Miniature Raman Spectrometer Research Articles

Development and Optimization of a SERS Method for On-site Determination of Nitrite in Foods and Water

A rapid, sensitive, and selective detection method of nitrite in foods and water was established by surface-enhanced Raman spectroscopy (SERS) based on the diazo reaction of nitrite with p-nitroaniline and 1-naphthylamine in acidic solution. The azo dye (4-(4-nitrophenyldiazenyl)naphthalene-1-aminium, NNA), which was derived from the diazo reaction, was determined by SERS using citrate-coated silver nanoparticles (AgNPs) as SERS substrates. The concentrations of nitrite in samples were finally calculated from the intensities of SERS signals generated by NNA in the testing solutions. By using the present method combined with a portable miniature Raman spectrometer, on-site determination of nitrite could be performed easily and efficiently. The effects of several experimental parameters on the intensity of SERS signals, such as the volume of AgNP solution and the mixing time of AgNPs and NNA, were investigated. The linear range of the method was 0.1–10.0 mg L−1. The limits of detection (LODs) were 0.01, 0.03, and 0.05 mg L−1 at 720, 1,459, and 1,609 cm−1, respectively. The method was applied to the determination of nitrite in real food and water samples, with recoveries in the range of 86.9–103.4 % and relative standard deviations (RSDs) less than 9.64 %.

Scytonin, a novel cyanobacterial photoprotective pigment: calculations of Raman spectroscopic biosignatures

The Raman spectrum of scytonin, a novel derivative of the parent scytonemin, is predicted from DFT calculations of the most stable, lowest energy, conformational structure. The diagnostic importance of this study relates to the spectral ability to discriminate between scytonemin and its derivatives alone or in admixture with geological matrices from identified characteristic Raman spectral signatures. The successful interpretation of biosignatures from a wide range of cyanobacterial extremophilic colonization in terrestrial and extraterrestrial scenarios is a fundamental requirement of the evaluation of robotic spectroscopic instrumentation in search for life missions. Scytonemin is produced exclusively by cyanobacterial colonies in environmentally stressed habitats and is widely recognized as a key target biomarker molecule in this enterprise. Here, the detailed theoretical analysis of the structure of scytonin enables a protocol to be established for the recognition of characteristic bands in its Raman spectrum and to accomplish the successful differentiation between scytonin and scytonemin as well as other scytonemin derivatives such as the dimethoxy and tetramethoxy compounds that have been isolated from cyanobacterial colonies but which have not yet been characterized spectroscopically. The results of this study will facilitate an extension of the database capability for miniaturized Raman spectrometers which will be carried on board search for life robotic missions to Mars, Europa, and Titan.

Hole arrayed metal-insulator-metal structure for surface enhanced Raman scattering by self-assembling polystyrene spheres

A fabrication process based on the self-assembling polystyrene spheres is proposed to obtain hole arrayed metal-insulator-metal (HA-MIM) structure for surface enhanced Raman scattering (SERS). The localized field enhancement aroused by the gap resonance in the HA-MIM structure is analyzed by finite-different time domain (FDTD) method. With reference to the theory result, the structure is experimentally fabricated and the Raman scattering spectrum of rhodamine 6G (R6G) is measured by a miniaturized Raman spectrometer. The results shows that the enhancement factor is 3.85 times higher than the control sample with single layered metal hole array. The fabrication process to obtain the HA-MIM SERS substrate is reproducible, fast, large area and low cost.

Raman Spectroscopic Analysis of Geological and Biogeological Specimens of Relevance to the ExoMars Mission

A novel miniaturized Raman spectrometer is scheduled to fly as part of the analytical instrumentation package on an ESA remote robotic lander in the ESA/Roscosmos ExoMars mission to search for evidence for extant or extinct life on Mars in 2018. The Raman spectrometer will be part of the first-pass analytical stage of the sampling procedure, following detailed surface examination by the PanCam scanning camera unit on the ExoMars rover vehicle. The requirements of the analytical protocol are stringent and critical; this study represents a laboratory blind interrogation of specimens that form a list of materials that are of relevance to martian exploration and at this stage simulates a test of current laboratory instrumentation to highlight the Raman technique strengths and possible weaknesses that may be encountered in practice on the martian surface and from which future studies could be formulated. In this preliminary exercise, some 10 samples that are considered terrestrial representatives of the mineralogy and possible biogeologically modified structures that may be identified on Mars have been examined with Raman spectroscopy, and conclusions have been drawn about the viability of the unambiguous spectral identification of biomolecular life signatures. It is concluded that the Raman spectroscopic technique does indeed demonstrate the capability to identify biomolecular signatures and the mineralogy in real-world terrestrial samples with a very high degree of success without any preconception being made about their origin and classification.

The Miniaturized Raman System and Detection of Traces of Life in Halite from the Atacama Desert: Some Considerations for the Search for Life Signatures on Mars

Raman spectroscopy is being adopted as a nondestructive instrumentation for the robotic exploration of Mars to search for traces of life in the geological record. Here, miniaturized Raman spectrometers of two different types equipped with 532 and 785 nm lasers for excitation, respectively, were compared for the detection of microbial biomarkers in natural halite from the hyperarid region of the Atacama Desert. Measurements were performed directly on the rock as well as on the homogenized, powdered samples prepared from this material-the effects of this sample preparation and the excitation wavelength employed in the analysis are compared and discussed. From these results, 532 nm excitation was found to be superior for the analysis of powdered specimens due to its high sensitivity toward carotenoids and hence a higher capability for their detection at relatively low concentration in bulk powdered specimens. For the same reason, this wavelength was a better choice for the detection of carotenoids in direct measurements made on the rock samples. The 785 nm excitation wavelength, in contrast, proved to be more sensitive toward the detection of scytonemin.

Feasibility of Raman microspectroscopic identification of biomarkers through gypsum crystals

The miniaturized Raman spectrometer is considered to be a candidate instrument for the Pasteur payload (the ExoMars mission scheduled for 2018). This mission will, for the first time, combine mobility and access to subsurface locations where organic molecules might be well preserved. Evaporitic crystals are among the potential protected habitats that have been postulated. Various concentrations of biomarkers (beta-carotene, glycine and phthalic acid) dispersed in a gypsum matrix were analyzed through transparent mineral (gypsum) plates of different thicknesses. By doing so, conditions were simulated in which biomarkers were trapped within evaporitic crystals. Using a long-working distance objective, all studied concentrations of biomarkers mixed in gypsum powder were detected. The characteristic Raman bands were easily observable for a 10% mixture of all chosen biomarkers not only through a 3.3 mm plate and but even through a 5.2 mm plate. It was possible to detect key Raman bands of 1% phthalic acid/gypsum mixture and 1% beta-carotene/gypsum mixture even through a 5.2 mm gypsum plate. The 1% beta-carotene/gypsum mixture was still clearly distinguishable through an 8.5 mm gypsum crystal due to the known resonance Raman effect of the molecule.

Raman spectroscopy study of lichens using three spectrometers under different experimental conditions: analyses of the results with relevance for extraplanetary exploration

We have carried out analyses on three extremophile lichens from the Tabernas Desert (Spain) under different experimental conditions: dry and wet samples in the laboratory and wet specimens in the field, using three spectrometers: one portable, one FT-Raman and one dispersive micro-Raman instruments. Apart from pigment characterization, the information obtained from the spectra is compared and the differences analysed. The fact that no results were achieved on dry lichens using the handheld spectrometer is of special relevance for those looking for life in hazardous environments. Since in some extreme habitats, life could be “dormant” and only activates under appropriate conditions, miniaturized instruments (such as those included for extraplanetary exploration missions) could not detect bio-markers and, then, life signals will not be noticed when, actually, life is there. Furthermore, we show in this work, using the miniaturized Raman spectrometer, that not only laser wavelength, spectral resolution and wavenumber region of the spectrum are important for the bio- or geo-marker recognition but also spot-size is of vital relevance for the unambiguous biomolecular characterization. In our opinion, mini-Raman instruments are useful for assessing in situ the eventual presence of bio-markers in complex natural samples, such as organisms, but are not accurate enough for precise molecular identification.

Using portable Raman spectrometers for the identification of organic compounds at low temperatures and high altitudes: exobiological applications

Organic minerals, organic acids and NH-containing organic molecules represent important target molecules for astrobiology. Here, we present the results of the evaluation of a portable hand-held Raman spectrometer to detect these organic compounds outdoors under field conditions. These measurements were carried out during the February-March 2009 winter period in Austrian Alpine sites at temperatures ranging between -5 and -25 degrees C. The compounds investigated were detected under field conditions and their main Raman spectral features were observed unambiguously at their correct reference wavenumber positions. The results obtained demonstrate that a miniaturized Raman spectrometer equipped with 785 nm excitation could be applied with advantage as a key instrument for investigating the presence of organic minerals, organic acids and nitrogen-containing organic compounds outdoors under terrestrial low-temperature conditions. Within the payload designed by ESA and NASA for several missions focusing on Mars, Titan, Europa and other extraterrestrial bodies, Raman spectroscopy can be proposed as an important non-destructive analytical tool for the in situ identification of organic compounds relevant to life detection on planetary and moon surfaces or near subsurfaces.

Understanding the Application of Raman Spectroscopy to the Detection of Traces of Life

Investigating carbonaceous microstructures and material in Earth's oldest sedimentary rocks is an essential part of tracing the origins of life on our planet; furthermore, it is important for developing techniques to search for traces of life on other planets, for example, Mars. NASA and ESA are considering the adoption of miniaturized Raman spectrometers for inclusion in suites of analytical instrumentation to be placed on robotic landers on Mars in the near future to search for fossil or extant biomolecules. Recently, Raman spectroscopy has been used to infer a biological origin of putative carbonaceous microfossils in Early Archean rocks. However, it has been demonstrated that the spectral signature obtained from kerogen (of known biological origin) is similar to spectra obtained from many poorly ordered carbonaceous materials that arise through abiotic processes. Yet there is still confusion in the literature as to whether the Raman spectroscopy of carbonaceous materials can indeed delineate a signature of ancient life. Despite the similar nature in spectra, rigorous structural interrogation between the thermal alteration products of biological and nonbiological organic materials has not been undertaken. Therefore, we propose a new way forward by investigating the second derivative, deconvolution, and chemometrics of the carbon first-order spectra to build a database of structural parameters that may yield distinguishable characteristics between biogenic and abiogenic carbonaceous material. To place Raman spectroscopy as a technique to delineate a biological origin for samples in context, we will discuss what is currently accepted as a spectral signature for life; review Raman spectroscopy of carbonaceous material; and provide a historical overview of Raman spectroscopy applied to Archean carbonaceous materials, interpretations of the origin of the ancient carbonaceous material, and a future way forward for Raman spectroscopy.

Raman spectra of organic acids obtained using a portable instrument at −5 °C in a mountain area at 2000 m above sea level

AbstractWell‐resolved Raman spectra of organic acids were obtained with 785 nm excitation using a portable Raman instrument (Ahura First Defender XL) under low temperature −5 °C atmospheric conditions at an altitude of 2000 m (Axamer Lizum, Innental, Austria). The portable Raman spectrometer tested in this setting permits fast and unambiguous detection of solid forms of these organic acids (formic, acetic, valeric, hexanoic, heptanoic, isophthalic, ascorbic and mellitic) under field conditions. This demonstrates the possibility to use a miniaturized Raman spectrometer as a key instrument for investigating the presence of organic compounds and biomolecules under low temperature conditions. These results are important for future missions focusing not only on Mars, where Raman spectroscopy will be a key non‐destructive analytical tool for the in situ identification of organic compounds relevant to life detection on planetary surfaces or near sub‐surfaces. Copyright © 2009 John Wiley & Sons, Ltd.

Raman Spectroscopy for the Undergraduate Teaching Laboratory: Quantification of Ethanol Concentration in Consumer Alcoholic Beverages and Qualitative Identification of Marine Diesels Using a Miniature Raman Spectrometer

Raman spectroscopy has steadily gained popularity as a powerful tool in both the analytical lab and the undergraduate classroom. The technique is attractive because it allows for rapid, nondestructive qualitative or quantitative analyses of many analytes with little or no sample preparation requirements. The introduction of less expensive, smaller, and more powerful diode laser excitation sources and the recent availability of rugged, red‐sensitive, charge‐coupled device–based miniature modular spectrometers has prompted the integration of Raman spectroscopy into the undergraduate curriculum. We have evaluated the analytical utility of a small, portable Raman instrument for the qualitative and quantitative analyses of two “real” samples. The experiments in this paper were designed to be used as a laboratory component for undergraduate education and include the quantification of ethanol in consumer alcoholic beverages and the qualitative identification of marine diesel fuels that had been spilled on surface waters. In the case of the liquor samples, the ethanol concentration in colorless, odorless alcoholic beverages could be determined very rapidly, but colored and heavily scented liquors proved more difficult and required pretreatment with activated carbon to remove fluorescence that masked the Raman signal. Similarly, a high‐intensity fluorescence background was observed to mask characteristic Raman bands of the diesel fuels. Some reduction in the intensity of the fluorescence was observed after carbon pretreatment of the fuels. The set of undergraduate experiments described in this paper treat the concepts of quantitative and qualitative analysis using portable instrumentation, instrumental calibration by the standard addition and external curve methods, and method development for the analysis of real consumer and environmental samples.

Raman and SEM analysis of a biocolonised hot spring travertine terrace in Svalbard, Norway.

BackgroundA profile across 8 layers from a fossil travertine terrace from a low temperature geothermal spring located in Svalbard, Norway has been studied using both Raman spectroscopy and SEM (Scanning Electron Microscopy) techniques to identify minerals and organic life signals.ResultsCalcite, anatase, quartz, haematite, magnetite and graphite as well as scytonemin, three different carotenoids, chlorophyll and a chlorophyll-like compound were identified as geo- and biosignatures respectively, using 785 and/or 514 nm Raman laser excitation wavelengths. No morphological biosignatures representing remnant microbial signals were detected by high-resolution imaging, although spectral analyses indicated the presence of organics. In contrast, in all layers, Raman spectra identified a series of different organic pigments indicating little to no degradation or change of the organic signatures and thus indicating the preservation of fossil biomarker compounds throughout the life time of the springs despite the lack of remnant morphological indicators.ConclusionWith a view towards planetary exploration we discuss the implications of the differences in Raman band intensities observed when spectra were collected with the different laser excitations. We show that these differences, as well as the different detection capability of the 785 and 514 nm laser, could lead to ambiguous compound identification. We show that the identification of bio and geosignatures, as well as fossil organic pigments, using Raman spectroscopy is possible. These results are relevant since both lasers have been considered for miniaturized Raman spectrometers for planetary exploration.

FT–Raman spectroscopic study of calcium-rich and magnesium-rich carbonate minerals

Calcium and magnesium carbonates are important minerals found in sedimentary environments. Although sandstones are the most common rock colonized by endolith organisms, the production of calcium and magnesium carbonates is important in survival strategies of organisms and as a source for the removal of oxalate ions. Extremophile organisms in some situations may convert or destroy carbonates of calcium and magnesium, which gives important information about the conditions under which these organisms can survive. The identification on the surface of Mars of ‘White Rock’ formations, in Juventae Chasma or Sabaea Terra, as possibly carbonate rocks makes the study of these minerals a prerequisite of remote Martian exploration. Here, we show the protocol for the identification by Raman spectroscopy of different calcium and magnesium carbonates and we present a database of relevance in the search for life, extinct or extant, on Mars; this will be useful for the assessment of data obtained from remote, miniaturized Raman spectrometers now proposed for Mars exploration.

Spectroscopic requirements for Raman instrumentation on a planetary lander: potential for the remote detection of biosignatures on Mars

The special characteristics of Raman spectroscopy (relative insensitivity to water, non-destructive detection, sensitivity to bio- and geosignatures, molecular structural composition information, etc.) together with the development of miniaturized Raman spectrometers make the consideration of this technique for future robotic landers on planetary surfaces, particularly Mars, a very interesting option. The development of light and rugged Raman spectrometers limits the possible scope of the instrumentation which has particular importance in the recognition of biomolecular and mineral signatures. In this work, we evaluate the spectral resolution and scan time parameters and the effect that they have on the Raman spectra of extremophilic biomolecules, together with the wavenumber ranges which are critical for the detection of life signals. This is of vital relevance for the design of miniaturized Raman spectrometer systems. From our results, we conclude that for extraterrestrial biological signatures unambiguous Raman spectral identification provided with a minimum of 16 cm−1 spectral resolution is required for the most significant biosignature wavenumber range in the 1700–700 cm−1 region.

Development of a simultaneous measurement system of x‐ray diffraction and raman spectra: Application to structural study of crystalline‐phase transitions of chain molecules

AbstractThe development of a bench‐top‐type system for simultaneous measurement of X‐ray diffraction and Raman spectra has been made to investigate structural changes in the phase transitions of chain molecules such as polyethylene, n‐alkane, and so forth from various viewpoints. For the X‐ray diffraction measurement an imaging plate or a charge‐coupled device camera was used as a two‐dimensional detector. For the Raman spectral measurement a miniature Raman spectrometer was used with optical fibers for the irradiation of incident laser beams and collection of scattered signals. For example, in the heating process of the n‐C30H62 sample, the phase transition from orthorhombic‐to‐hexagonal lattices could be detected clearly by the X‐ray and Raman measurements. By comparing these two different types of data in detail, an intimate relationship between conformational disordering and rotational motion of molecular chains is detected more clearly than before. Also, similar discussion can be made for the orthorhombic‐to‐hexagonal phase transition of the geometrically constrained polyethylene sample occurring immediately below the melting point. Another example concerns the structural change in the photoinduced solid‐state polymerization of cis,cis‐diethylmuconate single crystal. From the shifts in the X‐ray reflection position and Raman frequency characteristic of the produced polymer, it was found that the molecular deformation of the polymer chains and lattice strain was induced in the early stage of the polymerization reaction. For the ferroelectric‐phase transition of vinylidene fluoride copolymer, the simultaneous measurement was made for collecting triple information of small‐angle and wide‐angle X‐ray scatterings and Raman spectra to know the relationship between the structural change in the crystal lattice and the morphological change in the lamellar stacking mode. © 2002 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 40: 495–506, 2002; DOI 10.1002/polb.10112

Pigmentation as a survival strategy for ancient and modern photosynthetic microbes under high ultraviolet stress on planetary surfaces

Solar radiation is the primary energy source for surface planetary life, so that pigments are fundamental components of any surface-dwelling organism. They may therefore have evolved in some form on Mars as they did on Earth. Photosynthetic microbes are major primary producers on Earth, but are concurrently vulnerable to ultraviolet (UV) damage. Using non-intrusive laser Raman spectroscopy to recognize the component parts of biomolecules, we have shown not only the abundance of microbial photosynthetic and photoprotective pigments in situ, but also their spatial distribution within their microhabitat. This essential aspect of their screening or avoidance survival strategies is lost on extraction with solvents. This precise approach is eminently suited to analysis of epilithic (surface) and endolithic (within rocks) communities in Antarctic desert habitats, which are putative analogues of early Mars. Raman spectra for key biomolecules (e.g. the UV screen parietin and the antioxidant β-carotene in epilithic lichens) enable not only the detection of organics in light-stratified habitats, but also the characterization of unknown pigments. Typical biomarkers of astrobiological relevance in our Raman spectral database include scytonemin (a UV screen), chlorophyll (primary photosynthetic pigment), phycocyanin (accessory pigment for shade adaptation) and a hopanoid extracted from 2·5 Gya microbial stromatolite from Australia. This compound dates from the same time period when a wetter Mars could have had a potentially flourishing surface microbial community of its own. Analyses with a laboratory Raman instrument have been extended to a novel miniature Raman spectrometer, operating at the same optimal excitation wavelength (1064 nm) via an In-Ga-As detector. After evaluation in Antarctica, this instrument will be space-qualified for a proposed Mars rover mission to detect biomolecules in the near-surface sediment profile of palaeolakes, using experience with Antarctic biomarkers to interpret alien spectra of fundamental components, without the need for prior knowledge of the identity of the target compounds.

Why astrobiology needs collaboration

Antarctic astrobiological research has benefited from the affiliation of the UK Astrobiology Forum (UKAF) to the NASA Astrobiology Institute (NAI). NAI funding of a miniature Raman spectrometer for the characterization of biomolecules in Antarctic cold deserts as a prelude to its space qualification for a future Mars lander/rover mission is a practical example of collaboration between the UK and USA fostered by this agreement. Research by the British Antarctic Survey into cyanobacterial communities under stress from UV, desiccation, salinity stress and low temperature features in joint studies associated with NASA Ames Research Center, Montana State University and the International Space Station.

A Miniaturized, No-Moving-Parts Raman Spectrometer

A solid-state acousto-optic tunable filter (AOTF) is combined with krypton laser excitation (647 nm), holographic Raman filters, and photon-counting silicon avalanche photodiode (APD) detection to construct a miniaturized Raman spectrometer with no moving parts. The physically compact AOTF and the highly integrated APD provide a rugged, digitally controlled spectrometer of moderate spectral resolution and with a footprint comparable in size to a laboratory notebook. Instrument design details are considered and representative spectra are reported. Potential areas of application for this prototype Raman spectrometer are also discussed.