompanied by the π···π or CH···π interactions. The interaction strength was shown to be an important factor for the pronouncement of the ROA chirality transfer effect.

Measurement of Raman Optical Activity with High-Frequency Polarization Modulation.

Many chiroptical spectroscopic techniques have been developed to detect chirality in molecular species and probe its role in biological processes. Raman optical activity (ROA) should be one of the most powerful methods, as ROA yields vibrational and chirality information simultaneously and can measure analytes in aqueous and biologically relevant solvents. However, despite its promise, the use of ROA has been limited, largely due to challenges in instrumentation. Here, we report a new approach to ROA that exploits high-frequency polarization modulation. High-frequency polarization modulation, usually implemented with a photoelastic modulator (PEM), has long been the standard technique in other chiroptical spectroscopies. Unfortunately, the need for simultaneous spectral and polarization resolution has precluded the use of PEMs in ROA instruments. We combine a specialized camera system (the Zurich imaging polarimeter, or ZIMPOL) with PEM modulation to perform ROA measurements. We demonstrate performance similar to the current standard in ROA instrumentation while reducing complexity and polarization artifacts. This development should aid researchers in exploiting the full potential of ROA for chemical and biological analysis.

All-dielectric chiral-field-enhanced Raman optical activity

Raman optical activity (ROA) is effective for studying the conformational structure and behavior of chiral molecules in aqueous solutions and is advantageous over X-ray crystallography and nuclear magnetic resonance spectroscopy in sample preparation and cost performance. However, ROA signals are inherently minuscule; 3–5 orders of magnitude weaker than spontaneous Raman scattering due to the weak chiral light–matter interaction. Localized surface plasmon resonance on metallic nanoparticles has been employed to enhance ROA signals, but suffers from detrimental spectral artifacts due to its photothermal heat generation and inability to efficiently transfer and enhance optical chirality from the far field to the near field. Here we demonstrate all-dielectric chiral-field-enhanced ROA by devising a silicon nanodisk array and exploiting its dark mode to overcome these limitations. Specifically, we use it with pairs of chemical and biological enantiomers to show >100x enhanced chiral light–molecule interaction with negligible artifacts for ROA measurements.

Raman Optical Activity on a Solid Sample: Identification of Atropisomers of Perfluoroalkyl Chains Having a Helical Conformation and No Chiral Center.

Perfluoroalkyl (Rf) chains have a specific helical conformation due to the steric repulsion between the adjacent CF2 units. Although Rf chains have no chiral center, two chiral structures, i.e., the right-handed (R) and left-handed (L) helices, are available as the most stable conformations, which are atropisomers to each other. According to the stratified dipole array (SDA) theory, the helical structure about the chain axis plays a key role in the spontaneous molecular aggregation of Rf chains in a two-dimensional manner, and the Rf chains having the same chirality tend to be aggregated spontaneously to generate molecular domains. This implies that an Rf compound in a solid state should be a mixture of the R and L domains, and each domain should exhibit distinguishable optical activity. To identify molecular domains with different atropisomers, in this study, Raman optical activity (ROA) measurements were performed on a Raman imaging spectrometer. Through the ROA measurements of recrystallized solid samples of an Rf compound, each particle exhibits an apparent optical activity, and the two atropisomers were readily distinguished. As a result, an Rf compound with the same helicity is found to be spontaneously aggregated as expected by the SDA theory.

Raman Optical Activity Reveals Carotenoid Photoactivation Events in the Orange Carotenoid Protein in Solution.

The orange carotenoid protein (OCP) plays an important role in photoprotection in cyanobacteria, which is achieved by the photoconversion from the orange dark state (OCPO) to the red active state (OCPR). Using Raman optical activity (ROA), we studied the conformations of the carotenoid chromophore in the active sites of OCPO and OCPR. This ROA measurement directly observed the chromophore conformation of native OCP in solution, and the measurement of OCPR first demonstrated the ROA spectroscopy for the transient species. For OCPO, the spectral features of ROA were mostly reproduced by the quantum chemical calculation based on the crystal structure of the OCP. Within the spatial resolution (∼2 Å), a slight modification of the polyene-chain distortion improved the agreement between the observed and calculated ROA spectra. While the crystal structure of OCPR is not available, the ROA spectrum of OCPR was reproduced by using the crystal structure of red carotenoid protein (RCP), an OCPR proxy. The present results showed that the chromophore conformations in the crystal structures of OCP and RCP hold true for OCPO and OCPR in solution. Particularly, ROA spectroscopy of the native OCPR provides a direct support for the 12 Å translocation of chromophore in the photoactivation, which was proposed by X-ray crystallography using RCP [R. L. Leverenz, M. Sutter, et al. Science 2015, 348, 1463-1466].

Formation of Enhanced Uniform Chiral Fields in Symmetric Dimer Nanostructures.

Chiral fields with large optical chirality are very important in chiral molecules analysis, sensing and other measurements. Plasmonic nanostructures have been proposed to realize such super chiral fields for enhancing weak chiral signals. However, most of them cannot provide uniform chiral near-fields close to the structures, which makes these nanostructures not so efficient for applications. Plasmonic helical nanostructures and blocked squares have been proved to provide uniform chiral near-fields, but structure fabrication is a challenge. In this paper, we show that very simple plasmonic dimer structures can provide uniform chiral fields in the gaps with large enhancement of both near electric fields and chiral fields under linearly polarized light illumination with polarization off the dimer axis at dipole resonance. An analytical dipole model is utilized to explain this behavior theoretically. 30 times of volume averaged chiral field enhancement is gotten in the whole gap. Chiral fields with opposite handedness can be obtained simply by changing the polarization to the other side of the dimer axis. It is especially useful in Raman optical activity measurement and chiral sensing of small quantity of chiral molecule.

Exploring Raman optical activity for transition metals: From coordination compounds to solids

We briefly review the field of Raman optical activity (ROA) for transition-metal containing systems with a focus on coordination compounds and solids. In contrast to ROA measurements for optically active solids, ROA for chiral metal complexes is a relatively novel field with only a few measurements and calculations published yet. Their results indicate a great sensitivity of ROA for the elucidation of such compounds, even in case of structurally very similar geometrical isomers, and the differentiation of several types of chirality. Resonance with one or more electronic transitions can lead to intensity enhancement in the ROA spectrum, which provides additional advantages such as shorter measurement times, lower sample amounts and valuable information about involved excited electronic states. A widely unknown variant is magnetic ROA where an external magnetic field is applied. This form is applicable to both chiral and achiral systems and has, among others, been employed to probe Zeeman splittings and sign of g factors. This review shows that ROA is a very promising field for in-depth study and design of transition-metal containing compounds and materials. It is still in its infancy and possible next steps for future research, with an emphasis on computational developments, are outlined.

The development of biomolecular Raman optical activity spectroscopy

Following its first observation over 40 years ago, Raman optical activity (ROA), which may be measured as a small difference in the intensity of vibrational Raman scattering from chiral molecules in right- and left-circularly polarized incident light or, equivalently, the intensity of a small circularly polarized component in the scattered light using incident light of fixed polarization, has evolved into a powerful chiroptical spectroscopy for studying a large range of biomolecules in aqueous solution. The long and tortuous path leading to the first observations of ROA in biomolecules in 1989, in which the author was closely involved from the very beginning, is documented, followed by a survey of subsequent developments and applications up to the present day. Among other things, ROA provides information about motif and fold, as well as secondary structure, of proteins; solution structure of carbohydrates; polypeptide and carbohydrate structure of intact glycoproteins; new insight into structural elements present in unfolded protein sequences; and protein and nucleic acid structure of intact viruses. Quantum chemical simulations of observed Raman optical activity spectra provide the complete three-dimensional structure, together with information about conformational dynamics, of smaller biomolecules. Biomolecular ROA measurements are now routine thanks to a commercial instrument based on a novel design becoming available in 2004.

Ribifolin, an orbitide from Jatropha ribifolia, and its potential antimalarial activity.

A new orbitide named ribifolin was isolated and characterized from Jatropha ribifolia using mass spectrometry, NMR spectroscopy, quantitative amino acid analysis, molecular dynamics/simulated annealing, and Raman optical activity measurements and calculations. Ribifolin (1) and its linear form (1a) were synthesized by solid-phase peptide synthesis, followed by evaluation of its antiplasmodial and cytotoxicity activities. Compound 1 was moderately effective (IC50 = 42 μM) against the Plasmodium falciparum strain 3D7.

Observation of Raman Optical Activity by Heterodyne-Detected Polarization-Resolved Coherent Anti-Stokes Raman Scattering

We report the first observation of Raman optical activity (ROA) by coherent anti-Stokes Raman scattering. Thanks to the more freedom of polarization configurations in coherent anti-Stokes Raman scattering than in spontaneous Raman spectroscopy, the contrast ratio of the chiral signal to the achiral background has been improved markedly. For (-)-β-pinene, it is 2 orders of magnitude better than that in the reported spontaneous ROA measurement. This is also the first measurement of ROA signal using a pulsed laser source.

Comparative Study of Measured and Computed Raman Optical Activity of a Chiral Transition Metal Complex

On the ROA to somewhere: The first combined study of measured and computed Raman optical activity (ROA) of a transition metal complex under non-resonant scattering conditions is reported. ROA measurements of the two enantiomers of the dichloro[ethylenebis(4,5,6,7-tetrahydro-1-indenyl)] zirconium(IV) complex yield virtually mirror-image ROA spectra. The experimental spectra are directly comparable with predicted ROA spectra.

Difference Frequency Generation Spectroscopy as a Vibrational Optical Activity Measurement Tool

Vibrational optical activity (VOA) of chiral molecules in condensed phases can be studied by using vibrational circular dichroism and Raman optical activity measurement techniques. Recently, IR-vis sum frequency generation has shown to be an alternative VOA measurement method. Such a three-wave-mixing method employing a polarization modulation technique can be a potentially useful VOA measurement tool. Here, a theoretical description of difference frequency generation (DFG) employing circularly polarized visible radiations is presented. Frequency scanning to obtain a VOA-DFG spectrum is achieved by controlling the difference between the two electronically nonresonant incident radiation frequencies. If the two incident beams are linearly polarized and their polarization directions are perpendicular to each other, one can selectively measure the all-electric-dipole-allowed chiral component of the DFG susceptibility. In addition, by using circularly polarized beams and taking the DFG difference intensity signal, which is defined as the difference between left and right circularly polarized DFG signals, additional chiral susceptibility components originating from the electric quadrupole transition can be measured. The DFG as a novel VOA measurement technique for solution samples containing chiral molecules will therefore be a useful coherent spectroscopic tool for determining absolute configuration of chiral molecules in condensed phases.

Special issue “Advances in Chiroptical Methods”

Special issue “Advances in Chiroptical Methods”

Vibrational Raman optical activity of 1‐phenylethanol and 1‐phenylethylamine: Revisiting old friends

The samples used for the first observations of vibrational Raman optical activity (ROA) in 1972, namely both enantiomers of 1-phenylethanol and 1-phenylethylamine, have been revisited using a modern commercial ROA instrument together with state-of-the-art ab initio calculations. The simulated ROA spectra reveal for the first time the vibrational origins of the first reported ROA signals, which comprised similar couplets in the alcohol and amine in the spectral range approximately 280-400 cm(-1). The results demonstrate how easy and routine ROA measurements have become, and how current ab initio quantum-chemical calculations are capable of simulating experimental ROA spectra quite closely provided sufficient averaging over accessible conformations is included. Assignment of absolute configuration is, inter alia, completely secure from results of this quality. Anharmonic corrections provided small improvements in the simulated Raman and ROA spectra. The importance of conformational averaging emphasized by this and previous related work provides the underlying theoretical background to ROA studies of dynamic aspects of chiral molecular and biomolecular structure and behavior.

Observation of SERS effect in Raman optical activity, a new tool for chiral vibrational spectroscopy

AbstractA new tool for chiral vibrational spectroscopy is reported here. A surface enhanced effect was observed using Raman optical activity (ROA). This observation opens new possibilities for ROA as a tool for vibrational spectroscopy. The combination of surface enhanced effect (SE) and ROA into surface enhanced Raman optical activity (SEROA) takes this tool to another level, where a single molecule may be studied with respect to chirality, secondary structure and fold determination. ROA has been able to provide information about important dynamics in molecular understanding. Until recently, however, ROA measurements required a longer exposure and higher concentration of the sample. With SEROA these obstacles can be overcome because both studies on single molecule, i.e. very low concentration, and faster acquisition of the signal can be carried out. In the present, work silver colloids were mixed with solution, in which a pentapeptide, Met‐Enkephalin, was dissolved. SEROA signals were recorded and the results are reported here. Copyright © 2006 John Wiley & Sons, Ltd.

Surface-Enhanced Raman Optical Activity on Adenine in Silver Colloidal Solution

We report the collection of Raman optical activity (ROA) spectra of adenine in silver colloidal solution, that is, surface-enhanced Raman optical activity (SEROA) using considerably shorter data acquisition times, reduced excitation power, and lower concentration, as compared to classical ROA measurements on molecules of biological interest so far reported in the literature. These improvements in experimental parameters for ROA measurements can be explained by enhanced Raman signals in the local optical fields of the silver nanoparticles and by at least 1 order of magnitude higher values for circular intensity differences (CIDs), as compared to classical ROA that has been suggested before and theoretically discussed in terms of large field gradients near a metal surface. The measured ROA effect for adenine can be understood in terms of adsorption-induced chirality in the prochiral molecules on the silver nanoparticles. Surface-enhanced Raman optical activity offers potential capabilities for sensitive, rapid, stereochemical characterization of basic building blocks of biopolymers, such as amino acids and nucleosides, as well as biologically active molecules, in particular, also for probing organization and self-assembling of such molecules on metal surfaces.

Vibrational Raman Optical Activity of α-Lactalbumin: Comparison with Lysozyme, and Evidence for Native Tertiary Folds in Molten Globule States

Proteins in aqueous solution are now accessible to Raman optical activity (ROA) measurements, which provide an incisive new probe of secondary and tertiary structure illustrated here by a study of bovine α-lactalbumin. The room-temperature ROA spectrum of native bovine α-lactalbumin is similar to that of native hen egg-white lysozyme except for features attributable to differences in the loop regions: in particular, a positive ROA band at ∼1338 cm −1assigned to conformationally homogeneous loop structure, possibly with local order corresponding to 3 10-helix, has more than double the intensity in α-lactalbumin compared with lysozyme. This is consistent with the two proteins having similar secondary structure but different local details in the tertiary fold. ROA measurements on α-lactalbumin at pH 2.0 over a range of temperatures have provided a new perspective on the molten globule state. Thus at 35°C ROA reveals the presence of some secondary structure but an almost complete loss of the tertiary loop structure; whereas at 2°C the ROA spectrum is almost identical with that of the native protein, which is strong evidence that virtually all of the secondary structure and the tertiary backbone fold persist, albeit within a looser framework associated with increased solvent exposure and change of environment of many of the side-chains as evidenced by an increase in noise and bandwidth of some of the ROA signals together with aromatic fluorescence and near-UV circular dichroism signals characteristic of the molten globule state. Our sample of acid α-lactalbumin at 2°C therefore appears to be an archetypal example of Ptitsyn's "native-like" molten globule, having a fixed native-like tertiary fold but with loss of tight packing of the side-chains; whereas at 35°C it is a "disordered" molten globule. At 20°C the acid molten globule appears to retain highly native-like secondary structure but with most of the tertiary fold already lost. A calcium-free sample of α-lactalbumin at neutral pH displayed a broad cooperative transition between native and molten globule states at ∼15 °C, with the latter state showing similar but somewhat degraded tertiary loop ROA signatures to the native protein. In both the acid and apo molten globule states the ROA signatures of the secondary structure and the tertiary loops showed a gradual change with temperature.

Vibrational Raman optical activity of biopolymers

Vibrational Raman optical activity measurements can now provide a wealth of new information about the structure and conformation of biopolymers in aqueous solution. Typical results are exemplified here for proteins by bovine serum albumin in H2O and D2O solution, and for polysaccharides by laminarin.

Instrumentation for Raman optical activity measurements

We describe the latest versions of two current multichannel Raman optical activity (ROA) instruments from our laboratory. Both are based on an astigmatic single-grating spectrograph equipped with a holographic notch filter and a charge coupled device detector and employ the incident circular polarization (ICP) modulation strategy for ROA measurements, one in backscattering and the other in right-angle scattering. A selection of ICP ROA spectra is presented which demonstrates the excellent performance characteristics of both instruments, especially in the lower wavenumber region.

Recent developments in Raman optical activity instrumentation

We describe the latest versions of two current multichannel Raman optical activity (ROA) instruments from our laboratory. Both are based on an astigmatic spectrograph equipped with a holographic notch filter and a charge-coupled device detector and employ the incident circular polarization (ICP) modulation strategy for ROA measurements, one in backscattering and the other in right-angle scattering. A selection of ROA spectra is presented which demonstrates the excellent performance characteristics of both instruments, especially in the lower wavenumber region. We also discuss the design for a future ICP instrument for backscattering ROA measurements which uses state-of-the-art stigmatic spectrograph technology and which is currently under construction in our laboratory.