Ultrasensitive Fiber Enhanced UV Resonance Raman Sensing of Drugs

Fiber enhanced UV resonance Raman spectroscopy is introduced for chemical selective and ultrasensitive analysis of drugs in aqueous media. The application of hollow-core optical fibers provides a miniaturized sample container for analyte flow and efficient light-guiding, thus leading to strong light–analyte interactions and highly improved analytical sensitivity with the lowest sample demand. The Raman signals of the important antimalaria drugs chloroquine and mefloquine were strongly enhanced utilizing deep UV and electronic resonant excitation augmented by fiber enhancement. An experimental design was developed and realized for reproducible and quantitative Raman fiber sensing, thus the enhanced Raman signals of the pharmaceuticals show excellent linear relationship with sample concentration. A thorough model accounts for the different effects on signal performance in resonance Raman fiber sensing, and conclusions are drawn how to improve fiber enhanced Raman spectroscopy (FERS) for chemical selective analysis with picomolar sensitivity

Isotopomeric Peak Assignment for N₂O in Cross-Labeling Experiments by Fiber-Enhanced Raman Multigas Spectroscopy

Human intervention in nature, especially fertilization, greatly increased the amount of N2O emission. While nitrogen fertilizer is used to improve nitrogen availability and thus plant growth, one negative side effect is the increased emission of N2O. Successful regulation and optimization strategies require detailed knowledge of the processes producing N2O in soil. Nitrification and denitrification, the main processes responsible for N2O emissions, can be differentiated using isotopic analysis of N2O. The interplay between these processes is complex, and studies to unravel the different contributions require isotopic cross-labeling and analytical techniques that enable tracking of the labeled compounds. Fiber-enhanced Raman spectroscopy (FERS) was exploited for sensitive quantification of N2O isotopomers alongside N2, O2, and CO2 in multigas compositions and in cross-labeling experiments. FERS enabled the selective and sensitive detection of specific molecular vibrations that could be assigned to various isotopomer peaks. The isotopomers 14N15N16O (2177 cm–1) and 15N14N16O (2202 cm–1) could be clearly distinguished, allowing site-specific measurements. Also, isotopomers containing different oxygen isotopes, such as 14N14N17O, 14N14N18O, 15N15N16O, and 15N14N18O could be identified. A cross-labeling showed the capability of FERS to disentangle the contributions of nitrification and denitrification to the total N2O fluxes while quantifying the total sample headspace composition. Overall, the presented results indicate the potential of FERS for isotopic studies of N2O, which could provide a deeper understanding of the different pathways of the nitrogen cycle

Analysis of Fiber-Enhanced Raman Gas Sensing Based on Raman Chemical Imaging

Fiber enhanced Raman spectroscopy (FERS) is an arising new technique for versatile highly sensitive and selective multigas analysis in various applications, such as environmental monitoring and medical breath diagnosis. In this study, the performance of FERS was thoroughly studied with the help of a specially designed multichannel Raman chemical imaging. Several types of hollow core photonic crystal fibers were thoroughly analyzed in terms of their performance in light confinement and sensitive gas sensing. The optimal fiber length for Raman gas sensing was found to be 15 cm in our spectroscopic system. To separate the Raman scattering of the target gas molecules from the background generated by the silica microstructure of the fiber, the optimal diameter of a spatial filter was analyzed and quantified as Ø3.9 μm, which balances the suppression of the silica background and the attenuation of the gas signal, originating from different regions in the plane of the fiber end-face. To achieve an easy-to-use gas monitoring system with stable performance, an automated coupling-method was developed, to simplify the alignment of the FERS setup. The optimized design of the FERS setup has remarkable potential for highly sensitive, miniaturized, easy-to-use, and versatile gas sensing

Fast and Highly Sensitive Fiber-Enhanced Raman Spectroscopic Monitoring of Molecular H₂ and CH₄ for Point-of-Care Diagnosis of Malabsorption Disorders in Exhaled Human Breath

Breath gas analysis is a novel powerful technique for noninvasive, early-stage diagnosis of metabolic disorders or diseases. Molecular hydrogen and methane are biomarkers for colonic fermentation, because of malabsorption of oligosaccharides (e.g., lactose or fructose) and for small intestinal bacterial overgrowth. Recently, the presence of these gases in exhaled breath was also correlated with obesity. Here, we report on the highly selective and sensitive detection of molecular hydrogen and methane within a complex gas mixture (consisting of H₂, CH₄, N₂, O₂, and CO₂) by means of fiber-enhanced Raman spectroscopy (FERS). An elaborate FERS setup with a microstructured hollow core photonic crystal fiber (HCPCF) provided a highly improved analytical sensitivity. The simultaneous monitoring of H₂ with all other gases was achieved by a combination of rotational (H₂) and vibrational (other gases) Raman spectroscopy within the limited spectral transmission range of the HCPCF. The HCPCF was combined with an adjustable image-plane aperture pinhole, in order to separate the H₂ rotational Raman bands from the silica background signal and improve the sensitivity down to a limit of detection (LOD) of 4.7 ppm (for only 26 fmol H₂). The ability to monitor the levels of H₂ and CH₄ in a positive hydrogen breath test (HBT) was demonstrated. The FERS sensor possesses a high dynamic range (∼5 orders of magnitude) with a fast response time of few seconds and provides great potential for miniaturization. We foresee that this technique will pave the way for fast, noninvasive, and painless point-of-care diagnosis of metabolic diseases in exhaled human breath

Fiber-Enhanced Raman Multigas Spectroscopy: A Versatile Tool for Environmental Gas Sensing and Breath Analysis

Versatile multigas analysis bears high potential for environmental sensing of climate relevant gases and noninvasive early stage diagnosis of disease states in human breath. In this contribution, a fiber-enhanced Raman spectroscopic (FERS) analysis of a suite of climate relevant atmospheric gases is presented, which allowed for reliable quantification of CH₄, CO₂, and N₂O alongside N₂ and O₂ with just one single measurement. A highly improved analytical sensitivity was achieved, down to a sub-parts per million limit of detection with a high dynamic range of 6 orders of magnitude and within a second measurement time. The high potential of FERS for the detection of disease markers was demonstrated with the analysis of 27 nL of exhaled human breath. The natural isotopes ¹³CO₂ and ¹⁴N¹⁵N were quantified at low levels, simultaneously with the major breath components N₂, O₂, and ¹²CO₂. The natural abundances of ¹³CO₂ and ¹⁴N¹⁵N were experimentally quantified in very good agreement to theoretical values. A fiber adapter assembly and gas filling setup was designed for rapid and automated analysis of multigas compositions and their fluctuations within seconds and without the need for optical readjustment of the sensor arrangement. On the basis of the abilities of such miniaturized FERS system, we expect high potential for the diagnosis of clinically administered ¹³C-labeled CO₂ in human breath and also foresee high impact for disease detection via biologically vital nitrogen compounds

Ultrasensitive Detection of Antiseptic Antibiotics in Aqueous Media and Human Urine Using Deep UV Resonance Raman Spectroscopy

Deep UV resonance Raman spectroscopy is introduced as an analytical tool for ultrasensitive analysis of antibiotics used for empirical treatment of patients with sepsis and septic shock, that is, moxifloxacin, meropenem, and piperacillin in aqueous solution and human urine. By employing the resonant excitation wavelengths λ_exc = 244 nm and λ_exc = 257 nm, only a small sample volume and short acquisition times are needed. For a better characterization of the matrix urine, the main ingredients were investigated. The capability of detecting the antibiotics in clinically relevant concentrations in aqueous media (LODs: 13.0 ± 1.4 μM for moxifloxacin, 43.6 ± 10.7 μM for meropenem, and 7.1 ± 0.6 μM for piperacillin) and in urine (LODs: 36.6 ± 11.0 μM for moxifloxacin, and 114.8 ± 3.1 μM for piperacillin) points toward the potential of UV Raman spectroscopy as point-of-care method for therapeutic drug monitoring (TDM). This procedure enables physicians to achieve fast adequate dosing of antibiotics to improve the outcome of patients with sepsis

Direct Raman Spectroscopic Measurements of Biological Nitrogen Fixation under Natural Conditions: An Analytical Approach for Studying Nitrogenase Activity

Biological N₂ fixation is a major input of bioavailable nitrogen, which represents the most frequent factor limiting the agricultural production throughout the world. Especially, the symbiotic association between legumes and Rhizobium bacteria can provide substantial amounts of nitrogen (N) and reduce the need for industrial fertilizers. Despite its importance in the global N cycle, rates of biological nitrogen fixation have proven difficult to quantify. In this work, we propose and demonstrate a simple analytical approach to measure biological N₂ fixation rates directly without a proxy or isotopic labeling. We determined a mean N₂ fixation rate of 78 ± 5 μmol N₂ (g dry weight nodule)⁻¹ h^–1 of a Medicago sativa–Rhizobium consortium by continuously analyzing the amount of atmospheric N₂ in static environmental chambers with Raman gas spectroscopy. By simultaneously analyzing the CO₂ uptake and photosynthetic plant activity, we think that a minimum CO₂ mixing ratio might be needed for natural N₂ fixation and only used the time interval above this minimum CO₂ mixing ratio for N₂ fixation rate calculations. The proposed approach relies only on noninvasive measurements of the gas phase and, given its simplicity, indicates the potential to estimate biological nitrogen fixation of legume symbioses not only in laboratory experiments. The same methods can presumably also be used to detect N₂ fluxes by denitrification from ecosystems to the atmosphere