Forensic analysis of architectural finishes using fourier transform infrared and Raman spectroscopy, part II: white paint

White household paints are commonly encountered as evidence in the forensic laboratory but they often cannot be readily distinguished by color alone so Fourier transform infrared (FT-IR) microscopy is used since it can sometimes discriminate between paints prepared with different organic resins. Here we report the first comparative study of FT-IR and Raman spectroscopy for forensic analysis of white paint. Both techniques allowed the 51 white paint samples in the study to be classified by inspection as either belonging to distinct groups or as unique samples. FT-IR gave five groups and four unique samples; Raman gave seven groups and six unique samples. The basis for this discrimination was the type of resin and/or inorganic pigments/extenders present. Although this allowed approximately half of the white paints to be distinguished by inspection, the other half were all based on a similar resin and did not contain the distinctive modifiers/pigments and extenders that allowed the other samples to be identified. The experimental uncertainty in the relative band intensities measured using FT-IR was similar to the variation within this large group, so no further discrimination was possible. However, the variation in the Raman spectra was larger than the uncertainty, which allowed the large group to be divided into three subgroups and four distinct spectra, based on relative band intensities. The combination of increased discrimination and higher sample throughput means that the Raman method is superior to FT-IR for samples of this type.     

酚醛树脂结构对其热解成炭特性的影响

采用傅立叶红外光谱对固化前后的酚醛树脂进行结构表征,通过热重法对固化后酚醛树脂的热降解失重温度及其高温下的成炭率进行分析,研究酚醛树脂结构中取代基的相对含量与其热稳定性及成炭率的关系。结果表明,酚醛树脂在高温下的成炭率与其结构中取代基的含量有一定的对应关系,亚甲基含量高、羰基和醚键含量低的酚醛树脂固化物高温成炭率较高。在六种不同醛酚比(F/P)的树脂固化物中,亚甲基的含量以F/P为1.2时的固化物最大,羰基的含量则以F/P为1.2时最小。固化后的六种酚醛树脂中以PF1.2用于交联的有效取代基(亚甲基)数目最多,交联密度最高,具有最高的成炭率(73.78%)。     

Discrimination between nontumor bladder tissue and tumor by Raman spectroscopy

We have applied Raman spectroscopy to discriminate between nontumor and tumor bladder tissue and to determine the biochemical differences therein. Tissue samples from 15 patients were collected, and frozen sections were made for Raman spectroscopy and histology. Twenty-five pseudocolor Raman maps were created in which each color represents a cluster of spectra measured on tissue areas of similar biochemical composition. For each cluster, the cluster-averaged spectrum (CAS) was calculated and classified as tumor and nontumor in accordance to pathohistology. Unguided hierarchical clustering was applied to display heterogeneity between and within groups of nontumor and tumor CAS. A linear discriminant analysis model was developed to discriminate between CAS from tumor and nontumor. The model was tested by a leave-one-patient-out validation, 84 of the 90 CAS (93%) were correctly classified with 94% sensitivity and 92% specificity. Biochemical differences between tumor and nontumor CAS areas were analyzed by fitting spectra of pure compounds to the CAS. Nontumor CAS showed higher collagen content while tumor CAS were characterized by higher lipid, nucleic acid, protein, and glycogen content. Raman spectroscopy enabled effective discrimination between tumor and nontumor bladder tissue based on characterized biochemical differences, despite heterogeneity expressed in both tumor and nontumor CAS.     

Determination of omega-6 and omega-3 fatty acids in pork adipose tissue with nondestructive Raman and fourier transform infrared spectroscopy

In order to predict omega-6 and omega-3 fatty acids in the diet of humans, seventy-three pork back fat adipose tissue samples were measured with Raman spectroscopy directly on adipose tissue and on melted fat. Melted fat samples were, in addition, measured with Fourier transform infrared (FT-IR) spectroscopy. Gas chromatography analyses were conducted as the reference analysis. Partial least squares regression (PLSR) was used to calibrate and validate all models predicting omega-3 and omega-6 fatty acids contents from spectra. Omega-6 fatty acids in melted fat measured with FT-IR was predicted with a correlation coefficient (R) of 0.93 and a root mean square error of cross-validation (RMSECV) of 1.61% of the total amount of fatty acids. Raman spectra measured on melted fat gave a prediction of omega-6 fatty acids with R=0.97, and RMSECV=0.99% of total amount of fatty acids. Omega-6 fatty acids were predicted with R=0.94, and RMSECV=1.50% of the total amount of fatty acids using Raman spectra recorded on adipose tissue. For omega-3 fatty acids, the highest R=0.91, and lowest RMSECV=0.23% of the total amount of fatty acids were obtained from Raman spectra acquired on melted fat. FT-IR and Raman spectroscopy may be used as rapid, nondestructive methods to determine omega-6 and omega-3 fatty acids in melted fat. Raman spectroscopy can also be used directly on adipose tissue.     

Use of Raman spectroscopy to evaluate the selectivity of bifunctional anion exchange resins for perchlorate

Raman spectroscopy is used to evaluate the selectivity of two bifunctional anion exchange resins, Purolite A-530 and Amberlite PWA-2. It was found that the adsorption of anions on the resins is described by a Frumkin isotherm, which is determined by the ion pair constant, K, and the Frumkin parameter, g. The ion pair constant, K, is a measure of the strength of interaction between the resin and the anion and the Frumkin parameter, g, takes into account interactions between adsorbed anions. Although both resins have a polystyrene backbone and trihexylammonium and triethylammonium functional groups, the Purolite A-530 resin exhibits greater selectivity for perchlorate. The only discernable differences between the two resins is that the Amberlite PWA-2 has a higher trihexylamine content and the Purolite A-530 resin exhibits greater cross-linking. How the trihexylamine/triethylamine content and the degree of cross-linking affects the performance of these resins is discussed.     

Structural characterization of carbonyl compounds by IR spectroscopy and chemometrics data analysis

Although there are some distinctive peaks in mid-IR region of the electromagnetic spectrum for carbonyl compounds, it is very difficult to assign a FT-IR spectrum to a specific carbonyl functional group due to the presence of other functional groups, which change the position of the distinctive peaks. Here, we analyzed the FT-IR spectra of a large set of carbonyl compounds by chemometrics methods to differentiate between the different carbonyl functional groups. FT-IR spectra of 370 carbonyl compounds (149 carboxylic acids, 47 aldehydes, 110 esters and 64 ketones) were collected from the Spectral Database for Organic Compounds and then were converted to digital data using a home-made program. The extended canonical variate analysis combined with partial least squares discriminate analysis method (ECVA-PLS-DA) was employed as a supervised classification method. Classification analysis by ECVA-PLS-DA resulted in a suitable classification model such that one can discriminate between the different carbonyl compounds using FT-IR spectra with a small error. The classification errors (reported as percentage of misclassified compounds) were 1.8% and 7.8% for training and test sets, respectively. Considering high structural diversity of the studied compounds and the employment of different methods for acquiring the spectra (i.e., KBr disk, CCl₄ solution, liquid film and Nujol moll) there are acceptable errors. Thus, it is concluded that with the help of chemometrics methods, one is able to differentiate the carbonyl compounds using their IR spectra without need to extra spectroscopic information. This can be considered as a significant improvement in structural characterization of organic compounds using only IR spectroscopy.     

Rapid forensic analysis and identification of ''lilac'' architectural finishes using Raman spectroscopy

The potential of Raman spectroscopy to discriminate between architectural finishes (household paint) has been investigated using a test set of 51 'lilac' paints and three different excitation wavelengths. The spectra obtained with visible excitation typically displayed a series of intense Raman bands on a featureless fluorescence background but the spectra of all the paints studied had essentially identical bands. With 785 nm excitation, although the same bands that dominated the 514 nm spectra were still observed, other bands with comparable intensity also appeared. The two strongest scattering constituents were identified as a dioxazine dye, Violet 23 and beta-Cu(phthalocyanine). A scatter plot of the intensities of marker bands for these constituents (normalized to the strong rutile bands that were always present) showed that, despite the fact that the sample set spanned a wide range of rutile : dioxazine dye : phthalo- cyanine ratios, many of the samples had very similar ratios and could not be discriminated. However, all the samples (even those with similar relative proportions of the main constituents) could be discriminated on the basis of their minor constituents, either by manually measuring band intensities or through the creation and searching of spectral libraries.     

Characterization of Raman spectra measured in vivo for the detection of cervical dysplasia

Raman spectroscopy has been shown to have the potential for providing differential diagnosis in the cervix with high sensitivity and specificity in previous studies. The research presented here further evaluates the potential of near-infrared Raman spectroscopy to detect cervical dysplasia in a clinical setting. Using a portable system, Raman spectra were collected from the cervix of 79 patients using clinically feasible integration times (5 seconds on most patients). Multiple Raman measurements were taken from colposcopically normal and abnormal areas prior to the excision of tissue. Data were processed to extract Raman spectra from measured signal, which includes fluorescence and noise. The resulting spectra were correlated with the corresponding histopathologic diagnosis to determine empirical differences between different diagnostic categories. Using histology as the gold standard, logistic regression discrimination algorithms were developed to distinguish between normal ectocervix, squamous metaplasia, and high-grade dysplasia using independent training and validation sets of data. An unbiased estimate of the accuracy of the model indicates that Raman spectroscopy can distinguish between high-grade dysplasia and benign tissue with sensitivity of 89% and specificity of 81%, while colposcopy in expert hands was able to discriminate with a sensitivity and specificity of 87% and 72%.     

Cure characteristics of alkali catalysed cashew nut shell liquid-formaldehyde resin

Cashew nut shell liquid (CNSL) is a naturally occurring chemical monomer consisting four alkyl substituted phenols. Its phenolic nature makes it suitable for polymerisation into resins by formaldehyde using sodium hydroxide (NaOH) as a catalyst and hexamethylenetetramine (HMTA) employed as a hardener. There is intense interest in understanding the cure characteristics and properties of CNSL-based resins. In this work the DSC technique has been applied to study the change in the glass transition temperature of the oven-cured resin with and without HMTA in order to monitor the extent of cure. The glass transition temperature was found to rise when the alkaline catalysed resin was subjected to higher curing temperatures regardless of the concentration of formaldehyde used. The mode of cure of the NaOH-catalysed CNSL-formaldehyde resin has been found to be more regular with HMTA hardener. FT-IR spectroscopy has been used to study the neat CNSL and polymerised CNSL-formaldehyde resin with and without HMTA. The use of the DSC and FT-IR techniques to elucidate the extent of cure of CNSL resins is a valuable step towards the production of commercially successful CNSL-natural fibre composites.     

Surface modification of activated carbon fabric with ozone,part 1: Kinetics and oxidation degree

Two samples of activated carbon fabrics (ACF) with very high surface area (>1300 to >1800 m²/g) were reacted with ozone inside a closed flask under static conditions. The kinetics of ozone decomposition and reaction with the ACF surface was measured in the gas phase using Fourier transform infrared analysis (FT-IR) spectroscopy. The ozone consumption under these conditions was following the pseudofirst-order kinetics law and was accompanied by the production of CO₂ and CO. The ozone-oxidized ACF were studied by FT-IR spectroscopy following the growth of key oxidized functional groups, i.e. phenolic OH, ketone groups intended as carboxyl, lactone, and anhydride, as well as quinone groups as a function of the amount of ozone reacted. The weight uptake of the ACF reacted with ozone was followed gravimetrically. The ACF having >1800 m²/g was able to reach a weight increase of 25% of its original weight due to the formation of oxygenated surface functional groups. Raman spectroscopy was used for the evaluation of defective structures formed in ACF because of ozonization reaction.     

Discrimination of soil-borne fungi using fourier transform infrared attenuated total reflection spectroscopy

Fourier transform infrared (FT-IR) attenuated total reflection (ATR) spectroscopy was used to discriminate five commonly encountered soil-borne fungi that cause severe economic damage to agriculture: Colletotrichum, Fusarium, Pythium, Rhizoctonia, and Verticillium. Contrary to previous studies related to microorganism discrimination using FT-IR-ATR spectroscopy, the pathogen samples were not dried on the ATR crystal, which is a time-consuming operation. Rather, after removing some pathogen filaments from the solution using tweezers, these were placed directly on a flat ATR crystal and pressure was applied using a pressure clamp. Following water subtraction, baseline correction, and normalization of the spectra, principal component analysis was used as a data-reduction step and canonical variate analysis was used for discrimination. Discrimination was performed at the genus level and at the strain level for Colletotrichum. For discrimination between the five fungi at the genus level, the success rate for the validation samples ranged from 75% to 89%. For discrimination between the two Colletotrichum strains, the success rate was 78%. Comparison with spectra of similar fungi dried on the ATR crystal showed that both types of spectra were very similar, indicating that drying the samples on the ATR crystal is not required and can be replaced by mathematical post-processing of the spectra. For routine analyses that involve rapid screening of very large amounts of samples, this approach allows for increasing significantly the number of samples that can be analyzed daily.     

Quantitative Raman spectroscopy of highly fluorescent samples using pseudosecond derivatives and multivariate analysis

Intense luminescence backgrounds cause significant problems in quantitative Raman spectroscopy, particularly in multivariate analysis where background suppression is essential. Taking second derivatives reduces the background, but differentiation increases the apparent noise that arises on spectra recorded with CCD detectors due to random, but fixed, variations in the pixel-to-pixel response. We have recently reported a very general method for correcting CCD fixed-pattern response in which spectra are taken at two or more slightly shifted spectrometer positions and are then subtracted to give a derivative-like shifted, subtracted Raman (SSR) spectrum. Here we show that differentiating SSR data (which has inherently higher S/N than the undifferenced data) yields spectra that are similar to those that are obtained from the normal two-step differentiation process and can be characterized as pseudo-second-derivative, PSD, spectra. The backgrounds are suppressed in the PSD spectra, which means they can be used directly in multivariate data analysis, but they have significantly higher S/N ratios than do simple second derivatives. To demonstrate the improvement brought about by using PSD spectra, we have analyzed known samples, consisting of simple binary mixtures of methanol and ethanol doped with laser dye. When the background levels of all samples included in the models were < or =10x greater than the intensity of the strongest Raman bands, partial least-squares calibration of the PSD data gave a standard error of prediction of 3.2%. Calibration using second derivatives gave a prediction error which was approximately twice as large, at 6.5%; however, when data with background levels . approximately 100x larger than the strongest Raman bands were included, the noise on the second-derivative spectra was so large as to prevent a meaningful calibration. Conversely, the PSD treatment of these samples gave a very satisfactory calibration with a standard error of prediction (3.3%) almost identical to that obtained when the most fluorescent samples were excluded. This method clearly has great potential for general purpose Raman analytical chemistry, because it does not depend on specialized equipment, is computationally undemanding, and gives stable and robust calibrations, even for samples in which the luminescence background level fluctuates between the extremes of being practically zero and completely dominating the Raman signal.     

利用拉曼光谱鉴别优化处理宝玉石

通过对一些宝玉石及其优化处理样品的测试,证明拉曼光谱在检测优化处理宝玉石中具有无可替代的独特优势。拉曼光谱可以检测红外光谱无法透过的闷镶翡翠样品,并可以清楚地检测出充填处理翡翠中的环氧树脂峰;可以明显地将绿松石的上蜡和注胶区分开来,相比于红外光谱,两者的区别更为明显;还可以精确地定位祖母绿裂隙中的充填物质,明确地判定祖母绿是否经充胶或染色处理。拉曼光谱在珠宝检测中的作用越来越重要,将会成为珠宝检测的重要手段之一。     

A mass spectrometry and electron paramagnetic resonance study of photochemical and thermal aging of triterpenoid varnishes

Photochemical and thermal aging of triterpenoid dammar and mastic resins used as varnishes on paintings were studied using graphite-assisted laser desorption/ionization mass spectrometry. This extends an earlier study on similar materials (Zumbühl et al., Anal. Chem. 1998, 70, 707-715) that focused on photoaging. Progressive aging results in development of groups of signals spaced by 14 and 16 Da, indicating incorporation of oxygen as well as simultaneous loss of hydrogen. Oligomers up to tetramers are formed, while cleavage reactions lead to increased signal intensities in the mass ranges between the oligomers and below the monomers. No major differences were found between the mass spectra of samples aged in light or darkness, except that deterioration was faster in light. Electron paramagnetic resonance spectroscopy revealed similar and significant amounts of radicals in films of dammar stored either in light or in darkness. It is concluded that oxidative radical reactions also take place in darkness and that differences in light and dark aging pathways are minor, although rates may differ. These findings lead to a unified explanation for yellowing of natural resin varnishes, one of the major degenerative changes in the appearance of paintings. It is also shown that the commercially available, nominally fresh resins are already in an advanced stage of oxidation and degradation. Energy-rich substances are formed upon irradiation with sunlight and are believed to restart the autoxidative chain reactions, regardless of storage conditions. As a result, varnishes are oxidized quite quickly (months) even when kept in darkness.     

FT-IR spectroscopy of fluoro-substituted hydroxyapatite: strengths and limitations

Fluoro substituted hydroxyapatite (FHAp) samples were prepared by a cyclic pH method. Both calcined and uncalcined samples were subjected to elemental analysis (F, Ca, P) and X-ray diffraction (XRD) analysis to verify composition and phase purity. Good correlation between a-axis parameters and fluoride ion content was found for calcined samples, however, for uncalcined samples the fluoride ion content was higher than estimated from the a-axis values. Fourier transform infra red (FT-IR) spectroscopy analysis of the calcined samples showed OH band shifts and splitting in accordance with F-HO interactions affecting the OH vibration. We conclude that the OH libration (620–780 cm^-1 range) is more suited for estimation of fluoride ion content than the OH stretching. In contrast, uncalcined samples all displayed FT-IR spectra similar to that of hydroxyapatite (HAp) despite the presence of fluoride ions (18–73%). FT-IR emission spectroscopy was used to probe the changes occurring in the FT-IR spectra of HAp and FHAp samples upon heating. Interpretation of the spectral changes occurring during heating to 1,000 °C and subsequent cooling is given. Room temperature spectra of samples heated to various temperatures was used to determine the temperature necessary to produce FT-IR spectra displaying the expected OH bands. A model accounting for the combined observations is proposed.     

Comparison between infrared and Raman spectroscopic analysis of maturing rabbit cortical bone

The molecular composition of the organic and inorganic matrices of bone undergoes alterations during maturation. The aim of this study was to compare Fourier transform infrared (FT-IR) and near-infrared (NIR) Raman microspectroscopy techniques for characterization of the composition of growing and developing bone from young to skeletally mature rabbits. Moreover, the specificity and differences of the techniques for determining bone composition were clarified. The humeri of female New Zealand White rabbits, with age range from young to skeletally mature animals (four age groups, n = 7 per group), were studied. Spectral peak areas, intensities, and ratios related to organic and inorganic matrices of bone were analyzed and compared between the age groups and between FT-IR and Raman microspectroscopic techniques. Specifically, the degree of mineralization, type-B carbonate substitution, crystallinity of hydroxyapatite (HA), mineral content, and collagen maturity were examined. Significant changes during maturation were observed in various compositional parameters with one or both techniques. Overall, the compositional parameters calculated from the Raman spectra correlated with analogous parameters calculated from the IR spectra. Collagen cross-linking (XLR), as determined through peak fitting and directly from the IR spectra, were highly correlated. The mineral/matrix ratio in the Raman spectra was evaluated with multiple different peaks representing the organic matrix. The results showed high correlation with each other. After comparison with the bone mineral density (BMD) values from micro-computed tomography (micro-CT) imaging measurements and crystal size from XRD measurements, it is suggested that Raman microspectroscopy is more sensitive than FT-IR microspectroscopy for the inorganic matrix of the bone. In the literature, similar spectroscopic parameters obtained with FT-IR and NIR Raman microspectroscopic techniques are often compared. According to the present results, however, caution is required when performing this kind of comparison.     

Characterization of woody and herbaceous biomasses lignin composition with 1064 nm dispersive multichannel Raman spectroscopy

Biomass representing different classes of bioenergy feedstocks, including woody and herbaceous species, was measured with 1064 nm Raman spectroscopy. Pine, oak, poplar, kenaf, miscanthus, pampas grass, switchgrass, alfalfa, orchard grass, and red clover were included in this study. Spectral differences have been identified with an emphasis on lignin guaiacyl and syringyl monomer content and carotenoid compounds. The interpretation of the Raman spectra was correlated with (13)C-nuclear magnetic resonance cross-polarization/magic-angle spinning spectra of select biomass samples. Thioacidolysis quantification of guaiacyl and syringyl monomer composition and the library of Raman spectra were used as a training set to develop a principal component analysis model for classifying plant samples and a principal component regression model for quantifying lignin guaiacyl and syringyl composition. Raman spectroscopy with 1064 nm excitation offers advantages over alternative techniques for biomass characterization, including low spectral backgrounds, higher spectral resolution, short analysis times, and nondestructive analyses.     

Serum analysis method combining cellulose acetate membrane purification with surface‐enhanced Raman spectroscopy for non‐invasive HBV screening

A highly sensitive, non‐invasive, and rapid HBV (Hepatitis B virus) screening method combining membrane protein purification with silver nanoparticle‐based surface‐enhanced Raman scattering (SERS) spectroscopy was developed in this study. Reproducible serum protein SERS spectra were obtained from cellulose acetate membrane‐purified human serum from 94 HBV patients and 89 normal groups. Tentative assignments of serum protein SERS spectra showed that the HBV patients primarily led to specific biomedical changes of serum protein. Principal components analysis and linear discriminate analysis were introduced to analyse the obtained spectra, with the diagnostic sensitivity of 92.6% and specificity of 77.5% were achieved for differentiating HBV patients from normal groups.Inspec keywords: patient diagnosis, surface enhanced Raman scattering, proteins, biomembranes, principal component analysis, purification, silver, nanoparticles, nanomedicine, diseasesOther keywords: serum analysis method, cellulose acetate membrane purification, surface‐enhanced Raman spectroscopy, noninvasive HBV screening, rapid HBV screening method, Hepatitis B virus, membrane protein purification, silver nanoparticle‐based surface‐enhanced Raman scattering spectroscopy, reproducible serum protein SERS spectra, cellulose acetate membrane‐purified human serum, linear discriminate analysis, diagnostic sensitivity, HBV patient, principal components analysis     

Spectral characterization of eucalyptus wood

The main difficulties in wood and pulp analyses arise principally from their numerous components with different chemical structures. Therefore, the basic problem in a specific analytical procedure may be the selective separation of the main carbohydrate-derived components from lignin due to their chemical association and structural coexistence. The processing of the wood determines some structural modification in its components depending on the type of wood and the applied procedure. Fourier transform infrared (FT-IR) spectrometry and X-ray diffraction have been applied to analyze Eucalyptus g. wood chips and unbleached and chlorite-bleached pulp. The differences between samples have been established by examination of the spectra of the fractions obtained by successive extraction (acetone extractives, acetone free extractive samples, hemicelluloses, and lignins) by evaluating the derivative spectra, band deconvolution, etc. The energy and the hydrogen bonding distance have been evaluated. The relationship between spectral characteristics and sample composition has been established, as well as the variation of the degree of crystallinity after pulping and bleaching. The integral absorption and lignin/carbohydrate ratios calculated from FT-IR spectra of the IR bands assigned to different bending or stretching in lignin groups are stronger in the spectrum of eucalyptus chips than those from brown stock (BS) pulp spectra because of the smaller total amount of lignin in the latter. FT-IR spectra clearly show that after chlorite bleaching the structure of the wood components is partially modified or removed. Along with FT-IR data, the X-ray results confirmed the low content of lignin in the pulp samples by increasing the calculated values of the crystalline parameters. It was concluded that FT-IR spectroscopy can be used as a quick method to differentiate Eucalyptus globulus samples.     

New method for pollen identification by FT-IR spectroscopy

A new methodology for identification of pollen was developed based on FT-IR spectroscopy. Pollen samples of twenty different plant species were collected and the diffuse reflectance infrared Fourier transform (DRIFTS) and KBr pellet spectra were recorded. Libraries of spectra were created. Spectra of unknown plant origin pollen were recorded and compared with those of the corresponding pollen library and the match value was measured automatically using the appropriate software (OMINC ver. 3.1). From the same pollen samples, microscopic slides were prepared and the photographs of the pollen grains were used as a second comparison method. Using light microscopy, the pollen identification is usually limited to the family or generic name, while FT-IR spectroscopy can distinguish species belonging to the same genus. This method is simple and fast, and when the DRIFTS technique is used the sample is not destroyed.