Plasmonic features of free-standing chitosan nanocomposite film with silver and graphene oxide for SERS applications.

A scalable procedure of SERS substrates design was developed using a novel plasmonic structure based on a freestanding chitosan film, silver nanoparticles, and graphene oxide. Chitosan provides a uniform distribution of silver nanoparticles from a colloidal suspension and, therefore, a reproducible Raman signal from local areas of measurements of several tens of microns. The addition of graphene oxide (GO) to the colloidal solution of silver nanoparticles suppresses the tortuous background fluorescence signal from the analyte and leads to an increase in the signal-to-fluorescence background intensity ratio by up to 6 times as compared to structures without GO. The manufactured plasmonic polymer nanocomposite provides a detection limit of down to 100 pM for R6G using a laser wavelength of 532 nm through a portable ×10 objective. The high colloidal stability of GO in water and the use of an aqueous colloid of silver nanoparticles simplify the procedure for creating a substrate by applying the GO-silver composite on the surface of a chitosan film without a need to form a GO film. Therefore, our approach paves a promising avenue to provide more sensitive detection even for the fluorescent analytes with short-wavelength lasers (532, 633 nm) instead of IR (785, 1024 nm) and foster the practical application of the developed plasmonic composites on portable Raman spectrometers.

Molecular Immobilization and Resonant Raman Amplification by Complex-Loaded Enhancers (MIRRACLE) on copper (II)-chitosan-modified SERS-active metallic nanostructured substrates for multiplex determination of dopamine, norepinephrine, and epinephrine.

A unique approach based on Molecular Immobilization and Resonant Raman Amplification by Complex-Loaded Enhancers (MIRRACLE) on copper (II)-chitosan-modified SERS-active metallic nanostructured substrates is proposed for sensitive and rapid determination of the catecholamines (CA) dopamine, norepinephrine, and epinephrine. The ternary (CA)2Cu(4AAP)2 complexes were characterized by the appearance of new absorbance bands at 555, 600, and 500 nm for dopamine, norepinephrine, and epinephrine, respectively. The new absorbance band matched with a broad surface plasmon resonance band of utilized silver nanoparticles: 450-600 nm, and 633 excitation wavelength. We observed enhancement factors up to 3.6·106 due to the additional resonant enhancement. The multiplexing capabilities of quantitative spectral unmixing for Raman spectra of a group of CAs, which differ by only either hydroxy or methyl group, at the fingerprint region were successfully demonstrated with the direct classic least squares model. The achieved nM limits of detection with only 1.5 mW laser power and analysis of spiked human blood plasma samples proved the possibility of the multiplex determination of the catecholamines at the level of reference concentrations in the blood of healthy people as well as promise for the future facilitation in the precision diagnosis of neuroendocrine tumors and neurodegenerative diseases.

Silver-chitosan nanocomposite as a plasmonic platform for SERS sensing of polyaromatic sulfur heterocycles in oil fuel.

Herein, a silver-chitosan nanocomposite for application in surface enhanced Raman spectroscopy (SERS) sensing was proposed. It was shown that optically transparent chitosan coatings with 0.8 µm thickness allow penetration of target analytes to silver nanoparticles and the analysis in both polar and nonpolar solvents. Under the chosen conditions, chitosan formed continuously smooth films and coatings stabilizing rough nanostructured metallic surfaces and served as a suitable matrix for immobilization, uniform spreading, and preconcentration of the analytes. Polycyclic aromatic sulfur heterocycles were chosen as target analytes being one of the most important fuel quality markers, hazardous components, and the hardest-to-remove impurities. For the most effective immobilization and even distribution of the analytes onto a nanostructured metallic surface, an additional polymer layer of chitosan was found to be needed. The presence of thin films of chitosan resulted in higher reproducibility of SERS spectra as compared to bare nanostructured silver substrates. Additionally, the developed nanocomposite SERS sensors provided the rapid determination of dibenzothiophene and its derivatives in isooctane with the threshold of detection better than 0.1 µM. This approach was successfully applied in the analysis of real fuel samples and the results agreed well with independently measured FTIR and GC-MS data.

Crystallinity as a factor of SERS stability of silver nanoparticles formed by Ar+ irradiation.

The plasmonic sensors based on silver nanoparticles are limited in application due to their relatively fast degradation in the ambient atmosphere. The technology of ion-beam modification for the creation of monocrystalline silver nanoparticles (NPs) with stable plasmonic properties will expand the application of silver nanostructures. In the present study, highly-stable monocrystalline NPs were formed on the basis of a thin silver film by low-energy ion irradiation. Combined with lithography, this technique allows the creation of nanoparticle ensembles in variant forms. The characterization of the nanoparticles formed by ion-beam modification showed long-term outstanding for Ag nanoparticles stability of their plasmonic properties due to their monocrystalline structure. According to optical spectroscopy data, the reliable plasmonic properties in the ambient atmosphere are preserved for up to 39 days. The mapping of crystal violet dye via surface-enhanced Raman spectroscopy (SERS) revealed a strong amplification factor sustaining at least thrice as long as the one of similarly sized polycrystalline silver NPs formed by annealing. The plasmonic properties sustain more than a month of storage in the ambient atmosphere. Thus, ion-beam modification of silver film makes it possible to fabricate NPs with stable plasmonic properties and form clusters of NPs for sensor technology and SERS applications.

Plier Ligands for Trapping Neurotransmitters into Complexes for Sensitive Analysis by SERS Spectroscopy.

Catecholamines-dopamine, noradrenaline and adrenaline are important biomarkers of neurotransmitter metabolism, indicating neuroendocrine tumors and neurodegenerative diseases. Surface-enhanced Raman spectroscopy (SERS) is a promising analytical technique with unprecedented multiplexing capabilities. However, not all important analytes exhibit strong SERS signals on stable and robust nanostructured substrates. In this work, we propose a novel indicator system based on the formation of mixed ligand complexes with bispidine-based bis-azole ligands which can serve as pliers to trap Cu(II) ions and stabilize its complexes with catecholamines. Four synthesized ligands with different functional groups: carboxyl, amino, benzyl, and methoxybenzyl, were applied for forming stable complexes to shift maximum absorbance of catecholamines from the ultraviolet region to 570-600 nm. A new absorbance band in the visible range resonates with the local surface plasmon resonance (LSPR) band of metal nanoparticles and most used laser wavelengths. This match allowed use of Molecular Immobilization and Resonant Raman Amplification by Complex-Loaded Enhancers (MIRRACLE) methodology to measure intense Raman signals on a nanostructured silver-based SERS-active substrate. The synthesized plier-like ligands fixed and stabilized catecholamine complexes with Cu(II) on the SERS sensor surface, which facilitated the determination of dopamine in a 3.2 × 10-12-1 × 10-8 M concentration range.

Model of the SARS-CoV-2 Virus for Development of a DNA-Modified, Surface-Enhanced Raman Spectroscopy Sensor with a Novel Hybrid Plasmonic Platform in Sandwich Mode.

The recent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has posed a great challenge for the development of ultra-fast methods for virus identification based on sensor principles. We created a structure modeling surface and size of the SARS-CoV-2 virus and used it in comparison with the standard antigen SARS-CoV-2-the receptor-binding domain (RBD) of the S-protein of the envelope of the SARS-CoV-2 virus from the Wuhan strain-for the development of detection of coronaviruses using a DNA-modified, surface-enhanced Raman scattering (SERS)-based aptasensor in sandwich mode: a primary aptamer attached to the plasmonic surface-RBD-covered Ag nanoparticle-the Cy3-labeled secondary aptamer. Fabricated novel hybrid plasmonic structures based on "Ag mirror-SiO2-nanostructured Ag" demonstrate sensitivity for the detection of investigated analytes due to the combination of localized surface plasmons in nanostructured silver surface and the gap surface plasmons in a thin dielectric layer of SiO2 between silver layers. A specific SERS signal has been obtained from SERS-active compounds with RBD-specific DNA aptamers that selectively bind to the S protein of synthetic virion (dissociation constants of DNA-aptamer complexes with protein in the range of 10 nM). The purpose of the study is to systematically analyze the combination of components in an aptamer-based sandwich system. A developed virus size simulating silver particles adsorbed on an aptamer-coated sensor provided a signal different from free RBD. The data obtained are consistent with the theory of signal amplification depending on the distance of the active compound from the amplifying surface and the nature of such a compound. The ability to detect the target virus due to specific interaction with such DNA is quantitatively controlled by the degree of the quenching SERS signal from the labeled compound. Developed indicator sandwich-type systems demonstrate high stability. Such a platform does not require special permissions to work with viruses. Therefore, our approach creates the promising basis for fostering the practical application of ultra-fast, amplification-free methods for detecting coronaviruses based on SARS-CoV-2.

Dual-Purpose SERS Sensor for Selective Determination of Polycyclic Aromatic Compounds via Electron Donor-Acceptor Traps.

Toxic, carcinogenic, and mutagenic properties of polycyclic aromatic hydrocarbons (PAHs) and environmental pollution caused by polycyclic aromatic sulfur heterocycles (PASHs) postulate the importance of their selective and sensitive determination in environmental and oil fuel samples. Surface-enhanced Raman spectroscopy (SERS) opens up an avenue toward multiplex analysis of complex mixtures, however not every molecule gives high enhancement factors and, thus, cannot be reliably detected via SERS. However, the sensitivity can be drastically increased by additional resonant enhancement as a result of the analyte absorption band overlapping with the surface plasmon band of nanoparticles (NPs) and the laser excitation wavelength. Using this idea, we developed a dual-purpose SERS sensor based on trapping the target PAHs and PASHs into colored charge-transfer complexes (CTCs) with selected organic π-acceptor molecules on the surface of AgNPs. Studying, computing, and then comparing stability constants of the formed CTC served as a powerful explanation and prediction tool for a wise choice of π-acceptor indicator systems for the further silver surface modification. Moreover, we show that CTC formation can be effectively utilized for increasing both selectivity and sensitivity by simple liquid-liquid extraction prior to SERS measurements. For the first time, the dual-purpose SERS sensor allowed determination of two different classes of polycyclic aromatic fuel components down to 10 nM concentration, lower than that restricted by the ASTM regulation, and demonstrated multi-purpose capabilities of the developed approach.

Optically transparent chitosan hydrogels for selective sorption and fluorometric determination of dibenzothiophenes.

Solid-phase extraction of polycyclic aromatic sulfur heterocycles (PASHs) and their rapid determination in oil fuel without tedious sample pretreatment are of high interest. We propose porous and optically transparent hydrogels prepared from the covalently crosslinked chitosan (CS) as the basis for a sensor system for the rapid and robust monitoring of PASHs. We efficiently combined the ability of the crosslinked CS to sorb PASHs, the capacity of microcavities in a molecularly imprinted polymer to selectively recognize and trap analytes, and the optical transparency of CS materials for selective sorption and solid-phase fluorometric determination of dibenzothiophenes. For the screening of PASHs in organic nonpolar media, ortho-phtalic dialdehyde appeared to be the most appropriate crosslinker. Synthetic and analytical procedures performed in microplate mode allowed obtaining CS hydrogels with suitable reproducible properties and their further time- and labor-efficient applying in analysis (particularly, as little as 2 µM dibenzothiophene oxide can be determined).

Novel biosensing system for the simultaneous multiplex fluorescent determination of catecholamines and their metabolites in biological liquids.

A novel original biosensing system for the simultaneous multiplex determination of general markers of catecholamine-producing diseases - catecholamines (dopamine, epinephrine, norepinephrine) and their metabolites (homovanillic and vanillylmandelic acids) in biological liquids without preliminary separation of analytes, in the absence of specific antibodies and receptors and with minimum pretreatment of a samples has been developed. This outstanding approach includes the unique combination of obtaining highly fluorescent derivatives of the analytes as a result of their interaction with two different amines ̶ benzylamine and 1,2-diphenylethylenediamine in the presence of peroxidase as a catalyst, with the application of first-order derivative fluorescence spectroscopy for the resolution of their spectra. Fluorescence is measured in 96-well microplates, which wells contain a bio-recognizing film consisted of horseradish peroxidase immobilized in the polymer chitosan. Spectra of the solutions are recorded in the range 400-500 nm (λex ∼ 305-356 nm). The proposed procedures provide sensitive (in the range of 3-200 nM), selective, and reproducible (RSDs ≤ 1%, n = 6) multiplex determination of the catecholamines and their metabolites in biological liquids were successfully applied for the rapid simultaneous (20 samples per 15-30 min) screening of human urine and mice blood plasma. The validated results showed good linearity, precision, accuracy and selectivity of this method.

Novel Multilayer Nanostructured Materials for Recognition of Polycyclic Aromatic Sulfur Pollutants and Express Analysis of Fuel Quality and Environmental Health by Surface Enhanced Raman Spectroscopy.

A novel concept of advanced SERS (surface enhanced Raman spectroscopy) planar sensors is suggested for fast analysis of sulfur-containing hazardous oil components and persistent pollutants. The main advantage of the proposed sensors is the utilization of an additional preconcentrating layer of optically transparent chitosan gel, which is chemically modified with appropriate π-acceptor compounds to selectively form charge-transfer complexes (CTCs) at the interface with nanostructured silver coatings. The CTCs shift absorption bands of polycyclic aromatic sulfur heterocycles (PASHs) and other important analytes in a controllable way and thus provide a surplus enhancement of vibration modes due to resonant Raman scattering. This novel indicator system provides multiplex determination of PASHs in different forms in a small volume of oil without any tedious sample pretreatment steps. This approach opens new possibilities of increasing either spectral and concentration sensitivity or specificity of SERS-based sensors, allowing for new developments in the fields of ecology, advanced fuel analysis, and other prospective applications.