Impact on the Raman spectra of liquids when a polarized light source is used.

The polarization state of the excitation light used in two Raman systems was controlled to study its effect in the unpolarized Raman spectra of unstructured samples. Both systems work in different regions of the electromagnetic spectrum (NIR and visible). Four polarization states (linear, linear at 45° and 90°, and circular) were used to excite liquid samples (ethanol, acetone, and their mixture). The results show that the Raman peaks intensities' ratio varies according to the polarization state of the excitation light. Peaks related to functional groups and C-H stretching modes increase their intensity when circular polarization (CP) is applied. The latter may help to study liquid mixtures with low concentrations. Different polarizing light states give a more detailed spectroscopic analysis since it gathers more structural information of the samples tested in this work with an undefined structure.

Imaging of bee honey sugar crystals by second-harmonic generation microscopy.

Bee honey is an exceptionally nutritious food with unique chemical and mineral contents. This report introduces the use of the second-harmonic generation (SHG) microscopy for imaging honey sugar crystals' morphology as an alternative for its authentication process. The crystals and their boundaries are clearly observed with SHG compared with bright-field microscopy, where the liquid honey avoids the visualization of a sharp image. Four different honey samples of Mexico's various floral origins and geographical regions are analyzed in our study. These samples are representative of the diversity and valuable quality of bee honey production. The SHG image information is complemented with Raman spectroscopy (RS) analysis, since this optical technique is widely used to validate the bee's honey composition stated by its floral origin. We relate the SHG imaging of honey crystals with the well-defined fructose and glucose peaks measured by RS. Size measurement is introduced using the crystal´s length ratio to differentiate its floral origin. From our observations, we can state that SHG is a promising and suitable technique to provide a sort of optical fingerprint based on the floral origin of bee honey.

Spectroscopic study of honey from Apis mellifera from different regions in Mexico.

The objective of this study was to analyze by Raman and UV-Vis-NIR Spectroscopic techniques, Mexican honey from Apis Mellífera, using representative samples with different botanic origins (unifloral and multifloral) and diverse climates. Using Raman spectroscopy together with principal components analysis, the results obtained represent the possibility to use them for determination of floral origin of honey, independently of the region of sampling. For this, the effect of heat up the honey was analyzed in relation that it was possible to greatly reduce the fluorescence background in Raman spectra, which allowed the visualization of fructose and glucose peaks. Using UV-Vis-NIR, spectroscopy, a characteristic spectrum profile of transmittance was obtained for each honey type. In addition, to have an objective characterization of color, a CIE Yxy and CIE L*a*b* colorimetric register was realized for each honey type. Applying the principal component analysis and their correlation with chromaticity coordinates allowed classifying the honey samples in one plot as: cutoff wavelength, maximum transmittance, tones and lightness. The results show that it is possible to obtain a spectroscopic record of honeys with specific characteristics by reducing the effects of fluorescence.

Raman spectroscopic analysis of iron chromium oxide microspheres generated by nanosecond pulsed laser irradiation on stainless steel.

Iron chromium oxide microspheres were generated by pulsed laser irradiation on the surface of two commercial samples of stainless steel at room temperature. An Ytterbium pulsed fiber laser was used for this purpose. Raman spectroscopy was used for the characterization of the microspheres, whose size was found to be about 0.2-1.7 µm, as revealed by SEM analysis. The laser irradiation on the surface of the stainless steel modified the composition of the microspheres generated, affecting the concentration of the main elemental components when laser power was increased. Furthermore, the peak ratio of the main bands in the Raman spectra has been associated to the concentration percentage of the main components of the samples, as revealed by Energy-Dispersive X-ray Spectroscopy (EDS) analysis. These experiments showed that it is possible to generate iron chromium oxide microspheres on stainless steel by laser irradiation and that the concentration percentage of their main components is associated with the laser power applied.

Infrared nanosecond pulsed laser irradiation of stainless steel: micro iron-oxide zones generation.

Nanosecond-pulsed, infrared (1064 nm) laser irradiation was used to create periodic metal oxide coatings on the surface of two samples of commercial stainless steel at ambient conditions. A pattern of four different metal oxide zones was created using a galvanometer scanning head and a focused laser beam over each sample. This pattern is related to traverse direction of the laser beam scanning. Energy-dispersive X-ray spectroscopy (EDS) was used to find the elemental composition and Raman spectroscopy to characterize each oxide zone. Pulsed laser irradiation modified the composition of the stainless steel samples, affecting the concentration of the main components within each heat affected zone. The Raman spectra of the generated oxides have different intensity profiles, which suggest different oxide phases such as magnetite and maghemite. In addition, these oxides are not sensible to the laser power of the Raman system, as are the iron oxide powders reported in the literature. These experiments show that it is possible to generate periodic patterns of various iron oxide zones by laser irradiation, of stainless steel at ambient conditions, and that Raman spectroscopy is a useful punctual technique for the analysis and inspection of small oxide areas.

Effect of the finishing oil of acrylic fibers in the optical rotation of the Raman scattered light.

Polarized Raman spectra have been obtained from polyacrylonitrile copolymers fibers with vinyl acetate Poly(AN-co-VA), and methyl acrylate Poly(AN-co-MA) with finishing and without finishing, in order to show the effect of the finishing in the optical rotation of the Raman scattered light. The polarized Raman spectra were used to calculate the depolarization ratios for both fibers. These values reveal that there is antisymmetric Raman scattering in the form of anomalous depolarization for some bands due to a dipolar interaction between the polar headgroup of the finishing with the polar nitrile group of the fiber causing changes in the orientation of fiber polymer chains, or pseudo antisymmetric Raman scattering due to planar hydrocarbons of the oil finishing which are optically active and are aligned when they are applied to the acrylic fibers during the spinning process. Although the finishing should not affect the physical or chemical properties of the fibers, in this work is shown that the finishing could introduce optical activity in the different wavenumbers of the Raman signal and this effect is proportional to the finishing content. According to the results obtained in this work, Raman polarized spectroscopy can provide an express method to identify acrylic fibers with finishing and without finishing agents.

Use of Raman spectroscopy and chemometrics for the quantification of metal ions attached to Lactobacillus kefir.

AIMS: To set-up an experimental and analytical methodology to evaluate the feasibility of developing simple, accurate and quantitative models based on Raman spectroscopy and multivariate analysis for the quantification of metal ions adsorbed to the bacterial surface of Lactobacillus kefir. METHODS AND RESULTS: One millilitre cultures from two strains of Lact. kefir in the stationary phase were harvested and washed twice with ultra pure water. The bacterial pellets were resuspended into 1 ml solutions of Pb(+2), Cd(+2) or Ni(+2) ranging from 0 to 0·9 mmol l(-1). The suspensions were further incubated for 1 h at 30°C at pH 5·5. After centrifugation, the pellets were kept to register the Raman spectra and the supernatants were used for the analytical determination of Pb(+2) , Cd(+2) and Ni(+2). Micro-organisms nontreated with metal ions were used as controls. Principal component analysis (PCA) was performed over the preprocessed Raman spectra to evaluate whether the clusters obtained could be correlated with the concentration of metal ions attached to the bacterial biomass. After that, partial least squares (PLS) models were calibrated with the aim of quantifying the metal ions adsorbed to the bacterial surface. According to the analytical determinations, the maximum binding capacity of all the metals (q(max)) attained values that are comparable with those observed for other lactic acid bacteria (ca. 0·200 mmol g(-1)). The spectral analysis revealed that the main functional groups involved in the bacteria/metal interaction are carboxylates, phosphates and polysaccharides. In PCA, the first two principal components explain more than 72% variance of the spectral data set contained in the data structure, allowing a clear discrimination among samples of different concentrations. Based on this information and using as reference the results obtained by analytical methods, PLS prediction models were successfully defined for the quantification of Pb(+2), Cd(+2) and Ni(+2) attached to the bacterial surface. CONCLUSIONS: The calibration and validation of methods based on multivariate analysis allowed the definition of models for the quantification of Pb(+2), Cd(+2) and Ni(+2) attached to bacterial surfaces. The high percentages of explained variances in PCA gave a strong support to calibrate the prediction models, depicting very good correlations with the reference method (correlations ∼0·90 in all cases). SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus kefir CIDCA 8348 and JCM 5818 bind Pb(+2), Cd(+2) and Ni(+2) in an efficient way. This fact gives support for their potential use as sequestrants of traces of these metals in products addressed to human and animal consume. The prediction models developed would be useful for the determination of the investigated metal ions in unknown samples giving at the same time, structural information about this interaction. This is certainly the most important contribution of this work.

Raman spectroscopy for the identification of pigments and color measurement in Dugès watercolors.

Spectroscopic and colorimetric analysis of a representative set of Dugès watercolor paintings was performed. These paintings were the result of scientific studies carried out by the zoologist Alfredo Dugès, who recorded the fauna of the Mexican Republic between 1853 and 1910. Micro-Raman spectroscopy, with an excitation wavelength of 830 nm, and colorimetric techniques were employed in order to understand if different colors with the same hue were reproduced using the same pigments. The color coordinates of the measured areas were obtained in the CIEL*a*b* color space. Raman analysis showed that, in some cases, to reproduce colors with the same hue the pigment employed was not the same. Pigments identified in the watercolors were vermilion, carbon-based black, lead white, gamboge and chrome yellow, Prussian and ultramarine blue. Some of these pigments have been used since ancient times, others as Prussian blue, chrome yellow and synthetic ultramarine blue arrived to the market at the beginning of the 18th and 19th centuries, respectively. Furthermore, regarding the white color, instead of left the paper unpainted, lead white was detected in the eye of a bird. The green color was obtained by mixing Prussian blue with chrome yellow. The results of this work show the suitability of using Raman spectroscopy for watercolor pigment analysis and colorimetric techniques to measure the color of small areas (246 microm x 246 microm) that was the case for the lead white pigment.

Raman spectroscopy and multivariate analysis of serum samples from breast cancer patients.

Serum samples were studied using Raman spectroscopy and analyzed through the multivariate statistical methods of principal component analysis (PCA) and linear discriminant analysis (LDA). The blood samples were obtained from 11 patients who were clinically diagnosed with breast cancer and 12 healthy volunteer controls. The PCA allowed us to define the wavelength differences between the spectral bands of the control and patient groups. However, since the differences in the involved molecules were in their tertiary or quaternary structure, it was not possible to determine what molecule caused the observed differences in the spectra. The ratio of the corresponding band intensities were analyzed by calculating the p values and it was found that only seven of these band ratios were significant and corresponded to proteins, phospholipids, and polysaccharides. These specific bands might be helpful during screening for breast cancer using Raman Spectroscopy of serum samples. It is also shown that serum samples from patients with breast cancer and from the control group can be discriminated when the LDA is applied to their Raman spectra.

Application of principal component analysis and Raman spectroscopy in the analysis of polycrystalline BaTiO3 at high pressure.

The principal component analysis (PCA) was applied to Raman spectra of polycrystalline BaTiO(3) under pressure from atmospheric pressure to approximately 6.72 GPa. For the system utilized, PCA was able to distinguish spectral features and to determine the phase transition pressure: tetragonal to cubic at approximately 2.0 GPa. The present study demonstrates the potentialities of the application of PCA to the investigation on phase transitions at high pressure by Raman spectroscopy.

CdTe surface roughness by Raman spectroscopy using the 830 nm wavelength.

A Raman spectroscopic study was performed to detect the surface roughness of a cadmium telluride (CdTe) wafer sample, using the 514.5, 632.8 and 830.0 nm excitations wavelengths. To verify the relation between the roughness and the structure of Raman spectra, in certain zones of the sample, we measured their roughness with an atomic force microscopy. It was found that, using the 830 nm wavelength there is a direct correspondence between the spectrum structure and the surface roughness. For the others wavelengths it was found, however, that there is not a clearly correspondence between them. Our results suggest that, using the excitation wavelength of 830 nm the Raman spectroscopy can be used as an on-line roughness monitor on the CdTe growth.

Qualitative study of ethanol content in tequilas by Raman spectroscopy and principal component analysis.

Using Raman spectroscopy, with an excitation radiation source of 514.5 nm, and principal component analysis (PCA) was elaborated a method to study qualitatively the ethanol content in tequila samples. This method is based in the OH region profile (water) of the Raman spectra. Also, this method, using the fluorescence background of the Raman spectra, can be used to distinguish silver tequila from aged tequilas. The first three PCs of the Raman spectra, that provide the 99% of the total variance of the data set, were used for the samples classification. The PCA1 and PCA2 are related with the water (or ethanol) content of the sample, whereas the PCA3 is related with the fluorescence background of the Raman spectra.

The application of NIR Raman spectroscopy in the assessment of serum thyroid-stimulating hormone in rats.

Serum blood samples of euthyroid and thyroidectomized rats treated with thyrotropin-releasing hormone (TRH) were analyzed on aluminum substrates using the near-infrared Raman spectroscopy (830 nm). Spectra of thyroid-stimulating hormone (TSH), TRH and prolactin standards were obtained. Differences between Raman spectra profiles of control and Tx + TRH samples groups were found. These differences were confirmed by the linear discriminant analysis (LDA), which presents a good classification between groups. It is supposed that these differences are produced by the increment of TSH in the thyroidectomized rats.

Amino acid contents along the visual and equatorial axes of a pig lens by Raman spectroscopy.

Using near infrared Raman microspectroscopy with laser light of 830 nm, the distribution of amino acids along the visual and equatorial axes of a normal pig lens was studied. The classification of pig lens Raman spectra in these axes was performed using principal component analysis and linear discriminant analysis. The analysis of the scattered light selectively collected from point to point, along the visual axis, indicated that the tyrosine and tryptophan increases and then, at approximately 4 mm position, decreases. Moreover, in the equatorial plane, the nuclear part has the highest concentration of these amino acids. However, the phenylalanine content increases from anterior to posterior cortex of the lens as long as in the equatorial axis it slightly increases and then at approximately 2-2.3 mm position, decreases. The changes in amino acid conformation along the visual axis, similarly to the changes in protein conformation, may explain the refractive gradient of the lens.

Study of the protein distribution in the pig lens cross section by Raman spectroscopy.

The distribution of proteins in the cross section of a normal pig lens was studied by near-infrared Raman spectroscopy. The Raman spectra were measured in the visual and equatorial axes of this cross section and the protein peak intensities were determined. It was found that along each axis the protein intensities fluctuate. They have a considerable increment along the visual axis with the exception of the C-N bond peak intensities at 1087.2 cm(-1), which decrease, and along the equatorial axis the increment is slight. This increment in protein distribution along the visual axis is related with the refractive gradient of the lens. The classification of pig lens spectra in these axes was performed using principal component analysis (PCA) and linear discriminant analysis (LDA). Cross-validation shows an excellent group separation.